反相高效液相色谱法测定薄芝菌丝体中3种核苷类成分的含量

Determination of Nucleosides in Bo Zhi Mycelium by RP-HPLC

目的建立高效液相色谱（HPLC）梯度洗脱法同时测定薄芝菌丝体中3种核苷类成分（尿嘧啶、尿苷、腺苷）的含量。方法采用RP—HPLC法测定，Alltima C18柱（5μm，4．6mm×250mm）为色谱柱，流动相为乙腈-水，梯度洗脱（0～15min，乙腈-水比例为5：995，15～30min，比例为15：985），流速1．0ml·min^-1，检测波长262nm。结果3个核苷的线性范围分别为，尿嘧啶：0．1402～0．7008μg，（r=0．9998，n=5）；尿苷：0．1032～0．5156μg（r=0．9997，n=5）；腺苷：0．1245～0．6224μg（r=0．9994，n=5）。平均加样回收率分别为：尿嘧啶99．44％，尿苷100．08％，腺苷99．96％。结论该方法灵敏度高、操作简便、重复性好、效率高，为全面评价薄芝菌丝体质量提供一个可靠方法。

Objective To determine the contents of nucleosides in Bo Zhi Mycelium（ uracil, uridine, adenosine） by HPLC with gradient elution. Methods Uracil, uridine, adenosine were dealt with by HPLC with Alhima - C18 analytical column （5μm,250 mm × 4.6 mm）. The mobile plase consisted of acetonitrile - water, gradient elution （ 0 - 15 min, CH3 CN - water 5 ： 995,15 - 30 min, CH3CN -water 15：985 ）. The flow rate was 1.0 ml · min^-1. The UV detection was set at 260 nm. Results The linear ranges of uracil, uridine and adenosine were 0. 140 2 - 0. 700 8 μg （ r = 0. 999 8, n = 5） ; 0. 103 2 - 0. 515 6 μg （r = 0. 999 7, n = 5 ） ; 0. 124 5 -0. 622 4μg （r = 0. 999 4,n = 5 ） respectively. The average recovery was 99.44% ; 100.08% and 99.96% respectively. Conclusion This method is simple, reliable, efficient, and provides a reference standard for the quality control of Bo Zhi Mycelium.