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目的:建立测定盐酸二甲双胍原料中有关物质二甲胺的柱前衍生化反相高效液相色谱法。方法:样品经氯甲酸-9-芴甲酯(FMOC-Cl)试剂衍生化,RP-HPLC分析,采用Wonda Sil C18Superb(150 mm×4.6 mm,5μm)色谱柱,流动相A为0.1%甲酸溶液,流动相B为乙腈,梯度洗脱:0~13 min,B 55%;13~13.1 min,B 55%~80%;13.1~20 min,B 80%;20~20.1 min,B 80%~55%;20.1~30 min,B 55%,流速为1.0 m L·min-1,进样量为20μL,柱温为30℃,紫外检测波长为265 nm。结果:衍生化反应在短时间内完成,且衍生物稳定。二甲胺在0.126~3.789μg·m L-1范围内线性关系良好(r=0.999 7)。检测限为0.039μg·m L-1。加样回收率为97.17%~102.0%。方法专属性、进样精密度与重复性良好。结论:本方法准确、方便、快速,可作为准确控制盐酸二甲双胍原料中有关物质二甲胺含量的有效分析手段。
Objective: To establish a pre-column derivatization reversed-phase high performance liquid chromatography (HPLC) for the determination of dimethylamine in the raw material of metformin hydrochloride. Methods: The sample was derivatized with FMOC-Cl reagent and analyzed by RP-HPLC using Wonda Sil C18 Superb (150 mm × 4.6 mm, 5 μm) column. The mobile phase A was 0.1% formic acid , Mobile phase B was acetonitrile, gradient elution: 0-13 min, B 55%; 13-13.1 min, B 55% -80%; 13.1-20 min, B 80%; 20-20.1 min, B 80% 55%; 20.1 ~ 30 min, B 55%, flow rate 1.0 m L · min-1, injection volume 20μL, column temperature 30 ℃, UV detection wavelength of 265 nm. Results: The derivatization reaction was completed in a short time and the derivatives were stable. The linear range of dimethylamine was 0.126 ~ 3.789μg · m L-1 (r = 0.999 7). The detection limit was 0.039μg · m L-1. The recovery rate was 97.17% ~ 102.0%. Method specificity, good injection precision and reproducibility. Conclusion: The method is accurate, convenient and rapid, which can be used as an effective analytical method to accurately control the dimethylamine content of metformin hydrochloride raw materials.