超高效液相色谱质谱联用法测定重组人乳头瘤病毒中3-(N-吗啡啉)丙磺酸含量

Determination of 3-(N-morpholine)propionic sulfonic acid in recombinant human papillomavirus by ultra-high performance liquid chromatography-mass spectrometry

目的:建立超高效液相色谱质谱联用法(UPLC-MS)测定重组人乳头瘤病毒中3-(N-吗啡啉)丙磺酸(MOPS)含量。方法:采用Waters ACQUITY UPLC BEH HILIC色谱柱,流动相为20 mmol·L^-1乙酸铵(pH5.0)-乙腈,梯度洗脱;流速0.5 mL·min^-1,柱温40℃,进样量为1.0μL;Qda检测器采用质量数208的SIR负离子扫描模式,锥孔电压为15 V,时间为3 min。结果:UPLC-MS测定重组人乳头瘤病毒原液和九价疫苗成品分离度分别为3.86和2.76,MOPS在浓度为20~0.0097656μg·g-1时线性关系良好(r^2=0.9979),定量下限为0.0097656μg·g-1,检测下限为0.0048828μg·g-1,精密度试验的RSD均不大于2.0%。HPV原液和HPV九价疫苗平均加样回收率分别为99.4%和100.6%。样品中未检测到MOPS。结论:该方法简单快速、准确、灵敏,重复性好,适用于重组人乳头瘤病毒原液和疫苗成品中的MOPS含量检测。

Objective:To establish a UPLC-MS method to determine the content of 3-(N-morpholine)propane sulfonic acid(MOPS)in recombinant human papillomavirus.Methods:Waters ACQUITY UPLC BEH HILIC column was used with the chromatographic method was as follows:20 mmol·L-1 ammonium acetate and acetonitrile were adopted as mobile phase with gradient elution at 0.5 mL·min-1 of the flow rate;the injection volume at 1.0μL,the column temperature at 40℃,and SIR negative scanning mode with a mass number of 208 for the Qda detector.The tapered hole voltage was 15 V.Results:The resolution product by UPLC-QDa of HPV bulk and HPV-9 vaccine was 3.86 and 2.76,respectively.The linear concentration of MOPS ranged from 0.0097656μg·g^-1 to 20μg·g-1(r^2=0.9979).The quantition and detection limit were 0.0097656μg·g-1 and 0.0048828μg·g^-1,respectively.The RSD were below 2.0%in precision study.The average recovery of HPV bulk and HPV 9 vaccine were 99.4%and 100.6%,respectively.Mops in samples was not detected.Conclusion:The method is simple,rapid,accurate,and with high resolution and repeatability.It could be applicable for determining the MOPS content of HPV bulk and HPV vaccine product.