酶解液相色谱法测定硫酸软骨素钠含量

目的:建立酶解液相色谱法测定硫酸软骨素钠的含量。方法:使用硫酸软骨素ABC酶酶解硫酸软骨素钠,使用强阴离子交换高效液相色谱测定酶解产生的二糖,通过计算二糖峰面积总和确定硫酸软骨素钠的含量。色谱柱为Zorbax SAX(250 mm×4.6 mm,5μm);流动相为pH 3.5的水和pH 3.5的2 mol.L-1的氯化钠溶液,流速为1.0 mL.min-1,检测波长为232 nm,柱温为40℃。结果:硫酸软骨素钠的线性范围为0.25～1.50 mg.mL-1,r=0.9999;精密度RSD为0.3%;回收率为101.2%,RSD为1.6%(n=5);最低检测限为4.0μg.mL-1,定量限为11.9μg.mL-1。结论:该方法具有专属性好,灵敏度高,重现性好的优点。

Enzymolysis-HPLC determination of the content of chondroitin sulfate sodium

Objective:To establish enzymolysis-HPLC method for determining of the content of chondroitin sulfate sodium.Methods: Chondroitin sulfate sodium was first digested by chondroitinase ABC.The CS disaccharides produced were analyzed by SAX-HPLC and CS content was quantified by calculating the total peak areas of the disaccharides.Zorbax SAX column(250 mm×4.6 mm,5 μm)was adopted the mobile phase of water(pH 3.5) and 2 mol·L-1NaCl(pH 3.5) at 40 ℃ with flow rate of 1.0 mL·min-1,detecting wavelength being 232 nm.Results: The calibration curve was linear in the range of 0.25-1.50 mg·mL-1with correlation coefficient of 0.9999.The method precision RSD was 0.3%.The average recovery was 101.2% and its RSD was 1.6%(n=5).The low detection limit was 4.0 μg·mL-1 and the quantitative limit was 11.9 μg·mL-1.Conclusion: This method is reproducible,sensitive and suitable for the determination of the content of chondroitin sulfate sodium.