北京生物醫(yī)學(xué)工程

《整形手術(shù)用交聯(lián)透明質(zhì)酸鈉凝膠》行業(yè)標(biāo)準(zhǔn)中交聯(lián)劑含量的檢測方法研究

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

作者：付海洋于浩李敏姜愛莉付步芳王召旭

單位：中國食品藥品檢定研究院（北京102629）煙臺(tái)大學(xué)（山東煙臺(tái)264000）

關(guān)鍵詞：透明質(zhì)酸；交聯(lián)透明質(zhì)酸鈉凝膠； 1，4-丁二醇二縮水甘油醚；氣相色譜法；酶標(biāo)法

分類號(hào)：R318.04

出版年·卷·期（頁碼）：2021·40·3（288-294）

摘要：

目的改進(jìn)和優(yōu)化YY/T 0962-2014《整形手術(shù)用交聯(lián)透明質(zhì)酸鈉凝膠》行業(yè)標(biāo)準(zhǔn)中交聯(lián)劑1，4-丁二醇二縮水甘油醚（1,4-butanediol diglycidyl ether ，BDDE）含量的標(biāo)準(zhǔn)檢測方法，為該標(biāo)準(zhǔn)的修訂提供技術(shù)依據(jù)。方法在氣相色譜法中，為使BDDE更好地從凝膠內(nèi)部釋放出來，在測定之前增加樣品的酶解處理步驟，并用乙酸乙酯對(duì)BDDE進(jìn)行萃取。此外，在酶標(biāo)法中研究透明質(zhì)酸酶的影響，將透明質(zhì)酸酶添加到標(biāo)準(zhǔn)溶液中，從而去除透明質(zhì)酸酶對(duì)測定結(jié)果的影響。最后委托M、Q、X三家實(shí)驗(yàn)室對(duì)該方法進(jìn)行驗(yàn)證，對(duì)同一批次樣品進(jìn)行BDDE含量測定，對(duì)數(shù)據(jù)結(jié)果進(jìn)行雙因子方差分析，以檢驗(yàn)該方法是否具有可行性。結(jié)果氣相法專屬性、精密度以及重復(fù)性良好，平均回收率為105.02%，相對(duì)標(biāo)準(zhǔn)偏差（relative standard deviation ，RSD）為9.15%（n=6）。酶標(biāo)法精密度、重復(fù)性良好，BDDE濃度0.5~8 μg/mL范圍內(nèi)，BDDE濃度和熒光強(qiáng)度有良好的線性關(guān)系，線性方程為y=36.89 x+2.135(r=0.9989)，檢出限為0.106 μg/mL，平均回收率為97.43%，RSD為8.22%；采用雙因子方差分析，對(duì)M、Q和本課題組（Z）三家實(shí)驗(yàn)室的驗(yàn)證結(jié)果進(jìn)行統(tǒng)計(jì)（P＞0.05），認(rèn)為不同實(shí)驗(yàn)室采用同一方法對(duì)同批次樣品的交聯(lián)劑含量測定結(jié)果不存在顯著性差異。結(jié)論對(duì)于整形手術(shù)用交聯(lián)透明質(zhì)酸鈉凝膠中交聯(lián)劑BDDE含量的測定，氣相法操作簡單快捷，可作為BDDE的日常測定方法；酶標(biāo)法特異性更高，數(shù)據(jù)更為真實(shí)精確，可作為整形手術(shù)用交聯(lián)透明質(zhì)酸鈉凝膠產(chǎn)品中交聯(lián)劑含量的仲裁方法。

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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