HPLC法测定不同产地地龙中尿嘧啶、次黄嘌呤、尿苷、肌苷的含量

目的:测定不同产地、品种的地龙药材中尿嘧啶、次黄嘌呤、尿苷、肌苷的含量,建立地龙质量控制的客观指标。方法:用0.9%生理盐水超声提取地龙药材,采用反相高效液相色谱法,Waters Symmetry C₁₈色谱柱（150mm×4.6mm,5μm）,以0.01mol·L^-1磷酸二氢钾溶液和50%甲醇为流动相,流速为0.8ml·min^-1,梯度洗脱,检测波长254nm,柱温27℃,同时测定尿嘧啶、次黄嘌呤、尿苷、肌苷等4种成分的含量。结果:尿嘧啶的线性范围为0.75～24.00μg·ml^-1（r=0.9993）,平均回收率99.82%,RSD=2.34%（n=6）;次黄嘌呤的线性范围2.50～80.00μg·ml^-1（r=0.9992）,平均回收率101.93%,RSD=1.45%（n=6）;尿苷的线性范围1.25～40.00μg·ml^-1（r=0.9992）,平均回收率97.53%,RSD=1.73%（n=6）;肌苷的线性范围5.00～160.00μg·ml^-1（r=0.9991）,平均回收率102.06%,RSD=2.44%（n=6）。结论:该方法重复性,回收率好,可用于地龙药材中尿嘧啶等4种成分的含量测定。

Determination of Uracil, Hypoxantine, Uridine and Inosine in Earthworm from Different Habitats by HPLC

Objective： To establish an accurate and stable method for the determination of uracil, hypoxanthine, uridine and inosine in earthworm in order to effectively control the quality of the medicinal material. Method： Uracil, hypoxantine, uridine and inosine in earthworm were extracted with 0. 9% NaC1 by ultrasonic and determined by HPLC. The chromatographic conditions were：Waters Symmetry Cls column （150 mm ×4. 6 mm,5μm）,50% methanol and 0. 01 mol·L^-1 KH2PO4 as the mobile phase with a flow rate of 0. 8 ml·min^-1, the detection wavelength was 254 nm, and the cohann temperature was set at 27 ℃. Result： The linear range was within 0. 75- 24. 00 μg·m1^-1 （r =0. 999 3） for uracil,2. 50-80. 00 id,g＇m]-1 （r =0. 999 2） for hypoxantine, 1.25-40. 00 μg-ml-l （r =0. 999 2） for uridine, and 5.00-160. 00μg·ml^ - 1 （ r = 0. 999 1 ） for inosine. The average recovery was 99. 82% （ RSD = 2. 34%, n = 6） for uracil, 101.93 % （RSD = 1.45% ,n =6） for hypoxantine,97. 53% （RSD = 1.73% ,n =6） for uridine,and 102. 06% （RSD =2. 44% ,n =6） for inosine. Conclusion： The method is accurate, sensitive and reproducible. It can be used to control the quality of earthworm from different habitats.