两种RP－HPLC柱前衍生化快速测定核糖霉素方法的建立与比较

Establishment and Comparison of Two Methods for the Analysis of Ribostamycin by Pre-column Derivatization Rapid RP-HPLC

分别选用３，５－二硝基苯甲酰氯和２，４－二硝基氟苯作衍生化试剂，柱前衍生，采用高效细粒度短柱（４．６ｍｍ×３．３ｃｍ，３μｍ）快速分离，二极管阵列紫外检测器检测，外标法定量，建立了核糖霉素及杂质新霉胺的两种ＲＰ－ＨＰＬＣ快速测定方法，分析时间均低于５ｍｉｎ。对方法的重现性、灵敏度、准确度、线性范围等指标进行考察，结果均令人满意。运用统计配对ｔ检验法对两种方法的测定结果进行相关分析，结果表明：方法之间无显著性差异。

Two rapid RP-HPLC methods have been established for the analysis of ribostamycin and impurity neamine using 3,5-dinitrobenzoyl chloride (DNBCL) and 1-fluoro-2,4-dinitrobenzene (DNFB) pre-column derivatization. A high performance short column packed with small pellets(4.6 mm ×3.3cm, 3μm) has been used for rapid separation. A diode array ultraviolet detector has been employed for detection. The entire analytical time for each method is less than 5 min. Based on the validation of compendial methods in USP(ⅩⅩ Ⅱ), we have evaluated such performance characteristics as repeatability, sensitivity, accuracy and linear range etc. The results are satisfied. Furthermore, we have evaluated the correlativity of the tow methods using paired statistical t-test. No significant difference between the two methods has been shown.