LC-MS/MS Method for Rapid Quantification of Progesterone in Rabbit Plasma and Its Application in a Pharmacokinetic Study of the Transdermal Formulation

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.

Development of Electroanalytical Methods Using Different Electrodes for the Determination of Progesterone (P4) in Pharmaceutical Formulations and Human, Cow and Goat Milk

“…Progesterone (pregn-4-ene-3,20-dione, P4) is an endogenous steroid hormone secreted from the ovaries, testes, adrenal cortex, placenta. Progesterone releases mature oocytes that help in the implantation and maintenance of pregnancy and it stimulates the mammary gland in preparation for milk secretion for parturition [1][2][3]. P4 in serum exists as the free form, the orosomucoid-bound form, the albumin-bound form and the globulin-bound form (1), only the free form is active biologically [4].…”

“…P4 in serum exists as the free form, the orosomucoid-bound form, the albumin-bound form and the globulin-bound form (1), only the free form is active biologically [4]. The average serum P4 level ranges from 0.15 to 25 μg L À 1 , but it can reach 150 μg L À 1 during pregnancy and only 0.38 μg L À 1 during menopause [1,5,6]. Although it is considered to be a female hormone, it is also produced in the testicles and the adrenal glands and it is essential in small amounts for men and children to maintain wellness and health [7][8][9].…”

“…Progesterone releases mature oocytes that help in the implantation and maintenance of pregnancy and it stimulates the mammary gland in preparation for milk secretion for parturition [1][2][3]. P4 in serum exists as the free form, the orosomucoid-bound form, the albumin-bound form and the globulin-bound form (1), only the free form is active biologically [4]. The average serum P4 level ranges from 0.15 to 25 μg L À 1 , but it can reach 150 μg L À 1 during pregnancy and only 0.38 μg L À 1 during menopause [1,5,6].…”

Determination of progesterone (steroid drug) in the semi‐solid dosage form (vaginal gel) using a stability‐indicating method by RP‐HPLC/PDA detector

“…From the literature, we obtained various quality methods on progesterone drugs, such as (1) determination of progesterone in the tablet and capsule dosage forms (Avilas et al, 2013; Maliwal et al, 2009; Yucel et al, 2010); (2) determination of progesterone in poly(methyl methacrylate) nanoparticles synthesized via miniemulsion polymerization (Odinei et al, 2017); (3) quantification of progesterone in rabbit plasma using LC–MS/MS method (Cao‐son Tran et al, 2020); (4) determination of progesterone in human serum using HPLC/MS with atmospheric pressure chemical ionization (Zhuping et al, 2000); 5) progesterone API‐related substances method using HPLC (Huang et al, 2018); (6) determination of progesterone in the crude methanol extract of benalu duku leaves (Lazuardi et al, 2019); (7) determination of progesterone in commercial formulations and in nonconventional micellar systems (Vincenzo et al, 2003). The detailed information is presented in Table 1.…”