核磁共振氢谱法对3种低相对分子质量肝素中残留溶剂乙醇的定性和定量分析  

Qualitative and Quantitative Analysis of Residual Solvent Ethanol in Three Low Molecular Weight Heparins by Proton Nuclear Magnetic Resonance

作　　者：李馨白 徐翊雯 张雅军[1] 孙会敏[1] 吴先富[1] LI Xin-bai;XU Yi-wen;ZHANG Ya-jun;SUN Hui-min;WU Xian-fu(National Institutes for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院,北京102629

出　　处：《中国药学杂志》2023年第8期725-729,共5页Chinese Pharmaceutical Journal

摘　　要：目的 建立测定达肝素钠、依诺肝素钠、那屈肝素钙3种低相对分子质量肝素中残留溶剂乙醇含量的定量核磁共振氢谱法(quantitative proton nuclear magnetic resonance, qHNMR)。方法 采用Bruker AVANCE NEO 600 MHz核磁共振波谱仪,采集3种低相对分子质量肝素的核磁共振氢谱(^(1)H-NMR),以2,2,3,3-d4-3-(三甲基硅基)丙酸钠盐(TSP-d4)作为内标物,重水(D2O)为溶剂,90°脉冲,采样次数(NS)为32次,测试温度25℃采样时间(AQ)为4.59 s,弛豫延迟时间(D1)为40 s,对3种低相对分子质量肝素中的残留溶剂乙醇进行定性和定量分析。结果 样品的^(1)H-NMR谱在化学位移δ 1.18和δ 3.65分别出现甲基和亚甲基质子信号,结合耦合常数可以确定样品中含有残留溶剂乙醇。同时,乙醇和TSP-d4定量峰信号在^(1)H-NMR谱上分离度良好,乙醇在线性范围内线性关系良好(r>0.999 9),方法精密度和重复性良好,平均回收率(n=9)为101.55%,3种低相对分子质量肝素中的乙醇含量为0.025%~0.493%,测定结果与顶空气相色谱法测定结果基本一致。结论 建立的qHNMR方法可以准确识别低相对分子质量肝素样品中是否含有乙醇,同时可以对其进行准确定量,相比于经典的气相色谱方法更加简便快速,并且不破坏样品,拓宽了残留溶剂的检测手段,可为不同低相对分子质量肝素产品中残留溶剂乙醇测定提供参考。OBJECTIVE To establish a quantitative proton nuclear magnetic resonance method for the determination of residual solvent ethanol content in three low molecular weight heparins:dalteparin sodium,enoxaparin sodium,and nadroparin calcium.METHODS Proton nuclear magnetic resonance spectra(^(1)H-NMR)were acquired using a Bruker AVANCE NEO 600 MHz nuclear magnetic resonance spectrometer,with 2,2,3,3-d4-3-(trimethylsilyl)propionate sodium salt(TSP-d4)as the internal standard and heavy water(D2O)as the solvent,90°pulse was adopted,the sampling number(NS)was 32 s,the test temperature was 25℃,the sampling time(AQ)was 4.59 s,and the relaxation delay time(D1)was 40 s,and the ethanol in three kinds of low molecular weight heparins was quantitatively studied.RESULTS The ^(1)H-NMR spectrum of the sample showed methyl and methylene proton signals at chemical shiftsδ1.18 andδ3.65 respectively,which was confirmed by coupling constants that the sample contains residual solvent ethanol.The quantitative peak signals of ethanol and TSP-d4 were well separated in the ^(1)H-NMR spectrum,and ethanol had a good linear relationship in the tested range(r>0.9999),and the precision and repeatability of the method were good.The recovery was 101.55%(n=9).The contents of ethanol in the three kinds of low molecular weight heparins were 0.025%-0.493%,the determination results were basically consistent with the determination results of headspace gas chromatography.CONCLUSION The established qHNMR method is simple,rapid,and accurate.Compared with the classic gas chromatography method,it is simpler and faster,and does not destroy the sample.The determination results are accurate and reliable.

关 键 词：定量核磁共振氢谱法 残留溶剂 乙醇 低相对分子质量肝素 达肝素钠 那屈肝素钙 依诺肝素钠 

分 类 号：R917[医药卫生—药物分析学]