Osr6102, osr6202, osr6602




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Procedure:
ALBUMIN
OSR6102, OSR6202, OSR6602

This procedure is valid for the following chemistry analyzers:

  • AU400/AU400e

  • AU640/AU640e

  • AU480

  • AU680

  • AU600

  • AU2700®/AU5400



Prepared By

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PRINCIPLE:


Serum albumin measurements are used in the diagnosis of numerous diseases. Elevated serum albumin levels are usually the result of dehydration. Decreased serum albumin levels are found in a number of conditions including kidney disease, liver disease, infections, severe burns and cancer.1

INTENDED USE:


System reagent for the quantitative determination of Albumin in human serum or plasma on Beckman Coulter AU Clinical Chemistry analyzers.

Albumin reagent OSR6602 is for use on the AU2700 and AU5400 analyzers only.


METHODOLOGY:


In 1965, Rodkey2 introduced a convenient, direct method for determining albumin concentrations in serum utilizing a neutral buffered solution of bromocresol green (BCG) as the dye binding indicator. In 1971, Doumas et al.3 increased the sensitivity of the reaction by adding a nonionic surfactant to the reagent to prevent turbidity and improve linearity. The Beckman Coulter AU System Albumin method is a modification of the Doumas and Rodkey procedures utilizing a different buffering system.

At pH 4.2, bromocresol green reacts with albumin to form an intense green complex. The absorbance of the albumin-BCG complex is measured bichromatically (600/800nm) and is proportional to the albumin concentration in the sample.



pH 4.2
Albumin + Bromocresol Green Complex

SPECIMEN:

Patient Preparation:


None required.

Additional instructions for patient preparation as designated by this laboratory:

Type:


Serum, heparinized or EDTA plasma samples, free from hemolysis, are the recommended specimens. Separate from blood cells as soon as possible.

Additional type conditions as designated by this laboratory:

Handling Conditions:


Albumin is stable in serum for one week at room temperature (15 - 25°C) and for one month refrigerated (2 - 8°C).4

Additional handling conditions as designated by this laboratory:


EQUIPMENT AND MATERIALS:

Equipment:


Beckman Coulter AU400/AU400e, AU480, AU600, AU640/AU640e, AU680, AU2700, and AU5400 analyzers.

Materials:


Beckman Coulter AU System Albumin Reagent

Final concentration of reactive ingredients:



Succinate buffer (pH 4.2)

100 mmol/L

Bromocresol green

0.2 mmol/L

Also contains preservatives.

Reagent storage location in this laboratory:

Test tubes 12 -16 mm in diameter or sample cups (Cat No. AU1063).

Storage location of test tubes or sample cups in this laboratory:

Beckman Coulter Chemistry Calibrator (Cat. No. DR0070)

Storage location of the calibrator in this laboratory:

Precautions:


  1. For in vitro diagnostic use.

  2. WARNING! IRRITANT! May cause sensitization by skin contact. Avoid contact with skin. In case of external contact, rinse affected areas with plenty of water.

  1. WARNING: POTENTIAL BIOHAZARDOUS MATERIAL. The calibrator is manufactured from human serum. No test method can offer complete assurance that HIV- 1/2, HCV, Hepatitis B, or other infectious agents are absent from biological materials, all calibrator material should be handled at the Biosafety Level 2 as recommended for any infectious human serum or blood specimen in the CDC/National Institutes of Health manual, Biosafety in Microbiological and Biomedical Laboratories, 1993.

Preparation:


The Beckman Coulter AU System Albumin Reagent is liquid, ready for use. No preparation is needed.

For OSR6602, insert the pipe supplied into the 180mL AU reagent vials before use on the analyzer. Care must be taken when handling the pipe to avoid contamination. The pipe is for single use only. Do not remove the large cap on the 180mL bottles.



The Beckman Coulter Chemistry Calibrator reconstitution:

  • Remove the vials of calibrator and diluent from storage and let stand at room temperature (15-25C) for 5 minutes.

  • Remove the cap and stopper from the vials of the lyophilized serum and reconstituting diluent.

  • Using a volumetric pipette or a calibrated air-displacement pipettor, add exactly 5.0 mL of reconstituting diluent to DR0070 lyophilized serum vial. DO NOT pour directly from the reconstituting diluent vial.

  • Replace the cap and stopper to the vial of lyophilized serum immediately after adding the diluent

  • Allow the calibrator to stand for 5-10 minutes. Gently swirl the contents until completely dissolved.

Storage Requirements:


1. The unopened reagents are stable until the expiration date printed on the label when stored at 2 - 25°C.

2. Opened reagents are stable for 90 days when stored in the refrigerated compartment of the analyzer.



  1. Contamination after opening must be avoided.

  2. Un-reconstituted calibrator and diluent are stable until the expiration date stated on the label when stored at 2 - 8°C.

  3. For Albumin, reconstituted calibrator materials are stable for 7 days from the date of reconstitution when stored at 2 - 8°C. The materials should be capped and stored upright 2 - 8°C when not in use



Additional storage requirements as designated by this laboratory:

Indications of Deterioration:


Discoloration of the reagent, visible signs of microbial growth, turbidity or precipitation in reagent may indicate degradation and warrant discontinuance of use.

