Simultaneous Determination of Tacrolimus and Four Main Compounds of Wuzhi Capsule in Patients'Whole Blood by HPLC-MS/MS
A HPLC-MS/MS method to simultaneously determine the concentration of tacrolimus and four main components of wuzhi capsule(schisandrin,schisandrin B,schisandrin alcohol A and schisandrin ester A)in human whole blood was established,and used for the clinical drug concentration monitoring of immunosuppressant tacrolimus combined with hepatoprotective Chinese medicine Wuzhi capsule.A Waters Acquity UPLC® BEH C18 column(50×2.1 mm,1.7 μm)was used,with 5 mmol/L ammonium acetate-0.1％formic acid as the aqueous phase and acetonitrile as the organic phase.The flow rate was 0.3 mL/min,and the column temperature was 60 ℃.Tacrolimus isotope compound(13C,2H,)was used as internal standard(IS)for each component.Whole blood samples were treated with zinc sulfate at first,and then acetonitrile containing internal standard was added to precipitate protein.Finally,after centrifugation,the supernatant was taken for injection.Multiple reactive ion monitoring(MRM)mode,and positive ion scanning mode were used with electrospray ion source.The chromatographic responses of tacrolimus and the four main components of Wuzhi capsules were well correlated with the concentration in the range of 1-100 ng/mL,and the lower limits of quantification were all 1 ng/mL.The retention times of tacrolimus,schisandrin,schisandrin B,schisandrin alcohol A,schisandrin esteryl A and internal standard were 2.54,2.33,2.65,1.14,1.56,2.54 min,respectively.The accuracy,recovery and matrix effect after internal standard calibration were qualified.The stability data meet the relevant requirements.The method is sensitive,specific and accurate to be suitable for determination of concentration of tacrolimus,schisandrin,schisandrin B,schisandrin alcohol A and chisandrin esteryl A in human whole blood.
HPLC-MS/MSTacrolimusWuzhi capsuleTherapeutic drug monitoring
万方数据
维普
