北京生物医学工程

《整形手术用交联透明质酸钠凝胶》行业标准中交联剂含量的检测方法研究

Detecting methods for content of cross-linking agent in the standard of Cross-linked sodium hyaluronate gel for plastic surgery

作者：付海洋于浩李敏姜爱莉付步芳王召旭

单位：中国食品药品检定研究院（北京102629）烟台大学（山东烟台264000）

关键词：透明质酸；交联透明质酸钠凝胶； 1，4-丁二醇二缩水甘油醚；气相色谱法；酶标法

分类号：R318.04

出版年·卷·期（页码）：2021·40·3（288-294）

摘要：

目的改进和优化YY/T 0962-2014《整形手术用交联透明质酸钠凝胶》行业标准中交联剂1，4-丁二醇二缩水甘油醚（1,4-butanediol diglycidyl ether ，BDDE）含量的标准检测方法，为该标准的修订提供技术依据。方法在气相色谱法中，为使BDDE更好地从凝胶内部释放出来，在测定之前增加样品的酶解处理步骤，并用乙酸乙酯对BDDE进行萃取。此外，在酶标法中研究透明质酸酶的影响，将透明质酸酶添加到标准溶液中，从而去除透明质酸酶对测定结果的影响。最后委托M、Q、X三家实验室对该方法进行验证，对同一批次样品进行BDDE含量测定，对数据结果进行双因子方差分析，以检验该方法是否具有可行性。结果气相法专属性、精密度以及重复性良好，平均回收率为105.02%，相对标准偏差（relative standard deviation ，RSD）为9.15%（n=6）。酶标法精密度、重复性良好，BDDE浓度0.5~8 μg/mL范围内，BDDE浓度和荧光强度有良好的线性关系，线性方程为y=36.89 x+2.135(r=0.9989)，检出限为0.106 μg/mL，平均回收率为97.43%，RSD为8.22%；采用双因子方差分析，对M、Q和本课题组（Z）三家实验室的验证结果进行统计（P＞0.05），认为不同实验室采用同一方法对同批次样品的交联剂含量测定结果不存在显著性差异。结论对于整形手术用交联透明质酸钠凝胶中交联剂BDDE含量的测定，气相法操作简单快捷，可作为BDDE的日常测定方法；酶标法特异性更高，数据更为真实精确，可作为整形手术用交联透明质酸钠凝胶产品中交联剂含量的仲裁方法。

Objective To improve and optimize the standard detection method for the content of crosslinking agent 1,4-butanediol diglycidyl ether (BDDE) in the industry standard of YY/T 0962-2014 rosslinked Sodium Hyaluronate Gel for Plastic Surgery, and to provide technical basis for the revision of the standard. Methods In gas chromatography, in order to better release BDDE from the inside of the gel, the enzymatic hydrolysis step of the sample is added before the measurement, and the enzymatic hydrolysis step is added during the treatment, and the BDDE is extracted with ethyl acetate. In addition, in the enzyme-labeled method, the effect of hyaluronidase was studied, and hyaluronidase is added to the standard solution to deduct the effect of hyaluronidase on the measurement results. Finally, three laboratories, M, Q, and X, are entrusted to verify the method. BDDE content determination is performed on the same batch of samples, and the data results are subjected to two-factor analysis of variance to test whether the method is feasible. Results The gas phase method has good specificity, precision and repeatability, the average recovery rate is 105.02%, and the relative standard deviation (RSD) is 9.15% (n=6). The precision and repeatability of the enzyme-labeled method are good. Within the range of BDDE concentration 0.5-8 μg/mL, there is a good linear relationship between BDDE concentration and fluorescence intensity. The linear equation is y=36.89 x+2.135 (r=0.9989), the detection limit is 0.106 μg/mL, the average recovery rate is 97.43%, and the RSD is 8.22%. Using two-way analysis of variance, the verification results of the three laboratories of M, Q and this research group (Z) are counted, P>0.05. It is believed that there is no significant difference in the determination results of the crosslinking agent content of the same batch of samples by different laboratories using the same method. Conclusions For the determination of the cross-linking agent BDDE content in cross-linked sodium hyaluronate gel for plastic surgery, the gas phase method is simple and quick to operate, and can be used as the daily determination method of BDDE; the enzyme-labeled method has higher specificity and more accurate data ,it can be used as an arbitration method for the content of cross-linking agent in cross-linked sodium hyaluronate gel products for plastic surgery.

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