Hajdú P, Schmidt D, Bomm M, Hack L, Keller A
Arzneimittelforschung. 1984;34(10B):1431-5.
A highly sensitive and specific enzymatic assay for the quantitative determination of 2-[N-[(S)-1-ethoxycarbonyl-3-phenylpropyl]-L-alanyl]-(1S,3S, 5S)-2-azabicyclo[3.3.0]octane-3-carboxylic acid (Hoe 498) and its hydrolysis product Hoe 498-diacid in serum and a GLC method for the simultaneous determination of both compounds in urine are described. Both methods involve extraction from the respective biological fluid using disposable C 18 columns. In serum, the enzyme-inhibiting properties of Hoe 498-diacid (Hoe 498 can readily be converted to its hydrolysis product) are utilized for the determination of both compounds concurrently. The serum extract is dissolved in buffer solution and incubated with human serum as angiotensin converting enzyme (peptidyl-dipeptide-hydrolase, EC 3.4.15.1) source and hip-his-leu substrate. The hippuric acid liberated is quantitated by HPLC. The urine extract is treated with diazomethane followed by trifluoroacetic anhydride to convert Hoe 498 and Hoe 498-diacid to their methylester, trifluoroacetyl derivatives, which are then determined simultaneously by GLC using a nitrogen specific detector.
本文描述了一种高灵敏度和特异性的酶法，用于定量测定血清中的2-[[(S)-1-乙氧羰基-3-苯基丙基]-L-丙氨酰]-(1S,3S,5S)-2-氮杂双环[3.3.0]辛烷-3-羧酸（Hoe 498）及其水解产物Hoe 498-二酸，以及一种气相色谱法用于同时测定尿液中的这两种化合物。两种方法均涉及使用一次性C18柱从各自的生物流体中进行萃取。在血清中，利用Hoe 498-二酸的酶抑制特性（Hoe 498可轻易转化为其水解产物）同时测定这两种化合物。将血清提取物溶解于缓冲溶液中，并与人血清作为血管紧张素转换酶（肽基二肽水解酶，EC 3.4.15.1）来源以及马尿酰-组氨酰-亮氨酸底物一起孵育。释放出的马尿酸通过高效液相色谱法定量。尿液提取物先用重氮甲烷处理，然后用三氟乙酸酐处理，将Hoe 498和Hoe 498-二酸转化为它们的甲酯、三氟乙酰衍生物，然后使用氮特异性检测器通过气相色谱法同时测定。
