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YMC GENERAL CATALOG
5Columns and bulk packingsfor BIO-MACROMolecular Separations
Types and characteristics of columns and bulk packing for BIO-MACRO molecular separations
YMC-BioPro Ion Exchange columnsYMC-BioPro QA-F/YMC-BioPro SP-FYMC-BioPro QA/YMC-BioPro SPYMC-BioPro Ion Exchange mediaYMC-Pack DiolReversed-phase columns for bio moleculesYMC-Pack PROTEIN-RPYMC-Pack Diol-NPYMC-Pack Polyamine Ⅱ
102~105
106107
108,109110,111112,113
114~121122,123
124125
101
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Columns and bulk packings for BIO-MACRO molecular separationsIt is increasingly more important to separate bio molecules such as peptides, proteins and nucleic acids, due to progress of bio pharmaceuticals like monoclonal antibodies. To meet such demands, YMC offers abundant columns and bulk packings for ion exchange, size exclusion, reversed-phase and normal-phase chromatography.Depending on target compounds or scale of the separation, choose appropriate columns and packings.
Columns and bulk packings for BIO-MACRO separations (See P10, 11 for choosing appropriate column for target compounds.)
Separation mode Product name Base
materialPore size
(Å) Functional group Purpose compounds Page
Ion exchange
YMC-BioPro QA-F
Polymer
non-porous−CH2N+(CH3)3
ProteinsPeptidesNucleic acidsOligonucleotides
106 〜111
YMC-BioPro SP-F −CH2CH2CH2SO3−
YMC-BioPro QAporous
−CH2N+(CH3)3
YMC-BioPro SP −CH2CH2CH2SO3−
YMC-BioPro Q30YMC-BioPro Q75
porous−CH2N+(CH3)3
YMC-BioPro S30YMC-BioPro S75 −CH2CH2CH2CH2SO3
−
Size exclusion YMC-Pack Diol Silica gel 60,120, 200, 300 Dihydroxypropyl
ProteinsPeptidesNucleic acidsOligonucleotidesCarbohydrates
112,113
Reversed-phase
Triart C18 Hybrid silica 120
C18ProteinsPeptidesNucleic acidsOligonucleotidesNucleotidesNucleosidesNucleobasesCarbohydrates
114 〜121
YMC-Pack Pro C18
Silica gel
120Hydrosphere C18 120YMC-Pack Pro C18 RS 80YMC-Pack ODS-A 120, 200, 300YMC-Pack ODS-AQ 120, 200YMC-Pack C8 120, 200, 300 C8YMC-Pack C4 120, 200, 300 C4YMC-Pack Ph 120 PhenylYMC-Pack CN 120, 300 CyanopropylYMCbasic 200 C8YMC-Pack PROTEIN-RP 200 − 122,123YMC-Pack PolymerC18 Polymer − C18 114 〜121
Normal phase
YMC-Pack Diol-NP
Silica gel
60,120 Dihydroxypropyl PeptidesNucleotidesNucleosidesNucleobasesCarbohydrates
124
YMC-Pack Polyamine Ⅱ 120 Polyamine 125
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0 10 20 30 min
Transferrin
Albumin
N080313E
IgGmAU
10
0
YMC-BioPro QA 5 µm, 50×4.6 mmI.D.
0 10 20 30 40 50 60 min
N080616A
IgG
AlbuminTransferrin
mAU
150
100
0
50
YMC-Pack Diol-300 + Diol-200 5 µm, 300×8.0 mmI.D. ×2
Eluent : A) 20 mM Tris-HCl (pH 8.6) B) 20 mM Tris-HCl (pH 8.6) containing 0.5 M NaCl 0-30% B (0-15 min), 30-100% B (15-30 min)
Flow rate :0.5 mL/minTemperature :25 ℃Detection :UV at 280 nmInjection :20 µL (100 µL/mL)
Eluent :0.1 M KH2PO4-K2HPO4 (pH 7.0) containing 0.2 M NaClFlow rate :0.5 mL/minTemperature :ambient (25 ℃)Detection :UV at 280 nmInjection :20 µL (100 µL/mL)
Proteins in human serum are separated by the difference in the surface charge on ion exchange chromatography (IEC) and by the difference in the molecular weight on size exclusion chromatography (SEC).
Separation of proteins Comparison of separation in different modeHuman serum
Ion exchange Size exclusion
min2 4 6 8 10 12 14 160
NaN3
Mouse monoclonal IgG1(Anti-human IgG4)
mAU
0
1
2
3
4
5
P080220A
YMC-BioPro QA-F 5 µm, 30×4.6 mmI.D.
0 105 15 20 25 30 min
NaN3Mouse monoclonal IgG1(Anti-human IgG4)
0
10
20
30
40
mAU
P080530A
YMC-Pack Diol-200 5 µm, 300×4.6 mmI.D.
Eluent : A) 20 mM Tris-HCl (pH 8.1) B) 20 mM Tris-HCl (pH 8.1) containing 0.5 M NaCl 10-25% B (0-18 min)
Flow rate :1.0 mL/minTemperature :25 ℃Detection :UV at 220 nmInjection :10 µL (0.1 mg/mL)
Eluent :0.1 M KH2PO4-K2HPO4 (pH 7.0)Flow rate :0.17 mL/minTemperature :ambient (25 ℃)Detection :UV at 220 nmInjection :10 µL (0.05 mg/mL)
Mouse monoclonal antibody against human IgG4 is analyzed on ion exchange chromatography (IEC) and size exclusion chromatography (SEC). Several peaks possibly derived from isoform of antibody are observed in ion exchange mode, while a single peak is detected in size exclusion mode.
Mouse monoclonal IgG1 anti-human IgG4 (Purified by DEAE chromatography, containing NaN3)
1. Rabbit IgG2. Mouse IgG Fc fragment
min
mAU
0 5 10 15 20
0
25
50
R080623D
1
2
YMC-Pack Diol-200 5 µm, 300×8.0 mmI.D.
30 40352520151050 min
R080619B
Mouse IgG Fc fragmentmAU
0
10
20
30
YMC-Pack C4(300 Å)5 µm, 150×4.6 mmI.D.
Eluent :0.1 M KH2PO4-K2HPO4 (pH 6.9) containing 0.2 M NaClFlow rate :1.0 mL/minTemperature :ambient (27 ℃)Detection :UV at 220 nmInjection :5 µL (0.5 mg/mL)
Eluent : A) water/TFA (100/0.1) B) acetonitrile/TFA (100/0.1) 25-45% B (0-40 min)
Flow rate :1.0 mL/minTemperature :37 ℃Detection :UV at 220 nmInjection :5 µL (1.0 mg/mL)
Size exclusion chromatography (SEC) is useful for separation of substances which have distinct differences in molecular weight, like between IgG and its fragments. On the other hand, reversed-phase chromatography (RPC) is suitable for a precise analysis of peptides and proteins with a molecular weight of less than 100 kDa such as IgG Fc fragment.
Mouse IgG Fc fragment (Prepared from normal serum)
Ion exchange Size exclusion
Size exclusion Reversed-phase
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Columns and bulk packings for BIO-MACRO molecular separations
0 60 min2010 30 40 50
N080422B
*
0
10
20
mAU
YMC-BioPro QA 5 µm, 50×4.6 mmI.D.
Eluent : A) 20 mM Tris-HCl (pH 8.6) B) 20 mM Tris-HCl (pH 8.6) containing 0.5 M NaCl 0-15% B (0-30 min), 15-60% B (30-60 min)
Flow rate :0.5 mL/minTemperature :25 ℃Detection :UV at 220 nmInjection :20 µL
Separation of proteins Comparison of separation in different modeTryptic digests of BSA
Ion exchange
0 60 min2010 30 40 50
0
30
50
mAU
60
40
20
10
N080623L
105
102
103
104
MW
5
4
3
2
1
101*
YMC-Pack Diol-120 + Diol-60 5 µm, 500×8.0 mmI.D. ×2
These chromatograms show separation of tryptic digests of BSA (MW: 66,000) in ion exchange chromatography (IEC), size exclusion chromatography (SEC) and reversed-phase chromatography (RPC). The molecular weight of the digests is estimated to be approximately from 100 to 20,000 by SEC chromatogram. IEC and RPC chromatograms show many peaks of fragments which are separated by the difference in structure, charge and hydrophobicity.
Size exclusion
Eluent : 0.1 M KH2PO4-K2HPO4 (pH 7.0) containing 0.2 M NaCl/acetonitrile (70/30)
Flow rate :0.7 mL/minTemperature :ambient (25 ℃)Detection :UV at 220 nmInjection :5 µL
0 60 min2010 30 40 50
0
20
30
mAU
10
40
N080318B
*
* undigested BSA
YMCbasic 5 µm, 150×2.0 mmI.D.Reversed-phase
Eluent : A) water/TFA (100/0.1) B) acetonitrile/TFA (100/0.1) 5-35% B (0-50 min), 35-45% B (50-55 min), 45% B (55-60 min)
Flow rate :0.2 mL/minTemperature :37 ℃Detection :UV at 220 nmInjection :1 µL
Calibration curve of peptides and proteins1. Myoglobin (MW 17,000)2. Insulin (Bovine) (MW 5,700)3. Neurotensin (MW 1,672)4. Tetraglycine (MW 246)5. Glycine (MW 75)
20 min100G910607A
YMC-Pack ODS-A(120 Å)5 µm,150×4.6 mmI.D.
Separation of sugar chains Comparison of separation in different modePyridylamino (PA) -Sugar chains
Reversed-phase
20 min100G910620A
YMC-Pack Polyamine Ⅱ 5 µm,150×4.6 mmI.D.
