Determination of Biotin Content in Milk Powder by Immunoaffinity Column Purification-HPLC
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Abstract:
The Immunoaffinity column purification - high performance liquid chromatographic method was developed in this paper for the determination of biotin in milk powder. Sample was dissolved with phosphate buffer and purified through biotin immunoaffinity column with ethanol, and concentrated to a direct determination after enrichment. UV detector wavelength was of 215 nm and C18 column was used at 35 ° C. Mobile phase is 50 mmol/L phosphate buffer salt (pH 3.5)-methanol (85:15, v/v) at a flow rate of 1.0mL/min. Standard curve regression equation was y=343.2+220.797x with a correlation coefficient (r) being of 0.9958. The recovery was 90.7% and RSD was of 2.62%. The method detection limit was 1.0 μg/kg. Compared to the national standard method, the method had the advantages of easy sample handling, high sensitivity, good reproducibility, short analysis time to meet modulation milk powder and infant formula milk powder biotin content determination.
