Determination of Lappaconitin, Diterpene Alkaloide Obtained from Plants Aconitum leucostomum, and its Active Metabolite N-desacetyllappaconitin in Human Plasma and Blood

Abstract

Introduction. Lappaconitine is an alkaloid, contained into Aconitum leucostomum Vorosh. roots and herbs. The alkaloid is indicated to arrhythmia. The lappaconitine drugs are metabolized into eight pharmacologically active substances, but N-desacetyllappaconitine is the most effective. Drugs based on a lappaconitine has narrow therapeutic range and many kinds of side effects. Pharmacokinetics of lappaconitine should be more studied for safety medical use of lappaconitine drugs.Aim. The aim of this study is to develop method for the quantitative determination of lappaconitine and its active metabolite N-desacetyllappaconitine in human plasma and blood by high performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS). Materials and methods. Determination of lappaconitine and N-desacetyllappaconitine in plasma and blood was carried out by HPLC-MS/MS. The samples were processed by acetonitrile protein precipitation.Results and discussion. This method was validated by next parameters: selectivity, matrix effect, calibration curve, accuracy, precision, spike recovery, lower limit of quantification, carry-over effect and stability.Conclusion. The method of the quantitative determination of lappaconitine and N-desacetyllappaconitine in human plasma and blood was developed and validated by HPLC-MS/MS. The analytical range of the was 0.50-50.00 ng/ml for lappaconitine and 0.50-100.00 ng/ml for N-desacetyllappaconitine in biological matrix. Method could be applied to determination of lappaconitine and N-desacetyllappaconitine for PK studies.