A rapid and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for the determination of hydroxysafflor yellow A in human plasma: Application to a pharmacokinetic study

A sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS/MS) method has been developed and validated for the determination of hydroxysafflor yellow A (HSYA) in human plasma. HSYA was extracted from human plasma by using solid-phase extraction technique. Puerarin was used as the internal standard. A Shim-pack VP-ODS C18 (150 mm × 4.6 mm, 5 μm) column and isocratic elution system composing of methanol and 5 mM ammonium acetate (80:20, v/v) provided chromatographic separation of analytes followed by detection with mass spectrometry. The mass transition ion-pair was followed as m/z 611.19 → 491.19 for HSYA and m/z 415.19 → 295.10 for puerarin. The proposed method has been validated with a linear range of 1–1000 ng/ml for HSYA with a correlation coefficient ≥0.999. The lower limit of quantitation was 1 ng/ml. The intra-batch and inter-batch precision and accuracy were within 10%. The average extraction recovery was 81.7%. The total run time was 5.5 min. The validated method was successfully applied to the study on pharmacokinetics of HSYA in 12 healthy volunteers after a single oral administration of safflower oral solution containing 140 mg of HSYA.