Methods are described for the extraction and quantification of total lipids in cereal grains and other similar tissues, and for the determination of all the major classes of acyl lipid found in these extracts. Total lipids, obtained by direct solvent extraction or after acid hydrolysis, are quantified as fatty acid methyl esters (FAME) by gas chromatography (g.c.), using heptadecanoate (17 : 0) as internal standard. Individual lipid classes are separated by thin-layer chromatography; non-polar lipids and glycolipids are measured as FAME by g.c., while phospholipids are determined from phosphorus distribution. Crude lipid extracts are used to avoid losses during purification, and methanolysis of lipid classes is always performed without extracting the lipids from silica gel in order to minimise autoxidation, handling losses and contamination. Corrections are described for minor losses during experimental procedures, and factors are given for conversion of weights of FAME or phosphorus into weights of original lipid. In the authors' laboratory the precision of routine determinations (variations expressed as percentage of mean values) are usually well within the limits: total lipids, 1.5 %; major lipid classes, 1.5 %; minor lipid classes, 5 %.
The quantitative distribution of 23 acyl lipid classes and unsaponifiable matter in kernels of amylomaize, LG‐11 hybrid maize and waxy maize is described. LG‐11 and waxy maize were normal (oil content) varieties, containing 4.9% and 5.1% lipid, respectively, while amylomaize (9.3% lipid) was a high oil variety. The distribution of kernel lipids was 76–83% in germ, 1–2% in pericarp, 1% in tip cap, 1–11% in starch, and 13–15% in aleurone plus the nonstarch fraction of the starchy endosperm. Germ contained 39–47% lipid, which was nostly triglyceride (TG), with some steryl esters (SE) and diglycerides (DG), and small amounts of glycolipids (GL) and phospholipids (PL). Aleurone lipids appeared to be TG with some free fatty acids (FFA) and SE. The other nonstarch lipids in starchy endosperm were FFA with very small amounts of SE, DG, GL and PL. The starches had a little surface lipid (FFA) and true (internal) starch lipid (FFA, lyso‐PL) in quantities roughly related to amylose content (amylomaize =ca. 73% amylose, 1.0% lipid; LG‐11=23% amylose, 0.7% lipid; waxy maize =<5% amylose, 0.2% lipid). Pericarp lipids (0.8–2.5%) were mainly unsaponifiable matter, the acyl lipids being TG, SE, DG and FFA. Tip cap lipids (2.5–2.9%) had more TG, GL and PL than pericarp lipids, but were otherwise similar. Pericarp lipids and endosperm nonstarch lipids appeared to have suffered extensive degradation at some time during kernel development or after harvesting, while lipids in starch, germ and tip cap were evidently unaffected. FFA and lyso‐PL are regarded as normal components of maize starch (rather than degradation products) and may occur as amylose inclusion complexes.
Acyl lipids were quantified in the germ, endosperm and pericarp of LG‐11 maize kernels at eight stages of development from 9 to 87 days after pollination (DAP). Changes in the lipids are discussed in relation to morphological events in the developing kernel. Storage lipids (triglyceride, steryl ester) and membrane lipids (diacylphospholipids) accumulated in germ until 52–76 DAP, then decreased slightly without formation of lipid degradation products, lated in endosperm until 36–42 DAP and then decreased. Maximum values for galactosyldiglycerides and diacylphospholipids (nonstarch lipids) were reached at 16–23 DAP, and all decreased to very low values at maturity. Loss of these functional (membrane) lipids during the period of endosperm cell filling is unexpected. Starch contained 82% of the lysophospholipids and 64% of the free fatty acids in endosperm at 76 DAP. Endosperm lysophospholipids increased until 76 DAP and then decreased slightly, while free fatty acids increased continuously mostly inside starch granules at all stages of development, and any possible decrease after 76 DAP was masked by acids formed by hydrolysis of aleurone and endosperm nonstarch lipids from 52 DAP. In DAP, and phospholipids decreased after 42 DAP. Loss of these lipids is associated with senescence of most pericarp tissue. Triglycerides and steryl esters accumulated steadily to maturity, while the main accumulation of unsaponifiables occurred after 52 DAP about the time of suberin formation.
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