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

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

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（4．6mm×200mm）色谱柱上，以30mmol／L磷酸二氢钠（pH3．5）-5％～70％（体积分数）乙腈（含0．25mmol／L十二烷基磺酸钠）进行梯度洗脱，流速为1mL／min，柱温为30℃。电化学阵列检测器的检测电势依次为：-20，140，210，310，400，450，490，730，800，900mV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0．2～2ng；进样量在20～500ng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。

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 Hy- persil 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. 991 0 to 0. 999 8. This method is easy to perform and can be applied to determine secondary metabolites in tobacco.