Ocampo A P, Hoyt K D, Wadgaonkar N, Carver A H, Puglisi C V
Metabolism Research Department, American Cyanamid Co., Pearl River, NY 10965.
J Chromatogr. 1989 Nov 10;496(1):167-79. doi: 10.1016/s0378-4347(00)82563-3.
A gradient elution high-performance liquid chromatographic method is described for the analysis of the beta-lactamase inhibitor tazobactam (YTR-830H) and a semi-synthetic parenteral penicillin, piperacillin, in human plasma, serum, bile and urine. The assay for plasma, serum and bile involves deproteinization with acetonitrile and the removal of lipids with dichloromethane; urine is diluted with buffer. Separation and quantitation are achieved using a mobile phase based on ion-suppression chromatography on a C18 reversed-phase column with ultraviolet detection at 220 nm. The limit of quantitation for both compounds is 1.0 microgram/ml in plasma, serum and bile using a 0.2-ml sample and 50.0 micrograms/ml in urine using a 0.1-ml sample. The method has been validated by preparing and analyzing a series of fortified samples (range 1.0-200 micrograms/ml for each compound in plasma, serum and bile and 50.0-10,000 micrograms/ml for each compound in urine). Excellent linearity, accuracy, precision and recovery were obtained. The method was not interfered with by other endogenous components, nor by other commonly administered antibiotics such as amoxicillin, mezlocillin, cefometazole and cefotaxime. The assay has been successfully applied to the analysis of samples from pharmacokinetic studies in man and animals.
本文描述了一种梯度洗脱高效液相色谱法，用于分析人血浆、血清、胆汁和尿液中的β-内酰胺酶抑制剂他唑巴坦（YTR-830H）和一种半合成肠外青霉素哌拉西林。血浆、血清和胆汁的分析方法包括用乙腈进行蛋白沉淀，并用二氯甲烷去除脂质；尿液用缓冲液稀释。使用基于离子抑制色谱的流动相，在C18反相柱上进行分离和定量，检测波长为220nm。对于血浆、血清和胆汁中的两种化合物，使用0.2ml样品时定量限为1.0μg/ml，对于尿液中的两种化合物，使用0.1ml样品时定量限为50.0μg/ml。该方法通过制备和分析一系列加标样品（血浆、血清和胆汁中每种化合物的范围为1.0-200μg/ml，尿液中每种化合物的范围为50.0-10,000μg/ml）进行了验证。获得了出色的线性、准确性、精密度和回收率。该方法不受其他内源性成分的干扰，也不受其他常用抗生素如阿莫西林、美洛西林、头孢美唑和头孢噻肟的干扰。该测定法已成功应用于人和动物药代动力学研究样品的分析。
