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目的:建立高效液相色谱法测定皖产古尼虫草中核苷和碱基成分(尿嘧啶、虫草素、腺苷、腺嘌呤、尿苷、次黄嘌呤、肌苷和鸟苷)的含量,并比较其与天然冬虫夏草核苷和碱基成分的异同。方法:采用Zorbax SB-Aq(250 mm×4.6 mm,5μm)色谱柱,以水-乙腈为流动相,梯度洗脱,流速为0.8 mL.min-1,检测波长260 nm,柱温20℃。结果:尿嘧啶、虫草素、腺苷、腺嘌呤、尿苷、次黄嘌呤、肌苷和鸟苷进样浓度分别在0.38～191.08,0.37～188.48,0.37～189.72,0.37～190.92,0.37～188.55,0.50～168.55,0.37～190.36,0.24～125.00μg.mL-1范围内线性关系良好,R2为0.9998～0.9999;加样回收率分别为102.1%,102.1%,95.3%,96.2%,96.9%,94.1%,108.7%,101.8%,RSD未超过2.8%。皖产古尼虫草和冬虫夏草均含有核苷和碱基成分,其中:皖产古尼虫草含有较高的尿嘧啶、尿苷、腺苷,而冬虫夏草中肌苷含量较高;二者的次黄嘌呤、鸟苷、腺嘌呤的含量相近;在该检测限下,两者均未测得虫草素。结论:初步认为:皖产古尼虫草核苷和碱基成分与冬虫夏草具有一定的相似性,但尚有待扩大样本量作进一步的确证。
OBJECTIVE: To establish a HPLC method for the determination of nucleosides and bases (uracil, cordycepin, adenosine, adenine, uridine, hypoxanthine, inosine and guanosine) Comparison with natural Cordyceps nucleosides and nucleotide composition of the similarities and differences. Methods: A Zorbax SB-Aq (250 mm × 4.6 mm, 5 μm) column was used. The mobile phase consisted of water and acetonitrile with a gradient of 0.8 mL.min-1. The detection wavelength was 260 nm and the column temperature was 20 ℃. Results: The concentrations of uracil, cordycepin, adenosine, adenine, uridine, hypoxanthine, inosine and guanosine were 0.38 ~ 191.08,0.37 ~ 188.48,0.37 ~ 189.72,0.37 ~ 190.92,0.37 ~ 188.55 , The linear range was 0.50 ~ 168.55,0.37 ~ 190.36,0.24 ~ 125.00μg.mL-1, the R2 was 0.9998 ~ 0.9999; the recoveries were 102.1%, 102.1%, 95.3%, 96.2%, 96.9% 94.1%, 108.7%, 101.8%, RSD did not exceed 2.8%. Cordyceps and Cordyceps sinensis both contained nucleosides and bases, among which, Cordyceps mongolicum contained higher uracil, uridine and adenosine, while Cordyceps sinensis had a higher content of inosine; Purine, guanosine, adenine content similar to the detection limit, both were not measured cordycepin. Conclusion: It is preliminarily believed that the nucleoside and base composition of Cordyceps in Cordyceoplasty have some similarities with that of Cordyceps sinensis, but we still need to expand the sample size for further confirmation.