Analyzing Aristolochic Acids in Chinese Herbal Preparations Using LC/MS/MS

Chiu-Yu Huang ； Mu-Chuan Tseng ； Jer-Huei Lin

《藥物食品分析》 13卷2期 (2005/06) Pp. 125-131

並列摘要

This paper introduces a newly developed method of applying liquid chromatography-tandem mass spectrometry (LC/MS/MS) to the analysis of aristolochic acids (AAs) in Chinese herbal (medicinal) preparations. Sensitive and highly accurate readings were obtained for this study using both a photodiode array (PDA) detector and tandem mass spectrometer. The following optimized conditions for LC/MS/MS were set for this study: Separation was accomplished using a reverse phase C18 column (2.1×150 mm, 5 μm). The mobile phase comprised 35% acetonitrile and a 65% aqueous solution containing 0.1% formic acid and 0.1% ammonium acetate at a flow rate of 0.3 mL/min with a split ratio of 1:1 into the PDA detector and the tandem mass spectrometer MS/MS qualitative analysis was performed at 3.0 kV capillary voltage, with a collision energy level for aristolochic acid I (AA-I) of 10 eV and for aristolochic acid Ⅱ(AA-Ⅱ) of 12ev. The electrospray ionization source was operated in the positive mode AA-I [M+NH4](superscript +) ions at m/z 359 and AA-Ⅱ[M+NH4](superscript +) ions at m/z 329 were selected as precursor ions for daughter ion scanning. The characteristic daughter ion mass spectra for AA-I and AA-Ⅱ were generated and studied carefully. Quantification of results was done using the multiple reaction monitoring (MRM) method. The [M+NH4](superscript +) and [(M+NH4)-NH3-CO2](superscript +) ions of both AA-I and AA-Ⅱ were selected as precursor and daughter ions for the MRM analysis MS/MS detection limits were defined as 2.0 ng/mL of AA-I, and 2.8 ng/mL of AA-Ⅱ. The linear regression correlation coefficients of the calibration cure were 0.9992 of AA-I within the range of 0.02~16.00 μg/mL and 0.9988 of AA-Ⅱ within the range of 0.028~22.40 μg/mL. Relative standard deviations of 0.73% and 10.44% for AA-I, and 1.38% and 6.10% for AA-Ⅱ were determined for the intraday and interday tests. Recoveries for five differing concentrations of AA-I ranged from 99.0% to 106.9% and, for five differing concentrations of AA-Ⅱ, ranged from 92.0% to 104.5% Levels of AA-I and AA-Ⅱ detected in the 12 commercial Chinese medicinal preparation samples ranged from 11.1 to 3376.0μg/g and from 5.4 to 725.4 μg/g, respectively.

並列關鍵字

liquid chromatography-tandem mass spectrometry LC/MS/MS ； aristolochic acid ； piromidic acid ； electrospray ； multiple reaction monitoring MRM