单唾液酸四己糖神经节苷脂钠有关物质测定方法的研究

Study onmethods for related substances measurment in monosialotetrahexosyl ganglioside

目的:建立单唾液酸四己糖神经节苷脂钠(GM1)有关物质的检测方法。方法:采用COSMOSIL 5C8-MS柱(250 mm×4.6 mm,5μm),以乙腈-0.01 mol.L-1四丁基硫酸氢铵(80∶20)为流动相,流速1.0 mL.min-1,在205 nm波长处,应用外标法对GM1中已知杂质GD1a和GD3进行检查,其他未知杂质用不加校正因子的主成分自身对照法进行检查。结果:GD1a和GD3在15～240μg.mL-1浓度范围内线性关系良好,相关系数均为0.9999;检测限分别为1.40μg和1.41μg;定量限分别为4.61μg和4.60μg;方法重复性的RSD分别为0.45%和0.51%;该方法与现行标准的对比测定结果显示,现行标准测得的杂质总量和总数目明显低于该方法。结论:本方法专属性强,灵敏度高,精密度好,可用于质量控制以及GM1原料药中有关物质的研究。

Objective： To establish a method for monosialotetrahexosyl ganglioside （ GM1 ） measurement. Method： The known impurities GD1a and GD3 in GM1 were analyzed by external standard method on a COSMOSIL 5C8 - MS column （250 mm x 4. 6 mm ,5 p,m）with mobile phase consisted of acetonitrile -tetrabutylammonium acid （0.01 μg · mL^-1 ,80：20）at a flow rate of 1.0 mL · rain^-1 ,and the detection wave -length was set at 205 nm;meanwhile,the unknown impurities were examined by the self - compare method but without calibration factors. Results： Good lin- ear relations of GD1a and GD3 were obtained in the range of 15 -240 μg · mL^-1 , and both of the correlation coeffi- cients were 0. 9999. The limits of detection were 1.40 μg and 1.41 ixg,respectively. The limits of quality were 4. 61 μg and 4.60 Ixg, respectively. Meanwhile, the reproducibility of RSD were 0. 45% and 0. 51%, respectively. Com- pared with the results using current criteria, the total impurity amount was higher with this method. Conclusion： The established method is specific, sensitive and accurate, which can be used for quality control and research in re- lated substances of raw material GM1.