Albumin Reagent

BCG Method
PRODUCT SUMMARY Symbols in Product Labelling
Stability : Until Expiry at 2-25°C Authorized Representative Temperature Limitation
Linear Range : Up to 60 g/L (6.0 g/dL)
For in vitro diagnostic use Use by/Expiration Date
Specimen Type : Serum
Method : Endpoint Batch code/Lot number
CAUTION. Consult instructions
Reagent Preparation : Supplied ready to use. Catalogue number for use.
Manufactured by
Consult instructions for use
Exclamation Mark

INTENDED USE If eye irritation persists: Get medical advice/attention

This reagent is intended for the in vitro quantitative determination of Albumin Precautionary Statements - Storage
in human serum. None
Precautionary Statements - Disposal
CLINICAL SIGNIFICANCE1 None
Albumin is quantitatively the major single contributor to the plasma total protein Hazards not otherwise classified (HNOC)
and performs a number of functions including: Not applicable
(i) Regulation of the distribution of extracellular fluid Unknown Toxicity
(ii) Acts as a transport agent for a wide variety of substances such as hormones, 0.0185% of the mixture consists of ingredient(s) of unknown toxicity
lipids, vitamins, calcium and trace metals and Other information
(iii) Forms part of the amino acid pool. Harmful to aquatic life
Measurement of total protein levels alone may be misleading, and may be normal Interactions with Other Chemicals
in the face of quite marked changes in the constituent proteins. For example - a No information available.
fall in albumin may roughly be balanced by a rise in immunoglobulin levels. This Refer to the product Safety Data Sheet for additional information.
is quite a common combination.
True hyperalbuminaemia probably does not occur and an increase in albumin REAGENT PREPARATION
concentration is usually only encountered in dehydration due to the reduced The reagent is supplied ready to use.
plasma water content or artefactually, as a result of venous stasis during
venipuncture (Most common cause). STABILITY AND STORAGE
Hypoalbuminaemia occurs as a result of - When stored at 2 - 25°C the reagent is stable until the expiration date stated on the
(i) Overhydration due to water excess, bottle and kit box label.
(ii) Excessive protein loss through the skin after severe burns, from the kidney
in the nephrotic syndrome and through the intestine in protein losing Indications of Reagent Deterioration:
enteropathy, • Turbidity;
(iii) Decreased synthesis due to dietary deficiency, liver disease or malabsorption • Presence of a precipitate; and/or
or, • Failure to recover control values within the assigned range.
(iv) Increased catabolism.
SPECIMEN COLLECTION AND HANDLING
METHODOLOGY Serum: Use non-haemolysed serum collected without prolonged venous stasis.
Several procedures are currently available for the determination of albumin Storage: Specimens are stable for at least 20 days when stored at 4°C.
and include dye binding methods, electrophoresis, immunological and salt
fractionation. ADDITIONAL EQUIPMENT REQUIRED BUT NOT PROVIDED
The most commonly used procedures are the dye binding methods of which • If required, pipettes for accurately dispensing measured volumes.
Bromocresol Green (BCG) is the most popular. However, one of the major • A clinical chemistry analyzer capable of maintaining constant temperature (37°C)
drawbacks of this method is its lack of specificity. Despite the many published and measuring absorbance at 630nm.
modifications, existing BCG methods still tend to overestimate low concentrations • Analyzer specific consumables, eg: sample cups.
of albumin2,3 due to non specific reactions with other plasma proteins. • Normal and Abnormal assayed control material.
This kit is based on the method of Doumas et al4 in which albumin binds with • Calibrator or suitable aqueous Albumin standard.
BCG causing a shift in the absorption spectra of the dye. The dye -albumin
complex formed has an absorbance peak at 625nm which is proportional to the ASSAY PROCEDURE
concentration of albumin in the sample. These instructions are for manual instrumentation but can be adapted to most
automated instruments. Specific instructions are available upon request.
REAGENT COMPOSITION
Active Ingredients Concentration SYSTEM PARAMETERS
Succinate buffer 90 mmol/L Temperature 37°C
Bromocresol Green 0.26 mmol/L Wavelength 630nm
Assay Type Endpoint
pH 4.10 ± 0.1 at 20°C Direction Increase
Sample : Reagent Ratio 1:100
Reagent also contains surfactant and stabilizers necessary for optimum reagent eg: Sample Vol 3µL
performance. Reagent Vol 300µL
Incubation Time 90 seconds
Hazard Symbol: Exclamation Mark Reagent Blank Limits Low 0.0 AU
Signal Word: Warning (630nm, 1cm lightpath) High 2.0 AU
Hazard Statements Linearity 0 - 60 g/L (0.0 - 6.0 g/dL)
H319 Causes serious eye irritation (refer to linearity section)
Precautionary Statements - Prevention Analytical Sensitivity 0.03DAbs per g/L
Wash face, hands and any exposed skin thoroughly after handling (630nm, 1cm lightpath) (0.3DAbs per g/dL)
Wear protective gloves/protective clothing/eye protection/face protection
Wear eye/face protection
Precautionary Statements - Response Calculations
Eyes Results are calculated, usually automatically by the instrument, as follows:
IF IN EYES: Rinse cautiously with water for several minutes. Remove contact
lenses, if present and easy to do. Continue rinsing
 Absorbance of Unknown EXPECTED VALUES6
Albumin = ——————————— ­x Calibrator Value Ambulant Male 35-48g/L (3.5 - 4.8 g/dL)
Absorbance of Calibrator Ambulant Female 33-45g/L (3.3 - 4.5 g/dL)
Example:
Absorbance of calibrator = 1.2 In non ambulatory hospitalised patients the haemodilution of recumbency may reduce
Absorbance of unknown = 0.62 the albumin levels by up to 5g/L. The quoted values were derived from non selected
Value of calibrator = 40 g/L (4.0 g/dL) male (100) and female (100) blood donors and should serve as a guide only. It is
0.62 recommended that each laboratory verify this range or derives a reference interval
Albumin = —— x 40 = 21 g/L for the population that it serves.7
(g/L) 1.2
0.62 PERFORMANCE DATA
Albumin = —— x 4.0 = 2.1 g/dL The following data was obtained using the Albumin reagent on an automated clinical
(g/dL) 1.2 chemistry analyzer.

