超高效液相色谱-串联四极杆飞行时间质谱法测定壬苯醇醚分子量分布及n值

Molecular weight distribution and n value of nonoxynol were determined by ultra-high performance liquid chromatography with tandem quadrupole time-of-flight mass spectrometry

目的:研究并建立超高效液相色谱-串联四极杆飞行时间质谱法测定壬苯醇醚低聚物的分子量分布及n值,并用国内外多批次壬苯醇醚样品进行考察。方法:采用Techmate STV SUNSHELL Peptide C_(18)(100 mm×2.1 mm,2.6μm,300A)色谱柱,以0.1%三氟乙酸水溶液(A)-0.1%三氟乙酸乙腈溶液(B)为流动相,线性梯度洗脱,流速0.3 mL·min^(-1),柱温60℃,进样量5μL。采用串联四极杆飞行时间质谱检测器,离子源为Dual ESI,氩气为碰撞气,以正离子模式,采集范围m/z 50~3000进行检测。采用ChemDraw19.0软件绘制相关结构,通过MassHunter PCDL Manager软件建立数据库,并编辑计算公式。结果:用新建立的方法对国内外生产的6批壬苯醇醚原料药的重均分子量及数均分子量进行了测定,并计算得到结构中n值的范围,结果发现国内外产品之间以及同一企业不同批次产品之间的重均分子量及数均分子量均有一定差异;产品结构中的n值也各不相同。数均分子量范围在664.6~686.8(n值约为10);重均分子量范围在677.1~715.5(n值约为11)。结论:此类低聚物药物的分子量分布及n值亟需在生产过程及质量标准中增加控制;新建立的方法灵敏度高,精密度和重现性好,可以满足测定壬苯醇醚原料药的分子量分布及n值的需要,为此类低聚物分子量分布的测定提供了解决方案,同时为《中华人民共和国药典》标准提高及修订提供了依据。

Objective:To develop an ultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q TOF MS)approach to investigate the molecular weight distribution and n value of nonoxynol oligomers,and to study several batches of samples produced domestically or abroad.Methods:The mobile phase was 0.1%trifluoroacetic acid aqueous solution(A)-0.1%trifluoroacetic acid in acetonitrile(B)with linear gradient elution at a flow rate of 0.3 mL·min^(-1).The column was Techmate STV SUNSHELL Peptide C_(18)(100 mm×2.1 mm,2.6μm,300A)and installed in a 60℃column oven.Injection volume was 5μL.A tandem quadrupole time-of-flight mass spectrometer with Dual ESI as ion source and argon as collision gas were used for detection in positive ion mode with a collection range of m/z 50-3000.ChemDraw 19.0 software was used to draw the relevant structure,and MassHunter PCDL Manager software was used to build the database and edit the calculation formula.Results:The weight average molecular weight and number average molecular weight of 6 batches nonoxynol drug substance produced domestically or abroad were determined by the newly established method,and the range of n average value in the structure was obtained.The results showed that the weight average molecular weight and number average molecular weight were different between domestic and foreign products and also between different batches produced by same manufacturer.The n value of structure basis also varies.The weight average molecular weight ranges were 664.6 to 686.8(n value was about 10),and number average molecular weight ranges were 677.1 to 715.5(n value was about 11).Conclusion:The molecular weight distribution and n value of such oligomer drugs need to be controlled in the production process and quality specifications.The newly established method has high sensitivity,precision and reproducibility,can be qualified for the needs of determining molecular weight distribution and n value of nonoxynol,and provides insight in determination of molecular weight distribution of such oligomers.This study also provides a data supplement for the improvement and revision of the standards of Chinese Pharmacopoeia.