Objective: To establish a UPLC-MS/MS method for simultaneous detection of 6 aristolochic acids (aristolochic acid Ⅰ, aristolochic acid Ⅱ, aristolochic acid Ⅲa, aristolochic acid Ⅳa, aristolochic lactam Ⅰ and aristolochic lactam Ⅱ) in Zhengtian Pills. Methods: The separation was performed on a Waters ACQUITY UPLC BEH C18 column (100 mm×2.1 mm, 1.7 µm) by gradient program with acetonitrile-0.1% formic acid solution (including 5 mmol · L-1 ammonium formate) as mobile phase. The flow rate was set at 0.3 mL · min-1 and the column temperature was 40 ℃. Detection and quantification were performed by mass spectrometry in multiple reaction monitoring (MRM) mode with m/z 359.2→298.2 for aristolochic acid Ⅰ, m/z 329.2→268.2 for aristolochic acid Ⅱ,m/z 345.2→282.2 for aristolochic acid Ⅲa, m/z 375.2→312.2 for aristolochic acid Ⅳa, m/z 294.2→279.2 for aristolochic lactam Ⅰ and m/z 264.1→206.2 for aristolochic lactam Ⅱ. Results: The linear ranges of aristolochic acid Ⅰ, aristolochic acid Ⅱ, aristolochic acid Ⅲa, aristolochic acid Ⅳa, aristolochic lactam Ⅰ and aristolochic lactam Ⅱ were 1.614-103.3 ng · mL-1, 6.486-1 038 ng · mL-1, 3.293-1 054 ng · mL-1, 6.270-1 003 ng · mL-1, 6.180-988.8 ng · mL-1 and 3.098-991.2 ng · mL-1 respectively. All ingredients showed good linear relationship (r≥0.999 6). The average recoveries were 82.8%-104.6%, 90.7%-98.4%, 94.4%-105.1%, 91.5%-113.7%, 92.3%-99.9%, 85.9%-102.0%, respectively, with RSDs of 2.8%-10.8%, 3.0%-5.8%, 1.5%-2.5%, 1.7%-6.7%, 3.4%-6.2%, 3.3%-7.2%, respectively. In 12 batches of Zhengtian Pills, the contents of aristolochic acid Ⅰ were 52.35-70.15 ng · g-1, the contents of aristolochic acid Ⅳa were 2 394.29-3 281.55 ng · g-1, and the contents of aristolochic lactam Ⅰ were 139.85-172.20 ng · g-1, while aristolochic acid Ⅱ, aristolochic acid Ⅲa and aristolochic lactam Ⅱ were not detected. Conclusion: The established method is specific, sensitive and accurate, and can be used to determine the contents of six aristolochic acids in Zhengtian Pills, providing reference for the safe use and quality control of Zhengtian Pills and traditional Chinese medicine preparations containing aristolochic acid.
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