离子色谱法测定肝素钠总己糖胺中半乳糖胺含量的方法研究

Determination of Galactosamine in Total Hexosamine of Heparin Sodium by Ion Chromatography

目的:建立肝素钠总己糖胺中半乳糖胺含量的检测方法。方法:将标准品及供试品在100℃下水解6 h,使用戴安ICS5000型离子色谱仪,以14 mm氢氧化钠溶液为淋洗液在0.5 m L/min流速下,经戴安氨基酸捕集柱(50 mm×4 mm)、PA20型色谱柱(150 mm×3 mm)分离,进入配备Au工作电极、pH-Ag-AgCl参比电极的安培检测器中进行检测。结果:经不低于6 h水解,供试品充分水解,空白溶液无干扰峰,水解标准溶液色谱图中,理论板数、分离度、拖尾因子等均符合USP〈831〉的规定,半乳糖胺的检测限和定量限分别达到2 ng/mL、4 ng/mL,半乳糖胺在4~160 ng/mL浓度范围内线性良好,平均加样回收率达到101.2%,RSD为2.64%,水解标准溶液和水解供试品溶液放置24 h,检测结果的RSD为1.53%。结论:方法专属性、系统适用性、线性、灵敏度、回收率良好,适用于肝素钠总己糖胺中半乳糖胺含量的检测。

Objective:To establish a method for the determination of galactosamine content in heparin sodium total hexosamine.Methods:The standard and test samples are hydrolyzed at 100 C for 6 hours,and then analyzed by Dionex ICS5000 ion chromatograph,using 14 mm sodium hydroxide solution as eluent at a flow rate of 0.5 mL/min,passing through Dionex amino acid trap column(50 mm×4 mm)and PA20 column(150 mm×3 mm),and entering the amperometric detector equipped with Au working electrode and pH-Ag-AgCl reference electrode.Results:After hydrolysis for no less than 6 hours,the test samples are fully hydrolyzed,and there is no interference peak in the blank solution.In the chromatogram of the hydrolyzed standard solution,the theoretical plate number,separation degree,tailing factor,etc.all met the requirements of USP<831>.The detection limit and quantitation limit of galactosamine were 2 ng/mL and 4 ng/mL,respectively.Galactosamine had good linearity in the concentration range of 4~160 ng/mL,the average recovery rate was 101.2%,and the RSD was 2.64%.The RSD of the test results of the hydrolyzed standard solution and the hydrolyzed test sample solution placed for 24 hours is 1.53%.Conclusion:The method has good specificity,system suitability,linearity,sensitivity,and recovery rate,and is suitable for the determination of galactosamine content in heparin sodium total hexosamine.