大鼠血浆中千金藤素超高效液相色谱-串联质谱法的建立、验证及其药代动力学研究OA

Objective To establish and validate a sensitive and reliable ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for quantitative analysis of cepharanthine in rat plasma.Methods Plasma samples were treated using the protein precipitation method.Chromatographic separation was performed on column maintained at 50℃.Gradient elution was employed with mobile phases composed of(A)0.05%formic acid aqueous solution containing 1 mmol/L ammonium formate and(B)0.1%formic acid in acetonitrile at a flow rate of 0.40 mL/min.Mass spectrometric detection was carried out using an electrospray ionization(ESI)source in a positive-ion mode with multiple reaction monitoring(MRM).Mass transition m/z 607.3→576.2 was adopted for cepharanthine and m/z 260.2→116.1 for propranolol,the internal standard.This method was validated and used to quantify cepharanthine in plasma samples collected from rats following a single intragastric administration to investigate its pharmacokinetic profile.Results Cepharanthine proved to be linear in rat plasma over the concentration range of 0.1 to 80 ng/mL.The intra-and inter-batch accuracies ranged from-0.43%to 11.73%,and precisions from 3.04%to 13.83%.The recovery rates of quality control samples ranged from 86.70%to 88.48%.The accuracy and precision met the standards across six lots of the matrix.The quality control samples remained stable under the following conditions:24 hours of storage in the autosampler at 15℃,followed by three freeze-thaw cycles,and 10 days of storage at-20℃.All these met the requirements for bioanalytical analysis of samples.Pharmacokinetic studies showed that the time to maximum concentration(Tmax)was 6 to 12 h,the maximum concentration(Cmax)was(12.31±3.78)ng/mL,the terminal elimination half-life(t1/2)was(10.07±0.66)h,and the mean residence time(MRT0-t)was(24.80±1.98)h after intragastric administration of 3 mg/kg cepharanthine to rats.Conclusion This study has established a UPLC-MS/MS method for quantitative analysis of cepharanthine in the rat plasma matrix,which is simple and sensitive,with a lower limit of quantification of 0.1 ng/mL.