反相高效液相色谱法测定蛹虫草中腺苷和虫草素的含量

Analysis of Adenosine and Cordycepin in Cordyceps militaris (L.) Link by Reversed Phase-High Performance Liquid Chromatography

建立一种新的反相高效液相色谱方法,用于蛹虫草中腺苷和虫草素的定性定量分析。样品经水超声提取,以终浓度20mmol/L柠檬酸、40mmol/L醋酸、20mmol/L三乙胺的混合溶液-乙腈(体积比98:2)为流动相,流速1.0mL/min,色谱柱CapcellpakC18(150mm×4.6mm,5μm)分离,柱温40℃,进样量10μL,紫外检测波长260nm。在此条件下,腺苷在1～1000μg/mL范围内有良好的线性关系(Y=33574X+54.526,R2=0.9999),检测限0.2μg/mL,回收率93.75%～96.90%,RSD为0.116%;虫草素在2～1000μg/mL范围内有良好的线性关系(Y=34385X+23.103,R2=1.0000),检测限0.2μg/mL,回收率92.03%～97.35%,RSD为0.104%。该方法样品处理简便、分离度高、杂质干扰小、回收率高、重复性好,能够对蛹虫草中腺苷和虫草素进行准确的定性定量分析。

In this paper, a RP-HPLC method for determining the contents of adenosine and cordycepin in Cordyceps militaris （ L.） Link is described. The separation column was Capcellpak C18 （150 mm × 4.6 mm, 5μm）, which was held at 40 ℃. The mobile phase was composed of a mixed solution containing 20 mmol/L citric acid, 40 mmol/L acetic acid and 20 mmol/L triethylamine and acetonitrile （V/V, 98：2） at a flow rate of 1.0 mL/min. The sample loading volume was 10μL. UV detection was performed at 260 nm. The method showed a good linear relationship between peak area and concentration over the range from 1 to 1000μg/mL （R2 = 0.9999） for adenosine and from 2 to 1000μg/mL （R2 = 1.0000） for cordycepin. The spike recoveries of adenosine in 3 replicates determinations ranged from 93.75% to 96.90% and those of cordycepin from 92.03% to 97.35%.The limit of detection was 0.2 μg/mL for both adenosine and cordycepin. The RSD of precision was 0.116% for adenosine and 0.104% for cordycepin. This method is characterized by simplicity and high sensitivity and accuracy.