色谱

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高效液相色谱-电化学阵列检测器检测烟草中的绿原酸等六种次生代谢产物

王文霞1,2,李曙光1,赵小明1,林炳承1,杜昱光1

  

  1. 1. Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, Dalian 116023, China; 2.Graduate University of Chinese Academy of Sciences, Beijing 100049, China
  • 收稿日期:2007-06-08 修回日期:2007-09-10 出版日期:2007-11-30 发布日期:1984-12-25
  • 通讯作者: 杜昱光

Determination of Six Secondary Metabolites Including Chlorogenic Acid in Tobacco Using High Performance Liquid Chromatography with Coulometric Array Detection

WANG Wenxia1,2, LI Shuguang1, ZHAO Xiaoming1, LIN Bingcheng1, DU Yuguang1   

  1. 1. Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, Dalian 116023, China; 2.Graduate University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2007-06-08 Revised:2007-09-10 Online:2007-11-30 Published:1984-12-25
  • Contact: DU Yuguang

摘要:

建立了一种利用反相高效液相色谱-电化学阵列检测器同时检测烟草中主要次生代谢产物的方法。在Hypersil BDS C-18(4.6 mm×200 mm)色谱柱上,以30 mmol/L磷酸二氢钠(pH 3.5)-5%~70%(体积分数)乙腈(含0.25 mmol/L十二烷基磺酸钠)进行梯度洗脱,流速为1 mL/min,柱温为30 ℃。电化学阵列检测器的检测电势依次为:-20,140,210,310,400,450,490,730,800,900 mV时,可以较好地分离和检测烟草中常见的次生代谢产物绿原酸、咖啡酸、对羟基肉桂酸、莨菪葶、芦丁和烟碱。该方法的相对标准偏差为0.71%~15.31%,回收率为52.0%~85.2%,各种次生代谢产物的检测限为0.2~2 ng;进样量在20~500 ng范围内呈现良好的线性关系,线性相关系数为0.9910~0.9998;具有较高的准确度和精确度。方法简便,可应用于烟草中次生代谢产物的检测。

关键词: 次生代谢产物 , 电化学库仑阵列检测, 反相高效液相色谱, 烟草

Abstract:

A procedure utilizing reversed-phase high performance liquid chromatography coupled with a coulometric array detection system was developed for the characterization of several major secondary metabolites in tobacco. Chromatographic separation was achieved on a Hypersil BDS C-18 column (4.6 mm×200 mm), with linear gradient elution of 5%-70%(v/v) of acetonitrile in a buffer containing 30 mmol/L NaH2PO4, 0.25 mmol/L dodecyl sulfate sodium salt (pH 3.5) at a flow rate of 1 mL/min and column temperature of 30 ℃. Ten serial coulometric detectors were set on -20, 140, 210, 310, 400, 450, 490, 730, 800 and 900 mV. The method was reliable and sensitive. The relative standard deviations ranged from 0.71% to 15.31%. The recoveries were between 52.0% and 85.2%. The detection limits were 0.2-2 ng, and there were good linear correlations when injection amount was in a range from 20 ng to 500 ng, with the regression coefficient of standard calibration curves from 0.9910 to 0.9998. This method is easy to perform and can be applied to determine secondary metabolites in tobacco.

Key words: coulometric array detection, secondary metabolites , tobacco, reversed-phase high performance liquid chromatography(RP-HPLC)