Development of direct competitive enzyme-linked immunoassay for sildenafil analysis in health foods
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摘要: 建立了一种在保健食品中检测西地那非的直接竞争酶联免疫吸附分析法(dc ELISA)。经优化最佳反应条件为:包被浓度为100ng/m L,酶标抗体稀释倍数为70倍,标准品稀释液为p H=9,0.01mol/L PBST溶液,抗体竞争反应50min。本方法的半抑制浓度IC50为1.47ng/m L,线性范围(IC20~IC80)为0.12~17.65ng/m L,检测限IC15为0.0845ng/m L,批内批间变异系数均小于20%,保健食品补肾糖浆的添加回收率在86.0%~111.0%之间,满足实际样品的快速筛检需求。
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关键词:
- 西地那非 /
- 酶联免疫吸附分析法(dc ELISA) /
- 保健食品
Abstract: A direct competitive enzyme- linked immunoassay ( dc ELISA) was developed for determination of sildenafil. The results showed that the optimized parameters were as follows :100ng/m L for coating antigen concentration, 70 times dilution of enzyme conjugated antibody, dilution solution of sildenafil in 0.01mol/L PBST (p H =9) and reaction time for 50 min. This developed method presented an IC50 of 1.47ng/m L, a linear range (IC20~IC80) of 0.12 ~17.65ng/m L, a detection limit IC15 of 0.0845ng/m L. Intra-and inter-batch relative standard deviations (RSD) were below 20%. The recoveries of sildenafil in spiked samples ranged from 86.0%~111.0%.In conclusion, the dc ELISA was a practical method for trace detection of sildenafil in real samples.-
Keywords:
- sildenafil /
- dc ELISA /
- health food
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