Pyridylamino (PA) sugar chains are often analyzed for structural determination of sugar chain in glycoproteins and glycolipids. Separations of PA sugar chains in reversed-phase (RP) mode and normal-phase (NP) mode are shown. Two dimensional HPLC combining two different modes, such as RP mode and NP mode, is useful tool for structural determination of sugar chain.
Normal-phase
Eluent :methanol/20 mM NH4H2PO4 (5/95)Flow rate :1.0 mL/minTemperature :37 ℃Detection :FLS at Ex. 320 nm, Em. 400 nmInjection :2 µL (3.3 pmol/mL)Sample : PA-Sugar Chain Series,
manufactured by TAKARA SHUZO CO., LTD.
Eluent :methanol/20 mM NH4H2PO4 (80/20)Flow rate :1.0 mL/minTemperature :37 ℃Detection :FLS at Ex. 320 nm, Em. 400 nmInjection :3 µL (3.3 pmol/mL)Sample : PA-Sugar Chain Series,
manufactured by TAKARA SHUZO CO., LTD.
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N080212G
0 2010 30 min
0
50
100
mAU
YMC-BioPro QA-F 5 µm, 100×4.6 mmI.D.
Separation of nucleic acids Comparison of separation in different modeDNA fragments 1 Kb DNA ladder (75 - 12,216 bp)
Ion exchange
Eluent : A) 20 mM Tris-HCl (pH 8.1) containing 0.7 M NaCl B) 20 mM Tris-HCl (pH 8.1) containing 1.0 M NaCl 0-100% B (0-30 min)
Flow rate :0.5 mL/minTemperature :25 ℃Detection :UV at 260 nmInjection :20 µL
DNA fragments are analyzed with YMC-BioPro QA-F ion exchange column. 100 mm length column of YMC-BioPro QA-F is ideal for high-resolution analysis of nucleic acids.
YMC-BioPro QA-F 5 µm, 100×4.6 mmI.D.
0 10 20 25 min155
N080610F
Plasmid pBR322 Hae III digests (8-587 bp)
Plasmid pBR322 (4,361 bp)
10
20
30
40
mAU
0
Plasmid pBR322 restriction fragments
Ion exchange
Eluent : A) 20 mM Tris-HCl (pH 8.1) B) 20 mM Tris-HCl (pH 8.1) containing 1.0 M NaCl 70-85% B (0-20 min), 85% B (20-25 min)
Flow rate :0.5 mL/minTemperature :35 ℃Detection :UV at 260 nmInjection :10 µL
YMC-Pack Diol-300 + Diol-200 5 µm, 500×8.0 mmI.D. ×2
P080617A
Plasmid pBR322 Hae III digests (8-587 bp)
Plasmid pBR322 (4,361 bp)
10
20
30
40
mAU
0
0 20 403010 50 min
Size exclusion
Eluent :0.1 M KH2PO4-K2HPO4 (pH 7.0) containing 0.2 M NaClFlow rate :0.7 mL/minTemperature :ambient (25 ℃)Detection :UV at 260 nmInjection :10 µL
The separation of plasmid pBR322 restriction fragments (8-857 bp) is compared between in ion exchange mode and size exclusion mode. Ion exchange chromatography (IEC) is applicable to identification of each fragment requiring high resolution and size exclusion chromatography (SEC) is usable for characterization of molecular weight distribution.
5'-CCGCTCGAGCTAAAAAAAGCCTGTGTTACC-3' (30 mer)
0 5 10 15 20 min
UV
TIC
30 mer2928
27
F060213C-04
Hydrosphere C18 3 µm, 50×2.0 mmI.D.
Primer of DNA sequencing (30 mer)
Reversed-phaseEluent : A) 10 mM DBAA* (pH 6.0)
B) 10 mM DBAA* (pH 6.0)/acetonitrile (50/50) 58-62% B (0-20 min), 62% B (20-25 min)
Flow rate :0.2 mL/minTemperature :35 ℃Detection :UV at 269 nm and ESI negative-modeInjection :1 µL (10 pmol/component)* di-n-butylammonium acetate
This figure shows LC/MS analysis of a mixture of synthetic 27-30 mer oligonucleotides in reversed-phase mode. Hydrosphere C18 columns have been designed for enhanced retention and selectivity of highly polar compounds. They can achieve excellent separation by one-nucleotide difference and sufficient intensity in UV and ESI-MS.
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Ion Exchange Columns
YMC-BioPro Ion Exchange ColumnsYMC-BioPro ion exchange columns are specially designed for separation of proteins, peptides, and nucleic acids. YMC-BioPro ion exchange columns are available in QA and SP chemistries and are based on 5 µm porous and non-porous hydrophilic polymer beads with low nonspecific adsorption.
Ion exchange columns ideal for analysis of proteins, peptides, and nucleic acids
Features
● Ion exchange columns designed for analytical and laboratory-scale purification of proteins, peptides, and nucleic acids
● Newly developed hydrophilic polymer beads with low nonspecific adsorption
● Effective surface structure designed for maximum interaction with biomolecules
● Available in a strong anion exchanger (QA, quaternary ammonium) and a strong cation exchanger (SP, sulfopropyl)
● Non-porous type for increasing resolution and throughput
● Porous type for higher binding capacity and recovery
SpecificationsYMC-BioPro QA-F YMC-BioPro SP-F YMC-BioPro QA YMC-BioPro SP
Matrix Hydrophilic non-porous polymer Hydrophilic porous polymerParticle size 5 µmCharged group −CH2N+(CH3)3 −CH2CH2CH2SO3
− −CH2N+(CH3)3 −CH2CH2CH2SO3−
Counter ion Cl− Na+ Cl− Na+
Ion exchange capacity(meq/mL-resin)
0.075 - 0.110 0.230 - 0.290 0.075 - 0.100 0.070 - 0.095
Dynamic binding capacity(mg/mL-resin)
>12(BSA) >10(human-IgG) >110(BSA) >70(human-IgG)
Usable temperature 4 - 60 ℃Usable pH range 2.0 - 12.0Column material PEEK
SEM images of polymer beads of YMC-BioPro ion exchange columns
Porous polymer beadsNon-porous polymer beads
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Ion Exchange Columns
YMC-BioPro QA-F / YMC-BioPro SP-F● Ion exchange column based on non-porous polymer beads● High efficiency with low operating pressure● 30 mm length column for ultra high-throughput analysis● 100 mm length column for high-resolution analysis
Ion exchange columns for high-throughput and high-resolution analysis of proteins, peptides, and nucleic acidsYMC-BioPro QA-F/SP-F columns are ion exchange columns based on non-porous hydrophilic polymer beads with high chemical and mechanical stability, and low nonspecific adsorption of biomolecules. The short
columns (30 mm, 50 mm) are useful for the fast analysis at a higher flow rate, and the 100 mm length columns are best choice for the quality control assessment of biopharmaceuticals requiring a high-resolution.
■Matrix:Hydrophilic non-porous polymer beads■Usable pH range:2.0 〜 12.0
Ultra high-throughput analysis of proteins
YMC-BioPro SP-F 5 µm, 30×4.6 mmI.D.YMC-BioPro SP-F 5 µm, 30×4.6 mmI.D.
1 2
3
N071023K
1 2 3 4 min0
0.0
0.2
0.4
AU1. Ribonuclease A2. Cytochrome c 3. Lysozyme
The high mechanical stability of non-porous polymer beads and the short column length enable faster elution of proteins at a higher flow rate.
Eluent : A) 20 mM KH2PO4-K2HPO4 (pH 6.8) B) 20 mM KH2PO4-K2HPO4 (pH 6.8) containing 0.5 M NaCl 0-100% B (0-4 min)
Flow rate :1.5 mL/min (540 cm/hr)Temperature :25 ℃Detection :UV at 220 nmInjection :20 µLPressure :4.8-5.2 MPa
Packing material
Particle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
QA-F S-5 non-porous
4.6× 30 QF00S05-0346WP4.6× 50 QF00S05-0546WP4.6×100 QF00S05-1046WP
Packing material
Particle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
SP-F S-5 non-porous
4.6× 30 SF00S05-0346WP4.6× 50 SF00S05-0546WP4.6×100 SF00S05-1046WP
Ordering information
High-resolution analysis of nucleic acids
YMC-BioPro QA-F 5 µm, 100×4.6 mmI.D.YMC-BioPro QA-F 5 µm, 100×4.6 mmI.D.
N080212G
20 min100
50
mAU
0The separation of DNA fragments is shown. YMC-BioPro QA-F of 100 mm length column is good choice for high-resolution analysis of nucleic acids.
Eluent : A) 20 mM Tris-HCl (pH 8.1) containing 0.7 M NaCl B) 20 mM Tris-HCl (pH 8.1) containing 1.0 M NaCl 0-100% B (0-30 min)
Flow rate :0.5 mL/min (180 cm/hr)Temperature :25 ℃Detection :UV at 260 nmInjection :20 µL (0.25 mg/mL)
DNA fragments 1Kb DNA ladder (75 - 12,216 bp)
High-resolution analysis of proteins
YMC-BioPro QA-F 5 µm, 100×4.6 mmI.D.YMC-BioPro QA-F 5 µm, 100×4.6 mmI.D.
P080424A
2010 30 40 min5 15 25 35
P080424B
20100 30 40 min5 15 25 35
NaN3
NaN3
MAb Lot. A
MAb Lot. B MAb
MAb
mAU
0
1
2
3
4
mAU
0
1
2
3
4 Two different lots of commercially available MAb purified by DEAE chromatography, are analyzed with 100 mm length column of YMC-BioPro QA-F. The MAb is resolved into several peaks, and the lot-to-lot variability is observed. 100 mm length column of YMC-BioPro QA-F/SP-F, which has high efficiency, is ideal for characterization of glycoproteins such as monoclonal antibodies and for quality control assessment of biopharmaceuticals.
