Effect of method of conservation on the n-alkane C31 concentration of two legumes and three temperate grasses

Abstract

SummaryThe objective of this study was to compare five methods of forage preservation (ice, dry ice, liquid-N, drying in newspaper, and sundried) on the concentration of the n-alkane C31 in Medicago sativa cv Puebla 76 (lucerne/alfalfa), Trifolium repens latum (white clover), Lolium perenne (perennial ryegrass), Pennisetum clandestinum (kikuyu grass) and Dactylis glomerata (orchardgrass/cocksfoot). The sampled forages in ice and dry ice were kept in hermetically sealed plastic bags in a plastic cooler. The deep frozen samples were put into small plastic bags and submerged in liquid-N. The samples dried in newspaper were placed between sheets of newspaper, and put into hermetically sealed plastic bags. All these samples were taken to the laboratory and freeze-dried approximately 24 h later. The sun-dried samples were taken to the laboratory in newspaper and sun-dried for 48 h. The forage samples were ground and worked on in triplicate, extracted by the Soxhlet method (C34 n-alkane was used as standard and n-heptane as solvent), saponified, separated, purified and injected into a gas chromatograph. The data were analysed for variance as a randomised factorial (n=74). The comparison between means was by the Boneferroni test (P<0.05). The interaction between forage and method of conservation was significant P<0.0001). There were no differences between methods of conservation for white clover (mean 26mg C31/kg DM) or orchardgrass/cocksfoot (mean 31 mgC31/ kgDM). In comparison with liquid-N, (261 mgC31/kgDM), the other methods reduced the concentration of n-alkane in kikuyu grass by 27-35%. Sun and newspaper dried samples increased the concentration of C31 in alfalfa/lucerne by 6 and 15% respectively in comparison with freezing with liquid-N (291 mgC31/kgDM), while that frozen on ice reduced C31 by about 8%. This method, however, increased the ryegrass n-alkane by 12% in relation to liquid-N (169 mgC31/kgDM), but sun drying reduced it by about 40%. It is concluded that drying in liquid-N is the most reliable method for the conservation of samples for n-alkane analysis.