用于甜叶菊中甜菊糖苷的验证高效薄层色谱法。

Validated high-performance thin-layer chromatography method for steviol glycosides in Stevia rebaudiana.

作者信息

Jaitak Vikas, Gupta A P, Kaul V K, Ahuja P S

机构信息

Natural Plant Products Division, Institute of Himalayan Bioresource Technology (CSIR), Palampur 176061, H.P., India.

出版信息

J Pharm Biomed Anal. 2008 Aug 5;47(4-5):790-4. doi: 10.1016/j.jpba.2008.03.022. Epub 2008 Mar 28.

DOI:10.1016/j.jpba.2008.03.022

PMID:18456448

Abstract

A high-performance thin-layer chromatographic (HPTLC) method was developed and validated as per ICH (International Conferences on Harmonization) guidelines for simultaneous quantification of three steviol glycosides, i.e. steviolbioside, stevioside and rebaudioside-A in Stevia rebaudiana leaves. For achieving good separation, mobile phase of ethyl acetate-ethanol-water (80:20:12, v/v/v) on pre-coated silica gel 60 F254 HPTLC plates were used. The densitometric quantification of steviol glycosides was carried out at lambda=510 nm in reflection-absorption mode after spraying with acetic anhydride:sulphuric acid:ethanol reagent. The calibration curves were linear in the range of 160-960 ng/spot for steviolbioside, 1-6 microg/spot for stevioside and 0.5-3 microg/spot for rebaudioside-A with good correlation coefficients (0.998-0.999). The method was found to be reproducible for quantitative analysis of steviol glycosides in S. rebaudiana leaves collected from ten different locations and will serve as a quality control indicator to monitor the commercial production of stevioside and its allied molecules during different stages of its processing.

摘要

根据国际协调会议（ICH）指南，开发并验证了一种高效薄层色谱（HPTLC）方法，用于同时定量测定甜叶菊叶片中的三种甜菊糖苷，即甜菊双糖苷、甜菊糖苷和莱鲍迪苷A。为实现良好的分离效果，在预涂硅胶60 F254 HPTLC板上使用乙酸乙酯 - 乙醇 - 水（80:20:12，v/v/v）的流动相。用乙酸酐：硫酸：乙醇试剂喷雾后，在反射吸收模式下于λ = 510 nm处对甜菊糖苷进行密度定量。甜菊双糖苷的校准曲线在160 - 960 ng/斑点范围内呈线性，甜菊糖苷在1 - 6 μg/斑点范围内呈线性，莱鲍迪苷A在0.5 - 3 μg/斑点范围内呈线性，相关系数良好（0.998 - 0.999）。该方法对于从十个不同地点采集的甜叶菊叶片中甜菊糖苷的定量分析具有可重复性，并将作为质量控制指标，用于监测甜菊糖苷及其相关分子在加工不同阶段的商业生产。