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建立了一种测定蚂蟥药材中抗凝血活性多肽类化合物蚂蟥多肽含量的反相高效液相色谱(RP-HPLC)新方法。色谱柱采用Hypersil-C18柱(200×4.6mm,5μm),流动相为乙腈和水(体积比为2:98),加入质量分数0.03%KH2PO4,再用体积分数36%HAc调至pH 3.80,检测波长280 nm,流速1.0 mL/min,柱温35℃。当蚂蟥多肽的质量浓度为186~942 mg/L时,其峰面积与质量浓度的线性关系良好(r=0.9996);平均回收率为99.8%,RSD<2%。方法可作为蚂蟥药材或含有蚂蟥药材的中成药的质控方法。
A new RP-HPLC method was developed for the determination of the anti-thrombotic activity of leech polypeptide in leech. The column was separated on a Hypersil-C18 column (200 × 4.6 mm, 5 μm) using acetonitrile and water as the mobile phases (2:98 volume ratio), adjusted to pH 3.80 with a mass fraction of 0.03% KH2PO4, The detection wavelength was 280 nm, the flow rate was 1.0 mL / min and the column temperature was 35 ℃. When the concentration of leech polypeptide was 186-942 mg / L, the linear relationship between peak area and mass concentration was good (r = 0.9996). The average recovery was 99.8% and the RSD was less than 2%. The method can be used as a quality control method of leech medicine or Chinese patent medicine containing leech medicine.