Ingredient Analysis

Study on the detection and distribution of 6 aristolochic acids in Asari Radix et Rhizoma by UPLC-MS/MS

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  • Hebei Institute for Drug and Medical Device Control, Hebei Key Laboratory of Traditional Chinese Medicine Quality Evaluation and Standard Research, Shijiazhuang 050227, China

Received date: 2023-06-06

  Online published: 2024-06-24

Abstract

Objective: To establish an UPLC-MS/MS method for simultaneous detection of 6 aristolochic acids (aristolochic acids A, B, C, D, Ⅲ and Ⅶa) in Asari Radix et Rhizoma. The distribution of the components in different parts (root or rhizome) was investigated. Methods: The chromalographic separation was proformed on a C18 (100 mm×2.1 mm, 1.8 μm) column by gradient program with acetonitrile-0.1% formic acid solution as mobile phase. The flow rate was set at 0.3 mL·min-1 and the column temperature was 30 ℃. Detection and quantification were performed by mass spectrometry in multiple reaction monitoring(MRM) mode with m/z 359.0→298.0 for aristolochic acid A, m/z 329.0→268.0 for aristolochic acid B, m/z 345.0→284.0 for aristolochic acid C, m/z 375. 0→312.0 for aristolochic acid D, m/z 359.0→296.0 for aristolochic acid Ⅲ, and m/z 375.0→314.0 for aristolochic acid Ⅶa. Results: The calibration curves were linear within the ranges of 5.269 5-526.95 ng·mL-1, 5.543 3-554.33 ng·mL-1, 10.056-502.79 ng·mL-1, 4.723 1-472.31 ng·mL-1, 4.938 2-493.82 ng·mL-1, 5.042 7-504.27 ng·mL-1 for aristolochic acids A, B, C, D, Ⅲ and Ⅶa, respectively. All ingredients showed a good linear relationship (r≥0.999 6). The average recoveries were 98.2%, 100.2%, 103.1%, 101.7%, 96.2%, 104.3% with RSDs of 4.3%, 4.2%, 3.7%, 5.2%, 3.0%, 2.5%, respectively. Aristolochic acid B, C and Ⅲ were not detected in the roots or rhizoids of 12 batches of samples. The content of aristolochic acid A were 0.039 9-0.280 1 μg·g-1, 0.216 1-1.459 1 μg·g-1in the roots and rhizoids, respectively. The contents of aristolochic acid D in roots and rhizomes were 4.355 2-11.187 9 μg·g-1, 3.126 2-8.014 4 μg·g-1, respectively. The content of aristolochic acid Ⅶa in roots and rhizomes were 0-0.017 3 μg·g-1 and 0.008 4-0.070 3 μg·g-1, respectively. And the content of aristolochic acid D was the highest. Conclusion: This method is proved to be accurate and specific. The samples are with significantly regular pattern in quality and quantity of aristolochic acid analogues. It provides a more comprehensive scientific basis for the rational application of Asari Radix et Rhizoma and the limited determination of aristolochic acid components.

Cite this article

SUN Hui-zhu, WANG Xiao-lei, DUAN Ji-ping, WANG Chang-shun, LIU Yong-li . Study on the detection and distribution of 6 aristolochic acids in Asari Radix et Rhizoma by UPLC-MS/MS[J]. Chinese Journal of Pharmaceutical Analysis, 2023 , 43(7) : 1148 -1155 . DOI: 10.16155/j.0254-1793.2023.07.07

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