基于TLC及UPLC-Q-TOF-MS/MS的铁棒锤毒效成分定性及HPLC定量研究OA

Objective:To establish a qualitative method for the five active components(neoaconitine,napelline,benzoyl aconitine,hypaconitine,and aconitine)in Aconitum pendulum Busch.and to quantitatively analyze their primary toxicity components.Methods:Ultra-high-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS)was used to characterize the chemical composition of A.pendulum.Based on the characterization results of its main components,a qualitative analysis method was established by optimizing the conditions of thin-layer chromatography(TLC).Furthermore,the methodology of simultaneous quantification by high-performance liquid chromatography(HPLC)was established for the toxicity components benzoyl aconitine,and aconitine.Results:The UPLC-Q-TOF-MS/MS analysis identified five main alkaloid components:neoaconitine,napelline,benzoyl aconitine,hypaconitine and aconitine.The results of TLC analysis showed that the identification method established based on these main alkaloid components could effectively distinguish A.pendulum from its adulterant Curculigo orchioides.The quantitative analysis results showed that benzoyl aconitine and aconitine had a good linear relationship within the concentration range of 194.00-833.00 μg·mL-1(r≥0.999 6),respectively.The detection limits were 1.10 and 1.00 μg·mL-1,respectively,and the limits of quantification were 7.39 μg·mL-1 and 6.90 μg·mL-1,respectively.The average spiked recovery rates were 97.51%and 102.21%,respectively,with relative standard deviations(RSD)both below 3.0%.Conclusion:The qualitative and quantitative methods established in this study can be used to distinguish between A.pendulum and C.orchioides,and provide references for their quality control and supporting clinical drug safety.