Determination of Mitragynine in Mitragyna speciosa Raw Materials and Finished Products by Liquid Chromatography with UV Detection: Single-Laboratory Validation

Mitragyna speciosa (kratom) is a tree indigenous to Southeast Asia, and its leaves are used in herbal formulations because they contain indole alkaloids mitragynine and 7-hydroxy (7-OH) mitragynine. An HPLC method was developed, optimized, and validated using single-laboratory validation guidelines...

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Bibliographic Details
Published in:Journal of AOAC International 2017-01, Vol.100 (1), p.18-24
Main Authors: Mudge, Elizabeth M, Brown, Paula N
Format: Article
Language:English
Subjects:
Chemical properties
Chromatography, High Pressure Liquid
Composition
Dietary supplements
Identification and classification
Liquid chromatography
Methods
Mitragyna - chemistry
Opioids
Secologanin Tryptamine Alkaloids - analysis
Ultraviolet radiation
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Summary:Mitragyna speciosa (kratom) is a tree indigenous to Southeast Asia, and its leaves are used in herbal formulations because they contain indole alkaloids mitragynine and 7-hydroxy (7-OH) mitragynine. An HPLC method was developed, optimized, and validated using single-laboratory validation guidelines to quantify mitragynine in kratom raw materials and finished products. The method optimization evaluated several extraction parameters including solvent type, solvent volume, time, and extraction method. The separation of the mitragynine alkaloids was achieved in 18 min with a fused-core C18 EVO column using gradient separation with ammonium bicarbonate (pH 9.5) and acetonitrile. The calibration range for mitragynine was 1.0-500 μg/mL with correlation coefficients of ≥99.9% throughout method development and validation. The method detection limit and LOQ were 0.2 and 0.6 μg/mL, respectively for mitragynine. Eight test samples were obtained to evaluate method repeatability. RSDr ranged from 0.4 to 1.0%, whereas intermediate precision ranged from 3.7 to 7.3%, with HorRat values from 0.68 to 1.96. 7-OH mitragynine was below the LOQ for all samples, therefore, spikes repeatability sample RSD values were
ISSN:1060-3271
1944-7922
DOI:10.5740/jaoacint.16-0220