Simultaneous determination of etonogestrel and ethinyl estradiol in human plasma by UPLC‐MS/MS and its pharmacokinetic study

Abstract

A selective, sensitive and rapid ultra-performance liquid chromatography tandem mass spectrometry method was developed and validated for the simultaneous determination of etonogestrel (ENG) and ethinyl estradiol (EE) in human plasma. The analytes and their deuterated internal standards, ENG-d7 and EE-d4, were extracted from plasma samples by solid-phase extraction on HyperSep™ Retain PEP cartridges. The chromatographic analysis was performed on an Acquity UPLC HSS Cyano column, 100 Å (50 × 2.1 mm, 1.8 μm), column using gradient mobile phase, acetonitrile and 2.0 mm ammonium trifluoroacetate at 0-1.7 min (65:35, v/v) and 1.8-2.7 min (95:5, v/v) with 0.250 mL/min flow rate. Analytes and IS protonated precursor → product ion transitions (ENG, m/z 325.2 → 257.2; EE, m/z 530.2 → 171.2; ENG-d7, m/z 332.2 → 263.2; EE-d4, m/z 534.2 → 171.2) were monitored on a Triple Quadrupole Mass spectrometer (TQMS), operating in multiple reaction monitoring and positive ionization mode. The calibration curves were established at 10.00-2500 pg/mL for ENG and 1.500-150.0 pg/mL for EE with a correlation coefficient (r ) ≥0.9996 for both. The validated method was successfully applied to support a bioequivalence study of 0.15 mg ENG and EE 0.03 mg tablet formulation, administered in 24 healthy Indian females. Method reliability was assessed by reanalysis of 94 incurred study samples.

4‐Azido‐7‐nitrobenzoxadiazole as innovative clickable fluorescence probe for trace and selective quantification of ethinylestradiol in human plasma

“…Although solid phase extraction (SPE) was implemented in earlier reports [33], it was not given priority as it was very costly. The nonpolar nature of the EE makes it an ideal candidate for liquid-liquid extraction (LLE) that is cost effective compared with SPE and could produce not only purified, but also concentrated, samples for spectral analysis.…”

Liquid Chromatographic-Electron Spray Ionization-Mass Spectroscopy Technique for the Simultaneous Determination of Ethinyl Estradiol & Etonogestrel and Characterization of New Forced Degradation Compounds

“…The literature review conducted on ethinyl estradiol and etonogestrel indicated that only derivative spectroscopic analysis has been reported [12]. However, for ethinyl estradiol with other drugs, derivative spectroscopy [13], UPLC-MS [14][15][16], for fixed combination dosage form of Nuvaring and with other com-binational drugs on HPLC [17][18][19] are reported. To the best of our knowledge, there is no published data about the use of HPLC-MS for the analysis of the fixed combination of ethinyl estradiol and etonogestrel.…”

Novel reversed‐phase HPLC method for simultaneous determination of ethynodiol diacetate (EDA)/ethinyl estradiol (EE) in pharmaceutical dosage form

“…A literature survey revealed that only few research studies are available on the simultaneous determination of EDA and EE in different formulations (Amin & Hassenbach, 1979; Babják, Balogh, Gazdag, & Görög, 2002; Carignan, Lodge, & Skakum, 1982; Gluck & Shek, 1980; Havlíková, Nováková, Matysová, Šícha, & Solich, 2006; Koneru, Kimbahune, Mubeen, Lalitha, & Chakraborty, 2015; Kotra, Ramisetti, & Konidala, 2017; Maziarz, 2013; Nair et al, 2018; Patel, Rathod, & Patel, 2013; Prasad, Reddy, Prasad, & Mukkanti, 2001; Qi Gui & Nieuweboer, 1989; Strusiak, Hoogerheide, & Gardner, 1982; Suhendi, 2016; Summers & Fountain, 2013; Yamada, Kuwahara, Takamatsu, & Hayase, 2000; Zorica, 2006). This paper describes a sensitive and straightforward high‐performance chromatographic method for determining EDA and EE in tablets.…”