Eluent : A) 20 mM Tris-HCl (pH 8.1) B) 20 mM Tris-HCl (pH 8.1) containing 0.5 M NaCl 10-25% B (0-60 min)
Flow rate :1.0 mL/min (360 cm/hr)Temperature :25 ℃Detection :UV at 220 nmInjection :14 µL (0.1 mg/mL)Sample : Mouse monoclonal IgG1 anti-human IgG4
(Purified by DEAE chromatography, containing NaN3)
Monoclonal antibody (MAb) against human IgG4
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Ion Exchange Columns
YMC-BioPro QA / YMC-BioPro SP● Ion exchange column based on porous polymer beads● Excellent resolution● High binding capacity and high recovery of biomolecules● Suitable for laboratory-scale purification
Ion exchange columns for analysis and laboratory-scale purification of proteins, peptides, and nucleic acidsYMC-BioPro QA/SP columns are ion exchange columns based on porous hydrophilic polymer beads with low nonspecific adsorption of biomolecules. YMC-BioPro QA/SP columns have superior resolution, high binding
capacity and high recovery of various biomolecules, and they allow highly effective analysis and laboratory-scale purification of biopharmaceutical proteins such as antibodies.
■Matrix:Hydrophilic porous polymer beads■Usable pH range:2.0 〜 12.0
Excellent resolutions
The separation of standard protein mixture is compared between YMC-BioPro SP and a commercial porous polymer cation exchange column. Many impurities are resolved from the main peaks of proteins on YMC-BioPro SP with superior peak shapes.
Eluent : A) 20 mM KH2PO4-K2HPO4 (pH 6.8) B) 20 mM KH2PO4-K2HPO4 (pH 6.8) containing 0.5 M NaCl 0-100% B (0-60 min)
Flow rate : 0.5 mL/min (180 cm/hr)Temperature :25 ℃Detection :UV at 220 nmInjection : 20 µL
1. Ribonuclease A (0.5 mg/mL)2. Cytochrome c (0.5 mg/mL)3. Lysozyme (0.5 mg/mL)
0 20 40 60 min
0
0.5
AU
1 23
YMC-BioPro SP5 µm, 50×4.6 mmI.D.
1 2 3
N071016E
0 20 40 60 min
0
0.5
AU
1 23
Brand T (porous S type)7 µm, 50×4.6 mmI.D.
1 2 3
N071018F
0 10 20 30 min
mAU
10
0
IgG
Albumin
N080303G
Transferrin
0 10 20 30 min
mAU
10
0
Transferrin
Albumin
N080313E
IgG
0 10 20 30 min
mAU
10
0 N080304A
Transferrin
AlbuminIgG
YMC-BioPro QA5 µm, 50×4.6 mmI.D.
Brand G (porous Q type)10 µm, 50×5.0 mmI.D.
Brand T (porous Q type)10 µm, 50×4.6 mmI.D.
Eluent : A) 20 mM Tris-HCl (pH 8.6) B) 20 mM Tris-HCl (pH 8.6) containing 0.5 M NaCl 0-30% B (0-15 min), 30-100% B (15-30 min)
Flow rate :0.5 mL/min (180 cm/hr for 4.6 mmI.D., 150 cm/hr for 5.0 mmI.D.)Temperature :25 ℃Detection :UV at 280 nmInjection :20 µLSample :Human serum (100 µL/mL)
Separation of proteins in human serumComparison of separation on YMC-BioPro QA and commercial porous Q type columns
YMC-BioPro QA5 µm, 50×4.6 mmI.D.
Brand G (porous Q type)10 µm, 50×5.0 mmI.D.
Brand T (porous Q type)10 µm, 50×4.6 mmI.D.
0 2010 30 40 50 60 min
0 2010 30 40 50 60 min
0 2010 30 40 50 60 min
peaks = 50
peaks = 11
peaks = 28
mAU
30
20
0
10
mAU
30
20
0
10
mAU
30
20
0
10
N080422B
N080417G
N080417C
Eluent : A) 20 mM Tris-HCl (pH 8.6) B) 20 mM Tris-HCl (pH 8.6) containing 0.5 M NaCl 0-15% B (0-30 min), 15-60% B (30-60 min)
Flow rate :0.5 mL/min (180 cm/hr for 4.6 mmI.D., 150 cm/hr for 5.0 mmI.D.)Temperature :25 ℃Detection :UV at 220 nmInjection :20 µLSample :Tryptic digest of BSA
Peptide mappingComparison of separation on YMC-BioPro QA and commercial porous Q type columns
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Analytical columnsPacking material
Particle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
QA S-5porous
4.6× 30 QAA0S05-0346WP4.6× 50 QAA0S05-0546WP4.6×100 QAA0S05-1046WP
Packing material
Particle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
SP S-5porous
4.6× 30 SPA0S05-0346WP4.6× 50 SPA0S05-0546WP4.6×100 SPA0S05-1046WP
Ordering information
High loadabilityComparison of the effect of sample load on YMC-BioPro QA and commercial Q type column
1
2
N080206A
N080206B
N080207C
N080207B
500 µg
250 µg
100 µg
50 µg
Loading amount
20100 30 min
12
N080206D
N080206E
N080208C
N080208B
20100 30 min
500 µg
250 µg
100 µg
50 µg0
100
200
mAU
0
100
200
mAU
YMC-BioPro QA5 µm, 50×4.6 mmI.D.YMC-BioPro QA5 µm, 50×4.6 mmI.D.
Brand G (porous Q type)10 µm, 50×5.0 mmI.D.Brand G (porous Q type)10 µm, 50×5.0 mmI.D.
YMC-BioPro QA shows the excellent resolution and peak shapes even when the loading amount increases. The porous type YMC-BioPro columns are suitable for laboratory-scale purification of proteins.
Eluent : A) 20 mM Tris-HCl (pH 8.1) B) 20 mM Tris-HCl (pH 8.1) containing 0.5 M NaCl 10-80% B (0-30 min)
Flow rate : 0.5 mL/min (180 cm/hr for 4.6 mmI.D., 150 cm/hr for 5.0 mmI.D.)
Temperature :25 ℃Detection :UV at 280 nmInjection :100 µL
1. Ovalbumin2. Trypsin inhibitor
High binding capacity and recoveryComparison of dynamic binding capacity (DBC) and recovery for BSA
Dynamic binding capacity(mg/mL-resin, 10% breakthrough)
Eluted amount(mg/mL-resin)
Recovery*(%)
YMC-BioPro QA 126 120 95Brand T (porous Q type) 73 58 79Brand G (porous Q type) 100 35 35
*Recovery:(Eluted amount/Dynamic binding capacity)×100
YMC-BioPro QA gives the superior DBC and recovery compared with conventional porous polymer anion exchange columns. The surface structure of YMC-BioPro which is designed for maximum interaction with proteins provides high binding capacity, and the hydrophilic property of polymer beads remarkably reduces nonspecific adsorption of proteins
Column : YMC-BioPro QA 50×4.6 mmI.D. Brand T (porous Q type) 50×4.6 mmI.D. Brand G (porous Q type) 50×5.0 mmI.D.
Linear velocity :180 cm/hrEquilibration buffer :20 mM Tris-HCl (pH 8.6)Elution buffer :20 mM Tris-HCl (pH 8.6) containing 1.0 M NaClSample :1 mg/mL Bovine serum albumin (BSA) in equilibration bufferDetection :UV at 280 nm
Breakthrough curves
10% breakthrough
BSA amount loaded (mg/mL-resin)
0 20 40 60 80 100 120 140 160
Brand T
Brand G
BioPro QA
0
0.50
0.25
AU
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olecular Separations
Ion Exchange Media
YMC-BioPro Ion Exchange Media● Hydrophilic polymer beads with low nonspecific adsorption● Strong anion exchanger (Q type) and strong cation exchanger (S type)● Effective surface structure designed for maximum interaction with biomolecules● High binding capacity and recovery
Ion exchange media for capture and intermediate purification of biomoleculesYMC-BioPro ion exchange media are available in Q and S chemistries on 75 µm and 30 µm of hydrophilic porous polymer beads with low nonspecific adsorption and high binding capacity. YMC-BioPro ion exchange
media have similar retention selectivity to YMC-BioPro QA/SP columns, and it allows predictable scale up from analytical to preparative separation.
■Matrix:Hydrophilic porous polymer beads■Usable pH range:2.0 〜 12.0
SpecificationsYMC-BioPro Q30 YMC-BioPro Q75 YMC-BioPro S30 YMC-BioPro S75
Matrix Hydrophilic porous polymer beadsParticle size 30 µm 75 µm 30 µm 75 µmCharged group −CH2N+(CH3)3 −CH2CH2CH2CH2SO3
−
Usable pH range 2.0 - 12.0
High dynamic binding capacity (DBC) for proteins
Anion exchanger Particle size(µm)
Ion exchange capacity(meq/mL-resin)
DBC*1
(mg/mL-resin)YMC-BioPro Q75 75 0.13 183Brand G(porous Q type) 90 0.19 102
Cation exchanger Particle size(µm)
Ion exchange capacity(meq/mL-resin)
DBC*1
(mg/mL-resin)YMC-BioPro S75 75 0.12 192Brand G(porous S type) 90 0.13 80
YMC-BioPro ion exchange media have 1.4 to 1.8 times higher DBC of protein than commercial ion exchange media. YMC-BioPro ion exchange media are effective in protein purification from capture step requiring high capacity to intermediate step requiring high efficiency.
