HPLC最终方法.doc

Comparison between the two Validated HPLC methods for PMIDA purity determination 两种已验证的测定双甘膦纯度的HPLC方法的比较 For the purity determination of PMIDA by HPLC technique, two separated methods were developed and validated following procedure were defined in agreement with ICH (International Conference on Harmonization) rules “Validation of Analytical Method: Definition and Terminology” 针对HPLC技术测定双甘膦的纯度，研发并验证了两种不同的方法，按照ICH（国际协调会）条例“分析方法的验证：定义和术语”来确定以下步骤。 The two methods, respectively reported in Report 048/11 and 140/11, are based on different chromatographic columns and different eluent solutions. 分别报告在报告048/11 和 140/11中的这两种方法基于不同的色谱柱和不同的洗脱溶液（流动相）。 In both the methods, the time of the analysis is comparable and the parameters obtained from the validation are quite comparable. 在这两种方法中，分析时间是可对比的并且从验证中获得的参数非常有可比性。 A summary of analytical data collected for each of them are following reported: 收集的两种方法的分析数据汇总报告如下： Report 048/11 with Strong Ion Exchange column 报告048/11，使用的是强离子交换柱 Analytical Procedure 分析步骤 Chromatographic conditions 色谱条件 Instrument: HPLC Agilent 1100 仪器：HPLC 安捷伦 1100 Column: ZORBAX SAX 4.6 x 250 mm 5 μm 柱：ZORBAX SAX 4.6 x 250 mm 5 μm Mobile phase: 85 % (250mM KH2PO4 pH 2) ÷ 15 % (MeOH) mixed 流动相：85 % (250mM 磷酸二氢钾 pH 2) ：15 % (甲醇) 的混合物 Flow: 1.5 mL/min 流量：1.5 毫升/分钟 Volume injected: 20 ?L 注入体积：20 ?L Column temperature: 25°C 柱温：25°C Detector: UV-visible at 195nm 探测器：195纳米的可视紫外线 Analysis time: 25 min. 分析时间：25分钟 Blank solution 空白溶液 Solvent. 溶剂 (85 % (250mM KH2PO4 pH 2) ÷ 15 % (MeOH) mixed) 85 % (250mM 磷酸二氢钾 pH 2) ：15 % (甲醇) 的混合物 Reference soltution (S) 参比溶液（S） 20 mg of PMIDA standard, accurately weighted, are dissolved into 20 mL of solvent. 把精确称取的20mg双甘膦标样溶解在20ml溶剂中。 Sample Solution (C) 样品溶液（C） 20 mg on the sample, accurately weighted, are dissolved into 20 mL of solvent. 把精确称取的20mg样品溶解在20ml溶剂中。 Sequence of injections 注入顺序 The solutions have to be injected in the instrument in the following sequence: 溶液必须按照以下顺序注入到仪器中： Blank 3 injectio