RP-HPLC法测定青海不同产地冬虫夏草中核苷类成分的含量

Determination of Nucleosides in Cordyceps sinensis from Qinghai Province by RP-HPLC

目的:测定青海不同产地冬虫夏草中主要核苷类成分的含量并比较其差异。方法:采用反相高效液相色谱(RP-HPLC)法测定不同产地的9份样品中尿苷、鸟苷、肌苷、腺苷、虫草素的含量。色谱柱为Symmetry ShieldTMRp18(250mm×4.6mm,5μm),流动相为乙腈-水(梯度洗脱),流速为0.2～1.2mL·min-1,检测波长为260nm,柱温为30℃。结果:5种成分的进样量分别在0.331～1.796、0.375～2.046、0.256～1.389、0.338～1.844、0.384～2.110mg范围与各自峰面积积分值呈良好的线性关系(r分别为0.9999、0.9999、0.9998、0.9997、0.9998);平均回收率分别为98.72%、101.48%、99.74%、98.46%、98.09%,RSD分别为1.96%、1.68%、1.82%、1.57%、2.74%(n均为6)。青海不同产地间冬虫夏草主要核苷类成分含量有差异。结论:本方法能有效鉴别天然冬虫夏草中的核苷类成分,而冬虫夏草化学成分与生态因子的相关性还需要进一步研究。

OBJECTIVE：To determine the nucleosides in Cordyceps sinensis from Qinghai province and to study their difference. METHODS： RP-HPLC method was applied to determine uridine, guanosine, inosine, adenosine, and cordycepin in nine samples of C. sinensis from different parts of Qinghai province. The separation was performed on a Symmetry Shield^TMRp18（250mm×4.6mm,5μm）with detection wavelength of 260 nm. The mobile phase consisted of acetonitrile-water（gradient elution） with column temperature set at 30℃. Flow rate was from 0.2 mL·min^-1 to 1.2 mL·min^-1.RESULTS： The linear range of uridine, guanosine, inosine, adenosine, and cordycepin were 0.331-1.796 mg（r=0.999 9）, 0.375-2.046 mg（r=0.999 9）, 0.256-1.389 mg（r= 0.999 8）, 0.338-1.844 mg（r=0.999 7）, 0.38442.110 mg（r=0.999 8）,respectively. The mean recovery of five components were 98.72% （RSD= 1.96%, n=6）, 101.48% （RSD= 1.68%, n=6）, 99.74% （RSD=1.82%, n=6）, 98.46% （RSD= 1.57%, n=6）, 98.09% （RSD=2.74% ,n=6）,respectively. The contents of nucleosides in C. sinensis from different origins were different from one another. CONCLUSION： The method effectively distinguishes nucleosides in C. sinensis while further study should be performed on the relationship between chemical component of C. sinensis and ecological factor.