Objective: To develop an UPLC-MS/MS method for the simultaneous determination of seven active components in Huaihua powder, including rutin, (+)-R-pulegone, nobiletin, naringoside, quercitrin, neohesperidin and quercetin. Methods: An ACQUITY UPLC HSS T3C18 (100 mm×2.1 mm,1.8 μm) column was used with 0.1% formic acid in water (A) and methanol (B) as mobile phases. A gradient elution was employed with flow rate of 0.4 mL·min-1 and column temperature of 35 ℃. Electrospray ionization (ESI) source was used. Samples were determined in both positive and negative ion modes. MS parameters were 3 L·min-1 for nebuliser gas flow, 10 L·min-1 for heater gas flow, 45 ms for dwell time, 300 ℃ for source temperature, 400 ℃ for heater block temperature, 250 ℃ for desolvation tube temperature, and 10 L·min-1 for dry gas flow. Multiple-reaction monitoring (MRM) was used to perform mass spectrometric quantification. Rutin, (+)-R-pulegone and nobiletin were measured by precursor/product pairs of m/z 611.15/303.00, 153.15/81.10 and 403.15/373.10 in positive ion mode. Naringoside, quercitrin, neohesperidin and quercetin were measured by precursor/product pairs of m/z 579.15/271.15, 447.10/300.15, 609.15/301.15, 301.05/151.05 in negative ion mode. Results: Rutin, (+)-R-pulegone, nobiletin, naringoside, quercitrin, neohesperidin and quercetin had good linearity in their respective linear ranges with all linear correlation coefficients greater than 0.999 5 and RSDs of the precision, repeatability and stability less than 5.0%. The average recoveries were 97.7%, 103.3%, 92.3%, 100.7%, 101.5%, 101.0% and 100.2%, respectively, with RSDs lower than 5.0%. The contents ranges of rutin, (+)-R-pulegone, nobiletin, naringoside, quercitrin, neohesperidin and quercetin in 6 batches of Huaihua powder were 44.30-47.00, 2.01-2.10, 0.31-0.33, 18.53-19.80, 0.79-0.85, 17.64-18.57 and 1.29-1.35 mg·g-1. Conclusion: The proposed method is rapid, sensitive and specific, which could offer an analytical method for the quality control of Huaihua powder, reference and insights for its pharmacodynamic and pharmacokinetic studies.
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