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目的建立巯基化壳聚糖中巯基含量测定的方法。方法 pH值为4.5时,4,4'-二吡啶基二硫(4,4'-dithiodipyridine,DTDP)与巯基化合物反应,生成4-硫代吡啶酮,在波长324 nm处有最大吸收,通过测定反应产物的吸光度值计算巯基含量。结果 DTDP乙腈溶液稳定性好,标准曲线方程ΔA=21.611c+6.12×10-2(r=0.999 9),巯基浓度在3～60μmol.L-1内吸光度差值与巯基含量的线性关系良好。结论本法灵敏度高,准确可靠,比经典的Ellman试剂法更适用于巯基化壳聚糖中巯基的含量测定。
Objective To establish a method for the determination of sulfhydryl content in thiolated chitosan. Methods 4,4'-dithiodipyridine (4,4'-dithiodipyridine, DTDP) reacts with thiol at pH 4.5 to generate 4-thiopyridone with maximum absorption at 324 nm The absorbance of the reaction product was measured to calculate the thiol content. Results DTDP acetonitrile solution showed good stability. The standard curve equation was ΔA = 21.611c + 6.12 × 10-2 (r = 0.999 9). The linear relationship between the difference of absorbance and the content of thiol was observed when the concentration of DTDP was 3 ~ 60μmol.L-1. Conclusion The method is sensitive, accurate and reliable, and is more suitable for the determination of sulfhydryl content in thiolated chitosan than the classical Ellman reagent method.