刘明智 黄光磊 张钰祺 易徐航 袁娟丽

摘要：目的 建立水蛭胶囊薄层鉴别及次黄嘌呤含量测定方法。方法 采用薄层色谱法以水蛭对照药材对水蛭胶囊进行定性鉴别；以高效液相色谱法对其中的次黄嘌呤进行含量测定：采用Agilent ZORBAX Eclipse Plus C18色谱柱（4.6mm×250mm，5μm），以甲醇-水（5：95）为流动相，流速为1.0 ml/min，检测波长为254 nm，柱温为25 ℃。结果 水蛭胶囊供试品的薄层色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点，可以作为水蛭胶囊的鉴别方法；高效液相色谱法中，次黄嘌呤的进样浓度在2.67-85.40μg·mL-1（r =1.0000）范围内呈良好线性关系；次黄嘌呤的平均回收率（n=6）分别为98.3%（RSD%=0.5）；5个批次水蛭胶囊样品中次黄嘌呤的含量测定结果分别为100.0μg/袋，104.7μg/袋，97.6μg/袋、101.5μg/袋、103.7μg/袋。结论 本文建立的方法经方法学验证，结果准确、可靠，可用于评价水蛭胶囊质量。

关键字：水蛭胶囊，薄层色谱法，高效液相色谱法，次黄嘌呤，含量测定

[ABSTRACT] Objective To establish a qualitative identification method and a HPLC method for the determination of hypoxanthine in Shuizhi Capsule.Methods TLC was employed to make qualitative identification of Hirudo in Shuizhi Capsule and HPLC was used to determine the content of hypoxanthine in Shuizhi Capsule：The analytical column was Agilent ZORBAX Eclipse Plus C18（4.6×250mm，5μm），methanol-water（5：95）as mobile phase.The flow rate was 1.0ml/min and the detection wavelength was 254nm，The column temperature was 25℃.Results The spot in the chromatograms of Shuizhi Capsule were in the same location and color with those in the chromatograms of standard hirudo and the result showed that the method has good durable ability.The linear range for hypoxanthine was 2.67-85.40μg·mL-1（r =1.0000）. The average recoveries（n=6）were 98.3%（RSD%=0.5）for hypoxanthine.The contents in 5 sapmles of Shuizhi Capsule were 100.0μg/bag，104.7μg/ bag，97.6μg/ bag、101.5μg/ bag、103.7μg/ bag.Conlusion The method is accurate and reliable，and it can be applied to the quality control of Shuizhi Capsule.

[Key words] Shuizhi Capsule；TLC；HPLC；hypoxanthine；determination

水蛭最早收载于《神农本草经》[1]，是一味具有强效破血逐瘀的虫类药，对血液动力等方面有积极的改善作用[2-3]。因其疗效确切，为历版《中国药典》所收载[4]。《中国药典》收载的水蛭为水蛭科动物蚂蟥Whitmania pigra Whitman、水蛭Hirudo nipponica Whitman或柳叶蚂蟥Whitmannia acranulata Whitman的干燥全体[5]。水蛭含有多种有效成分，主要含有两类：一类是以水蛭素为代表的多肽及蛋白类大分子成分；另一类是蝶啶等小分子化学成分[6]。水蛭“夏、秋二季捕捉，用沸水烫死，晒干或低温干燥[5]。”但水蛭素在80℃条件下15min能将其破坏[7]，即水蛭饮片中水蛭素已被破坏。次黄嘌呤是水蛭所含的有效成分之一，是核酸的代谢产物，广泛存在于动植物中，具有平喘、舒张支气管、降压等药理作用，其作为有效成分指标测定其含量的研究广泛[8-9]，已有报道测定水蛭饮片中次黄嘌呤的含量[10]。……