Department of Pharmacy Practice, Jerry H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, Dallas, TX 75235, United States; Department of Pharmaceutical Science, Jerry H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, Dallas, TX 75235, United States; Clinical Pharmacology and Experimental Therapeutics Center, Jerry H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, Dallas, TX 75235, United States.
Department of Pharmacy Practice, Jerry H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, Dallas, TX 75235, United States; Clinical Pharmacology and Experimental Therapeutics Center, Jerry H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, Dallas, TX 75235, United States.
J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Nov 30;1159:122354. doi: 10.1016/j.jchromb.2020.122354. Epub 2020 Sep 2.
The purpose of this work was to develop and validate a single sensitive, selective and rapid bioanalytical method to determine ceftolozane and tazobactam concentrations in human plasma and urine and to use this method to analyze samples from a human clinical study. Human plasma and urine samples were prepared by protein precipitation using a solution of acetonitrile, water and formic acid. Following protein precipitation, samples were analyzed by liquid chromatography tandem mass spectrometry. Chromatographic resolution was achieved on a Kinetex PFP column using a gradient elution, a flow rate of 0.4 mL/min, and a total run time of 5 min. Positive electrospray ionization was employed and analytes were quantitated using multi-reaction monitoring mode. Method validation was conducted in accordance with Unites States Food and Drug Administration's regulatory guidelines for bioanalytical method validation. Calibration curves were determined to linear over the range of 0.1 to 40 µg/mL for ceftolozane and 0.05 to 20 µg/mL for tazobactam. The method was determined to be accurate (-6.24 to 12.53 percent relative error), precise (less than 13.28 percent standard deviation) and sensitive in both human plasma and urine. Ceftolozane and tazobactam were determined to be stable across a battery of stability studies including autosampler, benchtop, freeze/thaw and long-term stability. This validated method successfully applied to human clinical samples to determine the concentration versus time profiles of the intravenously administered combination of Zerbaxa (ceftolozane-tazobactam) in burn patients.
本工作旨在开发和验证一种灵敏、选择性和快速的生物分析方法，以测定人血浆和尿液中的头孢洛扎他和他唑巴坦浓度，并将该方法用于分析人体临床研究中的样品。人血浆和尿液样品通过使用乙腈、水和甲酸的溶液进行蛋白质沉淀来制备。在蛋白质沉淀后，通过液相色谱串联质谱法分析样品。色谱分离在 Kinetex PFP 柱上进行，采用梯度洗脱，流速为 0.4 mL/min，总运行时间为 5 min。采用正电喷雾电离，采用多反应监测模式定量分析物。方法验证符合美国食品和药物管理局关于生物分析方法验证的监管指南。头孢洛扎他的校准曲线在 0.1 至 40 µg/mL 范围内呈线性，他唑巴坦的校准曲线在 0.05 至 20 µg/mL 范围内呈线性。该方法在人血浆和尿液中均表现出准确性（-6.24 至 12.53 ％相对误差）、精密度（小于 13.28 ％标准差）和灵敏度。头孢洛扎他和他唑巴坦在一系列稳定性研究中表现出稳定性，包括自动进样器、台式、冻融和长期稳定性。该验证方法成功应用于人体临床样品，以测定烧伤患者静脉给予泽布他（头孢洛扎他-他唑巴坦）的组合的浓度-时间曲线。
