Development of efficient SPE–TLC method and evaluation of biological interactions of contraceptives with progesterone receptors

AbstractTLC–SPE methodologies were developed to ascertain biological interactions of norethindrone acetate and dydrogesterone contraceptives with plasma progesterone receptor proteins. TLC solvent system for plain and Cu(II) impregnated silica gel plates was n-hexane-n-butanol (90:10, v/v), which took 20min to run up to 10.0cm. The best separation was on Cu(II) impregnated plates due to maximum difference in Rf values and compact spots. The optimized SPE conditions were pH 2.0 and 3.0 of phosphate buffer (50mM) for norethindrone acetate and dydrogesterone, respectively. The flow rate of plasma and eluting solvent (methanol) through C18 cartridge was 0.10mL/min. The interactions of these contraceptives with progesterone receptor proteins were analysed by TLC–SPE results, which were supported by modelling using PyMOL and Autodoc4 softwares. The dydrogesterone was found to be bound strongly than norethindrone acetate. Attempts have been made to discuss the drugs’ interactions at chemo-supramolecular level