吖啶橙分光光度法测定硫酸软骨素含量

Spectrophotometric determination of chondroitin sulfate with acridine orange

目的建立一种测定硫酸软骨素含量的分光光度分析新方法。方法在pH4．0的Britton．Robinson（B-R）缓冲溶液中，-丫啶橙与硫酸软骨素发生相互作用生成生物超分子复合物。吖啶橙在492nm处有一特征吸收峰，加入硫酸软骨素后，最大吸收波长不变，但吸收峰强度降低。结果在最佳条件下，溶液吸收度值的降低与硫酸软骨素浓度成线性关系，线性范围为0．04—3．00mg／L，表观摩尔吸光系数（ε）=6600L／（mol·cm）。工作曲线为△A：0．0001＋0．0655C（mg／L）（n=8，r=0．998）。结论此法简单、准确、灵敏、选择性好。优化了结合反应的条件，建立了分光光度法测定硫酸软骨素的方法。

Purpose To establish a new method for the determination of the content of the chondroitin sulfate （CS）. Methods A bio-supramolecular complex was formed when acridine orange interacted with CS in the pH 4.0 Britton-Robinson （B-R） buffer solution. Acridine orange has a characteristic absorption peak at 492 nm. After the addition of CS to the above solution, it resulted in the decrease of the maximal absorbance value of acridine orange and did not have any new absorption peak, Results Under the optimal conditions, there was a linear relationship between the decrease of the absorbance value and the concentration of CS. The linear range for the determination of CS was in the range of 0.04-3,00 mg/L with the linear regression equation as △A = 0.000 1± 0.065 5℃（mg/L）（ n = 8, r = 0.998）. The molar absorptivity of the reaction solution was calculated as 6 600 L/（mol·cm）. Conclusion The method was sensitive, accurate and suitable for sample determination.