A design of experiments approach to production of medicinal bioactives in Mitragyna speciosa (Korth.) Havil.

Abstract

Context: Mitragyna speciosa (Korth.) commonly known as 'kratom' is a tropical tree native to southeast Asia, that produces psychoactive monoterpenoid indole alkaloids including mitragynine. Kratom products are growing in popularity for increasing energy, improving mood and alleviating minor pain, but availability of commercial plant material is limited in North America. Propagation relies primarily on shoot cuttings rooted in soils with high levels of humus and is relatively inefficient.

Objective: This study aimed to optimize in vitro shoot induction system for kratom using a Design of Experiments (DOE) approach and compare the metabolite profiles of in vitro and greenhouse-grown plants using metabolomics.

Materials and methods: A DOE approach was used to minimize the number of treatments necessary to optimize in vitro culture media to induce shoot and root morphogenesis in kratom explants. A clean mother stock of in vitro cultures was established and evaluated for medicinal bioactives via targeted and untargeted analyses by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS).

Results: We found no significant difference in the quantified levels of mitragynine, melatonin, and some related metabolites, between in vitro-grown and greenhouse-grown plants within the experimental conditions of this study.

Discussion and conclusions: The presence of similar specialized metabolite profiles in greenhouse and in vitro-grown kratom plants paves the way for further studies on biotechnological engineering and production of these valuable compounds. In vitro propagation is a viable approach to produce bioactive compounds in kratom plants.