To determine the sildenafil in adulterated functional foods, a rapid direct competitive enzyme-linked immunosorbent assay (ELISA) procedure employing a polyclonal antibody generated from sildenafil-KLH was established. The antibody showed high sensitivity and specificity in phosphate
buffer, with an IC50 of 6±0.1 ng/mL and the limit of detection was 0.6±0.02 ng/mL. The matrix effect of the functional food samples was easily eliminated by using methanol and distilled water for extraction and dilution with PBS, without any clean-up procedure such
as solid phase extraction or liquid–liquid extraction, much more simple and rapid than the other reports. For the validation of the established SIL–ELISA, the functional food sample spiked at three different concentrations, analysed by HPLC. The results showed good correlation
between the data of ELISA and HPLC (r
2=0.9832). It approved that HPLC methods show the same results with ELISA, and the developed immunoassay method is considered reliable for detection of sildenafil.
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Document Type: Research Article
Tianjin Key Laboratory of Food Nutrition and Safety, Faculty of Food Engineering and Biotechnology,Tianjin University of Science and Technology, Tianjin,300457, PR China
December 1, 2012
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