- In tube T, add 0.02 ml of serum sample and in blank add 0.02 ml of distilled
water. - Then add 4 ml of BCG solution in all the tubes and immediately read at 620 nm.
Reagents Blank S 1 S 2 S 3 S 4 S 5 T
Normal saline (ml) 1 1 1 1 1 1 1
BSA (ml) – 0.02 0.02 0.02 0.02 0.02 –
BSA concentration (g/dl) – 12345 –
BSA amount (mg) – 12345 –
Serum (ml) ––––––0.02
Distilled water (ml) 0.02 ––––––
BCG Bromo cresol green (BCG)
solution (ml)4 44444410.11 Calculations
Plot a graph between amount of BSA at x-axis and absorbance at y-axis. Then
extrapolate quantity of albumin in given serum sample from the graph. The albumin
concentration in serum sample can also be calculated by using the following
equation:
Amount of albumin in serum¼ODof testamount of standard mgðÞ 100
ODof standardvolume of sample mlðÞ 1000
¼xg=dl10.12 Clinical Significance
The normal levels of total proteins in serum are 5.5–8.0 g/dl. The total protein
concentration may be altered by changes in plasma volume. Increase in protein
concentration may occur in dehydration, due to increased globulin synthesis, acute
or chronic infection, and multiple myeloma. The decrease in protein levels occurs
due to low protein intake, starvation, malnutrition, and decreased synthesis due to
liver diseases or nephrotic syndrome.
The normal serum albumin value is 3.5–5.5 g/dl and globulin is 2.0–3.5 g/dl. The
albumin/globulin (A/G) ratio is 1.5:2.5 (serum globulin levels are measured by
subtracting serum albumin levels from total serum proteins). In a very low or high
A/G ratio, further testing must be performed to determine the cause and diagnosis. A
low A/G ratio may indicate autoimmune disease, multiple myeloma, cirrhosis, or
kidney disease, while high A/G ratio suggests genetic deficiencies or leukemia.
Albumin amount increases in dehydration state, the condition is counteracted in
vomiting and diarrhea, and this may impair intestinal absorption of amino acids, so,
10.12 Clinical Significance 47