SYNCHRONSystem(s) ChemistryInformationSheet...Value SD %CV Serum Control1 1 80 3.54g/dL 0.06 1.8 Serum Control2 1 80 7.52g/dL 0.05 0.7 CSF HumanPool 1 80 261.1mg/dL 3.1 1.2 Within-run

SYNCHRON System(s) TPmChemistry Information Sheet© 2014 Beckman Coulter, Inc. All rights reserved.

Total Protein465986

For In Vitro Diagnostic Use

Rx Only

ANNUAL REVIEW

Reviewed by Date Reviewed by Date

PRINCIPLEINTENDED USE

TPm reagent, when used in conjunction with UniCel® DxC 800 System and SYNCHRON® Systems Protein Calibrator,is intended for the quantitative determination of total protein in human serum, plasma or cerebrospinal uid (CSF).

CLINICAL SIGNIFICANCE

Total protein measurements are used in the diagnosis and treatment of diseases involving the liver, kidney or bonemarrow, as well as other metabolic or nutritional disorders.

METHODOLOGY

SYNCHRON System(s) determine total protein concentration by means of a rate biuret method.

A precise volume of sample (8.0 microliters serum or 60 microliters CSF) is injected into the reaction cup containingalkaline copper reagent. The ratio used is one part sample to 79 parts reagent for serum or one part sample to 11 partsreagent for CSF. Proteins in the sample combine with the reagent producing alkaline copper-protein chelate. The ratechange in absorbance is monitored by a detector at 545 nanometers. The observed rate of chelate formation is directlyproportional to the total protein concentration in the sample.

CHEMICAL REACTION SCHEME

Chemistry Information Sheet A18561 AK EN TPmAPRIL 2015 Page 1 of 12

SPECIMENTYPE OF SPECIMEN

Biological uid samples should be collected in the same manner routinely used for any laboratory test.1 Freshly drawnserum, plasma, or CSF are the preferred specimens. Acceptable anticoagulants are listed in the PROCEDURAL NOTESsection of this chemistry information sheet. Whole blood or urine is not recommended for use as a sample.

SPECIMEN STORAGE AND STABILITY

1. Tubes of blood are to be kept closed at all times and in a vertical position. It is recommended that the serum orplasma be physically separated from contact with cells within two hours from the time of collection.2

2. Separated serum or plasma should not remain at room temperature longer than 8 hours. If assays are notcompleted within 8 hours, serum or plasma should be stored at +2°C to +8°C. If assays are not completed within48 hours, or the separated sample is to be stored beyond 48 hours, samples should be frozen at -15°C to -20°C.Frozen samples should be thawed only once. Analyte deterioration may occur in samples that are repeatedlyfrozen and thawed.2

3. CSF specimens should be centrifuged and analyzed without delay.

4. Refer to References (3) for additional information on the effects of preanalytical variables on sample storage andstability. Each laboratory should determine if the recommended requirements are appropriate.

Additional specimen storage and stability conditions as designated by this laboratory:

SAMPLE VOLUME

A lled 0.5 mL sample cup is the optimum volume. For optimum primary sample tube volumes in primary tube samplesand minimum volumes, refer to the Primary Tube Sample Template for your system.

CRITERIA FOR UNACCEPTABLE SPECIMENS

Refer to the PROCEDURAL NOTES section of this chemistry information sheet for information on unacceptablespecimens.

Criteria for sample rejection as designated by this laboratory:

TPm EN Chemistry Information Sheet A18561 AKPage 2 of 12 APRIL 2015

PATIENT PREPARATION

Special instructions for patient preparation as designated by this laboratory:

SPECIMEN HANDLING

Special instructions for specimen handling as designated by this laboratory:

REAGENTSCONTENTS

Each kit contains the following items:

Two Total Protein Reagent Bottles (2 x 2 L)

VOLUMES PER TEST

Sample Volume

Serum/Plasma 8.0 µL

CSF 60 µL

Total Reagent Volume 630 µL

REACTIVE INGREDIENTS

REAGENT CONSTITUENTSCopper Sulfate 8.8 mmol/L

Also non-reactive chemicals necessary for optimal system performance.

