Preparation and Identification of Sudan I Immunoaffinity Column

Abstract

Abstract: A Sudan I immunoaffinity chromatographic column (IAC) was prepared by carbodiimide method, in which a modified Sepharose 4FF containing carboxyl group was used as the carrier to couple with a monoclonal antibody against Sudan I. A reversed-phase HPLC method was established to determine Sudan I using a mobile phase composed of 0.1% formic water-acetonitrile-0.1% formic acetonitrile-acetone (21.25:3.75:60:15, V/V) at a flow rate of 1 mL/min. The detection wavelength was 478 nm. The retention time of Sudan I was found to be 7.97 min. The prepared IAC column as a conjugate between 0.3 g of Sepharose 4FF and 1 mg of purified anti-Sudan I monoclonal antibody showed a capacity of 1.6μg. The average recovery rates of Sudan I from chili powder spiked with Sudan I standard at levels varying from 0.25 to 3 mg/kg was 44.52%～77.40% with relative standard deviation of 4.6%～8.3%. The detection limit was 15 ng/mL based on a signal-to-noise ratio of 3:1. Therefore, a Sudan I IAC column has been prepared successfully.

Key words: Sudan I, immunoaffinity column, high performance liquid chromatography (HPLC)

CLC Number:

O658
TS207.3

References

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