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I General Description of EnzymesA Chemical composition 1 Enzymes are proteins compounds of high molecular weight containing Carbon (C) hydrogen (H) oxygen (O) sulfur (S) nitrogen (N) 2 Possess primary secondary tertiary (sometimes quaternary) structure 3 Same properties as other proteins
a Charge (isoelectric point electrophoresis) b Solubility c Denaturation (extremes in heat pH chemical) d Non-specific or specific (antigenic) binding 4 Some enzymes have iosenzyme forms
a Different form of an enzyme due to differing arrangements of the polypeptide units in the quaternary structure
b Maintains specificity for the same chemical reaction c Possess unique diagnostic value show specificity to certain organ
systems 1) Isoenzymes of creatine kinase (CK) CK-MM to skeletal muscle CK-MB to cardiac tissue
CK-BB 2) Isoenzymes of lactate dehydrogenase (LD) LD1 and LD2 to cardiac tissue LD4 and LD5 to liver and skeletal muscle
B Biochemical Function
1 Catalyze chemical reactions accelerates specific chemical reactions
toward equilibriumS PSubstrate (S) + Enzyme (ES Product (P) +Enzyme
2 Lowers the energy of activation required for the reaction to proceed
3 Properties of enzymes
a Effective in small concentrationsb Remain unchanged in the reaction not
consumed in the reactionc Effect speed of reaction (not final
concentration of substrate orproducts)d Show greater degree of specificity than
chemical catalystse Unstable and easily destroyed
4 Cofactors substances associated with some enzymes that are needed for
maximal enzyme activity
a Coenzyme organic cofactors (NAD NADH) serve as second
substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++
Zn++ K+ Mg++)
C Importance of Enzymes in Health and Disease
1 Enzymes are intracellular2 Enzyme concentration differs dependent upon
type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and
salivary glands
Enzymes of Clinical SignificancebullCreatine Kinase
bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also
bullorderedbullb Heparinized plasma may be acceptable
bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)
bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored
bullat lower temps (-20oC)
Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
B Biochemical Function
1 Catalyze chemical reactions accelerates specific chemical reactions
toward equilibriumS PSubstrate (S) + Enzyme (ES Product (P) +Enzyme
2 Lowers the energy of activation required for the reaction to proceed
3 Properties of enzymes
a Effective in small concentrationsb Remain unchanged in the reaction not
consumed in the reactionc Effect speed of reaction (not final
concentration of substrate orproducts)d Show greater degree of specificity than
chemical catalystse Unstable and easily destroyed
4 Cofactors substances associated with some enzymes that are needed for
maximal enzyme activity
a Coenzyme organic cofactors (NAD NADH) serve as second
substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++
Zn++ K+ Mg++)
C Importance of Enzymes in Health and Disease
1 Enzymes are intracellular2 Enzyme concentration differs dependent upon
type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and
salivary glands
Enzymes of Clinical SignificancebullCreatine Kinase
bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also
bullorderedbullb Heparinized plasma may be acceptable
bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)
bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored
bullat lower temps (-20oC)
Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
2 Lowers the energy of activation required for the reaction to proceed
3 Properties of enzymes
a Effective in small concentrationsb Remain unchanged in the reaction not
consumed in the reactionc Effect speed of reaction (not final
concentration of substrate orproducts)d Show greater degree of specificity than
chemical catalystse Unstable and easily destroyed
4 Cofactors substances associated with some enzymes that are needed for
maximal enzyme activity
a Coenzyme organic cofactors (NAD NADH) serve as second
substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++
Zn++ K+ Mg++)
C Importance of Enzymes in Health and Disease
1 Enzymes are intracellular2 Enzyme concentration differs dependent upon
type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and
salivary glands
Enzymes of Clinical SignificancebullCreatine Kinase
bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also
bullorderedbullb Heparinized plasma may be acceptable
bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)
bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored
bullat lower temps (-20oC)
Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
3 Properties of enzymes
a Effective in small concentrationsb Remain unchanged in the reaction not
consumed in the reactionc Effect speed of reaction (not final
concentration of substrate orproducts)d Show greater degree of specificity than
chemical catalystse Unstable and easily destroyed
4 Cofactors substances associated with some enzymes that are needed for
maximal enzyme activity
a Coenzyme organic cofactors (NAD NADH) serve as second
substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++
Zn++ K+ Mg++)
C Importance of Enzymes in Health and Disease
1 Enzymes are intracellular2 Enzyme concentration differs dependent upon
type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and
salivary glands
Enzymes of Clinical SignificancebullCreatine Kinase
bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also
