The use of immobilized Candida antarctica lipase for simultaneous supercritical fluid extraction and in‐situ methanolysis of cis‐vaccenic acid in milkweed seeds

Abstract

cis-Vaccenate was determined in milkweed (Asclepias) seeds employing supercritical fluid extraction (SFE) with in-situ enzymatic methanolysis. Candida antarctica lipase type B immobilized onto two different types of carrier materials-a hydrophobic polymer (Novozyme 435) and a silica-based material (NovoSample 40013)-was investigated. Several SFE parameters were investigated to find the optimal conditions, including extraction time, temperature, methanol concentration, and water level. It was shown that 90 min of dynamic extraction/reaction time was required, and that the optimal methanol and water concentrations differed for the two enzyme preparations studied. Overall, Novozyme 435 demonstrated the fastest reaction kinetics and consistently gave the highest vaccenate yields. The optimized methodology was applied on 15 species of milkweed seeds, giving an average recovery of 105 ± 7% when compared to results obtained using a conventional methodology. The average relative standard deviations were 8% for the enzyme-based method and 4% for the reference method. The proposed methodology was faster, was less laborious, and consumed less organic solvent than the reference method. Hence, SFE with in-situ enzymatic methanolysis is a promising methodology for the analysis of FA in oilseeds.

Optimization of a Supercritical Fluid Extraction/Reaction Methodology for the Analysis of Castor Oil Using Experimental Design

“…In addition, when lipase-catalyzed transmethylation is achieved simultaneously with dynamic SFE of the oil, there is a considerable advantage with reduced sample handling and shorter analysis time. This has been demonstrated in an analytical context using immobilized Candida antarctica lipase type B (Novozyme 435) for the determination of fat composition in meat samples (18), oilseeds (19,20), and vegetable oil soapstocks (21).…”

“…The four experiment factors under study were methanol and water concentration, temperature, and pressure. Values for these parameters were chosen to bracket conditions known to produce FAMEs from oil via SFE (20). To test for curvature of the response, three levels of each experimental parameter were required: selected values were 1, 3, and 5 vol % methanol; 0.02, 0.10, and 0.18 vol % water; temperatures of 40, 60, and 80 °C; and pressures of 200, 300, and 400 bar.…”

Fatty Acid Composition of Seed Oil from <i>Fremontodendron californicum</i>

“…Cisvaccenate, (octadec-cis-11-enoate) (l.6%) is present as a trace component in many plant oils and is present in a few oils, e.g. milkweed, at higher levels [8]. The linoleic acid content (71.2%) is high, but similar to levels observed in corn, safflower and sunflower oil [9].…”

“…MeOH with immobilized lipase to produce methyl esters [8]. Use of SFE coupled with immobilized lipase-catalyzed transesterification is an especially valuable method for FAME production from oils containing reactive functional groups.…”

Development of novel biodegradable water chestnut starch/PVA composite film. Evaluation of plasticizer effect over physical, barrier, and mechanical properties

“…Crude protein, fiber, fat, and ash content were determined by approved methods of AOAC (Turner & McKeon, 2002). Amylose content of WCS was determined spectrophotometrically by following the method of McGrance et al (1998).…”