Determination of Sudan I by High Performance Liquid Chromatography with Immuno-Affinity Column Clean-Up

Abstract:

To prepare an immuno-affinity column (IAC) for Sudan I, Sepharose 4FF was activated by epichlorohydrin, and the density of epoxy is 80 μmol/g. Then it was coupled with 6-aminocaproic acid. The Sudan I immunoaffinity column was prepared by carbodiimide method in which modified Sepharose 4FF was used as carrier to couple with monoclonal antibody against Sudan I. A reversed-phase high performance liquid chromatographic method was used to determine the samples elution after the IAC cleaning up. Sudan I was determined by HPLC with a retention time of 5.82 min (flow rate of 1 ml/min). The column capacity is 1582.8 ng Sudan I when using 0.3 g Sepharose 4FF. Average recoveries of Sudan I from red chili powder spiked at levels of 0.25-3 mg/kg range from 44.52% to 77.40%. The coefficient of variation is 4.6% to 8.3%. The IAC method was found to be highly effective, sensitive and convenient for isolating the target analyte from sample.

• Periodical: Advanced Materials Research (Volumes 781-784)
• Pages: 1694-1699
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.781-784.1694
• Online since: September 2013
• Authors: Li Xin Zhu, Wei Meng, Xue Mei Qiu, Ren Rong Liu, Ling Xu, Fu Yong Xu
• Keywords: High Performance Liquid Chromatography (HPLC), Immuno-Affinity Column, Sudan i

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