Rane Shaligram S, Ajameri Alkesh, Mody Rustom, Padmaja P
Department of Applied Chemistry, Faculty of Technology and Engineering, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, India.
Intas Biopharmaceuticals Ltd., Ahmedabad, India.
J Pharm Anal. 2012 Apr;2(2):136-142. doi: 10.1016/j.jpha.2011.12.001. Epub 2011 Dec 30.
Rapid and sensitive reversed phase high performance liquid chromatography (RP-HPLC) and ultra performance liquid chromatography (RP-UPLC) method with UV detection has been developed and validated for quantification of parathyroid hormone (PTH) in presence of meta-cresol as a stabilizer in a pharmaceutical formulation. Chromatography was performed with mobile phase containing 0.1% Trifluoroacetic acid (TFA) in MilliQ water and 0.1% TFA in acetonitrile with gradient program and flow rate at 0.3 mL/min for HPLC and 0.4 mL/min for UPLC. Quantification was accomplished with internal reference standard (qualified against innovator product and National Institute for Biological Standards and Control (NIBSC) standard). The methods were validated for linearity (correlation coefficient=0.99), range, accuracy, precision and robustness. Robustness was confirmed by considering three factors; mobile phase composition, column temperature and flow rate/age of mobile phase. Intermediate precision was confirmed on different equipments, different columns and on different days. The relative standard deviation (RSD) (<2% for RP-HPLC and <1% for UPLC, =30) indicated a good precision. Retention time was found about 17 min and 2 min by HPLC and UPLC methods, respectively. Both methods are simple, highly sensitive, precise and accurate and have the potential of being useful for routine quality control.
已开发并验证了一种采用紫外检测的快速灵敏反相高效液相色谱法（RP-HPLC）和超高效液相色谱法（RP-UPLC），用于定量测定药物制剂中作为稳定剂的间甲酚存在下的甲状旁腺激素（PTH）。使用含有0.1%三氟乙酸（TFA）的超纯水和含有0.1% TFA的乙腈作为流动相，通过梯度程序进行色谱分析，HPLC的流速为0.3 mL/min，UPLC的流速为0.4 mL/min。采用内参标准品（与创新产品和国家生物标准与控制研究所（NIBSC）标准进行比对）进行定量。对方法的线性（相关系数 = 0.99）、范围、准确度、精密度和稳健性进行了验证。通过考虑三个因素确认了稳健性；流动相组成、柱温以及流动相的流速/使用时长。在不同设备、不同色谱柱以及不同日期确认了中间精密度。相对标准偏差（RSD）（RP-HPLC < 2%，UPLC < 1%，n = 30）表明精密度良好。通过HPLC和UPLC方法分别测得保留时间约为17分钟和2分钟。两种方法均简单、高度灵敏、精确且准确，具有用于常规质量控制的潜力。
