[固相萃取-高效液相色谱法测定烟草中的多酚类物质]

[Determination of polyphenols in tobacco by solid phase extraction and high performance liquid chromatography].

作者信息

Wang L, Fang R B, Li Z, Jiang C Q, Yang G Y

机构信息

Tobacco Chemical Lab, Yunnan Academy of Tobacco Science, Kunming 650106, China.

出版信息

Se Pu. 2001 Nov;19(6):564-6.

PMID:12545477

Abstract

A high performance liquid chromatographic (HPLC) method for the determination of polyphenols in tobacco was studied. Polyphenols can be extracted from tobacco sample by refluxing with 80% (volume fraction) methanol, then subjected to degrease by solid phase extraction with Sep-Pak-C18 column. Chlorogenic acid, rutin, scopoletin, caffeic acid, scopolin and other polyphenols can be separated on a Nova-Pak-C18 column (3.9 mm i.d. x 150 mm) gradiently eluted with methanol and 0.05 mol.L-1 potassium dihydrogen phosphate buffer solution as mobile phase at a flow-rate of 0.5 mL.min-1. Polyphenols were determined at the maximum wavelength of each polyphenol, for chlorogenic acid 326.1 nm, rutin 354.8 nm, scopoletin 344.0 nm, caffeic acid 323.7 nm and scopolin 365.2 nm by photodiode array detector. The main polyphenols in tobacco can be identified with UV spectroscopy. The recoveries of tobacco polyphenols were 94%-105%, and the relative standard deviations were 1.28%-1.49%. This method can be applied to the determination of polyphenols in tobacco with satisfactory results.

摘要

研究了一种用于测定烟草中多酚的高效液相色谱（HPLC）方法。多酚可通过用80%（体积分数）甲醇回流从烟草样品中提取，然后用Sep-Pak-C18柱进行固相萃取脱脂。绿原酸、芦丁、东莨菪素、咖啡酸、东莨菪苷等多酚可在Nova-Pak-C18柱（内径3.9 mm×150 mm）上分离，以甲醇和0.05 mol·L-1磷酸二氢钾缓冲溶液为流动相，流速为0.5 mL·min-1进行梯度洗脱。通过光电二极管阵列检测器在各多酚的最大波长处测定多酚，绿原酸为326.1 nm，芦丁为354.8 nm，东莨菪素为344.0 nm，咖啡酸为323.7 nm，东莨菪苷为365.2 nm。烟草中的主要多酚可用紫外光谱法鉴定。烟草多酚的回收率为94% - 105%，相对标准偏差为1.28% - 1.49%。该方法可用于烟草中多酚的测定，结果令人满意。