Development of an Enzyme Immunoassay Using a Monoclonal Antibody against the Psychoactive Diterpenoid Salvinorin A

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• 作者：Madan Kumar Paudel ; Osamu Shirota ; Kaori Sasaki-Tabata ; Hiroyuki Tanaka ; Setsuko Sekita ; Satoshi Morimoto
• 刊名：Journal of Natural Products
• 出版年：2013
• 出版时间：September 27, 2013
• 年：2013
• 卷：76
• 期：9
• 页码：1654-1660
• 全文大小：343K
• 年卷期：v.76,no.9(September 27, 2013)
• ISSN：1520-6025

文摘

Salvinorin A (1), the main active constituent in Salvia divinorum, is a highly selective kappa-opioid receptor agonist with hallucinogenic effects, which is regulated in several countries. In the present study, a monoclonal antibody (mAb) against 1 was prepared, and an indirect competitive enzyme-linked immunosorbent assay (icELISA) system was developed for the detection of salvinorins. To raise mAbs against 1, salvinorin B (2) hemisuccinate was synthesized and used to prepare the immunogen 2鈥揵ovine serum albumin conjugate. This technique was used to prepare a hybridoma cell line, 3D5, which secreted a mAb that recognized 1. The mAb was shown to have specificity for 1 and other salvinorins in cross-reactivity tests. The intra-assay calibration range by icELISA using the mAb against 1 was 0.0195鈥?.625 渭g/mL. After validating the icELISA using intra- and interassays, a recovery experiment and analysis of several plants in the family Lamiaceae, including S. divinorum, confirmed that the analytical method based on ELISA is not only simple but also precise, accurate, sensitive, and sufficiently reliable. The results indicate that icELISA is a useful tool in the identification of S. divinorum.