Extraction and quantitation of dolichol and dolichyl phosphate in soybean embryo tissue.
A procedure for the quantitative extraction of both dolichol and dolichyl phosphate (Dol-P) in plant tissue (soybean embryos) into diethyl ether from an alkaline saponification mixture is described. A complete and quantitative separation of total dolichol and total Dol-P is then obtained based on their respective solubilities in diethyl ether and water. After separation dolichol and Dol-P can both be analyzed and quantitated directly by reverse-phase HPLC on C18 columns without additional purification. The two major homologs of dolichol and Dol-P are those with 17 and 18 isoprene units. The total dolichol and total Dol-P contents of dry embryos were 96.3 +/- 0.8 and 5.3 +/- 0.1 micrograms/g, respectively. The post-HPLC recoveries for dolichol and Dol-P were 101 +/- 2 and 84 +/- 3% respectively, using [1-14C]dolichol and Dol-P containing 20 isoprene units as recovery standards. Dol-P estimations could be carried out on material equivalent to as little as 65 mg embryo tissue.
D002851:Chromatography, High Pressure Liquid; D004224:Diterpenes / Q000032:analysis*; D004288:Dolichol Phosphates / Q000032:analysis*; D004286:Dolichols / Q000032:analysis*; D004986:Ether; D010944:Plants / Q000032:analysis*; D011106:Polyisoprenyl Phosphates / Q000032:analysis*; D012995:Solubility; D013025:Glycine max / Q000032:analysis; D014867:Water
D004224:Diterpenes; D004288:Dolichol Phosphates; D004286:Dolichols; D011106:Polyisoprenyl Phosphates; D014867:Water; D004986:Ether; C028306:dolichol monophosphate