Hayeema Toveelah, Wungsintaweekul Juraithip
Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Prince of Songkla University, Hat Yai Campus, Songkhla, Thailand.
Phytochem Anal. 2025 Jan;36(1):296-306. doi: 10.1002/pca.3442. Epub 2024 Aug 28.
Kratom (leaves from Mitragyna speciosa Korth.; Rubiaceae) is a herbal medicine known for its analgesic properties and psychoactive effects. Kratom in Thailand is currently legal; however, it is prohibited in some countries and considered a narcotic plant.
Our aim was to establish a reliable, simple, and rapid method for quantifying mitragynine in Kratom leaves and related products through a combination of high-performance thin-layer chromatography (HPTLC) and densitometry.
A densitometric HPTLC method was developed and validated in terms of specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, precision, and robustness. The fingerprints of kratom leaves, Mitragyna spp., and related products were constructed.
For HPTLC, samples were applied to silica gel 60 F plates, and the mobile phase comprised n-hexane, ethyl acetate, and triethylamine (1:1:0.15, v/v/v). Densitometric detection was carried out under ultraviolet light at a wavelength of 226 nm. The validated method exhibited a range of 14.31-143.10 μg/mL, yielding a correlation coefficient of 0.9993. Spiked recovery rates were within a range of 98.3%-100.9%, and the LOD and LOQ were 3.80 and 11.53 μg/mL, respectively. Kratom samples were analyzed with the developed method, and the correlation coefficient was 0.9641, compared to the high-performance liquid chromatography-diode-array detection (HPLC-DAD) method. The HPTLC fingerprints displayed a distinctive pattern, facilitating discrimination among different plant parts and Mitragyna spp.
The established method offers the advantages of simplicity, ease of use, and speed of analysis, serving as a practical alternative for mitragynine quantification in kratom leaf and its related products.
kratom（大托叶帽柱木叶；茜草科）是一种以其止痛特性和精神活性作用而闻名的草药。kratom在泰国目前是合法的；然而，在一些国家被禁止，并被视为麻醉植物。
我们的目的是通过结合高效薄层色谱法（HPTLC）和密度测定法，建立一种可靠、简单且快速的方法来定量kratom叶及相关产品中的帽柱木碱。
开发了一种密度测定HPTLC方法，并在特异性、线性、检测限（LOD）、定量限（LOQ）、准确性、精密度和稳健性方面进行了验证。构建了kratom叶、帽柱木属植物及相关产品的指纹图谱。
对于HPTLC，将样品点样于硅胶60 F板上，流动相由正己烷、乙酸乙酯和三乙胺（1:1:0.15，v/v/v）组成。在波长226 nm的紫外光下进行密度测定检测。验证后的方法线性范围为14.31 - 143.10 μg/mL，相关系数为0.9993。加标回收率在98.3% - 100.9%范围内，LOD和LOQ分别为3.80和11.53 μg/mL。用所开发的方法对kratom样品进行分析，与高效液相色谱 - 二极管阵列检测（HPLC - DAD）法相比，相关系数为0.9641。HPTLC指纹图谱显示出独特的模式，便于区分不同的植物部位和帽柱木属植物。
所建立的方法具有简单、易用和分析速度快的优点，可作为定量kratom叶及其相关产品中帽柱木碱的实用替代方法。
