SEC for albumin solution

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prabhakarthota
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SEC for albumin solution

Hai ,
i am confused with the following statement,
"The area of the peak due to polymers and aggregates is not greater than 10 per cent of the total area of the chromatogram (corresponding to about 5 per cent of polymers and aggregates)".
can any one tell me the acceptance criteria for polymers and aggregates.
(For example if polymer and aggregate is 8% by area normalisation the sample complies or doesnt)
Here i am attaching the monograph of Albumin solution from BP2007
"Molecular size distribution

Liquid chromatography (2.2.29). Test solution Column: — size: l = 0.6 m, Ø = 7.5 mm, or l = 0.3 m, Ø = 7.8 mm, — stationary phase: hydrophilic silica gel for chromatography R, of a grade suitable for fractionation of globular proteins with relative molecular masses in the range 10 000 to 500 000. Mobile phase Flow rate Detection The peak due to polymers and aggregates is located in the part of the chromatogram representing the void volume. Disregard the peak due to the stabiliser. The area of the peak due to polymers and aggregates is not greater than 10 per cent of the total area of the chromatogram (corresponding to about 5 per cent of polymers and aggregates).

 Spectrophotometer at 280  nm.0.5 ml/min. Dissolve 4.873 g of  disodium hydrogen phosphate dihydrate R, 1.741 g of sodium dihydrogen phosphate monohydrate R, 11.688 g of sodium chloride R and 50 mg of sodium azide R in 1 litre of water R. Dilute the preparation to be examined with a 9  g/l solution of sodium chloride R to a concentration suitable for the chromatographic system used. A concentration in the range of 4-12 g/l and injection of 50-600 µg of protein are usually suitable.

prabhakarthota
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Hi all,

Hi all,
I am waiting for a answer for the question posted by me frm a long time,
Here I got a good answer

" the molecuar weight of dimer of albumin is 2 times of the monomer. and for polymer its multiple times of the monomer. So when area is 10% of total it means half of this is present as dimer because for the detection wavelenght of 280 nm, there is no difference between monomer and dimer and polymer. so they asume all dimer and polymer to be 2 time the monomer. Finally they say that 10% area right so our product passes this criteria because its 10% area and ours is 7.5% so product passes with 3.75% polymers."