*1 Dynamic binding capacities were determined at 10% breakthrough under following conditions:
Column : 50×4.6 mmI.D.Linear velocity :180 cm/hr
for anion-exchange resinEquilibration buffer :20 mM Tris-HCl (pH 8.6)Elution buffer :0.5 M NaCl in equilibration bufferSample :1.5 mg/mL BSA in equilibration bufferDetection :UV at 280 nm
for cation-exchange resinEquilibration buffer :20 mM Glycine-NaOH (pH 9.0)Elution buffer :0.5 M NaCl in equilibration bufferSample :1.5 mg/mL Lysozyme in equilibration bufferDetection :UV at 300 nm
160
180
200
220
240
260
0 200 400 600 800 1000 1200
YMC-BioPro S75Brand T(porous S type)Brand G(porous S type)
DBC(mg/mL-resin)
Linear velocity(cm/hr)
Dependency of DBC to linear velocity
YMC-BioPro ion exchange media show high DBC over a wide range of linear velocity, and the difference of DBC is less than 5% between 200 cm/hr and 1000 cm/hr. YMC-BioPro ion exchange media give increased productivity and reduced cost in biopharmaceutical production.
Column :50×5.0 mmI.D.Equilibration buffer :20 mM Glycine-NaOH (pH 9.0)Elution buffer :0.5 M NaCl in equilibration bufferSample :1.0 mg/mL Lysozyme in equilibration bufferDetection :UV at 300 nm
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Excellent durability under CIP condition with 1 M NaOH
Cleaning in place (CIP) is an important procedure for cleaning and sterilization of columns used for protein purification. The DBC and the selectivity of proteins are unaffected following 20 cycles of CIP with 1 M NaOH. The high chemical stability of BioPro ion exchange media allows effective cleaning with alkaline solution.
Conditions of DBC measurementColumn :YMC-BioPro S75 50×5.0 mmI.D.Linear velocity :800 cm/hrEquilibration buffer :20 mM Glycine-NaOH (pH 9.0)Elution buffer :0.5 M NaCl in equilibration bufferSample :1.0 mg/mL Lysozyme in equilibration bufferDetection :UV at 300 nm※DBC is determined at 10% breakthrough
Test protocols
Separation of standard proteins
Determination of DBC and recovery
CIP with 1 M NaOH(5 column volumes at 400 cm/hr)
DBC and recovery
Number of CIP with 1M NaOH
120
100
60
40
80
250
200
100
50
150
10 20
DBC
Recovery
2 4 6 8 12 14 16 18
DBC(mg/mL-resin)
Recovery(%)
initial
10th
20th
1 2 3
min10 20 30 40 50
Number of CIP
Separation of standard proteins
1. Ribonuclease A2. Cytochrome c3. Lysozyme
Purification of IgY from egg yolk extract
Egg yolk antibody (IgY) can be isolated with high purity more than 99% by two chromatographic purification steps, which consist of a capture step by ion exchange chromatography on YMC-BioPro Q75 and a polishing step by size exclusion chromatography on YMC-Pack Diol-200.
min
mV
0 5 10 15 20 25 30
0
25
50
75
100
YMC-Pack Diol-200 5 µm, 300×8.0 mmI.D.Sample : Fraction from capture purification by IEC
Polishing by size exclusion chromatography (SEC)
Analysis of purified fraction
YMC-Pack Diol-200 5 µm, 300×4.6 mmI.D.
IgY
min
mAU
0 5 10 15 20
0.0
2.5
5.0
7.5
97.2 kDa
66.4 kDa
45.0 kDa
29.0 kDa
20.1 kDa14.3 kDa
Marker
StandardIgY
Non-reduced SDS-PAGEIEC
fractionSEC
fractionCrudeextract
purity >99%
IgY
SEC
Fraction from polishing step by SEC
Eluent : 0.1 M KH2PO4-K2HPO4 (pH 6.9) containing 0.2 M NaCl
Flow rate :0.7 mL/minTemperature :ambientDetection :UV at 280 nmInjection :1 mL (ca. 0.45 mg IgY)
min
mV
0 10 20 30 40 50 60
0
100
200
300
400
500
YMC-BioPro Q75 75 µm, 50×4.6 mmI.D.Sample : Crude extract from egg yolk*
Capture purification by ion exchange chromatography (IEC)
Polishing step
Eluent : A) 20 mM Tris-HCl (pH 8.1) B) 20 mM Tris-HCl (pH 8.1) containing 0.5 M NaCl 10% B (0-15 min), 30% B (15-30 min), 90% B (30-40 min)
Flow rate :0.5 mL/min (180 cm/hr)Temperature :ambientDetection :UV at 280 nmInjection :1 mL (ca. 20 mg Protein)
*Courtesy of Pharma Foods International Co., Ltd.
Packing material Particle size(µm)
Productnumber
Volume50 mL 250 mL 1 L 5 L 25 L
YMC-BioPro Q30 30 QAA0S30 * * * * *YMC-BioPro S30 30 SPA0S30 * * * * *YMC-BioPro Q75 75 QAA0S75 * * * * *YMC-BioPro S75 75 SPA0S75 * * * * *
Ordering Information
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Silica gel SEC
YMC-Pack Diol● 5 µm silica-based column with high mechanical stability● Low-cost size exclusion chromatography (SEC) column● Useful for molecular weight determination of proteins and sugars
S i l i c a - b a s e d s i z e e x c l u s i o n c h r o m a t o g r a p h y ( S E C ) c o l u m nYMC-Pack Diol is a size exclusion chromatography column based dihydroxypropyl-bonded silica, and available in four different pore sizes. Diol-120, 200, and 300 are suitable for separation or molecular weight
determination of proteins with molecular weights of 5,000 to several hundred thousand. Diol-60 is the most suitable for separation of peptides or oligosaccharides whose molecular weights are 10,000 or less.
■Particle size:5 µm■Pore size:60, 120, 200, 300 Å■Usable pH range:5.0 〜 7.5
Column : YMC-Pack Diol 300×8.0 mmI.D.
Eluent :0.1 M KH2PO4-K2HPO4 (pH 7.0) containing 0.2 M NaClFlow rate :0.5 mL/minTemperature :25 ℃Detection :UV at 280 nm
Calibration curves of various proteins for three different pore sizes
5 131197
Elution volume (mL)
MW
104
105
106
Diol-300
Diol-200Diol-120
1
11
14 15
9
87
65
43
2
10
12
16
13
Diol-120, Diol-200 and Diol-300 are suitable for the separation or molecular weight determination of proteins with molecular weights of 5,000 to several hundred thousand.
MW 1. IgM 900,000 2. Thyroglobulin 670,000 3. IgA 390,000 4. Fibrinogen 340,000 5. γ-Globulin 158,000 6. IgG 150,000 7. Transferrin 75,000 8. HSA (human serum albumin) 66,000 9. α1-Antitrypsin 50,00010. Ovalbumin 45,00011. Carbonic anhydrase 30,00012. Trypsin inhibitor 20,10013. Myoglobin 17,00014. α-Lactalbumin 14,10015. Ribonuclease A 13,70016. Cytochrome c 12,400
Separation for standard protein markers
10 20 30
1
54
32
0 min
Diol-120
For molecular weight 10,000 to 500,000 compounds, Diol-200 is suitable for the separation.
Column : YMC-Pack Diol 500×8.0 mmI.D.
Eluent : 0.1 M KH2PO4-K2HPO4 (pH 7.0) containing 0.2 M NaCl
Flow rate :0.7 mL/minTemperature :ambientDetection :UV at 280 nm
MW1. Glutamate dehydrogenase 290,0002. Lactate dehydrogenase 142,0003. Enolase 67,0004. Adenylate kinase 32,0005. Cytochrome c 12,400
1
54
32
10 20 300 40min
Diol-200
1 543
2
10 20 300 40 min
Diol-300
Specifications
Column Base Functional groupPore size (Å)
Particle size
(µm)
Usable pH
rangeCharacteristics
Diol-60
Silica gel Dihydroxypropyl
60
5 5 〜 7.5
For molecular weight below 10,000
Diol-120 120 For molecular weight 5,000 to 100,000
Diol-200 200 For molecular weight 10,000 to ca. 500,000
Diol-300 300 For molecular weight ca. 50,000 to 1,000,000
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Separation for molecular weight below 10,000 peptides
Diol-60
10 20 30 40 min
12
34
0
For molecular weight below 10,000 peptides, Diol-60 is suitable for the separation.
MW1. Insulin (Bovine) 5,7002. Neurotensin 1,6723. Angiotensin II 1,0404. Glycine 75
Column : YMC-Pack Diol-60 500×8.0 mmI.D.
Eluent : 0.1 M KH2PO4-K2HPO4
(pH 7.0) containing 0.2 M NaCl/ acetonitrile (70/30)
Flow rate :0.7 mL/minTemperature :ambientDetection :UV at 215 nm
MW 1. Myoglobin 17,000 2. Ribonuclease A 13,700 3. Cytochrome c 12,400 4. Insulin (Bovine) 5,700 5. Insulin B chain 3,496 6. α-Mating factor 1,684 7. Neurotensin 1,672 8. Angiotensin I 1,296 9. CCK-Octapeptide 1,14310. Bradykinin 1,06011. Angiotensin II 1,04012. Oxytocin 1,00713. Angiotensin III 93114. Met-Enkephalin 57315. Leu-Enkephalin 55516. Tetraglycine 24617. Glycine 75
10 15 20
102
103
104
MW
Elution volume (mL)
1 2
3
4
5
7 68
911 13 10
12
1514
16
17
Separation of oligo- and polysaccharide
Diol-200
10 20 min
1
2
34
0
For separation or molecular weight determination of water-soluble oligo- and polysaccharides, Diol-60, Diol-120, Diol-200, and Diol-300 are useful individually or in combination.