GHS HAZARD CLASSIFICATION

Total Protein Reagent DANGER

H314 Causes severe skin burns and eye damage.

P280 Wear protective gloves, protective clothing and eye/faceprotection.


P303+P361+P353 IF ON SKIN (or hair): Rinse skin with water.

P305+P351+P338 IF IN EYES: Rinse cautiously with water for several minutes.Remove contact lenses, if present and easy to do. Continuerinsing.

P310 Immediately call a POISON CENTER or doctor/physician.

Sodium Hydroxide 0.1 - 1%

Safety Data Sheet is available at techdocs.beckmancoulter.com

EUROPEAN HAZARD CLASSIFICATION

Total Protein Reagent C;R35

R35 Causes severe burns.

S26 In case of contact with eyes, rinse immediately withplenty of water and seek medical advice.

S37/39 Wear suitable gloves and eye/face protection.

S45 In case of accident or if you feel unwell, seek medicaladvice immediately (show the label where possible).

MATERIALS NEEDED BUT NOT SUPPLIED WITH REAGENT KIT

SYNCHRON® Systems Protein CalibratorAt least two levels of control materialSaline

REAGENT PREPARATION

No preparation is required.

ACCEPTABLE REAGENT PERFORMANCE

The acceptability of a reagent is determined by successful calibration and by ensuring that quality control results arewithin your facility’s acceptance criteria.

REAGENT STORAGE AND STABILITY

TPm reagent stored unopened at room temperature is stable until the expiration date printed on the bottle label. TPmreagent is stable on instrument for 60 days, unless the expiration date is exceeded.

If the reagent is frozen in transit, thaw completely, warm to room temperature and mix thoroughly by gently inverting thebottle at least 10 times.

NOTICE

Excessive exposure to heat may cause irreversible deterioration of the reagent, asindicated by the appearance of dark brown precipitates. If such precipitation occurs,discard reagent.


Reagent storage location:

CALIBRATIONCALIBRATOR REQUIRED

SYNCHRON® Systems Protein Calibrator

CALIBRATOR PREPARATION

No preparation is required.

CALIBRATOR STORAGE AND STABILITY

SYNCHRON® Systems Protein Calibrator Levels 1 and 2 is stable until the expiration date printed on the calibrator bottleif stored unopened at -15°C to -20°C. Once opened, resealed calibrators stored at +2°C to +8°C are stable for 60 daysunless the expiration date is exceeded.

CAUTION

Because this product is of human origin, it should be handled as though capableof transmitting infectious diseases. Each serum or plasma donor unit usedin the preparation of this material was tested by United States Food and DrugAdministration (FDA) approved methods and found to be negative for antibodiesto HIV and HCV and nonreactive for HbsAg. Because no test method can offercomplete assurance that HIV, hepatitis B virus, and hepatitis C virus or otherinfectious agents are absent, this material should be handled as though capableof transmitting infectious diseases. This product may also contain other humansource material for which there is no approved test. The FDA recommends suchsamples to be handled as specied in Centers for Disease Control’s BiosafetyLevel 2 guidelines.4

Calibrator storage location:

CALIBRATION INFORMATION

1. The system must have valid calibration factors in memory before controls or patient samples can be run.

2. Under typical operating conditions the TPm assay must be calibrated every 14 days or with each new bottle ofreagent and also with certain parts replacements or maintenance procedures, as dened in the UniCel DxC 600/800Systems Instructions for Use (IFU) manual.

3. For detailed calibration instructions, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.


4. The system will automatically perform checks on the calibration and produce data at the end of calibration. In theevent of a failed calibration, the data will be printed with error codes and the system will alert the operator of thefailure. For information on error codes, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.

TRACEABILITY

For Traceability information refer to the Calibrator instructions for use.

QUALITY CONTROLAt least two levels of control material should be analyzed daily. In addition, these controls should be run with each newcalibration, with each new bottle of reagent, and after specic maintenance or troubleshooting procedures as detailed inthe appropriate system manual. More frequent use of controls or the use of additional controls is left to the discretion ofthe user based on good laboratory practices or laboratory accreditation requirements and applicable laws.

