LC-MS/MS法测定芒果苷在INS-1细胞中药代动力学

Pharmacokinetics of mangiferin in INS-1 cells determined by LC-MS/MS

目的建立LC-MS/MS法测定芒果苷(mangiferin,MGF)在INS-1细胞的含量,考察不同剂量芒果苷在INS-1细胞药代动力学特征,及同剂量下正常和模型组的药动学参数。方法以芦丁为内标,细胞样品采用乙腈∶乙酸(40∶1)液液萃取。色谱柱为C18柱(150×2.1 mm,3.5μm),1%冰乙酸-甲醇为流动相,在电喷雾离子化和正离子多反应模式下运用QTRAP 6500+型液质联用仪测定。INS-1细胞给药剂量为50、100、500μmol·L^-1 MGF;用140μmol·L^-1 H2O2损伤INS-1细胞1 h后给药100μmol·L^-1 MGF;进行MGF药动学研究。结果在2~500μg·L^-1范围内,MGF线性关系良好。日内、日间RSD均≤14.49%,准确度在93.97%~103.54%。细胞给药MGF低中高剂量后,AUC(0-t)和Cmax随给药剂量增加而增加;正常和造模组INS-1细胞Cmax分别为454.83±45.46和334.00±35.00μg·L^-1,AUC(0-t)分别为13 289.23±1 080.58和9 349.89±1 101.36μg·h·L^-1。结论该方法简单、特异、灵敏、可靠,适用于测定INS-1细胞中MGF。在50~500μmol·L^-1剂量内,MGF在INS-1细胞内未表现出线性药动学特征;模型组INS-1细胞Cmax和AUC(0-t)明显低于正常组。

Aim To establish a LC-MS/MS method for the determination of mangiferin(MGF)in INS-1 cells,and to investigate the pharmacokinetics of mangiferin in INS-1 cells at different doses,as well as the normal and model groups of drugs at the same dose.Methods Rutin was used as internal standard(IS).Analytes were extracted from INS-1 cells samples with acetonitrile:acetic acid(40∶1).Chromatographic separation was performed on a C 18 column(150×2.1 mm,3.5μm)by gradient elution with 1%glacial acetic acid-methanol.The QTRAP 6500+LC/MS/MS was used for determination with the positive ion multiple reaction monitoring mode.Pharmacokinetics were studied in INS-1 cells after administration of 50,100 and 500μmol·L^-1 MGF,as well as in model INS-1 cells injured one hour with 140μmol·L^-1 H 2O 2 at dosage of 100μmol·L^-1.Results Excellent linearity was observed in concentration range of MGF 2~500μg·L-1.The intra-day and inter-day precision RSD was≤14.49%,with the accuracy ranging from 93.97%to 103.54%.After administration of low,medium and high doses of MGF,AUC(0-t)and C max in INS-1 cells increased in a dose-dependent manner.In normal and model INS-1 cell groups,the C max was 454.83±45.46 and 334.00±35.06μg·L^-1;AUC(0-t)was 13289.23±1080.58 and 9394.89±1101.36μg·L^-1,respectively.Conclusions Being simple,specific,sensitive and reliable,the method can be applied to the determination of MGF in INS-1 cells.In the dose range of 50~500μmol·L^-1,the pharmacokinetics of MGF in INS-1 cells does not meet linear pharmacokinetic characteristics.The C max and AUC(0-t)of MGF in model INS-1 cells are significantly lower than those in normal cells.