Effect of Various Anticoagulants on Ethinyl Estradiol and Levonorgestrel Analysis in Human Plasma in Vitro by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

Yahdiana Harahap, Firda Zakiatun Nufus, Sunarsih

Research output: Contribution to journalArticlepeer-review

Abstract

Objective: This research objective is to evaluate the different types of anticoagulant to ethinyl estradiol and levonorgestrel analysis in plasma using ultra performance liquid chromatography tandem mass spectrometry. Method: Chromatography condition was obtained with Acquity® UPLC BEH C18 column (1.7 μm; 50 x 2.1 mm); mobile phase consisting 0.1% formic acid in water-acetonitrile under gradient elution ; flow rate of 0.3 mL/minute; column temperature of 40°C; injection volume of 10.0 μL; 5-minute analysis time and prednisone as internal standard. Sample preparation used protein precipitation followed by liquid-liquid extraction. Results: There was a linear result ranging from 5-500 pg/mL concentration of ethinyl estradiol and 100-10, 000 pg/ml concentration of levonorgestrel. There was no significant difference for stability and recovery of ethinyl estradiol and levonorgestrel in citrate, heparin, and EDTA plasma (p > 0.05; ANOVA). However, significant difference for peak area ratio (p < 0.05; Kruskal Wallis), between citrate, EDTA, and heparin plasma was observed. Conclusion: Citrate and heparin plasma analysis had better result than EDTA plasma analysis.

Original languageEnglish
Pages (from-to)42-50
Number of pages9
JournalJournal of Global Pharma Technology
Volume10
Issue number12
Publication statusPublished - 2018

Keywords

  • Citrate
  • EDTA
  • Ethinyl estradiol
  • Heparin
  • Levonorgestrel
  • Prednisone

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