AOAC SPDS Set 2 ERP

Method Review Form

AOAC INTERNATIONAL Standards Development

Name of Reviewer: Christine R. Casey Title of Method: Identification and Characterization of Indole and Oxindole Alkaloids from Leaves of Mitragyna speciosa Hroth Using Liquid Chromatography - Accurate QTof Mass Spectrometry

AOAC Candidate Method Number: KRA-03

Applicable SMPR : AOAC SMPR 2015.008

I.

SUMMARY OF METHOD

The analytical method was developed to characterize and qualitatively determine the alkaloids from various M. speciose samples. The method uses two separate extraction procedures: a simple methanol extraction and an acid-base extraction procedure. Qualitative determination was performed via a RP C8 column with a water-acetonitrile formic acid mobile phase followed by high resolution mass spectrometry. The overall objective of the method was to develop a single qualitative LC-MS/MS ( QTof)method for the separation, characterization, and chemical profiling of indoles and oxindole alkaloids in Mirtagynine speciosa. REVIEW OF THE METHOD ONLY: 1. Does the applicability of the method support the applicability of the SMPR? If no, please explain what is missing. Overall, the method does not meet all the requirements to support the AOAC SMPR 2015.008. Presently, the method does not preform quantitation for 7-hydroxy mitragynine, mitragynine, system suitability, repeatability, and reproducibility. However, the method does separate 7- hydroxy mitragynine, mitragynine, and other relevant indoles of Mitragynine speciose.

II.