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基于离子液体微波辅助萃取和硅胶整体柱,采用高效液相色谱法快速测定猫爪草中氧化吲哚生物碱。

Rapid determination of oxindole alkaloids in cat's claw by HPLC using ionic liquid-based microwave-assisted extraction and silica monolithic column.

作者信息

Chang Chih-Wei, Yeh Yu-Ying, Chang Li-Ching, Hsu Mei-Chich, Wu Yu-Tse

机构信息

School of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan.

Department of Occupational Therapy, I-Shou University, Kaohsiung, Taiwan.

出版信息

Biomed Chromatogr. 2017 Aug;31(8). doi: 10.1002/bmc.3925. Epub 2017 Jan 24.

Abstract

Cat's claw is a large woody vine with hook-like thorns, and has been traditionally used to treat inflammatory disorders in South and Central America. In this study, a rapid, validated high-performance liquid chromatographic (HPLC) method using a silica monolithic column was developed for the simultaneous determination of oxindole alkaloids, namely rhynchophylline, pteropodine, isomitraphylline and isopteropodine, in cat's claw. The ionic liquid-based microwave-assisted extraction (ILMAE), considered as an environmentally friendly and powerful tool, was first applied in the extraction of oxindole alkaloids. To optimize the HPLC method, the stationary phases, pH values of mobile phase and flow rates were investigated. The validated HPLC method using a Monolithic RP column (100 × 4.6 mm) enables these analytes to be separated almost twice as fast as with a conventional particulate column (~16 vs ~30 min) with limits of quantification and detection of 0.5 and 0.15 μg/mL, respectively. The ILMAE conditions were optimized by the Taguchi orthogonal array design. In comparison with conventional water boiling extraction, ILMAE offers almost four times higher yields within an extremely short extraction time. The developed HPLC coupled with ILMAE method could be efficient and practical for rapid determination of oxindole alkaloids in cat's claw.

摘要

猫爪草是一种长有钩状刺的大型木质藤本植物,在南美洲和中美洲传统上被用于治疗炎症性疾病。在本研究中,开发了一种使用硅胶整体柱的快速、经过验证的高效液相色谱(HPLC)方法,用于同时测定猫爪草中的氧化吲哚生物碱,即钩藤碱、翅果定碱、异钩藤碱和异翅果定碱。基于离子液体的微波辅助萃取(ILMAE)被认为是一种环保且强大的工具,首次应用于氧化吲哚生物碱的萃取。为了优化HPLC方法,研究了固定相、流动相的pH值和流速。使用整体反相柱(100×4.6mm)的经过验证的HPLC方法能够使这些分析物的分离速度几乎比传统颗粒柱快一倍(约16分钟对约30分钟),定量限和检测限分别为0.5和0.15μg/mL。通过田口正交阵列设计对ILMAE条件进行了优化。与传统的水煮萃取相比,ILMAE在极短的萃取时间内产量几乎高出四倍。所开发的HPLC与ILMAE联用方法对于快速测定猫爪草中的氧化吲哚生物碱可能是高效且实用的。

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