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目的建立基于核磁共振(nuclear magnetic resonance,NMR)技术检测C群脑膜炎球菌荚膜多糖(group C meningococcal capsular polysaccharide,GCMP)、Y群脑膜炎球菌荚膜多糖(GYMP)和W135群脑膜炎球菌荚膜多糖(GWMP)化学结构的方法。方法建立1H-NMR检测方法:使用布鲁克600 MHz超导傅里叶核磁共振谱仪,配置5 mm BBO正向宽带高分辨液体探头。脉冲程序为水峰压制(zgpr),温度为298 K(25℃)。具体参数设置如下:中心频率约为4.7 ppm;图谱宽度为7 211 Hz;采样时间为2.272 s;弛豫延迟时间为2 s;扫描次数为512次;信号经傅里叶转换后加入1.0 Hz的窗口函数,检测结果使用Topspin 3.0软件进行处理。验证该方法的专属性、重复性及中间精密性。结果 5批GCMP、GYMP和GWMP检测结果的CV值均小于0.35%;3个浓度GCMP、GYMP和GWMP在相同的操作条件下以及不同工作日间、不同实验员间的检测结果的CV值均小于0.35%。结论建立的1H-NMR检测方法简便、快速、准确,检测GCMP、GYMP和GWMP具有极高的专属性、重复性和中间精密性,适用于疫苗生产研究中GCMP、GYMP和GWMP样品的定性结构鉴定。
OBJECTIVE To establish a method for the detection of group C meningococcal capsular polysaccharide (GCMP), group Y meningococcal capsular polysaccharide (GYMP) and group W135 meningococcal pod based on nuclear magnetic resonance (NMR) Membrane polysaccharide (GWMP) chemical structure of the method. Methods 1H-NMR detection method was established: Brück 600 MHz superconducting Fourier nuclear magnetic resonance spectrometer was used to configure 5 mm BBO forward broadband high resolution liquid probe. The pulse program was zgpr and the temperature was 298 K (25 ° C). The specific parameters are set as follows: the center frequency is about 4.7 ppm; the width of the spectrum is 7 211 Hz; the sampling time is 2.272 s; the relaxation delay time is 2 s; the number of scanning times is 512 times; Window functions, test results using Topspin 3.0 software for processing. Verify the method’s specificity, repeatability and intermediate precision. Results The CV values of GCMP, GYMP and GWMP were all less than 0.35% in 5 batches. The CV values of GCMP, GYMP and GWMP among 3 batches of GCMP, GYMP and GWMP under different operating conditions and different working days were all less than 0.35%. Conclusion The established 1H-NMR detection method is simple, rapid and accurate. The detection of GCMP, GYMP and GWMP has high specificity, repeatability and intermediate precision. It is suitable for the qualitative structure identification of GCMP, GYMP and GWMP samples in vaccine production research .