Separation of chloroplast polar lipids and measurement of galactolipid metabolism by high-performance liquid chromatography

✍ Scribed by Johan W.M. Heemskerk; Gerard Bögemann; Martin A.M. Scheijen; J.F.G.M. Wintermans

• Publisher: Elsevier Science
• Year: 1986
• Tongue: English
• Weight: 514 KB
• Volume: 154
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(86)90499-9

✦ Synopsis

Procedures are described for the separation of polar lipids from plant chloroplasts by high-performance liquid chromatography, using a polar-modified silica column. Glycolipids and phospholipids were eluted with a gradient of 2-propanol/n-hexane (80:55, v/v) and 2-propanol/n-hexane/water/methanol (80:55:15:10, v/v). The lipids were detected by uv absorbance at 202 nm. Diacylglycerol and mono-, di-, and trigalactosyldiacylglycerol and phosphatidylcholine were separated on a LiChrosorb NH2 column (7-microns particles, Merck, FRG), but acidic lipids were retained. These lipids could be quantified from their 202-nm absorbance recording. The absorption coefficients obtained depended on the mean number of double bonds in the different lipid classes. The separation was applied for a rapid monitoring of the lipid composition in thylakoids and in fractionated inner and outer envelopes. The activities of galactosyltransferases involved in galactolipid metabolism, UDPGal:diacylglycerol galactosyltransferase and galactolipid:galactolipid galactosyltransferase, could be measured quantitatively in specific assays for both enzymes.