LC/ESI-MS分离和测定冬虫夏草及其代用品的有效成分

Separation and Determination of Active Components in Cordyceps Sinensis and Its Substitutes with LC/ESI-MS

本文报道采用电喷雾离子化(ESI),用高效液相色谱质谱联用法同时分离和测定冬虫夏草及其代用品中主要活性成分尿苷和腺苷的方法。本法使用Shimadzu VP ODS色谱柱,以水甲醇甲酸(90∶9.0∶1.0,V/V)为流动相,2 氯腺苷为内标,选择性离子检测(SIM)模式作定量分析。结果表明:尿苷和腺苷线性范围分别为2.0~110.0μg·mL-1和1.0~120.0μg·mL-1,检出限分别为0.3μg·mL-1和0.2μg·mL-1,标准加入回收率为97.0%~103.3%。该法已成功地用于冬虫夏草及其代用品中尿苷和腺苷的分析。

A simple,sensitive and reproducible high performance liquid chromatography(HPLC)-electrospray ionization mass spectrometry (ESI-MS) method for simultaneous separation and determination of adenosine and uridine is developed.The analytical column is a 2.0×150 mm Shimadzu VP-ODS column and the mobile phase is water (90.0%),methanol(9.0%) and formic acid (1.0%).(2-Chloroadenosine) is used as internal standard for this assay.Selective ion monitoring (SIM) mode was used and SIM ions at m/z 267 for uridine and m/z 268 for adenosine are chosen for quantitative analysis.The regression equations and liner range are Y=0.078 54X+0.006 27 and 2.0~110.0 μg·mL^(-1) fo uridine,Y=0.146 1X+0.012 4 and 1.0~120.0 μg·mL^(-1) for adenosine,respectively. The limits of detection (LOD) are 0.3 μg·mL^(-1) for uridine and 0.2 μg·mL^(-1) for adenosine, respectively.The recoveries of three constituents are from 97.0% to 103.3%.This method has been successfully applied for the determination of uridine and adenosine in various natural Cordyceps sinensis and its substitutes.