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.