Augey V, Grosse P Y, Albert G, Audran M, Bressolle F
Laboratoire de Pharmacocinétique, Faculté de Pharmacie, Montpellier, France.
J Chromatogr B Biomed Appl. 1996 Jun 28;682(1):125-36. doi: 10.1016/0378-4347(96)00049-7.
A high-performance liquid chromatographic (HPLC) method with ultraviolet (UV) absorbance was developed for the analysis of piperacillin-tazobactam (tazocillin), in plasma and urine. The detection was performed at 218 nm for tazobactam and 222 nm for piperacillin. The procedure for assay of these two compounds in plasma and of piperacillin in urine involves the addition of an internal standard (ceftazidime for tazobactam and benzylpenicillin for piperacillin) followed by a treatment of the samples with acetonitrile and chloroform. To quantify tazobactam in urine, diluted samples were analysed using a column-switching technique without internal standard. The HPLC column, LiChrosorb RP-select B, was equilibrated with an eluent mixture composed of acetonitrile-ammonium acetate (pH 5). The proposed technique is reproducible, selective, and reliable. The method has been validated, and stability tests under various conditions have been performed. Linear detector responses were observed for the calibration curve standards in the ranges 5-60 micrograms/ml for tazobactam, and 1-100 micrograms/ml for piperacillin and spans what is currently though to be the clinically relevant range for tazocillin concentrations in body fluids. The limit of quantification was 3 micrograms/ml for tazobactam and 0.5 microgram/ml for piperacillin in plasma and urine. Extraction recoveries from plasma proved to be more than 85%. Precision, expressed as C.V., was in the range 0.4-18%.
建立了一种采用紫外吸光法的高效液相色谱（HPLC）方法，用于分析血浆和尿液中的哌拉西林 - 他唑巴坦（他唑西林）。他唑巴坦在218 nm处进行检测，哌拉西林在222 nm处进行检测。血浆中这两种化合物以及尿液中哌拉西林的测定程序包括加入内标（他唑巴坦用头孢他啶，哌拉西林用苄青霉素），然后用乙腈和氯仿处理样品。为了定量尿液中的他唑巴坦，使用柱切换技术对稀释后的样品进行分析，无需内标。HPLC柱LiChrosorb RP - select B用由乙腈 - 醋酸铵（pH 5）组成的洗脱液混合物平衡。所提出的技术具有可重复性、选择性和可靠性。该方法已经过验证，并进行了各种条件下的稳定性测试。对于校准曲线标准品，观察到线性检测器响应，他唑巴坦的范围为5 - 60微克/毫升，哌拉西林的范围为1 - 100微克/毫升，涵盖了目前认为的体液中他唑西林浓度的临床相关范围。血浆中他唑巴坦的定量限为3微克/毫升，哌拉西林为0.5微克/毫升。血浆的提取回收率超过85%。以变异系数（C.V.）表示的精密度在0.4 - 18%范围内。
