Spahn-Langguth H, Podkowik B, Stahl E, Martin E, Mutschler E
Pharmakologisches Institut für Naturwissenschaftler, Johann Wolfgang Goethe-Universität, Frankfurt am Main, Germany.
J Anal Toxicol. 1991 Nov-Dec;15(6):327-31. doi: 10.1093/jat/15.6.327.
For the enantiospecific assay of propranolol in biological material, formation of diastereomeric derivatives is one possible approach. The aim of the present study was the development and optimization of three analytical methods based on different chiral reagents: phenylethylisocyanate and the acyl chloride as well as the isocyanate that are derived from the fluorescent S-flunoxaprofen. Pronethalol is used as internal standard in all three procedures and improves the coefficients of variation significantly. After extraction from human plasma or urine, propanolol is reacted with one of these compounds in anhydrous organic solvents with addition of triethylamine. The diastereomeric derivatives are then resolved on an octadecylsilane column using mixtures of water and methanol with or without addition of glacial acetic acid. Good resolutions of the diastereomeric derivatives are found under these conditions. Conjugates are cleaved prior to analysis using beta-glucuronidase-arylsulfatase and assayed as parent propranolol enantiomers. All three procedures were suitable for analysis of propranolol enantiomers in biological samples in the lower nanogram range (1-2 ng/mL). A preliminary clinical study confirmed the known enantiospecificity in the pharmacokinetics of propranolol and showed high concentrations of conjugates with R/S ratios that were similar to those of the parent enantiomers.
对于生物材料中普萘洛尔的对映体特异性分析，形成非对映体衍生物是一种可行的方法。本研究的目的是开发和优化三种基于不同手性试剂的分析方法：苯乙基异氰酸酯、酰氯以及由荧光S-氟洛芬衍生的异氰酸酯。在所有三个程序中均使用普萘洛尔作为内标，显著提高了变异系数。从人血浆或尿液中提取后，普萘洛尔在无水有机溶剂中与这些化合物之一反应，并加入三乙胺。然后使用水和甲醇的混合物（添加或不添加冰醋酸）在十八烷基硅烷柱上分离非对映体衍生物。在这些条件下，发现非对映体衍生物具有良好的分离度。在分析前使用β-葡萄糖醛酸酶-芳基硫酸酯酶裂解缀合物，并将其作为母体普萘洛尔对映体进行测定。所有三个程序均适用于分析纳克范围内（1-2 ng/mL）生物样品中的普萘洛尔对映体。一项初步临床研究证实了普萘洛尔药代动力学中已知的对映体特异性，并显示出高浓度的缀合物，其R/S比值与母体对映体相似。
