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：---------------------------------------------------------------------------------------
1 LING Pei-xue(凌沛学),WANG Feng-shan(王凤山).Advance on Research of Biological Drugs(生化药物研究进展).Beijing(北京):Peoples Medical Publishing House(人民卫生出版社),2004.403
2 FAN Hui-hong(范慧红).The revision for the quality standard of polysaccharide drug in Chinese Pharmacopoeia 2010(2010年版《中国药典》中多糖类药物标准的修订).Chin Pharm J(中国药学杂志),2010,45(17):1294
3 USP32.2009.980
4 EP6.0.2008.1525.
5 BP2010.2010.489.
6 ChP(中国药典).2010.Vol Ⅱ(二部):984
7 Tatsuya Y,Hidehiko S,Osami H. Purification and properties of bacterial chondroitinases and chondrosulfatases. J Biol Chem, 1968, 243(7): 1523
8 Hidehiko S, Tatsuya Y,Sakaru S. Enzymatic methods for the determination of small quantities of isomeric chondroitin sulfates. J Biol Chem, 1968, 243(7): 1536
9 Rafael J,Jack B, Henry I. Purification and properties of the enzyme chondroitinase. J Biol Chem, 1959, 234(9): 2236
欢迎阅读《药物分析杂志》！您是该文第 1810位读者！
