Development and verification of an ion pair reversed phase ultra-performance liquid chromatography for purity detection of mRNA vaccine bulk solution
10.13200/j.cnki.cjb.004674
- VernacularTitle:mRNA疫苗原液纯度离子对反相超高效液相色谱检测方法的建立及验证
- Author:
Wenyan CUI
- Publication Type:Journal Article
- Keywords:
mRNA vaccine;
Purity;
Quality control;
Ion pair reversed phase ultra-performance liquid chromatography(IPRP-UPLC)
- From:
Chinese Journal of Biologicals
2026;39(08):959-965
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo develop and verify an ion pair reversed phase ultra-performance liquid chromatography(IPRPUPLC) method for the determination of the purity of mRNA vaccine bulk solution, so as to provide a reliable, effective and stable detection method for the quality control of mRNA vaccine purity.MethodsThermo DNA Pac RP column(50 mm ×2.1 mm, 4 ??m) was used for IPRP-UPLC detection, and the mobile phase A ion pair reagent type [N,N-diisopropylethylamine(DIEA) + 1,1,1,3,3,3-hexafluoro-2-propanol(HFIP), triethylammonium acetate(TEAA)], mobile phase A ion pair reagent concentration(100, 200, 300, 400 mmol/L) and mobile phase B(acetonitrile, methanol), as well as column flow rate(0.4 and 0.5 mL/min) and column temperature(60 and 80 ℃) of the method were optimized.The specificity, applicability,linear range, limit of detection(LOD) and limit of quantitation(LOQ), precision and stability of the method were verified.In addition, the purity of six batches of GL2288.2 mRNA vaccine bulk solutions was detected using the established method.ResultsThe optimum chromatographic conditions were mobile phase A ion pair reagent of TEAA with a concentration of300 mmol/L and methanol as mobile phase B at a flow rate of 0.4 mL/min and a temperature of 80 ℃.Under the optimized chromatographic conditions, the resolution between the test solution, impurity reference solution, impurity reference mixed solution and the main components met the requirements.The retention time of degradation impurities produced by mRNA vaccine bulk solution under acid, alkali, oxidation, light, high temperature and other damage conditions was basically the same as that of poly(A) tail-deleted impurities, and the material balance was good.GL2288.2 reference substance had a good linear relationship with the peak area in the concentration range of 0.004 8 to 1.45 mg/mL, with linear equation of y =92.497 x-2.0393, R~2=0.997.The LOD and LOQ were 0.96 and 4.80 ??g/mL, respectively.The RSDs of precision verification were less than 1.0%.The RSD of peak purity of the test solution stored at 4 ℃ for 0-12 h was 0.17%, and the RSD of retention time was 0.19%.The purity RSD of the six batches of GL2288.2 mRNA vaccine bulk solutions detected by the established method was 0.07%, with good inter-batch consistency.ConclusionThe established IPRP-UPLC method demonstrates good specificity, precision, applicability and stability, and can be used for the quality control of mRNA vaccine bulk solution.
