Determination of abundance of dexamethasone related metabolic enzymes and transporter proteins in the placenta of Chinese pregnant women by LC-MS/MS method
Objective:To establish an LC-MS/MS method for the determination of the abundance data of dexamethasone-related metabolic enzymes and transporters in the placenta of Chinese pregnant women.Methods:A Shim-pack GISS-HP C18(100 mm ×2.1 mm,1.9 μm)chromatographic column was used,and the mobile phases were consisted of 0.2％formic acid-water(phase A)and 0.2％formic acid-acetonitrile(phase B)with gradient elution at a flow rate of 0.2 mL·min-1.The post-column phase C consisted of 0.5％ethylene glycol-acetonitrile was added at a flow rate of 0.1 mL·min-1,and the ESI source positive ion MRM mode was used for quantitative analysis.The established method was investigated methodologically and the expres-sion levels of 11 β hydroxy steroid dehydrogenase 1(11β-HSD1),11 β hydroxysteroid dehydrogenase 2(11 β-HSD2),cytochrome P450 3A4 enzyme(CYP3A4)and P-glycoprotein protein(P-gp)were quantitatively analyzed in the placenta of Chinese pregnant women in the third trimester.Results:The established LC-MS/MS method had a linear range of 0.1-100 nmol·L-1(r＞0.999).The precision and accuracy results met the re-quirements of the biological sample analysis method verification in Chinese Pharmacopoeia(RSD≤15％),and the stability results showed that the samples were stability.The protein abundance of 11 β-HSD2,11 β-HSD1,CYP3A4 and P-gp were(84.46±59.97)pmol·g-1,(11.44±3.73)pmol·g-1,(8.83±2.78)pmol· g-1 and(7.94±4.10)pmol·g-1,respectively.Besides,the results of the study also showed that there was no significant difference in the distribution of related metabolic enzymes and transporters in different parts of the placenta.However,there was a significant difference in the abundance of P-gp in the placenta between Chinese people(7.94±4.10)pmol·g-1 and white people(4.41±2.46)pmol·g-1.Conclusions:The LC-MS/MS method established in this study has high accuracy and sensitivity and is suitable for detecting the abundance values of dexamethasone-related metabolic enzymes and transporters in human placenta.
LC-MS/MSCYP3A4P-gp11β hydroxysteroid dehydrogenaseprotein abundanceplacenta of Chinese pregnant womendexamethasone
万方数据
维普
