Immunoaffinity Column Cleanup with Liquid Chromatography Using Postcolumn Bromination for the Determination of Aflatoxins in Black and White Sesame Seed: Single-Laboratory Validation

Liu G., Zhu Z., Cheng J., Senyuva H. Z.

JOURNAL OF AOAC INTERNATIONAL, vol.95, no.1, pp.122-128, 2012 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 95 Issue: 1
  • Publication Date: 2012
  • Doi Number: 10.5740/jaoacint.11-150
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.122-128
  • Middle East Technical University Affiliated: Yes


A single-laboratory validation was conducted to establish the effectiveness of an immunoaffinity column cleanup procedure followed by LC with fluorescence detection for the determination of aflatoxins B-1, B-2, G(1), and G(2) in sesame seeds. The sample is homogenized with 50% water (w/w) to form a slurry, then the test portion is extracted with methanol-water (60 + 40, v/v) using a high-speed blender. The sample extract is filtered, diluted with 15% Tween 20 in phosphate-buffered saline solution, and applied to an immunoaffinity column. Aflatoxins are removed with neat methanol, then directly determined by RP-LC with fluorescence detection using postcolumn bromination (Kobra cell). Test portions of blank white sesame seed slurry were spiked with a mixture of aflatoxins to give total levels of 4 and 10 mu g/kg. Recoveries for individual and total aflatoxins ranged from 92.7 to 110.3% for spiked samples. Based on results for spiked sesame paste (triplicates at two levels), the RSD for repeatability (RSDr) averaged 1.1% for total aflatoxins and 1.4% for aflatoxin B-1. The method was demonstrated to be applicable to naturally contaminated samples of black and white sesame seeds obtained from local markets in China.