The following controls should be prepared and used in accordance with the package inserts. Discrepant quality controlresults should be evaluated by your facility.

NOTICE

Do not use controls containing diethylamine HCl.

Table 1.0 Quality Control Material

CONTROL NAME SAMPLE TYPE STORAGE

TESTING PROCEDURE(S)1. If necessary, load the reagent onto the system.

2. After reagent load is completed, calibration is required.

3. Program samples and controls for analysis.

4. After loading samples and controls onto the system, follow the protocols for system operations.

For detailed testing procedures, refer to the UniCel DxC 600/800 System Instructions For Use (IFU) manual.

CALCULATIONSThe SYNCHRON System(s) performs all calculations internally to produce the nal reported result. The system willcalculate the nal result for sample dilutions made by the operator when the dilution factor is entered into the systemduring sample programming.

REPORTING RESULTSEquivalency between the SYNCHRON LX and UniCel DxC 800 Systems has been established. Chemistry resultsbetween these systems are in agreement and data from representative systems may be shown.


REFERENCE INTERVALS

Each laboratory should establish its own reference intervals based upon its patient population. The reference intervalslisted below were taken from literature and a study performed on SYNCHRON Systems.5

Table 2.0 Reference intervals

INTERVALS SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITSSerum (Adult Ambulatory) 6.4 – 8.3 g/dL 64 – 83 g/L

Serum (Adult Recumbent) 6.0 – 7.8 g/dL 60 – 78 g/L

Literature

CSF 15 – 45 mg/dL 150 – 450 mg/L

SYNCHRON Serum 6.1 – 7.9 g/dL 61 – 79 g/L

INTERVALS SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITSLaboratory

Refer to References (6, 3, 7) for guidelines on establishing laboratory-specic reference intervals.

Additional reporting information as designated by this laboratory:

PROCEDURAL NOTESANTICOAGULANT TEST RESULTS

1. If plasma is the sample of choice, the following anticoagulants were found to be compatible with this method basedon a study of 20 healthy volunteers:

Table 3.0 Compatible Anticoagulants

ANTICOAGULANTLEVEL TESTED FOR IN VITRO

INTERFERENCEAVERAGE PLASMA-SERUM

BIAS (g/dL)Ammonium Heparin 14 Units/mL NSIa

Lithium Heparin 14 Units/mL NSI

Sodium Heparin 14 Units/mL NSIa NSI = No Signicant Interference (within ±0.4 g/dL or 4%).

2. The following anticoagulants were found to be incompatible with this method:


Table 4.0 Incompatible Anticoagulants

ANTICOAGULANTLEVEL TESTED FOR IN VITRO

INTERFERENCE PLASMA-SERUM BIAS (g/dL)a

Potassium Oxalate/SodiumFluoride

2.0 / 2.5 mg/mL -1.3

a Bias is based on worst case instead of average. Plus (+) or minus (-) signs in this column signify positive or negative bias.

LIMITATIONS

1. Plasma samples generally yield values slightly higher than serum samples due to the presence of brinogen in theplasma. Experimental data showed an average increase of 0.3 g/dL with a range 0.1 to 0.5 g/dL.

2. Samples at a Lipemia Index Level of 4 and above should be ultracentrifuged and the analysis performed on theinfranate.

3. Samples with high immunoprotein levels may falsely decrease total protein results. These samples should bediluted one part sample plus one part saline and reanalyzed. The result should be multiplied by two.

4. CSF samples showing evidence of hemolysis should not be used. Hemolysis may cause falsely elevated results.

INTERFERENCES

1. The following substances were tested for interference with this methodology:

Table 5.0 Interferences - Serum/Plasma

SUBSTANCE SOURCE LEVEL TESTED OBSERVED EFFECTa

Bilirubin (unconjugated) Bovine 30 mg/dL NSIb

Hemoglobin RBC hemolysate 500 mg/dL NSI

Lipemia Human Lipemia Index 4 -0.4 g/dL

Carbenicillin Carbenicillin Disodium Salt 2500 µg/mL +0.2 g/dL

MethylbenzethoniumChloride

NAc 0.5 mg/dL -0.2 g/dL

a Plus (+) or minus (-) signs in this column signify positive or negative interference.b NSI = No Signicant Interference (within ±0.4 g/dL or 4%).c NA = Not applicable.

