Huang Lan-Fang, Liang Yi-Zeng, Guo Fang-Qiu, Zhou Zhi-Feng, Cheng Ben-Mei
Research Center of Modernization of Chinese Herbal Medicines, College of Chemistry and Chemical Engineering, Central South University, Changsha 410083, PR China.
J Pharm Biomed Anal. 2003 Dec 4;33(5):1155-62. doi: 10.1016/s0731-7085(03)00415-1.
A simple and rapid isocratic LC/MS coupled with electrospray ionization (ESI) method for simultaneous separation and determination of adenine, hypoxanthine, adenosine and cordycepin in Cordyceps sinensis (Cs) and its substitutes was developed. 2-Chloroadenosine was used as internal standard for this assay. The optimum separation for these analytes was achieved using the mixture of water, methanol and formic acid (85:14:1, v/v/v) as a mobile phase and a 2.0 x 150 mm Shimadzu VP-ODS column. Selective ion monitoring (SIM) mode ([M+H]+ at m/z 136, 137, 268, 252 and 302) was used for quantitative analysis of above four active components. The regression equations were liner in the range of 1.4-140.0 microg ml(-1) for adenine, 0.6-117.5 microg ml(-1) for hypoxanthine, 0.5-128.5 microg ml(-1) for adenosine and 0.5-131.5 microg ml(-1) for cordycepin. The limits of quantitation (LOQ) and detection (LOD) were, respectively 1.4 and 0.5 microg ml(-1) for adenine, 0.6 and 0.2 microg ml(-1) for hypoxanthine, 0.5 and 0.1 microg ml(-1) for adenosine and cordycepin. The recoveries of four constituents were from 93.5 to 107.0%. The nucleoside contents of various types of natural Cs and its substitutes were determined and compared with this developed method.
建立了一种简单快速的等度液相色谱/质谱联用（LC/MS）方法，该方法结合电喷雾电离（ESI），可同时分离和测定冬虫夏草及其替代品中的腺嘌呤、次黄嘌呤、腺苷和虫草素。2-氯腺苷用作该测定的内标。以水、甲醇和甲酸（85:14:1，v/v/v）的混合物为流动相，使用2.0×150 mm的岛津VP-ODS柱，实现了这些分析物的最佳分离。采用选择性离子监测（SIM）模式（m/z 136、137、268、252和302处的[M+H]+）对上述四种活性成分进行定量分析。腺嘌呤的回归方程在1.4 - 140.0 μg ml(-1)范围内呈线性，次黄嘌呤在0.6 - 117.5 μg ml(-1)范围内，腺苷在0.5 - 128.5 μg ml(-1)范围内，虫草素在0.5 - 131.5 μg ml(-1)范围内。腺嘌呤的定量限（LOQ）和检测限（LOD）分别为1.4和0.5 μg ml(-1)，次黄嘌呤为0.6和0.2 μg ml(-1)，腺苷和虫草素为0.5和0.1 μg ml(-1)。四种成分的回收率为93.5%至107.0%。使用该开发的方法测定并比较了各种天然冬虫夏草及其替代品的核苷含量。
