柱前衍生化RP-HPLC法测定盐酸二甲双胍原料中有关物质二甲胺  

Determination of N-methylmethanamine in metformin hydrochloride bulk drug using pre-column derivatization and RP-HPLC

作　　者：冯超[1,2] 张友谊[3] 徐雪[1,2] 孙晓敏[1,2] 张峻颖[4] 严芳[1,2] 冯芳[1,2] 吴春勇[1,2] 

机构地区：[1]药物质量与安全预警教育部重点实验室(中国药科大学),南京210009 [2]中国药科大学药物分析教研室,南京210009 [3]广州医科大学卫生职业技术学院,广州510925 [4]中国药科大学中药制剂教研室,南京211198

出　　处：《中国新药杂志》2015年第10期1191-1196,共6页Chinese Journal of New Drugs

基　　金：教育部高等学校博士学科点专项科研基金(20120096120004);江苏省自然科学基金(BK2012358);药物质量与安全预警教育部重点实验室开放课题(MKLDP2013MS05)

摘　　要：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 reversed-phase high performance liquid chromatographic method with 9-fluorenylmethyl chloroformate （FMOC-C1） pre-column derivatization for the determination of N-methylmethana- mine, a potential related substance in metformin hydrochloride bulk drug. Methods： After N-methylmethanamine was derivatized with FMOC-C1, the separation was carried out on a WondaSil C18 Superb column （150 mm × 4.6 mm,5 μm） with a 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%. The mobile phase was consisted of 0.1% formic acid solution （A） and acetonitrile （ B）, and the flow rate was 1.0 mL·min - 1. The injection volume was 20 μL and the column temperature was 30 ℃. The UV detector was set at 265 nm. Results ： The reaction of derivatization was very fast and the derivative was stable. The calibration curve was linear in the range over 0. 126 ~ 3. 789 μg·mL-1 （r =0. 999 7）. The limit of detection （LOD） was 0. 039 μg· mL-1. The recovery was in the range of 97.17% ~ 102.0%. The method showed good specificity, injection precision and repeatability. Conclusion： The proposed method is sensitive, accurate, and convenient for the determination of the potential related substance N-methyl- methanamine in metformin hydrochloride bulk drug

关 键 词：盐酸二甲双胍 有关物质 二甲胺 柱前衍生化法 反相高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]