Table 6.0 Interferences - CSF

SUBSTANCE SOURCE LEVEL TESTED OBSERVED EFFECTBilirubin Bovine 1 mg/dL Results suppressed

Methicillin NAa 5000 µg/dL +5 mg/dL

Rifampin NA 0.5 mg/dL +5 mg/dLa NA = Not applicable.

2. Refer to References (8,9,10) for other interferences caused by drugs, disease and preanalytical variables.

PERFORMANCE CHARACTERISTICSANALYTIC RANGE

The SYNCHRON System(s) method for the determination of this analyte provides the following analytical ranges:


Table 7.0 Analytical Range

SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITSSerum or Plasma 1.0 – 12.0 g/dL 10 – 120 g/L

CSF 10 – 1500 mg/dL 100 – 15,000 mg/L

Samples with concentrations exceeding the high end of the analytical range should be diluted (one part sample plus onepart saline) and reanalyzed. The result should be multiplied by two.

REPORTABLE RANGE (AS DETERMINED ON SITE):

Table 8.0 Reportable Range

SAMPLE TYPE CONVENTIONAL UNITS S.I. UNITS

SENSITIVITY

Sensitivity is dened as the lowest measurable concentration which can be distinguished from zero with 95% condence.Sensitivity for TPm determination is 1.0 g/dL (10 g/L) for serum or plasma, and 10.0 mg/dL (100 mg/L) for CSF.

EQUIVALENCY

Equivalency was assessed by Deming regression analysis of patient samples to accepted clinical methods.

Serum or Plasma (in the range of 1.3 to 11.6 g/dL):Y (UniCel DxC Systems) = 0.992X + 0.08

N = 191

MEAN (UniCel DxC Systems) = 6.8

MEAN (SYNCHRON LX Systems) = 6.8

Correlation Coefcient(r)

= 0.996

CSF (in the range of 17 to 1192 g/dL):Y (UniCel DxC Systems) = 1.023X - 1.83

N = 91

MEAN (UniCel DxC Systems) = 426

MEAN (SYNCHRON LX Systems) = 418

Correlation Coefcient(r)

= 1.000

Refer to References (11) for guidelines on performing equivalency testing.

PRECISION

A properly operating SYNCHRON System(s) should exhibit imprecision values less than or equal to the maximumperformance limits in the table below. Maximum performance limits were derived by an examination of the imprecisionof various methods, prociency test summaries, and literature sources.


Table 9.0 Maximum Performance Values

TYPE OFPRECISION SAMPLE TYPE 1 SD CHANGEOVER VALUEa % CV

Serum or Plasma 0.2 g/dL 2.0 g/L 10.0 g/dL 100.0 g/L 2.0Within-run

CSF 5.0 mg/dL 50 mg/L 100 mg/dL 1000 mg/L 5.0

Serum or Plasma 0.3 g/dL 3.0 g/L 10.0 g/dL 100.0 g/L 3.0Total

CSF 7.5 mg/dL 75 mg/L 100 mg/dL 1000 mg/L 7.5a When the mean of the test precision data is less than or equal to the changeover value, compare the test SD to the SD guideline given above to

determine the acceptability of the precision testing. When the mean of the test precision data is greater than the changeover value, compare thetest % CV to the guideline given above to determine acceptability. Changeover value = (SD guideline/CV guideline) x 100.

Comparative performance data for a SYNCHRON LX® System evaluated using the NCCLS Proposed Guideline EP5-T2appears in the table below.12 Each laboratory should characterize their own instrument performance for comparisonpurposes.

Table 10.0 NCCLS EP5-T2 Precision Estimate Method

EP5-T2 CalculatedPoint EstimatesTYPE OF

IMPRECISION SAMPLE TYPENo.

