Narola Bhavil, Singh A S, Mitra M, Santhakumar P R, Chandrashekhar T G
Ranbaxy Research Laboratories, Plot No 20, Sector-18, Udyog Vihar Industrial Complex, Gurgaon-122001, Haryana, India.
Anal Chem Insights. 2011;6:7-14. doi: 10.4137/ACI.S5953. Epub 2011 Feb 23.
This paper deals with development and validation of a high performance liquid chromatographic method for the quantitative determination of disodium EDTA (Ethylenediaminetetraacetic acid) in Meropenem active pharmaceutical ingredient (API). EDTA was derivatized with Ferric chloride solution by heating at 70 °C in water bath for about 20 minutes and the chromatographic separation achieved by injecting 100 μL of the derivatized mixture into a Waters HPLC system with photodiode array detector using a Phenomenex Luna C18(2) column (250 × 4.6 mm), 5 μ. The mobile phase consisting of 5% methanol and 95% of 0.7 g/L solution of Tetra butyl ammonium bromide and 4.6 g/L solution of sodium acetate trihydrate in water (pH adjusted to 4.0 with the help of acetic acid glacial) and a flow rate of 1 milliliter/minute. EDTA eluted at approximately 6 minutes. The method was suitably validated with respect to specificity, linearity of response, precision, accuracy, ruggedness, stability in analytical solution, limit of quantitation and detection and robustness for its intended use.
本文论述了一种高效液相色谱法的开发与验证，该方法用于定量测定美罗培南活性药物成分（原料药）中的乙二胺四乙酸二钠（Ethylenediaminetetraacetic acid）。通过在70°C水浴中加热约20分钟，使乙二胺四乙酸（EDTA）与氯化铁溶液进行衍生化反应。将100μL衍生化混合物注入配备光电二极管阵列检测器的沃特世高效液相色谱系统中，使用菲罗门Luna C18(2) 柱（250×4.6 mm，5μm）实现色谱分离。流动相由5%甲醇和95%的0.7 g/L四丁基溴化铵溶液以及4.6 g/L三水合乙酸钠溶液（用冰醋酸将pH调至4.0）的水溶液组成，流速为1毫升/分钟。EDTA在约6分钟时洗脱。该方法针对特异性、响应线性、精密度、准确度、耐用性、分析溶液中的稳定性、定量限和检测限以及其预期用途的稳健性进行了适当验证。
