Determination of Four Aristolochic Acids in Asarum by Ultra-high Performance Liquid Chromatography-tandem Triple Quadrupole Tandem Mass Spectrometry
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Abstract:
In this study, an analytical method for the simultaneous determination of four aristolochic acids in asarum by ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry (UPLC-QQQ-MS/MS) was established. The samples were dissolved with 70% methanol, extracted ultrasonically, and separated on a Waters ACQUITY UPLC HSST3 chromatographic column (2.1 mm×100 mm×1.8 μm) using 0.1% formic acid in water -methanol as the mobile phase in gradient elution. The flow rate was 0.3 mL/min, and the column temperature was 40 ℃. Aristolochic acid A, aristolochic acid B, aristolochic acid C and aristolochic acid D were analyzed using electrospray ionization (ESI), positive ion mode scanning and multiple reaction monitoring mode (MRM). Results showed that aristolochic acid A, aristolochic acid B, aristolochic acid C and aristolochic acid D showed good linear relationships within the concentration range of 4~100 μg/L (r≥0.995). The detection limits were 3.00 μg/kg, 5.00 μg/kg, 25.00 μg/kg and 20.00 μg/kg, respectively, with the spiked recoveries ranging from 91.97% to 106.40% and RSD≤4.24% (n=9). The method had high specificity, accuracy, precision and wide linear range, and is suitable for simultaneous determination of aristolochic acids A, aristolochic acid B, aristolochic acids C, aristolochic acids D in asarum. The test results show that asarum and asarum-containing Chinese traditional medicine had certain amounts of aristolochic acid A and aristolochic acid D, but did not contain aristolochic acid B and aristolochic acid C. The data provide a scientific basis for safe daily consumption and medicinal use of asarum, and the quality control of asarum-containing Chinese traditional medicines.