SystemsNo. DataPointsa

Test MeanValue SD %CV

Serum Control 1 1 80 3.54 g/dL 0.06 1.8


CSF Human Pool 1 80 261.1 mg/dL 3.1 1.2

Within-run

CSF Control 1 80 22.9 mg/dL 1.1 4.6



CSF Human Pool 1 80 261.1 mg/dL 4.7 1.8

Total

CSF Control 1 80 22.9 mg/dL 1.4 5.9a The point estimate is based on the pooled data from one system, run for twenty days, two runs per day, two observations per run on an instrument

operated and maintained according to the manufacturer’s instructions.

NOTICE

These degrees of precision and equivalency were obtained in typical testing procedureson a SYNCHRON LX® System and are not intended to represent the performancespecications for this reagent.

ADDITIONAL INFORMATIONFor more detailed information on UniCel DxC Systems, refer to the appropriate system manual.

Beckman Coulter, the Beckman Coulter Logo, Synchron, UniCel and DxC are trademarks of Beckman Coulter, Inc andare registered in the USPTO.

SHIPPING DAMAGE

If damaged product is received, notify your Beckman Coulter Clinical Support Center.


REVISION HISTORY

Revision AE

Revised Quality Control section, and removed the sodium azide warning.

Revision AF

Updated corporate address; updated European Hazard Classication and removed EDTA as an AcceptableAnticoagulant claim.

Revision AG

Added Revision History.

Revision AJ

Removed references to CX and LX systems as they are discontinued effective 12/2013.

Added Beckman Coulter trademark statement and disclaimer.

Revision AK

Added GHS Classication information


REFERENCES1. Tietz, N. W., "Specimen Collection and Processing; Sources of Biological Variation", Textbook of Clinical

Chemistry, 5th Edition, W. B. Saunders, Philadelphia, PA (2005).

2. National Committee for Clinical Laboratory Standards, Procedures for the Handling and Processing of BloodSpecimens Approved Guideline, NCCLS publication H18-A, Villanova, PA (1990).

3. National Committee for Clinical Laboratory Standards, How to Define, Determine, and Utilize Reference Intervalsin the Clinical Laboratory Approved Guideline, NCCLS publication C28-A, Villanova, PA (1995).

4. CDC-NIH, Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, (Washington, D.C.: U.S.Government Printing Office, 2009). (CDC 21-1112)

5. Tietz, N. W., Clinical Guide to Laboratory Tests, 3rd Edition, W. B. Saunders Company, Philadelphia, PA (1995).

6. Tietz, N. W., ed., Fundamentals of Clinical Chemistry, 6th Edition, W. B. Saunders, Philadelphia, PA (2007).

7. Henry, J. B., Clinical Diagnosis and Management by Laboratory Methods, 22nd Edition, W. B. Saunders Company,Philadelphia, PA (2006).

8. Young, D. S., Effects of Drugs on Clinical Laboratory Tests, 5th Edition, AACC Press, Washington, D. C. (2000).

9. Friedman, R. B., Young, D. S.,Effects of Disease on Clinical Laboratory Tests, 4th Edition, AACC Press,Washington, D.C. (2001).

10. Young, D. S., Effects of Preanalytical Variables on Clinical Laboratory Tests, 3rd Edition, AACC Press,Washington, D. C. (2007).

11. National Committee for Clinical Laboratory Standards, Method Comparison and Bias Estimation Using PatientSamples Approved Guideline, NCCLS publication EP9-A, Villanova, PA (1995).

12. National Committee for Clinical Laboratory Standards, Precision Performance of Clinical Chemistry DevicesTentative Guideline, 2nd Edition, NCCLS publication EP5-T2, Villanova, PA (1992).

Beckman Coulter Eurocenter S.A., 22, rue Juste-Olivier. Case Postale 1044, CH - 1260 Nyon 1, SwitzerlandTel: +41 (0)22 365 36 11

Beckman Coulter, Inc., 250 S. Kraemer Blvd., Brea, CA 92821 U.S.A.


Documents

SYNCHRONSystem(s) ChemistryInformationSheet...Value SD %CV Serum Control1 1 80 3.54g/dL 0.06 1.8 Serum Control2 1 80 7.52g/dL 0.05 0.7 CSF HumanPool 1 80 261.1mg/dL 3.1 1.2 Within-run