Trivedi Harshal Kanubhai, Kshtri Nayan, Patel Mukesh C
Analytical Research Lab, Cadila Pharmaceutical Ltd, Dholka-387 810, Gujarat, India. ; P. S. Science and H. D. Patel Arts College, S. V. Campus, Kadi-382 715, Gujarat, India.
Sci Pharm. 2013 Jan-Mar;81(1):151-65. doi: 10.3797/scipharm.1208-20. Epub 2012 Nov 17.
The present work reports a rapid reversed-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous determination of 12 beta-lactam components for cleaning validation and cross-contamination. A strategic experimental approach was implemented for the method development. The desired chromatographic separation was achieved on a Symmetry C18 (4.6 X 75 mm, 3.5 μm) column using gradient elution. The optimized mobile phase consisted of the buffer tetrabutylammonium hydroxide pH-6.8 and acetonitrile. The eluted compounds were monitored at 215 nm and 254 nm wavelength using a photodiode array detector. The developed method separated 12-beta-lactam compounds from each other within a run time of 50 min. The method is effective for the determination of cross-contamination of penicillin and cephalosporin production blocks. The present method is specific and a lower limit of quantification was determined on the basis of the signal-to-noise ratio method; it is 1 μg/mL for all components. The developed RP-HPLC method was validated according to the International Conference on Harmonization (ICH) guidelines.
本研究报告了一种快速反相高效液相色谱（RP-HPLC）方法，用于同时测定12种β-内酰胺成分，以进行清洁验证和交叉污染检测。该方法开发采用了一种策略性实验方法。在Symmetry C18（4.6×75 mm，3.5μm）色谱柱上使用梯度洗脱实现了所需的色谱分离。优化后的流动相由氢氧化四丁基铵pH-6.8缓冲液和乙腈组成。使用光电二极管阵列检测器在215 nm和254 nm波长处监测洗脱的化合物。所开发的方法在50分钟的运行时间内将12种β-内酰胺化合物彼此分离。该方法对于测定青霉素和头孢菌素生产批次的交叉污染有效。本方法具有特异性，基于信噪比法确定了较低的定量限；所有成分的定量限均为1μg/mL。所开发的RP-HPLC方法根据国际协调会议（ICH）指南进行了验证。
