色谱

色谱

• 研究简报 • 上一篇    下一篇

高效液相色谱法测定血管紧张素转化酶抑制剂的活性

吴琼英,马海乐,骆琳,吴守一   

  1. College of Biology and Environment Engineering, Jiangsu University, Zhenjiang 212013, China
  • 收稿日期:2004-02-25 修回日期:1900-01-01 出版日期:2005-01-30 发布日期:1989-03-25
  • 通讯作者: 吴琼英
  • 基金资助:

Determination of Angiotensin Converting Enzyme Inhibitor Activity by High Performance Liquid Chromatography

WU Qiongying, MA Haile, LUO Lin, WU Shouyi   

  1. College of Biology and Environment Engineering, Jiangsu University, Zhenjiang 212013, China
  • Received:2004-02-25 Revised:1900-01-01 Online:2005-01-30 Published:1989-03-25

PDF

793

被引次数

198

摘要： 建立了体外直接测定血管紧张素转化酶抑制剂活性的高效液相色谱分析方法。以马尿酰-组氨酰-亮氨酸为反应底物,血管紧张素转化酶为催化剂,反应所生成的马尿酸为测定指标,未加酶抑制剂的反应为空白对照。使用ZORBAX SB-C18色谱柱(4.6 mm i.d.×150 mm,填料粒径5　μm),柱温25 ℃,流动相为乙腈-超纯水(体积比为25∶75,各含0.05%（体积分数）三氟乙酸及0.1%（体积分数）三乙胺),流速0.5 mL/min,检测波长228 nm。在马尿酸浓度为0.005～1.000 mmol/L时,马尿酸浓度与其峰面积呈良好的线性关系(r＝0.9999),最小检测限为0.50 μmol/L;该方法对马尿酸的回收率为99.48%～105.64%,相对标准偏差(RSD)为2.20%(n＝6)。该方法可适用于血管紧张素转化酶抑制剂活性的体外测定,具有操作简便、精密度和准确性高的特点,为降血压药物的研制提供了方便可靠的检测手段。

关键词: 高效液相色谱法, 马尿酸 , 血管紧张素转化酶抑制剂

Abstract: A high performance liquid chromatographic method to determine angiotensin-converting enzyme inhibitor activity in vitro was established by using N-hippuryl-His-Leu tetrahydrate as the reaction substrate and hippuric acid as the reaction product. The chromatographic conditions were as follows: column, ZORBAX SB-C18 (4.6 mm i.d.×150 mm, 5 μm); column temperature, 25 ℃; mobile phase, acetonitrile-distilled water (25∶75, v/v, both containing 0.05% (v/v) trifluoroacetic acid and 0.1% (v/v) triethylamine); flow rate, 0.5 mL/min; detection wavelength, 228 nm. An excellent linearity over the range of 0.005-1.000 mmol/L (r＝0.9999) was observed. The detection limit was 0.50 μmol/L. The recoveries of hippuric acid ware 99.48%-105.64%, with a relative standard deviation (RSD) of 2.20%(n＝6). It is a simple, precise and reliable assay method for developing antihypertension drugs.

Key words: angiotensin-converting enzyme inhibitor, hippuric acid , high performance liquid chromatography