bullorderedbullb Heparinized plasma may be acceptable
bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)
bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored
bullat lower temps (-20oC)
Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
4 Cofactors substances associated with some enzymes that are needed for
maximal enzyme activity
a Coenzyme organic cofactors (NAD NADH) serve as second
substrates for enzyme reactionsb Activators inorganic cofactors (Ca++ Fe++
Zn++ K+ Mg++)
C Importance of Enzymes in Health and Disease
1 Enzymes are intracellular2 Enzyme concentration differs dependent upon
type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and
salivary glands
Enzymes of Clinical SignificancebullCreatine Kinase
bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also
bullorderedbullb Heparinized plasma may be acceptable
bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)
bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored
bullat lower temps (-20oC)
Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
C Importance of Enzymes in Health and Disease
1 Enzymes are intracellular2 Enzyme concentration differs dependent upon
type of tissuea AST high concentration in liverb Amylase high concentration in pancreas and
salivary glands
Enzymes of Clinical SignificancebullCreatine Kinase
bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also
bullorderedbullb Heparinized plasma may be acceptable
bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)
bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored
bullat lower temps (-20oC)
Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
Enzymes of Clinical SignificancebullCreatine Kinase
bull1 Specimen collection and handlingbulla Serum is preferred especially if fractionationelectrophoresis also
bullorderedbullb Heparinized plasma may be acceptable
bullMany anticoagulants (fluoride EDTA sodium citrate) inhibitbullenzyme activity and required activators (Ca2+ Mg2+)
bullc No hemolysis intracellular components interfere with chemicalbullreaction in laboratory measurements (NADH ATP HK etc)bulld Avoid lipemia especially for spectrophotometric analysisbulle Relatively unstable analyze within 4 hours of collection elsebullstable at 2-6oC for up to one week stability increases when stored
bullat lower temps (-20oC)
Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
Wide tissue distribution injury tissue necrosis will cause elevation of thisenzyme in plasmaseruma Skeletal muscleb Heart tissuec Brain nerve tissue
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
Method of Measurment
3 Physiologic reaction CK is a transferase ATP storage and generation incontractile or transport systems predominantly in muscle cells
Creatine + ATP CK1048774 creatine phosphate + ADP4 Method of measurement
Most common method modified Rosalki-Oliver methodcreatine phosphate + ADP CK creatine + ATPATP + glucose HK ADP + glucose-6-phosphate
Glucose-6-phosphate + NADP+ G6PD 6-phosphogluconate +NADPH
(Increased ABS at 340 nm)
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
Isoenzymes clinical relevance of CK activity often depends more onisoenzyme fractionation than on total levelsa Isoenzymes occur as a dimer1) B polypeptide units2) M polypeptide unitsb Three isoenzyme forms numbered by their anodal (+)electrophoresis mobility with lsquofraction 1rsquo migrating fastest towardsthe anode1) CK1 = CK-BB (brain type) Normal ~02) CK2 = CK-MB (hybrid heart) Normal lt53) CK3 = CK-MM (muscle type) Normal 95-100
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
c Immunologic measurement of CK-MB1 )Immunoinhibition of subunit M (anti-M)
2 )Measure CK before and after inhibition3 )Disadvantage falsely elevated if CK-BB activity present
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
d Enzyme immunoassay measurement of CK-MB (mass)
A lsquosandwich techniquersquo that uses 2 antibodies one directedagainst the lsquoMrsquo subunit the other against the lsquoBrsquo subunit which isalso labeled with a reagent enzyme that catalyzes the conversion ofsubstrate to product
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
6 Reference Ranges affected by gender
bullmuscle mass exercise agebullTotal CK
bullMale 15-160 IULbullFemale 15-130 IULbullCK-MB lt5 of total CK
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
7 Clinical significancea Acute Myocardial Infarction (AMI)1) Increased total CK increased CK-MB2) Total CK increase 7-12 times normal withrise apparent at 4-6 hours post MIpeaks at 18-24 hours andreturns to normal by 72 hours3) The rise peak and fall of CK-MB parallels that of total CKbut a little sooner4) Other cardiac events (traumainflammation) variable results
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
b Skeletal muscle disease1) Increased total CK normal CK-MB (increased CK-MM)2) Muscular dystrophy especially Duchenne type may seetotal CK 50x ULN3) Inflammation of muscle especially viral also malignanthyperthermia4) Normal CK activity in neurogenic muscle disordersmultiple sclerosis myasthenia gravis poliomyelitis
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
c Central nervous system disorders
1) Increased total CK normal CK-MB (increased CK-BB)
2) Trauma or pathology indicated when CK-BB is present
3) Cerebral vascular disease encephalopathy stroke (CK-BB )may be normal
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
d Other
1) Neonates normally have 1-3 CK-BB2) Hypothyroidism increased total CK increased
CK-MM3) CK activity is NORMAL in liver disease4) Effect of hemolysis is procedural and not due
