荧光分光光度法测定交联透明质酸凝胶中交联剂的残留量

Determination of free 1,4-butanediol diglycidyl ether in cross-linked hyaluronic acid gel by fluorospectrophotometry

目的:建立测定交联透明质酸凝胶中交联剂1,4-丁二醇二缩水甘油醚(BDDE)残留量的方法。方法:利用BDDE与烟酰胺产生的强荧光物质,在λex=370 nm/λem=430 nm检测其荧光强度,建立测定BDDE残留量的荧光分光光度法,并与气相色谱法进行比较。结果:在0.5～8.0μg.mL-1范围内,BDDE浓度和荧光强度有良好的线性关系,回归方程为y=76.729x-2.9517,r2=0.998;定量限为0.53μg.mL-1;平均回收率为99.4%,RSD=2.5%;市售样品中BDDE平均残留量为(1.47±0.41)μg.mL-1。气相色谱法定量限为1.85μg.mL-1,灵敏度和线性相关性均较差。结论:荧光分光光度法简单准确,灵敏度高,更适用于交联透明质酸凝胶的质量控制。

Objective：To establish a method of determining free 1,4-butanediol diglycidyl ether（BDDE） which is used as a cross-linking agent and a leftover remained in the cross-linked hyaluronic acid gel.Methods：Making use of strong fluorescent substance produced by BDDE and nicotinamide where excitation wavelength and emission wavelength were located at 370 nm and 430 nm respectively to detect its fluorescence intensity,to establish fluorospectrophotometry of testing BDDE residues and to make a comparison with gas chromatography.Results：The method had an excellent linear range of 0.5-8.0 μg·mL-1,y=76.729x-2.9517,r2=0.998.The quantification limit was 0.53 μg·mL-1.The average recovery of this method was 99.4%,RSD=2.5%.The content of BDDE in the cross-linked Hyaluronic acid gel produced in our lab was（1.47±0.41）μg·mL-1.The quantification limit of gas chromatography was 1.85 μg·mL-1,the sensitivity and linear correlation were worse than the fluorospectrophotometry.Conclusions：The fluorospectrophotometry was simple,accurate and highly sensitive.It was more suitable to the quality control of cross-linked hyaluronic acid gel.