Development of Immunochemical Methods for Purification and Detection of the Steroid Drug Medroxyprogesterone Acetate

Abstract

An immunochemical sol-gel-based immunoaffinity purification (IAP) method for purification and detection of the progestin drug medroxyprogesterone acetate (MPA) was developed. A polyclonal antibody (Ab) for MPA was generated, and two competitive (indirect and direct) sensitive enzyme-linked immunosorbent assays (ELISAs) for its detection were developed and implemented to determine the recovery and efficiency of the sol-gel based IAP method. The detection limits of the assays were 1.4 ± 0.2 ng·mL^-1 (<i>n</i> = 4) and 4.0 ± 0.4 ng·mL^-1 (<i>n</i> = 25) for the indirect and direct ELISAs, respectively. The Abs did not exhibit cross-reactivity with any other progestin or steroid hormone, with the exception of megestrol acetate, with which the Ab exhibited 76% cross-reactivity. The sol-gel IAP method successfully eliminated serum interference to a degree that enabled ELISA analysis of spiked serum samples. This method was also found fully compatible with subsequent chemical analytical methods, such as liquid chromatography followed by mass spectrometry (LC-MS/MS). The approaches developed in this study form a basis for analysis of MPA in biological samples and may be further used to study population exposure to MPA and to monitor MPA contamination in water samples

Teh Hijau Menurunkan Kadar Koleterol Total Pada Ibu Akseptor Kb Depomedroxyprogesterone Acetate (Dmpa)

“…DMPA merupakan salah satu metode kontrasepsi yang paling efektif yang saat ini banyak digunakan (Abasiattai, Udoma, & Ukeme, 2010). DMPA mengandung hormon progestogen sintetik (progestin) yang mempunyai struktur kimia seperti progesterone (Bronshtein, Krol, Schlesinger, & Altstein, 2012). Mekanisme kerja DMPA sebagai kontrasepsi adalah dengan mencegah ovulasi.…”

Development of an enzyme‐linked immunosorbent assay and a beta‐1 adrenergic receptor–based assay for monitoring the drug atenolol

“…In the past few years, we have developed highly effective and reproducible IAP technology, based on the entrapment of Abs in a ceramic SiO 2 matrix termed sol-gel, which facilitates efficient, singlestep cleanup and concentration of analyzates from large volumes of crude samples. The applicability of this approach was tested with Abs raised in our laboratory against a variety of PPs such as the steroid hormones levonorgestrel, ethinylestradiol, and nortestosterone [24], medroxyprogesterone acetate [25], the nonsteroid anti-inflammatory drug indomethacin [26], several other pesticides (pyrethroids and atrazine) [34,35], forensic compounds (TNT) [36], and environmental contaminants [37,38]. Implementation of the Abs for the development of a sol-gel based ATL IAP method is under development.…”

“…In previous studies, we developed ELISAs and immunoaffinity purification methods for some PPs: steroid hormones, levonorgestrel, ethinylestradiol and nortestosterone [24], medroxyprogesterone acetate [25], and the nonsteroid antiinflammatory drug indomethacin [26]. In the present study, we used two different approaches for ATL monitoring: an immunochemical approach based on ELISA, and an innovative diagnostic environmental assay based on a receptor-binding assay that can monitor the interaction between ATL and its target receptor b1AR.…”