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 expression levels of 11β hydroxysteroid dehydrogenase 1(11β-HSD1), 11β hydroxysteroid dehydrogenase 2(11β-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 requirements 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 11β-HSD2, 11β-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.
LI Hui, GUO Xuan, HOU Zhe, LIU Dong-yang, DONG Jing, HANG Tai-jun, ZHANG Lei
. Determination of abundance of dexamethasone related metabolic enzymes and transporter proteins in the placenta of Chinese pregnant women by LC-MS/MS method*[J]. Chinese Journal of Pharmaceutical Analysis, 2024
, 44(1)
: 76
-88
.
DOI: 10.16155/j.0254-1793.2024.01.08
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