development of immunochemical methods for purification and detection of the steroid drug medroxyprogesterone acetate发展的免疫化学方法净化和类固醇药物醋酸甲羟孕酮的检测.pdfVIP

Journal of Environmental Protection, 2012, 3, 624-639 /10.4236/jep.2012.37076 Published Online July 2012 (http://www.SciRP.org/journal/jep) Development of Immunochemical Methods for Purification and Detection of the Steroid Drug Medroxyprogesterone Acetate 1 2 2 1 Alisa Bronshtein , Alex Krol , Haim Schlesinger , Miriam Altstein 1Department of Entomology, Institute of Plant Protection, The Volcani Center, Bet Dagan, Israel; 2Analyst Research Laboratories, Rehovot, Israel. Email: vinnie2@.il Received March 12th, 2012; revised April 17th, 2012; accepted May 19th, 2012 ABSTRACT An immunochemical sol-gel-based immunoaffinity purification (IAP) method for purification and detection of the pro- gestin 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 detec- tion limits of the assays were 1.4 ± 0.2 ng·mL−1 (n = 4) and 4.0 ± 0.4 ng·mL−1 (n = 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