Abstract
A rapid and effective method using QuEChERS-based sample preparation procedure and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis has been developed and validated to determine progesterone in rabbit plasma. The analyte was extracted from plasma by acetonitrile with phase partitioning by a mixture of magnesium sulfate and sodium chloride. The supernatant was then directly injected into LC-MS/MS in a positive electrospray ionization mode and quantified using progesterone-d9 as the internal standard. The method linearity was in the range from 1 ng/mL (LOQ) to 200 ng/mL. Method recovery was from 86.0% to 103%, and repeatability was lower than 5.5%. The plasma sample was stable for 12 weeks stored at 18 ± 2°C. This method was applied to quantify progesterone in rabbit plasma in a pharmacokinetic study of two transdermal formulations: a reference drug and a eutectic-hydrogel system. The data indicate that the eutectic-hydrogel system's bioavailability was 1.5 times better than that of the reference drug, and the transdermal system is a potential drug delivery system for progesterone.
Highlights
Progesterone (Figure 1) is an endogenous steroid hormone secreted from the ovaries, testes, adrenal cortex, and placenta
Intending to develop a method that is quick and uses less solvent as well as to evaluate the pharmacokinetic study of progesterone from the transdermal route, this paper presents a rapid QuEChERS-based method coupled with LC-MS/MS for the quantification of progesterone in rabbit plasma
Optimization of Mass Spectrometry Condition. e ion transitions of progesterone and progesterone-d9 were obtained by directly infusing the standard solution of 1 μg/mL into the mass spectrometer. e precursor ions were obtained when the molecular ion combined with a proton. e optimal collision energies were selected for two product ions (Table 1)
Summary
Progesterone (Figure 1) is an endogenous steroid hormone secreted from the ovaries, testes, adrenal cortex, and placenta. Some pharmacokinetic studies have evaluated the effectiveness of various formulations of progesterone [12,13,14], including the transdermal application [11]. Abraham et al developed an RIA method to determine progesterone in plasma: the sensitivity varied from 10 to 25 pg, and the recovery was 84.2 ± 4.8% [15]. Zhang et al used solid-phase extraction with Oasis HLB cartridge prior to LC-MS/MS analysis to determine 17α-hydroxyprogesterone caproate, 17α-hydroxyprogesterone, and progesterone in human plasma with medroxyprogesterone acetate as the internal standard. Intending to develop a method that is quick and uses less solvent as well as to evaluate the pharmacokinetic study of progesterone from the transdermal route, this paper presents a rapid QuEChERS-based method coupled with LC-MS/MS for the quantification of progesterone in rabbit plasma
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