Quality Assessment of selected liriodenine bearing plants endemic to Thailand

Liriodenine, an aporphine isoquinoline alkaloid, has many biological activities, including anti-platelet, anti-fungal and anti-microbial actions. Additionally previous studies revealed that it had potent cytotoxicity against a number of cancer cell lines. This study aimed to determine liriodenine content in selected Thai medicinal plants by TLC image analysis compared to HPLC. Twenty-eight plant materials in Magnoliaceae, Annonaceae and Nelumbonaceae were collected from natural sources in Thailand. Crude extracts were prepared by Soxhlet extraction with 95% ethanol. Validations of both methods were performed including linearity, accuracy, precision and sensitivity based on International Conference of Harmonization (ICH) guideline. Top three highest contents of liriodenine were found in Michelia longifolia (bark), Michelia champaca (bark) and Nelumbo nucifera (leaf) which are also used as crude drug in various Thai traditional recipes. Fifteen different locations throughout Thailand of each medicinal plant were then examined for pharmacognostic specification to establish their standardization for quality assessment. Pharmacognostic parameters from Michelia longifolia bark revealed that the acid-insoluble ash, total ash, loss on drying and water content should be not more than 3.49, 5.74, 7.17 and 6.67 % of dry weight respectively; while ethanol and water-soluble extractive should be not less than 3.85 and 8.25 % of dry weight respectively. Michelia champaca bark showed that the acid-insoluble ash, total ash, loss on drying and water content should be not more than 2.98, 6.25, 5.62 and 7.37 % of dry weight respectively; while ethanol and water-soluble extractive should be not less than 6.71 and 12.38 % of dry weight respectively. Pharmacognostic parameters of Nelumbo nucifera leaves revealed that the acid-insoluble ash, total ash, loss on drying and water content should be not more than 2.61, 9.62, 7.69 and 7.06 % of dry weight respectively; while ethanol and water-soluble extractive should be not less than 6.24 and 9.51 % of dry weight respectively. In summary, calibration showed good linear correlation coefficients (R2 > 0.995) over the range of concentration 5-200 µg/mL. Method validation of both methods was successfully developed performing reliability and sensitivity. TLC image showed a well-defined fluorescent spot of liriodenine at the Rf value of 0.75 under UV 365 nm, while HPLC chromatogram indicating liriodenine peak at 11 min of retention time. Both proposed methods could be used as a tool for the quantification of liriodenine in medicinal plants. There was no significant difference between the results of both analytical methods (p>0.05). TLC image could be applied for analysis of the content of this compound because it is simple, rapid and inexpensive. Moreover, the fluorescent coloring spot which was a dominant characteristic of liriodenine on TLC plate allowed TLC image analysis to be more suitable and convenient for further development.鹅掌楸碱（Liriodenine）是一种阿朴菲类异喹啉生物碱，具备多种生物学活性，包括抗血小板、抗真菌及抗微生物作用。此外，既往研究表明其对多种癌细胞系具有强效细胞毒性。 本研究旨在通过薄层色谱（TLC）图像分析法，并与高效液相色谱（HPLC）法对比，测定选定泰国药用植物中的鹅掌楸碱含量。 本研究从泰国本土自然生境中采集了木兰科、番荔枝科及莲科的28份植物样品，采用95%乙醇通过索氏提取法制备粗提物。依据国际人用药品注册技术协调会（ICH）指南，对两种方法开展了包括线性、准确度、精密度及灵敏度在内的方法学验证。 鹅掌楸碱含量排名前三的样品分别为长叶含笑（Michelia longifolia，树皮）、黄兰（Michelia champaca，树皮）及莲（Nelumbo nucifera，叶），上述原料亦被用于多种泰国传统方剂中作为原料药。随后，针对每种药用植物在泰国全境的15个不同采集点的样品进行了生药学规格检测，以建立其质量评价的标准化体系。 长叶含笑树皮的生药学参数显示：其酸不溶性灰分、总灰分、干燥失重及水分含量分别不得超过干重的3.49%、5.74%、7.17%及6.67%；而乙醇浸出物与水浸出物含量分别不得少于干重的3.85%及8.25%。 黄兰树皮的生药学参数为：酸不溶性灰分、总灰分、干燥失重及水分含量分别不得超过干重的2.98%、6.25%、5.62%及7.37%；乙醇浸出物与水浸出物含量分别不得少于干重的6.71%及12.38%。 莲叶的生药学参数为：酸不溶性灰分、总灰分、干燥失重及水分含量分别不得超过干重的2.61%、9.62%、7.69%及7.06%；乙醇浸出物与水浸出物含量分别不得少于干重的6.24%及9.51%。 综上，在5~200 μg/mL的浓度范围内，校准曲线的线性相关系数良好（R²>0.995）。两种方法的方法学验证均顺利完成，具备可靠的稳定性与灵敏度。 薄层色谱图像显示，在365nm紫外光下，鹅掌楸碱在比移值（Rf）0.75处呈现出清晰的荧光斑点；而高效液相色谱图则显示鹅掌楸碱的保留时间为11分钟。所建立的两种方法均可作为药用植物中鹅掌楸碱含量定量分析的工具，且二者检测结果无显著性差异（p>0.05）。 薄层色谱图像分析法因操作简便、快速且成本低廉，可用于该化合物的含量分析。此外，鹅掌楸碱在薄层色谱板上的标志性特征为荧光显色斑点，这使得薄层色谱图像分析法更适于后续开发与应用。

搜集汇总

背景与挑战

背景概述

该数据集是关于泰国特有植物中liriodenine（一种具有抗癌等生物活性的异喹啉生物碱）的质量评估研究。研究采用薄层色谱（TLC）图像分析和高效液相色谱（HPLC）方法，对来自泰国自然来源的28种植物样本（涵盖木兰科、番荔枝科和莲科）进行liriodenine含量测定，并建立了三种高含量植物（Michelia longifolia树皮、Michelia champaca树皮和Nelumbo nucifera叶子）的生药学标准化参数，以支持其质量评估。