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| Determination of Sildenafil in Health-care Foods by Ultrasensitive Chemiluminescence Enzyme Immunoassay |
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| DOI： |
| KeyWord:sildenafil direct competitive chemiluminescence enzyme immunoassay（dcCLEIA） health-care food horseradish peroxidase （HRP） |
| Author | Institution |
| CHENG Zhi，ZHANG Yong-yi，YANG Jin-yi，XU Zhen-lin，WANG Hong，LEI Hong-tao，SUN Yuan-ming，SHEN Yu-dong |
College of Food Science，South China Agricultural University，Guangdong Provincial Key Laboratory of Food Quality and Safety，Guangzhou ，China |
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| Abstract: |
| To address the issues of sildenafil adulteration in health-care foods，a direct competition chemiluminescence enzyme immunoassay method（dcCLEIA） was developed for the detection of sildenafil in so-called health-care foods in this work．Horseradish peroxidase（HRP） and luminol were used as signal output system．Based on specific polyclonal antibody，the optimal assay conditions were finalized as follows： the concentration of coating antigen was 41.67 ng/mL，the concentration of antibody was 1.25 μg/mL，the content of Tween－20 in blocking stock solution and washing buffer was 0.05%，the content of methanol in sildenafil dilution solution was 5% and the competitive reaction time was 40 min．Under the optimal conditions，the dcCLEIA method for sildenafil detection was established．The assay showed a half-inhibition concentration（IC50） of 0.17 ng/mL，with a linear detection range（IC20－IC80） of 0.024－1.21 ng/mL，and the limit of detection （LOD，IC10） was 0.008 ng/mL．Meanwhile，the cross-reactivity of dcCLEIA towards tadalafil analogues was less than 0.1%．The recovery test results showed that the average recoveries of the developed dcCLEIA for sildenafil in samples ranged from 82.0% to 114%，with relative standard deviation（RSD） less than 15%．The detection results for random blind samples by dcCLEIA had a good correlation with those of high performance liquid chromatography tandem mass spectrometry（HPLC－MS/MS）．The developed dcCLEIA method was accurate and reliable，and it was suitable for the rapid detection of sildenafil in real samples. |
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