Column :YMC-Pack Diol, 500×8.0 mmI.D.Eluent :waterFlow rate :1.0 mL/minTemperature :ambientDetection :RI
MW 1. Pullulan (P-800) 853,000 2. Pullulan (P-400) 380,000 3. Pullulan (P-200) 186,000 4. Pullulan (P-100) 100,000 5. Pullulan (P-50) 48,000 6. Pullulan (P-20) 23,700 7. Pullulan (P-10) 12,200 8. Pullulan (P-5) 5,800 9. Maltopentadecaose (G15) 2,44810. Maltoundecaose (G11) 1,80011. Maltoheptaose (G7) 1,15212. Maltopentaose (G5) 82413. Maltotriose (G3) 50414. Maltose (G2) 34215. Glucose (G1) 180
MW1. P-800 853,0002. P-50 48,0003. P-20 23,7004. P-5 5,800
10 15 20 25
102
103
104
105
106
MW
Elution volume (mL)
1011
1213
1415
9
8
7
6
5
43
2
1
Diol-60
Diol-300
Diol-200
Diol-120
Analytical columns(Stainless columns)Packing material
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
Diol-60 60 4.6×300 DL06S05-3046WT8.0×300 DL06S05-3008WT8.0×500 DL06S05-5008WT
Diol-120 120 4.6×300 DL12S05-3046WT8.0×300 DL12S05-3008WT8.0×500 DL12S05-5008WT
Diol-200 200 4.6×300 DL20S05-3046WT8.0×300 DL20S05-3008WT8.0×500 DL20S05-5008WT
Diol-300 300 4.6×300 DL30S05-3046WT8.0×300 DL30S05-3008WT8.0×500 DL30S05-5008WT
For preparative columns with an inner diameter of 20 mm or more, see page 153.
Guard columns(Stainless columns)Packing material
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
Diol-60 60 8.0×30 DL06S05-0308WTG
Diol-120 120 8.0×30 DL12S05-0308WTG
Diol-200 200 8.0×30 DL20S05-0308WTG
Diol-300 300 8.0×30 DL30S05-0308WTG
Analytical columns(Glass columns)Diol-60 60 8.0×300 DL06S05-3008FG
8.0×500 DL06S05-5008FG
Diol-120 120 8.0×300 DL12S05-3008FG8.0×500 DL12S05-5008FG
Diol-200 200 8.0×300 DL20S05-3008FG8.0×500 DL20S05-5008FG
Diol-300 300 8.0×300 DL30S05-3008FG8.0×500 DL30S05-5008FG
Ordering information
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Reversed-phase columns
Reversed-phase columns for BIO-MACRO molecular separations● Abundant gel types● Excellent peak shapes● High resolution
Reve r s ed - p ha se co l umns f o r p ep t i d e s , p r o t e i n s a nd n uc l e i c a c i d sFor BIO-MACRO molecular separations, YMC has many choices for reversed-phase HPLC.A listing of the most popular column is found below.
Specifications For products other than below listed, see the "Reversed-phase C18 columns (ODS)" and "Reversed-phase columns (other than ODS)" sections.
Separation for molecular weight below 5,000 (pore size 80 Å, 120 Å)
Column Base Functional group
Pore size(Å)
Particle size(µm)
Carbon content (% )
Usable pH range Characteristics
Triart C18 Hybrid silica C18 120 2, 3, 5 20 1.0 〜12.0 ・Excellent chemical durability
・Superior peak shapes in every condition
Pro
ser
ies
Pro C18
Silica gel
C18
120 2*, 3, 5,10 162.0 〜 8.0
・ Processed with YMC's advanced endcapping technology・ Superior separation for basic compounds
Hydrosphere C18 120 2*, 3, 5 12 ・Superior separation for hydrophilic compounds
・Can be used with 100% water mobile phase
Pro C18 RS 80 3, 5 22 1.0 〜10.0
・Highly durable ODS・ Superior separation for basic compounds
and hydrophobic compounds
YMC-
Pack
serie
s ODS-A 120 3, 5,10,15, 20, 50
172.0 〜 7.5
・Currently in use worldwide・Available from analytical to preparative
ODS-AQ 120 14 ・Superior separation for hydrophilic compoundsPh Phenyl 120 3, 5,10,15, 20 9 ・ Reversed-phase packing material with π electronsPolymerC18 Polymer C18 − 6,10 − 2.0 〜13.0 ・Polymer based ODS
* Materials for 2 µm is YMC-UltraHT products.
Separation for molecular weight 5,000 - 100,000 (pore size 200 Å, 300 Å)
Column Base Functional group
Pore size(Å)
Particle size(µm)
Carbon content (% )
Usable pH range Characteristics
Wid
e po
re c
olum
n
ODS-A
Silica gel
C18200 5,10,
15, 2012
2.0 〜 7.5
・ODS with wide pore size・For separation of peptides and proteins300 7
ODS-AQ C18 200 5,10,15 10 ・Low carbon ODS
C8 C8200 5,10,
15, 207 ・C8 with wide pore size
・For separation of relatively highly hydrophobic compounds300 4
C4 C4200 5,10,
15, 205 ・C4 with wide pore size
・Superior separation for proteins300 3
CN cyano propyl 300 5 3 ・CN with wide pore size
・Unique selectivity due to cyano group
YMCbasic C8 200 3, 5 7 ・ Superior separation for proteins and peptides, especially for insulin
PROTEIN-RP − 200 5 4 1.5 〜 7.5 ・ Specialized column with excellent acid resistance for separation of proteins and peptides
How to select reversed-phase columnsTo separate proteins or peptides, select columns based on the molecular weight of the compounds to be separated is important. As shown in the table on the right, the C18 column with 120 Å pore size is generally suitable for small peptides up to MW 5,000. In the case of large peptides or small proteins up to MW 20,000, the C8 column with 200 Å pore size often gives the best column efficiency. Furthermore, most of proteins are eluted effectively by the C4 column with 300 Å. Separation may also be influenced by the hydrophobicity of the analyte and the type of the functional group as well as molecular weight. If the sufficient separation is not achieved with columns marked with a double circle, perform optimization as indicated by the arrows shown in the table. In addition to columns C18, C8, and C4 shown in the table, PROTEIN-RP and CN type columns with different selectivity are also useful.
Molecular weightof sample
Functionalgroup
Pore sizeC18 C8 C4
120 Å ◎ ○ ○
200 Å ○ ◎ ○
300 Å ○ ○ ◎
5,000
20,000
100,000
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Separation of peptides (MW 573 - 3,465)
Generally, the conventional C18 column with 120 Å pore size is suitable for analysis of small peptides up to 5,000 in molecular weight. Especially Pro series ODS columns, which are processed with advanced endcapping technology, are ideal for separation of basic peptides. As shown in the above, Hydrosphere C18, a Pro series column, exhibits excellent separations and superior peak shapes of basic peptides (peak 1 and 7), in contrast to the commercial ODS column for hydrophilic compounds, Brand E2.
Eluent : A) water/TFA (100/0.1) B) acetonitrile/TFA (100/0.1) 20-40% B (0-15 min), 40% B (15-20 min)
Flow rate :1.0 mL/minTemperature :37 ℃Detection :UV at 220 nm
1. BAM-12P (MW 1,425)2. [D-Ala2,Met5]-Enkephalinamide (MW 587)3. α-Endorphin (MW 1,746)4. Met-Enkephalin (MW 573)5. [D-Ala2,Met5]-Enkephalin (MW 588)6. γ-Endorphin (MW 1,899)7. β-Endorphin (MW 3,465)
Hydrosphere C18(120 Å)5 µm,150×4.6 mmI.D.Hydrosphere C18(120 Å)5 µm,150×4.6 mmI.D.
1
23
45
6
7
0 5 10 15 20 min
A000308B
Excellent peak shapes for basic peptides
Brand E2(100 Å)5 µm,150×4.6 mmI.D.(ODS column for hydrophilic compounds)
Brand E2(100 Å)5 µm,150×4.6 mmI.D.(ODS column for hydrophilic compounds)
1
2 34
5 6
7
0 5 10 15 20 min
A000313D
Separation of peptides and proteins (MW 4,300 - 17,000)
For proteins and peptides with molecular weight of 4,300 to 17,000, separation characteristics are compared using columns with different pore size and functional group. In accordance with the table on the previous page, the suitable column is C8, 200 Å for groups of compounds with a molecular weight within this range. If either pore size or functional group of the packing material is not optimized, peak broadening and poor resolution are observed. By using the most suitable column (C8, 200 Å) for the target compounds, sharp peak shapes and excellent separation are achieved.
Column :5 µm, 150×4.6 mmI.D.Eluent : A) water/TFA (100/0.1)
B) acetonitrile/TFA (100/0.1) 25-60% B (0-20 min)
Flow rate :1.0 mL/minTemperature :37 ℃Detection :UV at 220 nm
1. Cytochrome c (MW 12,400)2. Insulin (Bovine) (MW 5,700)3. Amyloid β-protein (MW 4,300)4. Lysozyme (MW 14,300)5. α-Lactalbumin (MW 14,100)6. Myoglobin (MW 17,000)
C8, 300 Å
C8, 120 Å
04073006.D
min0 5 10 15
04080204.D
1 2
56
4
3
12
35
64
Pore size
Comparison of separation on columns with different pore size and functional group
C4, 200 Å
C18, 200 Å
min0 5 10 1504073002.D
04072910.D
12
35
64
1,2
3 5
64
Functional group
1
2
35
64
min0 5 10 15
04082002.D
C8, 200 Å
Optimized combination of pore size and functional group
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Reversed-phase columns
Reversed-phase columns for BIO-MACRO molecular separations
Separation of proteins (MW 66,000 - 96,000)
Gradient elution of water and acetonitrile containing TFA are often employed in an analysis of proteins and peptides. In some cases, addition of a “third solvent” is effective for change in selectivity and separation. The above example shows the resolution between high-molecular weight proteins (peak 1 and 2) is improved by adding 2-propanol into the standard mobile phase of acetonitrile/water/TFA.