to CKactivity in RBC
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
B Lactate dehydrogenase (LD) 1 Specimen collection and handling
a Serum is preferred especially if fractionationelectrophoresis alsoorderedb Heparinized plasma may be acceptable many anticoagulants
(fluoride EDTA sodium citrate )inhibit enzyme activityc No hemolysis LD-1 activity in the RBC is 150x greater than thatin plasmad Avoid lipemia especially for spectrophotometric analysise Relatively unstable analyze within 4 hours of collection elsestable at 2-6oC for up to one week stability increases when storedat lower temps (-20oC)f LD-4 and LD-5 are especially labile stable when stored at roomtemperature do not store in refrigerator or freezer unlessglutathione is added as preservative
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
2 Wide cellulartissue distribution
a Skeletal muscleb Cardiac muscletissuec Liverd RBCe Kidney lung tumor cells
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
3 Physiologic reaction
bull LD is an oxidoreductase catalyzes thebull interconversion of lactic and pyruvic acidsbull Lactate + NAD+ LD Pyruvate + NADH +
H+
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
4 Method of measurement
Forward reactionLactate + NAD+ ---LD (pH 83-89) Pyruvate + NADH + H+(Increased ABS at 340 nm)
Reverse reactionPyruvate + NADH ---LD (pH 71-74) Lactate + NAD+(Decreased ABS at 340 nm)
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
Both the forward and reverse reactions have been used in the clinical lab
with the rate of the reverse reaction being ~3 times faster allowing
measurement of smaller sample volumes and shorter reaction times
However the forward reaction has better linearity at this time
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
5 Isoenzymes clinical relevance of total LD activity often depends more on
isoenzyme fractionation correlating to total LD activitya Isoenzymes occur as a tetramer1) H polypeptide units (heart)2) M polypeptide units (muscle)
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
b Five isoenzyme forms numbered by their anodal electrophoresis
6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
mobility with lsquofraction 1rsquo migrating fastest towards the anode)+(
1) LD1 = HHHH Heart RBC kidney2) LD2 = HHHM3) LD3 = HHMM4) LD4 = HMMM Skeletal muscle liver5) LD5 = MMMM6 Reference RangesTotal LD 100-200 IULLD-1 14-26 of total LDLD-2 29-39 of total LDLD-3 20-26 of total LDLD-4 8-16 of total LDLD-5 6-16 of total LD
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
7 Clinical significance
a Hepatocellular disease1) Increased total LD (7-10 x ULN) increased LD-4 LD-52) Toxic and viral hepatitis infectious mononucleosis3) Normal or slightly elevated in obstructive liver
disordersb Skeletal muscle disease1) Increased total LD increased LD-4 and LD-52) Muscular dystrophy especially Duchenne typeSkeletal muscle liver
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
c Acute myocardial infarction (AMI)
1) Increased total LD increased LD-1 and LD-2 with aflipped ratio2) Total LD increases 3-7 times normal withrise apparent at 8-12 hours post MIpeaks around 72 hours andreturns to normal by 7-12 days3) Not useful for diagnosis of AMI4) Other cardiac events (traumainflammation) variable
results
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
d Hemolytic anemia
1) Increased total LD 20-50 times normal increased LD-1 and
LD-2 with a false flipped ratio (not due to MI)2) Untreated Pernicious Anemia Folate
MagaloblasticAnemia3) Hemolytic anemia
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
C Aspartate Transaminase (AST) Historical Serum glutamate oxaloacetate transaminase (SGOT)
1 Specimen collection and handlinga Serum preferred heparinized plasma acceptable
other anticoagulants inhibit enzyme activityb Avoid hemolysis RBC contain ASTc Avoid lipemia especially for spectrophotometric
analysisd Relatively stable analyze within 8 hours of
collection else store at2-4oC for up to 4 days
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
2 Wide cellulartissue distribution
a Skeletal muscle
b Cardiac muscletissue
c Liver
d Kidney pancreas RBC
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
3 Physiologic reaction
AST is a transferase catalyzes the transfer of an amino group between aspartate and alpha-keto acids Important in the
synthesis and degradation of amino acids ketoacids formed are used in the TCA cycle to provide a source of energy
Aspartate + α-ketoglutarate AST Oxaloacetate + Glutamate
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
4 Method of measurement Karmen method
Aspartate + alpha-ketoglutarate AST Oxaloacetate + Glutamate
Oxaloacetate + NADH + H+ 1048774 Malate + NAD+(decreased ABS at 340 nm)
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
5 Reference Range 0-56 IUL
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
6 Clinical significance
bull a Liver disease hepatocellular parenchymal disease
bull 1) Inflammation toxic or viral hepatitis infectiousbull mononucleosis activity may reach up to 50-100 x
normalbull 2) Obstructive conditions (cholestasis some
stages ofbull cirrhosis) generally show normal or slightly
increased AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
b Skeletal muscle disease
1) Muscular dystrophy especially Duchenne type2) Inflammation of the muscle malignant
hyperthermiacrushing muscle injury3) Neurogenic muscle disorders such as multiple
sclerosismyasthenia gravis poliomyelitis show normal AST
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