Development of indirect competitive ELISA for quantification of mitragynine in Kratom (Mitragyna speciosa (Roxb.) Korth.)

Supattra Limsuwanchote, Juraithip Wungsintaweekul, Niwat Keawpradub, Waraporn Putalun, Satoshi Morimoto, Hiroyuki Tanaka

研究成果: Contribution to journalArticle査読

11 被引用数 (Scopus)

抄録

Monoclonal antibody (MAb) against mitragynine (MG), an analgesic alkaloid from Kratom leaves (Mitragyna speciosa), was produced. MG was coupled to carrier proteins employing either 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS), a zero-length cross linker or a 5-carbon length glutaraldehyde cross linker. To confirm the immunogenicity, the hapten numbers were determined using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Preparation of the MAb was accomplished by the electrofusion method. Hybridoma 1A6 that was constructed from the fusion between splenocytes of EDC/NHS conjugate immunized mice and SP2/0-Ag14 myeloma cells was selected, cloned twice and expanded. The cross-reactivities (CRs) of this MAb 1A6 with a series of indole alkaloids were 30.54%, 24.83% and 8.63% for speciogynine, paynantheine and mitraciliatine, respectively. Using this MAb, an indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed with a measurement range of 32.92-250μg/mL. Quantitative analysis of the MG contents in plant samples by icELISA correlated well with the standard high performance liquid chromatography method (R2=0.994). The MAb against mitragynine provided a tool for detection of MG in Kratom preparations.

本文言語英語
ページ(範囲)70-77
ページ数8
ジャーナルForensic Science International
244
DOI
出版ステータス出版済み - 1 1 2015

All Science Journal Classification (ASJC) codes

  • 病理学および法医学

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