Column : YMC-Pack C4 (5 µm, 300 Å) 150×4.6 mmI.D.
Eluent : 30-75% B (0-15 min), 75% B (15-20 min)
Flow rate :1.0 mL/minTemperature :37 ℃Detection :UV at 220 nm
1. BSA (MW 66,000)2. Conalbumin (MW 77,000)3. Lipoxidase (MW 96,000)
Optimization of eluent conditions (C4, 300 Å)
Separation characteristics of proteins with molecular weight of 66,000 to 96,000 are compared using columns with different pore size and functional group. The columns with smaller pore size, which have the same C4 functional groups, provide broader peak shapes and poor separations. In comparison among the 300 Å pore columns with different functional groups, the longer alkyl chain such as C18 and C8 results in poor resolution. It is important to choose optimal pore size and functional group depending on molecular weight of proteins for better peak shapes and resolutions. Proteins with molecular weight of 20,000 to 100,000 are separated effectively by the C4 column with 300 Å pore size.
1. BSA (MW 66,000)2. Conalbumin (MW 77,000)3. Lipoxidase (MW 96,000)
Comparison of separation on columns with different pore size and functional group
Column :5 µm, 150×4.6 mmI.D.Eluent : A) water/TFA (100/0.1)
B) acetonitrile/2-propanol/TFA(50/50/0.1) 30-75% B (0-15 min), 75% B (15-20 min)
Flow rate :1.0 mL/minTemperature :37 ℃Detection :UV at 220 nm
1
3
2
A) water/TFA(100/0.1)B) acetonitrile/TFA(100/0.1)
05011405.D
0 5 10 15 min
3
1
2
A) water/TFA(100/0.1)B) acetonitrile/2-propanol/TFA(50/50/0.1)
04093007.D
0 5 10 15 min
2-propanoladdition
12
3
12 3
C4, 200 Å
C4, 120 Å
0 4 8 12 16 min
Pore size
05011714.D
05011711.D
1
2
3
C4, 300 Å
04093007.D
12 16 min840
1 23
1 23
0 4 8 12 16 min
C8, 300 Å
C18, 300 Å
Functional group
05011708.D
05011802.D
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Separation of nucleic acid bases and nucleotides
0S991029A
10 20 minA000922A0 5 10 15 20 min
Nucleic acid bases Nucleotides
Hydrosphere C18 is designed for separation of hydrophilic compounds and can be used with 100% aqueous mobile phase. Hydrosphere C18 is favorable to the analyses of highly polar compounds such as nucleic acid bases and nucleotides.
Column : Hydrosphere C18 (5 µm, 120 Å) 150×4.6 mmI.D.
Eluent :20 mM KH2PO4-K2HPO4 (pH 6.9)Flow rate :1.0 mL/minTemperature :37 ℃Detection :UV at 254 nm
Column : Hydrosphere C18 (5 µm, 120 Å) 150×4.6 mmI.D.
Eluent :100 mM KH2PO4-K2HPO4 (pH 5.5)Flow rate :1.0 mL/minTemperature :30 ℃Detection :UV at 260 nm
In analytical scale, many impurities could be separated from the target compound by one-nucleotide difference on Hydrosphere C18. Even in purification scale, YMC-Actus gave superior separation and recovery.YMC-Actus Hydrosphere C18 is useful for purification of hydrophilic compounds such as oligonucleotides, organic acids, oligosaccharides and glycosides.
Eluent : A)10 mM DBAA*(pH 6.0)/methanol(60/40) B)10 mM DBAA*(pH 6.0)/methanol(20/80) 10-35% B(0-30 min)
Temperature :ambientDetection :UV at 269 nmSample :synthetic oligonucleotide(100 µM)* di-n-butylammonium acetate
5'-CCGCTCGAGCTAAAAAAAGCCTGTGTTACC-3'
Purification of highly polar compounds -oligonucleotide-Crude synthetic 30 mer oligonucleotide
Hydrosphere C18 5 µm,50×4.6 mmI.D.
YMC-Actus Hydrosphere C18 5 µm,50×20 mmI.D.
Brand I1 5 µm,50×4.6 mmI.D.
Brand I1 5 µm,50×19 mmI.D.
Analysis Purification1.0 mL/min, 5 µL injection 19 mL/min, 100 µL injection
30 mer
30 mer
Impurities
Recovery 56%■ purity>99%
0 10 20 30 min 20 22.5 25 27.5 min
Recovery 35%■ purity>99%
Impurities
7.5 10 12.50 10 20 30 min min
Separation of oligonucleotides
0 5 10 15 20 25 30 min
T2 T10
T20
T2-20
T10
T20
T20T10
T2
T2
J050214A
J050209B
J050210E
J050209A
Hydrosphere C18, 10 mM TEAA
Pro C18, 10 mM TEAA
Brand I1, 10 mM TEAA
Brand I1, 100 mM TEAA Hydrosphere C18 shows strong retention and excellent resolution of oligonucleotides even at a low concentration of triethylamine compared to ordinary C18 columns.
Column :5 µm, 150×4.6 mmI.D.Eluent : A) 10 mM or 100 mM TEAA* (pH 6.0)
B) 10 mM or 100 mM TEAA* (pH 6.0)/acetonitrile (80/20) 55-61% B (0-30 min)
Flow rate :1.0 mL/minTemperature :35 ℃Detection :UV at 269 nm* triethylammonium acetate
Oligonucleotides d(pT)2-20
1. 5'-UTP 2. 5'-UDP 3. 5'-CMP 4. 5'-UMP 5. 5'-dCMP 6. 5'-GMP 7. 5'-ATP 8. 5'-IMP 9. 5'-ADP10. 5'-TMP11. 5'-AMP
See P138-141 for details of YMC-Actus series.
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Reversed-phase columns
Reversed-phase columns for BIO-MACRO molecular separations
YMC-Pack ODS-A
Analytical columnsParticle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
S-5 200 4.6×150 AA20S05-1546WT4.6×250 AA20S05-2546WT
300 1.0×150 AA30S05-1501WT1.0×250 AA30S05-2501WT1.5×150 AA30S05-15P5WT1.5×250 AA30S05-25P5WT2.0× 75 AA30S05-L502WT2.0×150 AA30S05-1502WT2.0×250 AA30S05-2502WT4.6× 75 AA30S05-L546WT4.6×100 AA30S05-1046WT4.6×150 AA30S05-1546WT4.6×250 AA30S05-2546WT4.6×300 AA30S05-3046WT6.0×100 AA30S05-1006WT6.0×150 AA30S05-1506WT6.0×250 AA30S05-2506WT6.0×300 AA30S05-3006WT
S-10 300 4.6×100 AA30S11-1046WT4.6×150 AA30S11-1546WT4.6×250 AA30S11-2546WT4.6×300 AA30S11-3046WT6.0×100 AA30S11-1006WT6.0×150 AA30S11-1506WT6.0×250 AA30S11-2506WT6.0×300 AA30S11-3006WT
Semi-preparative columnsS-5 200 20×150 AA20S05-1520WT
20×250 AA20S05-2520WT
300 10×150 AA30S05-1510WT10×250 AA30S05-2510WT10×300 AA30S05-3010WT20×150 AA30S05-1520WT20×250 AA30S05-2520WT
S-10 200 20×150 AA20S11-1520WT20×250 AA20S11-2520WT
300 10×150 AA30S11-1510WT10×250 AA30S11-2510WT10×300 AA30S11-3010WT20×150 AA30S11-1520WT20×250 AA30S11-2520WT
See P145 for preparative columns other than those listed above.
Columns shown in above are only wide pore size columns. Details of pore sizes for 80 Å and 120 Å columns, see the pages of reversed-phase columns.
Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack)Particle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
S-5 200 4.0×23 AA20S05-G304CC
300 1.0×10 AA30S05-0101CC1.5×10 AA30S05-01P5CC2.0×10 AA30S05-0102CC4.0×23 AA30S05-G304CC
A guard cartridge holder will need to be purchased separately before using this product for the first time.
Cartridge holderSemi-micro guard cartridge holder(inner diameter: 1.0, 1.5, or 2.0 mm)
XPCHSMW
Cartridge holder(inner diameter: 4.0 mm)
XPCHW
YMC guard cartridge packS-5 200 4.0×23 AA20S05-G304CCPK
300 4.0×23 AA30S05-G304CCPK
A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK).
Guard columnsS-5 200 4.0×10 AA20S05-0104WFG
20×50 AA20S05-0520WTG
300 4.0×10 AA30S05-0104WFG5.0×10 AA30S05-0105WFG10×30 AA30S05-0310WTG20×50 AA30S05-0520WTG
S-10 200 20×50 AA20S11-0520WTG
300 4.0×10 AA30S11-0104WFG5.0×10 AA30S11-0105WFG10×30 AA30S11-0310WTG20×50 AA30S11-0520WTG
Ordering information
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YMC-Pack ODS-AQ
Analytical columnsParticle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
S-5 200 4.6×150 AQ20S05-1546WT4.6×250 AQ20S05-2546WT
Semi-preparative columnsS-5 200 20×150 AQ20S05-1520WT
20×250 AQ20S05-2520WT
S-10 200 20×150 AQ20S11-1520WT20×250 AQ20S11-2520WT
See P146 for preparative columns other than those listed above.
Guard cartridge columns (three-pack)S-5 200 4.0×23 AQ20S05-G304CC
A cartridge holder will need to be purchased separately before using this product for the first time.
Cartridge holderCartridge holder(inner diameter: 4.0 mm)
XPCHW
YMC guard cartridge packS-5 200 4.0×23 AQ20S05-G304CCPK
A guard cartridge pack includes a cartridge holder (1 set), a guard cartridge column, and a column coupler (produced by PEEK).
