Pelegrini-da-Silva Adriana, Prado Wiliam A, Juliano Maria A, Wilk Sherwin, Martins Antonio R
Department of Pharmacology, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Av Bandeirantes 3900, 14049-900 Ribeirão Preto, São Paulo, Brazil.
J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Nov 25;780(2):301-7. doi: 10.1016/s1570-0232(02)00542-1.
We describe here a gradient HPLC procedure for the separation, and quantification by UV absorption of renin tri- and tetradecapeptide substrates, angiotensins I, II, III, IV and V, angiotensin-derived peptides, and peptidase inhibitors including amastatin, bestatin, pepstatin, lisinopril, a renin peptide inhibitor, Z-Pro-prolinal, N-[1-(R,S)-carboxy-2-phenylethyl]-L-Ala-L-Ala-L-Phe-p-aminobenzoate, and phosphoramidon. Most peptides and peptidase inhibitors were baseline-resolved within 32 min. The overall intra- and inter-assay precisions ranged from 0.8 to 5.9 (n=6) and 2 to 13% (n=6), respectively. There was a linear relationship (correlation coefficients> or =0.9660) between peak height and peptide amount injected. In conclusion, the present method when combined with a peptidase-inhibitor paradigm can lead to the identification of renin-angiotensin system metabolizing enzymes, and when combined with radioimmunoassay can enhance the specificity of angiotensin measurement.
我们在此描述一种梯度高效液相色谱法，用于分离肾素三肽和十四肽底物、血管紧张素I、II、III、IV和V、血管紧张素衍生肽以及肽酶抑制剂（包括抑氨肽酶素、贝他汀、胃蛋白酶抑制剂、赖诺普利、一种肾素肽抑制剂、Z-脯氨酰脯氨醛、N-[1-(R,S)-羧基-2-苯乙基]-L-丙氨酰-L-丙氨酰-L-苯丙氨酰对氨基苯甲酸和膦甲脒），并通过紫外吸收进行定量分析。大多数肽和肽酶抑制剂在32分钟内实现基线分离。测定内和测定间的总体精密度分别为0.8%至5.9%（n=6）和2%至13%（n=6）。峰高与注入的肽量之间存在线性关系（相关系数≥0.9660）。总之，本方法与肽酶抑制剂模式相结合可用于鉴定肾素-血管紧张素系统代谢酶，与放射免疫测定相结合可提高血管紧张素测量的特异性。
