软骨素酶ABC酶解高效液相法测定鱼翅中的透明质酸

 建立了一种测定鲨鱼翅中透明质酸的酶解 高效液相方法。在37℃,0 2mol/LTris HCl缓冲溶液中,鱼翅中的透明质酸被硫酸软骨素酶ABC酶解成透明质酸二糖。条件为:ZORBAX糖分析柱(4 6mmi d ×250mm,5μm)柱;紫外检测波长226nm;流动相为乙腈 0 5%磷酸(体积比为2∶98),流速1mL/min;进样量10μL。透明质酸二糖的线性范围为25g/L～600g/L。将此法应用到鱼翅的实际样品检测中,取得了良好的结果,透明质酸在鱼翅样品中的质量分数为0 86%～1 96%。

Zymohydrolysis with Chondroitinase ABC and High Performance Liquid Chromatography Used for the Determination of Hyaluronic Acid in Shark Fin

A method for the determination of hyaluronic acid in shark fin by high performance liquid chromatography (HPLC) is described. At 37 degrees C, with 0.2 mol/L Tris-HCl buffer solution, hyaluronic acid was converted to the hyaluronic acid disaccharide by zymohydrolysis with chondroitinase ABC. The chromatographic conditions were as follows: ZORBAX carbohydrate analysis column (4.6 mm i.d. x 250 mm, 5 microns); room temperature; UV-VIS detector set at 226 nm; mobile phase V(acetonitrile): V(0.5% phosphoric acid) = 2:98; 0.45 micron filter membrane, pumping filter; injection volume 10 microL; flow rate 1 mL/min. The calibration curve for the hyaluronic acid disaccharide was linear over the range of 25 g/L-600 g/L. This method was applied to the analysis of shark fin with satisfactory results. The hyaluronic acid contents in different shark fins were from 0.86% to 1.96%.