Guard columnsS-5 200 4.0×10 AQ20S05-0104WFG
20×50 AQ20S05-0520WTG
S-10 200 20×50 AQ20S11-0520WTG
Columns shown in above are only wide pore size columns. Details of pore sizes for 80 Å and 120 Å columns, see the pages of reversed-phase columns.
YMC-Pack CN
Analytical columnsParticle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
S-5 300 1.0×150 CN30S05-1501WT1.0×250 CN30S05-2501WT1.5×150 CN30S05-15P5WT1.5×250 CN30S05-25P5WT2.0×150 CN30S05-1502WT2.0×250 CN30S05-2502WT4.6× 75 CN30S05-L546WT4.6×100 CN30S05-1046WT4.6×150 CN30S05-1546WT4.6×250 CN30S05-2546WT4.6×300 CN30S05-3046WT6.0×100 CN30S05-1006WT6.0×150 CN30S05-1506WT6.0×250 CN30S05-2506WT6.0×300 CN30S05-3006WT
Semi-preparative columnsS-5 300 20×150 CN30S05-1520WT
20×250 CN30S05-2520WTSee P151 for preparative columns other than those listed above.
Guard cartridge columns (three-pack)S-5 300 4.0×23 CN30S05-G304CC
A cartridge holder will need to be purchased separately before using this product for the first time.
Cartridge holderCartridge holder(inner diameter: 4.0 mm)
XPCHW
Guard columnsS-5 300 4.0×10 CN30S05-0104WFG
5.0×10 CN30S05-0105WFG10×30 CN30S05-0310WTG20×50 CN30S05-0520WTG
Ordering information
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Reversed-phase columns
Reversed-phase columns for BIO-MACRO molecular separations
YMC-Pack C8
Analytical columnsParticle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
S-5 200 4.6×150 OC20S05-1546WT4.6×250 OC20S05-2546WT
300 1.0×150 OC30S05-1501WT1.0×250 OC30S05-2501WT1.5×150 OC30S05-15P5WT1.5×250 OC30S05-25P5WT2.0×150 OC30S05-1502WT2.0×250 OC30S05-2502WT4.6× 75 OC30S05-L546WT4.6×100 OC30S05-1046WT4.6×150 OC30S05-1546WT4.6×250 OC30S05-2546WT4.6×300 OC30S05-3046WT6.0×100 OC30S05-1006WT6.0×150 OC30S05-1506WT6.0×250 OC30S05-2506WT6.0×300 OC30S05-3006WT
S-10 300 4.6×100 OC30S11-1046WT4.6×150 OC30S11-1546WT4.6×250 OC30S11-2546WT4.6×300 OC30S11-3046WT6.0×100 OC30S11-1006WT6.0×150 OC30S11-1506WT6.0×250 OC30S11-2506WT6.0×300 OC30S11-3006WT
Semi-preparative columnsS-5 200 20×150 OC20S05-1520WT
20×250 OC20S05-2520WT
300 10×150 OC30S05-1510WT10×250 OC30S05-2510WT10×300 OC30S05-3010WT20×150 OC30S05-1520WT20×250 OC30S05-2520WT
S-10 200 20×150 OC20S11-1520WT20×250 OC20S11-2520WT
300 10×150 OC30S11-1510WT10×250 OC30S11-2510WT10×300 OC30S11-3010WT20×150 OC30S11-1520WT20×250 OC30S11-2520WT
See P148 for preparative columns other than those listed above.
Columns shown in above are only wide pore size columns. Details of pore sizes for 80 Å and 120 Å columns, see the pages of reversed-phase columns.
Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack)Particle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
S-5 200 4.0×23 OC20S05-G304CC
300 1.0×10 OC30S05-0101CC1.5×10 OC30S05-01P5CC2.0×10 OC30S05-0102CC4.0×23 OC30S05-G304CC
A cartridge holder will need to be purchased separately before using this product for the first time.
Cartridge holderSemi-micro guard cartridge holder(inner diameter: 1.0, 1.5, or 2.0 mm)
XPCHSMW
Cartridge holder(inner diameter: 4.0 mm)
XPCHW
Guard columnsS-5 200 4.0×10 OC20S05-0104WFG
20×50 OC20S05-0520WTG
300 4.0×10 OC30S05-0104WFG5.0×10 OC30S05-0105WFG10×30 OC30S05-0310WTG20×50 OC30S05-0520WTG
S-10 200 20×50 OC20S11-0520WTG
300 4.0×10 OC30S11-0104WFG5.0×10 OC30S11-0105WFG10×30 OC30S11-0310WTG20×50 OC30S11-0520WTG
Ordering information
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YMC-Pack C4
Analytical columnsParticle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
S-5 200 4.6×150 BU20S05-1546WT4.6×250 BU20S05-2546WT
300 1.0×150 BU30S05-1501WT1.0×250 BU30S05-2501WT1.5×150 BU30S05-15P5WT1.5×250 BU30S05-25P5WT2.0×150 BU30S05-1502WT2.0×250 BU30S05-2502WT4.6× 75 BU30S05-L546WT4.6×100 BU30S05-1046WT4.6×150 BU30S05-1546WT4.6×250 BU30S05-2546WT4.6×300 BU30S05-3046WT6.0×100 BU30S05-1006WT6.0×150 BU30S05-1506WT6.0×250 BU30S05-2506WT6.0×300 BU30S05-3006WT
S-10 300 4.6×100 BU30S11-1046WT4.6×150 BU30S11-1546WT4.6×250 BU30S11-2546WT4.6×300 BU30S11-3046WT6.0×100 BU30S11-1006WT6.0×150 BU30S11-1506WT6.0×250 BU30S11-2506WT6.0×300 BU30S11-3006WT
Semi-preparative columnsS-5 200 20×150 BU20S05-1520WT
20×250 BU20S05-2520WT
300 10×150 BU30S05-1510WT10×250 BU30S05-2510WT10×300 BU30S05-3010WT20×150 BU30S05-1520WT20×250 BU30S05-2520WT
S-10 200 20×150 BU20S11-1520WT20×250 BU20S11-2520WT
300 10×150 BU30S11-1510WT10×250 BU30S11-2510WT10×300 BU30S11-3010WT20×150 BU30S11-1520WT20×250 BU30S11-2520WT
See P149 for preparative columns other than those listed above.
Columns shown in above are only wide pore size columns. Details of pore sizes for 80 Å and 120 Å columns, see the pages of reversed-phase columns.
Guard cartridge columns (with inner diameter 1.0, 1.5, or 2.0 mm: 2/pack, 4.0 mm: 3/pack)Particle size(µm)
Pore size(Å)
Column sizeinner diameter×length(mm) Product number
S-5 200 4.0×23 BU20S05-G304CC
300 1.0×10 BU30S05-0101CC1.5×10 BU30S05-01P5CC2.0×10 BU30S05-0102CC4.0×23 BU30S05-G304CC
A cartridge holder will need to be purchased separately before using this product for the first time.
Cartridge holderSemi-micro guard cartridge holder(inner diameter: 1.0, 1.5, or 2.0 mm)
XPCHSMW
Cartridge holder(inner diameter: 4.0 mm)
XPCHW
Guard columnsS-5 200 4.0×10 BU20S05-0104WFG
20×50 BU20S05-0520WTG
300 4.0×10 BU30S05-0104WFG5.0×10 BU30S05-0105WFG10×30 BU30S05-0310WTG20×50 BU30S05-0520WTG
S-10 200 20×50 BU20S11-0520WTG
300 4.0×10 BU30S11-0104WFG5.0×10 BU30S11-0105WFG10×30 BU30S11-0310WTG20×50 BU30S11-0520WTG
See P76 for YMCbasic.See P122, 123 for YMC-Pack PROTEIN-RP.
Ordering information
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Reversed-phase columns
YMC-Pack PROTEIN-RP● Improved recovery of proteins or peptides● Improved durability when used with TFA solution● Enables elution of high molecular weight proteins
R e v e r s e d - p h a s e c o l u m n f o r s e p a r a t i o n o f p r o t e i n s o r p e p t i d e sYMC-Pack PROTEIN-RP is a reversed-phase column utilizing a silica gel base. It contains a stationary phase, specifically designed for separation of proteins or peptides. Problems that are associated with
conventional reversed-phase columns with short alkyl chain lengths are minimized.Robust column lifetime and excellent recovery of hydrophobic proteins are typically possible on this phase.
■Carbon content:4%■Usable pH range:1.5 〜 7.5
Recovery of various proteins and peptides is shown.YMC-Pack PROTEIN-RP gives greater recovery than competitive C4 columns.
Improved recovery of proteins or peptidesRecovery of proteins or peptides
SampleRecovery(%)
YMC-PackPROTEIN-RP
C4 column,company A
C4 column,company B
Ribonuclease A 93 95 86Cytochrome c 94 89 95Lysozyme 98 93 76Myoglobin 97 85 93BSA 94 92 86Ovalbumin 90 73 88Transferrin 94 98 88Insulin(bovine) 97 73 92Insulin B chain 82 76 70α-Mating factor 93 82 81Leu-Enkephalin 92 84 91Gly-Gly-Gly-Gly 95 86 70
Improved durability when used with TFA solution
110
100
90
80
70
60
50
40
30
20
10
0100 200 300 400
C4 column, Company B
C4 column, Company A
(Peak split)
YMC-Pack PROTEIN-RP
×
Solvent flow in hours
0.1%TFA condition
%
retention
of
diphenyl
The left graph shows the test result of the stability of the stationary phase with 0.1% aqueous TFA. Retention of diphenyl on C4 columns manufactured by other companies greatly decreases as time passes. This is caused by detachment of butyl groups from the packing material due to acid hydrolysis.Retention of diphenyl on YMC-Pack PROTEIN-RP is shown to be stable after 500 hours of mobile phase flow.
