A method combining solvent and gel extraction for isolation and preliminary purification of steroids in tissues

A method for the combined extraction and purification of steroids from testicular tissue is described. The tissue is homogenized and extracted with n-hexane/isopropyl alcohol, and the column to which a Sep-Pak C18 cartridge is attached. Following a wash of the Lipidex/Sep-Pak beds with water to remove inorganic and polar organic substances, steroids are eluted with 85% aqueous methanol. Most of the nonpolar lipids and phospholipids remain on the Lipidex/Sep-Pak. The steroid fraction is acidified with acetic acid, diluted to 70% methanol, and passed through a small bed of Lipidex 5000 to remove cholesterol. Recoveries of testosterone and progesterone are about 90%.