低分子肝素残留溶剂检测方法建立

GC Detection of Residual Solvents in Low Molecular Weight Heparin

目的建立不同低分子肝素中甲醇、乙醇、二氯甲烷残留溶剂的检测方法。方法采用顶空气相色谱法测定。色谱柱为DB-624毛细管柱(6%氰丙基苯基-94%二甲基聚硅氧烷),规格:0.53 mm×30 m,1μm;进样口温度:140℃;柱温:程序升温,起始温度40℃,维持4 min,5℃·min-1升至80℃,维持2 min;流速:2.0 mL·min-1;检测器:FID检测器,温度230℃;载气:氮气;分流比:2∶1;顶空条件:顶空瓶温度:80℃;顶空平衡时间:30 min;顶空循环时间:40min;定量环温度:90℃;Transfer Line温度:100℃。结果依诺肝素钠、达肝素钠及那曲肝素钙3种低分子肝素中的残留溶剂均可以得到有效检测,分离度良好,方法精密度及回收率较为理想。结论新建方法准确、灵敏,可用于不同低分子肝素甲醇等残留溶剂的检测。

OBJECTIVE To establish a method for determining methanol, ethanol and methylene dichloride residues of low molecular weight heparin（LMWH） by gas chromatography. METHODS DB - 624 capillary column （0. 53 mm × 30 m, 1 μm） was used, the injector temperature was 140 ℃, the column temperature was raised by program：the initial temperature was 40 %, maintained for 4 min was raised to 80 ℃ with a rate of 5 ℃ · min^-1 , maintained for 2 min, and the flow rate of carried gas N2 was 2.0 mL · min^-1, the FID detector temperature was 230 ℃, the ratio was 2： 1. The heated temperature of the headspace oven was 80 ℃, the heated time lasted 30min, and the cycle time was 40 rain, the injector temperature was 90 ℃, the transfer line temperature was 100 %. RESULTS Under the above conditions, each solvent of three LMWH was basically completely separated in chromatogram obtained from a systemic suitability test, and it has good precision and accuracy. CONCLUSION The method is simple and accurate, so it is useful for testing methanol and other solvents of LMWH.