NOTES IMPRECISION:
1. The reagent and sample volumes may be altered proportionally to Imprecision was evaluated using two levels of commercial control and following the
accommodate different spectrophotometer requirements. NCCLS EP5-T procedure.8
2. The temperature of the reaction is not critical, however the temperature of
the spectrophotometer should be held constant. Within Run LEVEL I LEVEL II
3. Final absorbance readings should be taken less than 90 seconds after Number of data points 80 80
sample addition. Mean (g/L / g/dL) 28 / 2.8 44 / 4.4
4. Decreasing the sample volume to reagent volume ratio to 1:200 will increase SD (g/L / g/dL) 0.47 / 0.05 0.6 / 0.06
the observed linearity to 100 g/L or 10 g/dL. Subsequently, sensitivity will CV (%) 1.7 1.5
be reduced. Between Day LEVEL I LEVEL II
5. Unit conversion: g/L x 0.1 = g/dL. Number of data points 80 80
Mean (g/L / g/dL) 28 / 2.8 40 / 4.0
CALIBRATION SD (g/L / g/dL) 0.6 / 0.06 1.1 / 0.11
Calibration is required. A suitable bovine or human Albumin Standard(s) or a CV (%) 2.1 2.5
serum based calibrator, with an assigned value traceable to a primary standard
(eg NIST or IRMM) is recommended. For calibration frequency on automated METHOD COMPARISON
instruments, refer to the instrument manufacturers specifications. Comparison studies were carried out using another commercially available BCG
However, calibration stability is contingent upon optimum instrument performance method for Albumin as a reference. Normal and abnormal human serum samples
and the use of reagents which have been stored as recommended in the stability were assayed in parallel and the results compared by least squares regression. The
and storage section of this package insert. Recalibration is recommended at following statistics were obtained.
anytime if one of the following events occurs:-
• The Lot number of reagent changes. Number of sample pairs 55
• After preventative maintenance is performed or a critical component is Range of results 7 - 48 g/L (0.7 - 4.8 g/dL)
replaced. Slope 0.935
• Control values have shifted or are out of range and a new vial of control Intercept 1.7 g/L (0.17 g/dL)
does not rectify the problem. Correlation coefficient 0.979

QUALITY CONTROL LINEARITY

To ensure adequate quality control, normal and abnormal control with assayed When run as recommended the assay is linear to 60 g/L (6.0 g/dL).
values should be run as unknown samples:-
• At least every eight hours. ANALYTICAL SENSITIVITY
• When a new bottle of reagent is used. When run as recommended the sensitivity of the assay is 0.03DA per g/L (0.3DA
• After preventative maintenance is performed or a critical component is per g/dL).
replaced.
Control results falling above the upper limit or below the lower limit of the REFERENCES
established ranges indicate the assay may be out of control. 1. “Plasma Proteins and Immunoglobulins” in Clinical Chemistry in Diagnosis and
The following corrective actions are recommended in such situations:- Treatment. Lloyd-Luke 1979; Chap XIV:305-10.
• Repeat the same controls. 2. Ferreria P, Price CP. Clin Chem Acta 1974;55:259.
• If repeated control results are outside the limits, prepare fresh control serum 3. Webster D, Clin Chem 1974;53:109.
and repeat the test. 4. Doumas BT, Arends RL, Pinto PC in Standard Methods of Clinical Chemistry
• If results are still out of control, recalibrate with fresh calibrator, then repeat 1972; 7:175-189
the test. 5. Young DS, Effects of Drugs on Clinical Laboratory Tests. Third Edition 1990;12-
• If results are still out of control, perform a calibration with fresh reagent, then 6.
repeat the test. 6. Tietz Textbook of Clinical Chemistry and Molecular Diagnosis (4th Ed.) Burtis,
• If results are still out of control, contact Technical Services or your local Ashwood & Bruns (Eds), Elsevier Saunders, 2005; 2254.
distributor. 7. Wachtel M et al, Creation and Verification of Reference Intervals. Laboratory
Medicine 1995; 26:593-7.
LIMITATIONS 8. National Committee of Clinical Laboratory Standards. User evaluation of Precision
1. Studies to determine the level of interference from haemoglobin, bilirubin Performance of Clinical Chemistry Devices NCCLS 1984; NCCLS publication
and lipaemia were carried out and the following results were obtained: EP5-T.
Haemoglobin: No interference from haemoglobin up to 540mg/dL.
Bilirubin: No interference from bilirubin up to 340µmol/L (20mg/dL).
Lipaemia: No interference from lipaemia, measured as triglycerides, up to
15.7 mmol/L (1380mg/dL).
2. Young DS5 has published a comprehensive list of drugs and substances
which may interfere with this assay.

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TR36026/1105-500 2x 250 mL
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