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

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。

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

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 x 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 degrees C. 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.