title: Development of an enzyme immunoassay using a monoclonal antibody against the psychoactive diterpenoid salvinorin A.
abstract: 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-bovine 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 0.0195-0.625 μg/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.
authors.affiliation: Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University , 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
authors.forename: Madan Kumar
authors.initials: MK
authors.lastname: Paudel
authors.forename: Osamu
authors.initials: O
authors.lastname: Shirota
authors.forename: Kaori
authors.initials: K
authors.lastname: Sasaki-Tabata
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Setsuko
authors.initials: S
authors.lastname: Sekita
authors.forename: Satoshi
authors.initials: S
authors.lastname: Morimoto
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D004224:Diterpenes; D045785:Diterpenes, Clerodane / Q000737:chemistry / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008297:Male; D008807:Mice, Inbred BALB C; D015394:Molecular Structure; D018515:Plant Leaves / Q000737:chemistry; D027544:Salvia / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D004224:Diterpenes; D045785:Diterpenes, Clerodane; C508456:salvinorin B; C090499:salvinorin A