〈Prior to flow〉1
2
3
4
0 5 10 min
Theoretical plate number (4) = 14,300
〈After 500 hours〉
1
2
3
4
0 5 10 min
Theoretical plate number (4) = 13,800
1. Uracil2. Benzene3. Naphthalene4. Diphenyl
〈Flow conditions〉Eluent :water/TFA (100/0.1)Flow rate :1.0 mL/minTemperature :ambient
〈Measurement conditions〉Column :YMC-Pack PROTEIN-RP 250×4.6 mmI.D.Eluent :acetonitrile/water (40/60)Flow rate :1.0 mL/minTemperature :30 ℃Detection :UV at 254 nm, 0.32 AUFS
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0 10 20 30 min
12
3
Proteins1. Cytochrome c (MW 12,400)2. BSA (MW 66,000)3. Ferritin (MW 440,000)
Column :YMC-Pack PROTEIN-RP 250×4.6 mmI.D.Eluent : A) water/TFA (100/0.1)
B) acetonitrile/TFA (100/0.1) 30-90% B (0-45 min, linear)Flow rate :1.0 mL/minTemperature :ambientDetection :UV at 280 nm, 0.04 AUFS
A p p l i c a t i o n
1
23 4 5
67
8
9
1011
12
13
403020100 min
Proteins and peptides 1. Met-Enkephalin (MW 573) 2. Leu-Enkephalin (MW 555) 3. Oxytocin (MW 1,007) 4. Bradykinin (MW 1,060) 5. Angiotensin I (MW 1,296) 6. Ribonuclease A (MW 13,700) 7. α-Mating factor (MW 1,684) 8. Insulin(bovine) (MW 5,700) 9. Cytochrome c (MW 12,400) 10. Lysozyme (MW 14,400) 11. BSA (MW 66,000) 12. β-Lactoglobulin (MW 18,400) 13. Ovalbumin (MW 45,000)
Column :YMC-Pack PROTEIN-RP 250×4.6 mmI.D.Eluent : A) water/TFA (100/0.1)
B) acetonitrile/TFA (100/0.1) 10-90% B (0-60 min, linear)Flow rate :1.0 mL/minTemperature :ambientDetection :UV at 220 nm, 0.32 AUFS
A p p l i c a t i o n
Analytical columnsParticle size
(µm)Column size
inner diameter×length(mm) Product number
S-5 2.0×150 PR99S05-1502WT2.0×250 PR99S05-2502WT4.6×150 PR99S05-1546WT4.6×250 PR99S05-2546WT
Semi-preparative columnsS-5 10×250 PR99S05-2510WT
20×150 PR99S05-1520WT20×250 PR99S05-2520WT
Guard cartridge columns (three-pack)S-5 4.0×23 PR99S05-G304CC
A cartridge holder will need to be purchased separately before using this product for the first time.
Cartridge holderCartridge holder(inner diameter: 4.0 mm)
XPCHW
Guard columnsS-5 4.0×10 PR99S05-0104WFG
10×30 PR99S05-0310WTG20×50 PR99S05-0520WTG
Ordering information
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Normal-phase columns
YMC-Pack Diol-NP● Silica gel bonded with dihydroxypropyl group● Two pore sizes of 120 Å and 60 Å are available● Normal-phase separation using non-polar
solvents
● Useful for hydrophilic interaction chromatography (HILIC)
● Different separation characteristics from bare silica gel
Diol column for non-aqueous Normal-Phase (NP) and Hydrophilic Interaction Chromatography (HILIC) YMC-Pack Diol-NP shows retention behavior of normal-phase chromatography when it is used with low-polarity mobile phases. Hydroxy groups on the surface of the stationary phase act as polar groups. YMC-Pack Diol-NP is as widely applicable to normal-phase separation as silica
gel. It is also useful in cases where separation optimization is difficult to achieve using bare silica gel. In addition, YMC-Pack Diol-NP is usuable as a HILIC mode column when it is used with a mobile phase consisted of a high concentration of organic solvent in water or an aqueous buffer.
■Pore size:120 Å, 60 Å■Usable pH range:2.0 〜 7.5
Separation of highly hydrophilic compounds in HILIC mode
0 105 min15
1
5
4
32
6
R090116P
YMC-Pack Diol-NP shows superior performance as a HILIC mode column when it is used with a mobile phase consisted of a mixture of organic solvent/aqueous solution.
Column : YMC-Pack Diol-NP (5 µm, 120 Å) 150×2.0 mmI.D.
Eluent :water/acetonitrile (10/90) containing 10 mM CH3COONH4
Flow rate :0.2 mL/minTemperature :30 ℃Detection :UV at 254 nm
1. Uracil 2. Uridine 3. Adenosine 4. Cytosine 5. Cytidine 6. Guanosine
NH
HN
O
O
N
HN
OH
O
OH
HOH2CO
O
N
N
N
N
OH
O
OH
HOH2C
NH2
NH
N
O
NH2
N
N
OH
O
OH
HOH2CO
NH2
HN
N
N
N
O
OH
O
OH
HOH2C
H2N
Nucleic acid bases and Nucleosides
Separation of hydrophilic compounds which are hardly retained on reversed-phase column
0 2 4 6 min
A991115E
YMC-Pack Diol-NP(5 µm, 120 Å) 50×4.6 mmI.D.
1
2
3
45
Comparison of peptide separation on YMC-Pack Diol-NP and YMC-Pack ODS-A is shown. YMC-Pack Diol-NP offers favorable resolution of between peak 1 and peak 2, highly hydrophilic peptides which cannot be separated in reversed-phase mode with ODS column. An interaction between hydroxyl groups on the Diol-NP stationary phase and highly hydrophilic compounds provides enhanced retention and superior resolution.
Eluent : A) acetonitrile/TFA/TEA (100/0.1/0.1) B) water/TFA/TEA (100/0.1/0.1) 0-50% B (0-4 min), 50% B (4-5 min)
Flow rate :3.0 mL/minTemperature :ambientDetection :UV at 220 nm
Eluent : A) water/TFA (100/0.1) B) acetonitrile/TFA (100/0.1) 0-30% B (0-4 min), 30% B (4-5 min)
Flow rate :3.0 mL/minTemperature :ambientDetection :UV at 220 nm
1. TRP-GLY (MW 261)2. GLY-GLY-PHE (MW 279)3. Angiotensin I (MW 1,296)4. Substance P (MW 1,348) 5. Insulin (bovine) (MW 5,700)
2,1
3
4
5
0 2 4 6 8 min
A991127I
YMC-Pack ODS-A(5 µm, 120 Å)50×4.6 mmI.D.
※Shipping solvent for Diol-NP is hexane/2-propanol (99.5/0.5). In case of using mobile phase including water, take care of miscibility.※For details and ordering information, see P86, 87.
Peptides
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Normal-phase columns
YMC-Pack Polyamine Ⅱ
A m i n o c o l u m n w i t h i m p r o v e d d u r a b i l i t yYMC-Pack Polyamine Ⅱ is a silica-based packing bonded with polyamine. It is particularly useful for separation of sugars. The column lifetime of YMC-Pack Polyamine Ⅱ in aqueous mobile phase is longer than conventional silica-based amino columns, and thus is
applicable to separation of oligosaccharides using mobile phase with relatively higher water content. In addition, YMC-Pack Polyamine Ⅱ can be used to separate ionic compounds with a combination of normal-phase mode and weak anion exchange mode.
■Pore size:120 Å■Usable pH range:2.0 〜 7.5
● Silica gel chemically bonded with polyamine● The most suitable column for separation of sugars● Solves durability problem of conventional
silica-based amino columns
● Useful for separation of hydrophilic compounds including vitamins
● Useful for separation of fat-soluble compounds using nonaqueous mobile phase
Suitable column for separation of sugars
1
23
10 20 300 minF030618H
4
5
Column : YMC-Pack Polyamine Ⅱ 250×4.6 mmI.D.
Eluent :acetonitrile/water (75/25)Flow rate :1.0 mL/minTemperature :25 ℃Detection :RI, 32×10-6 RIU/FS
※For details and ordering information, see P88, 89.
Elution volume of sugars and sugar alcohols
Arabitol
65
60
55
50
45
40
35
30
25
20
15
10
5
080%75% 85%
Acetonitrile concentration (%)
※Not detected with 85% acetonitrile
Elu
tion
volu
me
(mL)
Inositol
Palatinose
SucraloseSucralose
FucoseFucose
Fructose
Mannitol
Glucose
Galactose(※)
Mannose(※)
Sucrose
Maltitol
Raffinose
Maltose
Sucrose
Maltitol
Raffinose
Maltose
LactoseLactose
Palatinit
Lactitol
Sorbitol
Mannitol
Glucose
Sorbitol
XyloseXylose
Xylitol
GlycerolGlycerol
RhamnoseRhamnose
ErythritolErythritol
Ribitol
Column : YMC-Pack Polyamine Ⅱ 250×4.6 mmI.D.
Temperature :25 ℃
Excellent durability
120
100
80
60
40
20
00 500 1000 1500 2000 2500 3000 3500 4000
Polyamine Ⅱ
Column volumes
Silica based amino column
Rat
e of
initi
al r
eten
tion
of L
acto
se (%
)
YMC-Pack Polyamine Ⅱ shows little change in retention time of lactose after passing 3,000 column volumes of 75% acetonitrile mobile phase. On the other hand, retention time on conventional silica-based amino column decreases to 80% of the initial value continuously from the beginning.
Durability in 75% acetonitrile mobile phase condition
1. Fructose2. Glucose3. Sucrose4. Maltose5. Lactose
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Recommended