Department of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology (IHBT), Palampur, HP, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, Uttar Pradesh (H.P.), India.
Phytochem Anal. 2022 Oct;33(7):1121-1134. doi: 10.1002/pca.3164. Epub 2022 Jul 6.
Aconitum spp. are prime medicinal plants rich in alkaloids and have been used as the main constituents of traditional medicine in India and China. The whole plant can be toxic and creates pathophysiological conditions inside the human body. Therefore, simultaneous quantification of alkaloids within plant parts and herbal medicines associated with this genus is essential for quality control.
We aimed to develop and validate methods using ultra-high-performance liquid chromatography-diode array detector-quadrupole time-of-flight ion mobility mass spectrometry (UHPLC-DAD-QTOF-IMS) and to develop an analytical strategy for the identification and quantification of alkaloid compounds (aconitine, hypaconitine, mesaconitine, aconine, benzoylmesaconitine, benzoylaconine, bulleyaconitine A, and deoxyaconitine) from Aconitum heterophyllum.
We developed a simultaneous identification and quantification method for eight alkaloids using UHPLC-DAD-QTOF-IMS. The method was validated as per International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines and also in IMS mode.
The developed method has good linearity (r = 0.997-0.999), LOD (0.63-8.31 μg/mL), LOQ (0.63-2.80 μg/mL), recovery (86.01-104.33%), reproducibility, intra- and inter-day variability (<3.25%), and stability. Significant qualitative and quantitative variations were found among different plant parts (flower, leaf, stem, root, and tuber) and five market products of A. heterophyllum. Furthermore, a total of 21 metabolites were also profiled based on the fragmentation pattern of MS using the validated method.
An appropriate mobile phase using acetonitrile and water in a gradient elution gave a satisfactory chromatographic separation of eight Aconitum alkaloids with their adjacent peaks. Therefore, this method could provide a scientific and technical platform for quality control assurance.
乌头属植物是富含生物碱的主要药用植物，在中国和印度被用作传统药物的主要成分。整株植物都可能有毒，并在人体内造成病理生理状况。因此，对该属植物部分和草药中生物碱的同时定量对于质量控制至关重要。
我们旨在开发和验证使用超高效液相色谱-二极管阵列检测器-四极杆飞行时间离子淌度质谱（UHPLC-DAD-QTOF-IMS）的方法，并开发一种用于鉴定和定量乌头属植物中生物碱化合物（乌头碱、次乌头碱、新乌头碱、乌头原碱、苯甲酰乌头原碱、苯甲酰新乌头碱、野牛刁碱 A 和脱氧乌头碱）的分析策略。
我们开发了一种使用 UHPLC-DAD-QTOF-IMS 同时鉴定和定量八种生物碱的方法。该方法按照国际人用药品注册技术协调会（ICH）指南进行了验证，并在 IMS 模式下进行了验证。
所开发的方法具有良好的线性（r=0.997-0.999）、LOD（0.63-8.31μg/mL）、LOQ（0.63-2.80μg/mL）、回收率（86.01-104.33%）、重现性、日内和日间变异性（<3.25%）和稳定性。不同植物部位（花、叶、茎、根和块茎）和五种 A.heterophyllum 市售产品之间存在显著的定性和定量差异。此外，还根据经验证的方法对 MS 的碎片模式进行了 21 种代谢物的分析。
使用乙腈和水在梯度洗脱中的适当流动相实现了八种乌头生物碱及其相邻峰的满意色谱分离。因此，该方法可以为质量控制提供科学和技术平台。
