酶解液相色谱法测定硫酸软骨素钠含量
Enzymolysis-HPLC determination of the content of chondroitin sulfate sodium
分类号：
出版年·卷·期（页码）：2012,32 (7):0-0
DOI:
10.16155/j.0254-1793.2017.01.01
-----摘要：-------------------------------------------------------------------------------------------
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,5 μm);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4.0 μg·mL-1,定量限为11.9 μg·mL-1。 结论: 该方法具有专属性好,灵敏度高,重现性好的优点。
-----英文摘要：---------------------------------------------------------------------------------------
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.
-----参考文献：---------------------------------------------------------------------------------------
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