基于核壳色谱技术快速测定冬虫夏草中麦角甾醇的含量

Rapid Determination of Ergosterol in Chinese Cordyceps by Core-shell Column

目的采用核壳色谱技术建立冬虫夏草中麦角甾醇的快速定量分析方法。方法样品经甲醇超声提取后,用高效液相色谱测定。色谱柱为Poroshell 120 SB-C_(18)(30 mm×4.6 mm,2.7μm),流动相为甲醇-水(94∶6),流速为1 mL·min^(-1),柱温为35℃,检测波长为283 nm。结果本方法可在15 min内完成冬虫夏草中麦角甾醇的测定(10 min样品提取和5 min色谱分析)。麦角甾醇在7.869~94.43μg·mL^(-1)范围内的线性关系良好,相关系数为1.0000;精密度、重复性良好,RSD均小于3.0%;加样回收率为97.2%(RSD=3.3%,n=9);对照品溶液和供试品溶液在室温下放置24 h内稳定。15批不同来源冬虫夏草的麦角甾醇含量在0.133%~0.279%。结论建立的色谱分析方法简便、快速、准确,有助于提升冬虫夏草样品检测效率。

OBJECTIVE To develop a HPLC method for rapid determination of ergosterol in Cordyceps by core-shell column.METHODS The samples were prepared by ultrasonic extraction with methanol.HPLC separation was operated on a Poroshell 120 SB-C_(18) column(30 mm×4.6 mm,2.7μm)by methanol-water(94∶6)at the flow rate of 1 mL·min^(-1).The column temperature was 35℃and the detection wavelength was 283 nm.RESULTS The analysis of ergosterol was finished in 15 min,which included 10 min for sample extraction and 5 min for HPLC separation.The ergosterol had a good linear relationship in the range of calibration curve of 7.869-94.43μg·mL^(-1) and the correlation coefficient was 1.0000.The precision and repeatability were good(RSD<3.0%).The recovery was 97.2%(RSD=3.3%,n=9).The reference compound and sample solution were stable in 24 hours at room temperature.The ergosterol content in 15 batches of Chinese Cordyceps samples were 0.133%-0.279%.CONCLUSION The developed method is simple,rapid and accurate,which is helpful to improve the detection efficiency of Chinese Cordyceps samples.