Screening of Vitamin D activity (VDA) of Solanum glaucophyllum leaves measured by radioimmunoassay (RIA)

The ingestion of Solanum glaucophyllum (SG) causes a calcinosis of cattle named Enteque Seco (ES). The toxic principle is the 1,25-(OH)₂D₃, mainly conjugated as glycoside. This study aims to validate a simple novel method of evaluation of the VDA of SG leaves. Aqueous extracts of SG were purified using C₁₈ minicolumns and assayed by RIA with an antibody raised in rabbits by injection of the acid—C22, 1-(OH)Vitamin D₃. Data were expresed as glycoside equivalent to 1,25-(OH)₂D₃ in ng/g of dry leaves. We compared this data with 1,25-(OH)₂D₃ levels measured, in the same samples, by liquid chromatography (HPLC) after enzyme cleavage. This procedure involved the incubation of SG leaves with rumen fluid, followed by C₁₈-OH solid phase extraction. The 1,25-(OH)₂D₃ fraction was run by HPLC and detection was achieved using a photodiode array detector. Data were expressed as micrograms of 1,25-(OH)₂D₃/g dry leaves. A significant regression of 1,25-(OH)₂D₃ levels (Y) as a function of glycoside RIA 1,25-(OH)₂D₃ equivalents (X) was found: Y = 12.02 + 0.35X R = 0.81; P = 0,0002; N = 15], allowing us to conclude that this novel assay could be used to estimate the amount of this active principle contained in SG leaves.