PERFORMANCE PARAMETERS:


The following data was obtained using this Albumin Reagent on Beckman Coulter AU analyzers according to established procedures. Results obtained at individual facilities may differ.

Precision:9


Estimates of precision, based on CLSI recommendations8, are consistent with typical performance. The within run precision is less than 3%CV and total precision is less than 3%CV. Assays of control sera were carried out and data reduced following CLSI guidelines.

N=100

Within run

Total

Mean, g/dL

SD

CV%

SD

CV%

2.9

0.02

0.8

0.04

1.5

5.35

0.04

0.8

0.07

1.3

Method Comparison:9


Patient samples were used to compare this Albumin Reagent. Representative performance data on AU analyzers is shown in the next table.



Y Method

AU640/AU640e

X Method

AU600

Slope

0.982

Intercept

-0.02

Correlation Coeff. (r)

0.9988

No. of Samples (n)

182

Range (g/dL)

1.5-5.4

Sensitivity:


Typical change in absorbance for 1 g/dL of Albumin is 138 Absorbance.

CALIBRATION:

Standard Preparation:


The frequency of calibration is every 30 days. Calibration of this albumin procedure is accomplished by use of the Beckman Chemistry Calibrator (Cat No DR0070), which is traceable to the College of American Pathology (CAP) Reference Preparation for Serum Protein # 4.

Calibration Procedure:


Recalibration of this test is required when any of these conditions exist:

  1. A reagent lot number has changed or there is an observed shift in control values.

  2. Major preventative maintenance was performed on the analyzer.

3. A critical part was replaced.

QUALITY CONTROL:


During operation of the Beckman Coulter AU analyzer at least two levels of an appropriate quality control material should be tested a minimum of once a day. In addition, controls should be performed after calibration, with each new lot of reagents, and after specific maintenance or troubleshooting steps described in the appropriate AU User’s Guide. Quality control testing should be performed in accordance with regulatory requirements and each laboratory’s standard procedure.

Location of controls used at this laboratory.


ANALYZER PARAMETERS:


A complete list of test parameters and operating procedures can be found in the appropriate User’s Guide and at www.beckmancoulter.com.

CALCULATIONS:


For SI units (g/L), multiply the results by 10.

REPORTING RESULTS:

Reference Ranges:


New Born4: 2.8 - 4.2 g/dL

Recumbent Adult4: 3.5 - 5.0 g/dL

Ambulatory Female4: 3.7 - 5.3 g/dL

Ambulatory Male4: 4.2 - 5.5 g/dL

Beckman Determined Range7: 3.5 - 5.7 g/dL

Expected values may vary with age, sex, diet and geographical location. Each laboratory should determine its own expected values as dictated by good laboratory practice.



Expected reference ranges in this laboratory:

Procedures for Abnormal Results:


Abnormal results are flagged by the listed analyzers according to the normal values entered by the user into the instrument parameters.

Reporting Format:


Results are automatically printed for each sample in g/dL at 37°C.

Additional reporting information as designated by this laboratory:

LIMITATIONS:


The Beckman Coulter AU System Albumin procedure is linear from 1.5 to 6.0 g/dL. Samples exceeding the upper limit of linearity should be diluted and repeated. The sample may be diluted, repeated and multiplied by the dilution factor automatically utilizing the AUTO REPEAT RUN.

Interfering Substances:


Results of studies5 show that the following substances interfere with this albumin procedure.

The criteria for no significant interference is recovery within 10% of the initial value.



Bilirubin:

No significant interference up to 40 mg/dL Bilirubin

Hemolysis:

No significant interference up to 450 mg/dL Hemolysate

Lipemia:

No significant interference up to 800 mg/dL Intralipid*

* Intralipid, manufactured by KabiVitrium Inc., is a 20% IV fat emulsion used to emulate extremely turbid samples.

The information presented is based on results from Beckman Coulter studies and is current at the date of publication. Beckman Coulter Inc., makes no representation about the completeness or accuracy of results generated by future studies. For further information on interfering substances, refer to Young6 for a compilation of reported interferences with this test.



Laboratory specific procedure notes:


REFERENCES:


1. Friedman, R.B. and Young, D.S., Effects of Disease on Clinical Laboratory Tests, Third Edition, AACC Press, 1997.

2. Rodkey, F.L., Clin Chem, 2: 478; 1965.

3. Doumas, B.T., Watson, W.A. and Biggs, H.G., Clin Chem Acta 31: 87-96, 1971.

4. Tietz, N.W., Clinical Guide to Laboratory Tests, Second Edition, W.B.Saunders, 1990.

5. CLSI/NCCLS, Interference Testing in Clinical Chemistry, EP7-P, 1986.

6. Young, D.S., Effects of Drugs on Clinical Laboratory Tests, Fifth Edition, AACC Press, 2000.

7. Beckman Coulter Inc. data on samples collected from 200 blood donors in North Texas.

8. CLSI/NCCLS, Guideline EP5-T2, 1992.



9. Data on file for specific AU analyzers.

© Beckman Coulter, Inc. 2010 Page of
All printed copies are considered to be copies of the electronic original. Rev #:1, Dec 01, 10


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