磺基水杨酸处理对高效液相色谱法检测人纤维蛋白原中蔗糖含量的影响

Influence of sulfosalicylic acid treatment on the determination of sucrose content in human fibrinogen by high performance liquid chromatography

目的研究磺基水杨酸处理样品对高效液相色谱法检测人纤维蛋白原中蔗糖含量的影响。方法通过比较未经或经磺基水杨酸处理的蔗糖对照品溶液色谱图,确认磺基水杨酸处理对蔗糖水解的影响。蔗糖对照品溶液和人纤维蛋白原样品分别经磺基水杨酸处理2、3、4、5 h,研究处理时间对蔗糖水解的影响。用未经和经磺基水杨酸处理的蔗糖对照品溶液分别建立标准曲线,计算3批人纤维蛋白原中蔗糖含量,并与理论添加量比较,评价磺基水杨酸处理对人纤维蛋白原中蔗糖含量检测的影响。结果经磺基水杨酸处理的蔗糖对照品溶液色谱图比未经磺基水杨酸处理的增加2个水解峰,其中1个为葡萄糖峰;蔗糖对照品溶液和人纤维蛋白原样品经磺基水杨酸处理2、3、4、5 h,水解峰面积逐步增加;用未经和经磺基水杨酸处理的蔗糖对照品溶液分别建立的标准曲线计算3批人纤维蛋白原中蔗糖含量,计算结果与理论添加量的比例分别为93.1%±1.7%和97.7%±1.8%,单因素方差分析的F值为10.78,P值为0.03,差异有统计学意义。结论蔗糖经磺基水杨酸处理会发生水解,且处理时间延长,水解程度增加。蔗糖对照品溶液经磺基水杨酸处理后建立的标准曲线更适合人纤维蛋白原中蔗糖含量检测。

Objective To study the effect of sulfosalicylic acid treatment on determination of sucrose content in human fibrinogen by high performance liquid chromatography(HPLC).Methods The hydrolysis of sucrose was confirmed by comparing the chromatograms of sucrose solution treated with and without sulphosalicylic acid.The effect of treatment time on the hydrolysis of sucrose was studied by treating sucrose control solution and human fibrinogen with sulfosalicylic acid for 2,3,4 and 5 h,respectively.In order to evaluate the effect of sulfosalicylic acid on the determination of sucrose content in human fibrinogen,the sucrose content in 3 batches of human fibrinogen was calculated by different standard curves in which the sucrose control solution was treated not or treated with sulfosalicylic acid,and the results were compared with theoretical value.Results The chromatogram of sucrose control solution treated with sulfosalicylic acid showed two more hydrolytic peaks than that without sulfosalicylic acid,and one of them was glucose peak.The area of hydrolysis peak of sucrose control solution and human fibrinogen containing sucrose treated with sulfosalicylic acid for 2,3,4 and 5 h increased gradually.The sucrose content of 3 batches of human fibrinogen calculated using the standard curves of sucrose control solution not treated or treated with sulfosalicylic acid was compared with theoretical addition value and the ratio were 93.1%±1.7%and 97.7%±1.8%respectively,with statistically significant difference(F=10.78,p=0.03).Conclusions Sucrose can be hydrolyzed by sulfosalicylic acid,and the degree of hydrolysis increases with the prolongation of treatment time.The standard curve of sucrose control solution treated with sulfosalicylic acid is more suitable for the detection of sucrose content in human fibrinogen.