Extraction and quantitation of dolichol and dolichyl phosphate in soybean embryo tissue

✍ Scribed by Jack W. Rip; Kenneth K. Carroll

• Publisher: Elsevier Science
• Year: 1987
• Tongue: English
• Weight: 498 KB
• Volume: 160
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(87)90059-5

✦ Synopsis

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 Crs 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 ?I 0.8 and 5.3 f 0.1 ILplg, respectively. The post-HPLC recoveries for dolichol and Dol-P were 101 f 2 and 84 ? 3% respectively, using [ I-r4C]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. 0 1987 Academic PIVSS, IX.