Simultaneous determination of 9 active ingredients in Tongfu Jingyaotong tincture by UPLC-MS/MS
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-reac-tion 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 pre-cision,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,genkwa-nin,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.
Tongfu Jingyaotong tinctureultra-high performance liquid chromatography tandem mass spectrome-tryLC-MSactive ingredientsContent determinationquality control
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