In order to establish Ciona intestinalis as a new bioresource for n-3 fatty acids-rich marine lipids, the animal was fractionated into tunic and inner body tissues prior to lipid extraction. The lipids obtained were further classified into neutral lipids (NL), glycolipids (GL) and phospholipids (PL) followed by qualitative and quantitative analysis using GC-FID, GC-MS, H-1 NMR, 2D NMR, MALDI-TOF-MS and LC-ESI-MS methods. It was found that the tunic and inner body tissues contained 3.42-4.08 % and 15.9-23.4 % of lipids respectively. PL was the dominant lipid class (42-60 %) irrespective of the anatomic fractions. From all lipid fractions and classes, the major fatty acids were 16:0, 18:1n-9, C20:1n-9, C20:5n-3 (EPA) and C22:6n-3 (DHA). The highest amounts of long chain n-3 fatty acids, mainly EPA and DHA, were located in PL from both body fractions. Cholestanol and cholesterol were the dominant sterols together with noticeable amounts of stellasterol, 22 (Z)-dehydrocholesterol and lathosterol. Several other identified and two yet unidentified sterols were observed for the first time from C. intestinalis. Different molecular species of phosphatidylcholine (34 species), sphingomyelin (2 species), phosphatidylethanolamine (2 species), phosphatidylserine (10 species), phosphatidylglycerol (9 species), ceramide (38 species) and lysophospholipid (5 species) were identified, representing the most systematic PL profiling knowledge so far for the animal. It could be concluded that C. intestinalis lipids should be a good alternative for fish oil with high contents of n-3 fatty acids. The lipids would be more bioavailable due to the presence of the fatty acids being mainly in the form of PL.
Cellulose nanocrystals (CNs) were prepared from tunicate by enzymatic hydrolysis (ECN), TEMPOmediated oxidation (TCN) and acid hydrolysis (ACN). They were cast alone or blended with glucomannan (GM) from konjac or spruce to prepare films. Different CNs were obtained with a yield of ECN > TCN > ACN with corresponding order of decreased My but increased crystallinity. The CNs' diameters were on the nanometre scale, with lengths of ECN > TCN > ACN. For CN-films, TCN and ACN fibrils were stretched and parallel to each other due to surface charges. For CN-GM films, both components interacted strongly with each other, resulting in changes of crystallinity, specific surface area, fibril diameter and contact angle compared with CN films. The composite films had good thermal, optical and mechanical properties; the last ones are apparently better than similar films reported in the literature. This is the first systematic study of different tunicate CN-GM nanocomposite films and the first ever for spruce GM.
Alkaline extracted and untreated wheat straw were ball-milled with liquid nitrogen cooling rendering them completely soluble in the solvent system dimethylsulfoxide-aqueous tetrabutylammonium hydroxide for subsequent fractionation into two lignin-carbohydrate complex fractions termed glucan-lignin and xylan-lignin according to their preferred association with glucan or arabinoxylan, respectively. This is the first description using this fractionation protocol for wheat straw. Eventually, acidolysis lignins were prepared from both lignin-carbohydrate complexes and structurally characterized using wet chemistry and NMR spectroscopy methods. Using the novel procedure we could reveal differences regarding wheat straw lignin association with polysaccharides, p-hydroxycinnamic acids and tricin as well as in their monomer composition. In glucan-lignin the lignin moiety was found to be linked mainly to glucan but also to branched arabinoxylan. Xylan-lignin, however, was rich in structures creating cross-links between lignin and linear arabinoxylan via ether-ester bridges by bi-functional ferulic acid. Inter-molecular ether-ester-linkages by ferulic acid connecting the lignin moieties of the two LCC fractions glucan-lignin and xylan-lignin were proposed. Alkaline extraction of the straw resulted in a strikingly lower recovery of xylan-lignin in the subsequent fractionation which was attributed to cleavage of ester linkages between ferulic acid and arabinoxylan. Structural characteristics indicated glucan-lignin and xylan-lignin deriving from different morphological origins of the cell wall.
Dioxane lignin was isolated from ball-milled wheat straw by neutral dioxane-water extraction and subsequently molar-mass-fractionated by flash chromatography in dimethylformamide. The eluted lignin containing material was pooled into six distinct molar mass fractions that were in turn structurally characterized in detail by wet chemistry and NMR spectroscopy methods. Fractions of higher molar mass were found to be enriched in p-hydroxyphenyl units and contained more p-hydroxycinnamic acid units. They were found mainly associated with linear arabinoxylan, while in low-molar-mass fractions additional glucan contributions were found. Fractions of lower molar mass consisted of relatively more guaiacyl units and showed exclusive association with tricin units. One distinct lignin fraction with lower lignin content supposedly contained high ratios of arabinoxylan chains esterified by ferulic acid and cross-linked via ferulic acid dimerization, structures which were considered to represent lignin nucleation sites. High abundance of dibenzodioxocin type structures in this fraction indicated that they could be involved in cross-linking hemicelluloses networks with lignin moieties.
A unique process for the fractionation of lignin from wheat straw is proposed: Ball milling for 8 h, followed by a direct and an acidolysis-assisted dioxane-water extraction. Four distinctly different lignin structures were thus obtained: (1) one free non-core lignin, which is a cellulose-lignin with lowest molar mass and highest contents of p-hydroxycinnamic acids, condensed phenolic hydroxyl groups and tricin moieties, and with a detectable amount of cinnamyl alcohols; (2) two core xylan-lignins differing in their degree of branching as indicated by their xylose/arabinose ratios of >4 and similar to 2, respectively; (3) one core cellulose-lignin which is the residual fraction resistant to all extractions. Based on the mass balance of Klason lignin the yields of these four fractions are 13.8%, 18.1%, 37.5% and 12.5%, respectively, thus accounting for 82% of the total KL in wheat straw. Therefore, the isolated lignin fractions could be considered as highly representative allowing a good insight into the different features of wheat straw lignin.