基于UPLC-Q Exactive Plus Orbitrap MS和UPLC-MS的益脑膏化学成分定性和定量研究OA

Objective To establish qualitative and quantitative analysis methods for the chemical constituents of Yinao Ointment(益脑膏,YNO)and clarify its material basis.Methods Chromatographic separation was performed on a Thermo Fisher Scientific Accucore Phenyl Hexyl column(2.1 mm×100 mm,2.6 μm)with gradient elution using 0.1％formic acid aqueous solution-acetonitrile as the mobile phase at a flow rate of 0.2 mL/min.The column temperature was maintained at 35 ℃.Using a heated electrospray ionization source,chemical constituents in YNO were identified through Full Scan/ddMS² mode,combined with comparisons of retention times of reference substances,mass spectrometry ion information,and relevant mass spectrometry data from databases and literature.Further-more,an UPLC-MS method was developed for the simultaneous determination of 24 components in YNO using multiple reaction monitor-ing(MRM)mode scanning.Results Through comparison with reference substances and high-resolution mass spectrometry analysis,116 components were identified and their fragmentation patterns were elucidated.On this basis,an UPLC-MS quantitative method was established for the simultaneous determination of 24 components in YNO.These 24 components showed good linear relationships within their respective detection concentration ranges(r>0.9992).The RSDs for precision,repeatability,and stability were all less than 6.46％,and the average recoveries ranged from 96％to 103.90％(all RSDs≤5.50％).Conclusion The established qualitative method can rapidly,sensitively,and accurately identify the chemical constituents in YNO.The established quantitative method is sensitive,reliable,and specific,capable of comprehensively reflecting the contents of various components in YNO,thus providing a reference basis for quality evaluation and improvement of YNO.