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ZORBAXSB C18色谱柱(4 6mmi d ×150mm,填料粒径5μm),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 5mL/min,检测波长228nm。在马尿酸浓度为0 005～1 000mmol/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 50μmol/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。
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-C_(18) (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=(0.999 9)) 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.
出处
《色谱》
CAS
CSCD
北大核心
2005年第1期79-81,共3页
Chinese Journal of Chromatography
基金
江苏省高技术研究项目(项目编号:BG2003319).
关键词
高效液相色谱法
血管紧张素转化酶抑制剂
马尿酸
high performance liquid chromatography
angiotensin-converting enzyme inhibitor
hippuric acid
