HPLC测定蝙蝠蛾拟青霉培养物5种核苷类成分的含量

Determination of 5 nucleosides components in culture of Paecilomyces hepialid by HPLC

建立HPLC同时测定蝙蝠蛾拟青霉培养物中尿嘧啶、尿苷、鸟苷、腺嘌呤和腺苷5种核苷类成分含量的方法。采用Waters Sun Fire C18（4.6 mm×250 mm,5μm）色谱柱;以甲醇-水为流动相,梯度洗脱,柱温30℃,检测波长260 nm。结果尿嘧啶的线性范围为10.00~200.00 mg·L-1（r=0.999 4）,尿苷的线性范围为10.10~202.00 mg·L-1（r=0.999 2）,鸟苷的线性范围为10.00~200.00 mg·L-1（r=0.999 1）,腺嘌呤的线性范围为10.30~206.00 mg·L-1（r=0.999 2）,腺苷的线性范围为10.45~209.00 mg·L-1（r=0.999 1）;尿嘧啶、尿苷、鸟苷、腺嘌呤和腺苷的精密度RSD分别为0.032%,0.035%,0.039%,0.049%,0.000 80%;尿嘧啶、尿苷、鸟苷、腺嘌呤和腺苷的平均加样回收率分别为97.34%,99.10%,101.6%,98.61%,100.2%,RSD分别为1.3%,2.1%,0.96%,0.95%,1.3%。该方法灵敏度高、选择性好,线性关系及重复性好,适合于蝙蝠蛾拟青霉培养物及其提取物中核苷类成分的含量分析。

The concentration of 5 nucleosides,uracil,uridine,guanidine,adenine and adenosine in culture of Paecilomyces hepialid was determined by the developed method of HPLC. The HPLC method was performed on a Waters Sun Fire C18（ 4. 6 mm × 250 mm,5 μm） column with methanol-water gradient elution as the mobile phase. The detection wavelength was 260 nm and the colunmn temperature was controlled at 30 ℃. The linear range was 10. 00-200. 00 mg·L-1（ r = 0. 999 4） for uracil,10. 10-202. 00 mg·L-1（ r = 0. 999 2） for uridine,10. 00-200. 00 mg·L-1（ r = 0. 999 1） for guanidine,10. 30-206. 00 mg·L-1（ r = 0. 999 2） for adenine and 10. 45-209. 00 mg · L-1（ r = 0. 999 1） for adenosine,respectively. The RSD of precision was 0. 032%,0. 035%,0. 039%,0. 049%,0. 000 80%,respectively. The average recoveries of uracil,guanidine,adenine,and adenosine were 97. 34%,99. 10%,101. 6%,98. 61% and 100. 2% with RSD of 1. 3%,2. 1%,0. 96%,0. 95%,and 1. 3% respectively. The method showed high sensitivity,good selectivity,linearity and repeatability,which was suitable for the content analysis of 5 nucleosides components in P. hepialid and its extracts.