For the Determination of Albumin in Human Serum or Plasma
At a certain pH value, albumin is specifically combined with bromcresol green, to produce a coloured complex, which is photometrically measured
- R 1 :
- Phosphate buffer pH 4.0 —– 70 mmol/L
- BCG ———— 0/25 mmol/L
- Standard: ——- 4.0 g/dL
Test Requirements :
- Test Tubes
- Bio-Chemistry analyzer
- SGPT Test Kit
Reagent Preparation and Stability :
All Reagents are Ready to use reagents
Specimen collection and handling:
Analyzer Parameter Required :
|Reaction Type||End point|
|Cuvette Temp||37°C, 20 – 25°C|
|Zero Setting||reagent blank|
|Light Path||1 cm|
Pipette into clean dry test tube labeled as Sample (S), Calibrator Label (C)m Blank as (B) and QC Label (QC) or as your required:
|Specimin||(B)||(S)||(C) / (QC)|
|R1||1.0 ml (1000 µL )||1.0 ml (1000 µL )||1.0 ml (1000 µL )|
|Calibrator or QC||–||–||10 µL|
|Dist. water||10 µL||–||–|
|Mix well and incubate for 3 min at 37 ºC or 5 min at room temperature (20 – 25 °C). After incubation, zero the Photometer with the reagent blank. Read and record the incubated standard and samples.|
CALCULATION : With standard or calibrator
Albumin g/dL = ∆A Sample / ∆A Std/Cal x 4(Conc std/cal g/dL)
NOTE: Samples having a very low or high activity show a very low initial absorbance as most of the NADH is consumed prior to the start of the measurement. If this is suspected then dilute the sample and repeat the assay
The procedure is linearity is 7.5 g/dl and depends on KIT.
if the activity exceeds this limits dilute the sample with normal saline (NaCl 0.9 %) and multiply result by dilution factor.
QUALITY CONTROL :
For accuracy it is necessary to run known controls with every assay.
Normal Value :
Serum : 3.5 to 5.3 g/dL.
Note: Each Laboratory or Each Company Kits should establish it’s own normal range.
Possible References Used