Objective: To establish an ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for the simultaneous determination of daidzein, formononetin, genkwanin, quercetin, rutin, apigenin, kaempferol, luteolin and puerarin in Tongfu Jingyaotong tincture. Methods: The Waters ACQUITY UPLC HSS T3 Column (100 mm×2.1 mm,1.8 μm) was used with methanol and 0.1% formic acid (gradient elution) as the mobile phase at a flow rate of 0.3 mL·min-1. Column temperature was 45 ℃, and injection volume was 2.0 μL. Electrospray ion source was adopted with positive and negative ion modes and multi-reaction monitoring and acquisition. Results: Apigenin, genkwanin and daidzein had good linearity in the range of 0.2-1 000 ng·mL-1, puerarin, rutin, luteolin, kaempferol and formononetin in the range of 0.2-2 000 ng·mL-1, and quercetin in the range of 1.0-5 000 ng·mL-1. The resolution was good, and the RSDs of precision, repeatability, reproducibility and stability test were all below 5% (n=6). The average recoveries ranged from 82.7% to 111.8% with RSDs (n=6) of 2.0%-5.4%. The contents of daidzein, formononetin, genkwanin, quercetin, rutin, apigenin, kaempferol, luteolin and puerarin in 6 batches of Tongfu Jingyaotong tincture were 8.46-17.97 μg·mL-1, 0.62-1.67 μg·mL-1, 0.03-0.08 μg·mL-1, 1.34-2.22 μg·mL-1, 0.70-1.14 μg·mL-1, 0.48-0.99 μg·mL-1, 0.20-0.90 μg·mL-1, 0.10-0.16 μg·mL-1 and 167.95-227.75 μg·mL-1, respectively. Conclusion: This method has strong specificity, wide linear range, accuracy and efficiency, and can detect the contents of 9 active ingredients in Tongfu Jingyaotong tincture at the same time, which offers an effective quality control method for Tongfu Jingyaotong tincture.
