Objective: To establish an HPLC-MS/MS method for simultaneous determination of 11 components (harpagide, salidroside, nuezhenide, lobetyolin, wedelolactone, harpagoside, vaccarin, 6-gingerol, atractylenolide Ⅲ, atractylenolide Ⅱ, and atractylenolide Ⅰ) in Gengnianning. Methods: HPLC assay was performed on C18 column(100 mm×2.1 mm, 1.9 μm) with a mixture of methanol and 0.1% formic acid as the mobile phase in gradient elution at a flow rate of 0.3 mL·min-1.The column temperature was 25 ℃ and the injection volume was 1 μL. Detection was carried out on a triple quadrupole mass spectrometer in positive ion mode (harpagide, salidroside, nuezhenide, lobetyolin, wedelolactone and harpagoside) and negative ion mode(vaccarin, 6-gingerol, atractylenolide Ⅲ, atractylenolide Ⅱ, and atractylenolide Ⅰ) using an electrospray ion source(ESI). Multiple reaction monitoring(MRM)mode was employed. Results: The calibration curves were linear within the ranges of 1.485-29.71 μg·mL-1, 1.620-32.40 μg·mL-1, 7.801-156.0 μg·mL-1, 0.518-10.35 μg·mL-1, 0.167-3.333 μg·mL-1, 0.359-7.179 μg·mL-1, 1.455-29.10 μg·mL-1, 1.520-30.40 μg·mL-1, 0.160-3.205 μg·mL-1, 0.143-2.864 μg·mL-1 and 0.157-3.136 μg·mL-1 for harpagide, salidroside, nuezhenide, lobetyolin, wedelolactone, harpagoside, vaccarin, 6-gingerol, atractylenolide Ⅲ, atractylenolide Ⅱ, and atractylenolide Ⅰ, respectively. All 11 components showed good linearity (r≥0.998 0). The average recoveries(n=6)were in the range of 95.9%-102.6% with RSDs within 0.90%-3.0%. The contents of harpagide, salidroside, nuezhenide, lobetyolin, wedelolactone, harpagoside, vaccarin, 6-gingerol, atractylenolide Ⅲ, atractylenolide Ⅱ and atractylenolide Ⅰ in 10 tested samples from 5 manufactures were 14.8-104.5, 37.6-288.5, 335.8-1 332.8, 6.2-10.1, 6.6-61.8, 13.7-75.1, 57.4-132.8, 16.9-70.6, 11.8-33.9, 3.4-15.4 and 6.5-12.9 μg·g-1. Conclusion: The developed method is accurate and sensitive. It can be used in quality control of Gengnianning.
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