Qualitative and Quantitative Analysis of Aristolochic Acids in Aristolochia cinnabarina Dried Root Tubers by UPLC-QTOF-MS/MS / 中国实验方剂学杂志

ABSTRACT

Objective:To establish a qualitative and quantitative method for the determination of aristolochic acids in <italic>Aristolochia cinnabarina</italic> dried root tubers. Method:The dried root tubers of <italic>A. cinnabarina </italic>was qualitative and quantitative analysis by ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry （UPLC-QTOF-MS/MS）. The analysis was performed on Waters ACQUITY UPLC-BEH C₁₈ column （ 2.1 mm×100 mm， 1.7 μm） with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-1 min， 10%B； 1-9 min， 10%-30%B； 9-11 min， 30%-50%B； 11-15 min， 50%-90%B）. The flow rate was 0.45 mL·min^-1， column temperature was 35 ℃， and the detection wavelength was 250 nm. Mass spectral data was acquired in positive mode of electrospray ionization （ESI）. At the same time， the UPLC fingerprints of aristolochic acids in 21 batches of <italic>A. cinnabarina</italic> dried root tubers were established， and the contents of 5 aristolochic acids in <italic>A. cinnabarina</italic> dried root tubers from different producing areas and different harvesting periods were determined. Result:A total of 17 compounds， including 8 aristolochic acids， 7 aristololactams and 2 4，5-dioxoaporphine alkaloids， were identified from <italic>A. cinnabarina</italic> dried root tubers by mass spectrometry data and bibliographic information. Ten common peaks were identified in the UPLC fingerprint， and they were tuberosinone-<italic>N</italic>-<italic>β</italic>-<italic>D</italic>-glucoside， aristolactam Ⅰa-<italic>N</italic>-<italic>β</italic>-<italic>D</italic>-glucoside， aristolochic acid Ⅳa-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucoside， aristolactam Ⅲa-<italic>N</italic>-<italic>β</italic>-<italic>D</italic>-glucoside， aristolactam Ⅰ-<italic>N</italic>-<italic>β</italic>-<italic>D</italic>-glucoside， aristolochic acid Ⅲa， aristolochic acid Ⅳa， aristolochic acid Ⅱ， aristolactam Ⅰ and aristolochic acid Ⅰ. According to the quantitative analysis， the results exhibited that aristolochic acid Ⅲa， aristolochic acid Ⅳa， aristolochic acid Ⅱ， aristolactam Ⅰ and aristolochic acid Ⅰ had good linear relationships in the linear range. The relative standard deviations （RSDs） of precision， stability and reproducibility tests were all less than 3.0%， the recovery was 97.06%-101.84% （RSD<3.0%）. The contents of aristolochic acid Ⅰ， aristolochic acid Ⅱ， aristolochic acid Ⅲa， aristolochic acid Ⅳa， and aristolactam Ⅰ in 21 batches of <italic>A. cinnabarina</italic> dried root tubers were 0.938 6-3.567 5， 1.377 6-3.688 1， 0.056 3-0.527 7， 0.108 8-0.305 5， 0.021 0-0.081 7 mg·g^-1， respectively. Conclusion:The content of aristolochic acids in <italic>A. cinnabarina</italic> dried root tubers has a certain difference， the contents of aristolochic acid Ⅰ and Ⅱ are higher than other aristolochic acids. The established method is rapid， simple， accurate and reliable， which can provide reference for the quality control and evaluation of <italic>A. cinnabarina</italic> dried root tubers.