Lillie Cavonius

Oxidation of Cod Liver Oil during Gastrointestinal in Vitro Digestion

Oxidation of cod liver oil rich in long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) was investigated during a gastrointestinal (GI) in vitro digestion. The digestion stimulated TBA-reactive substances (TBARS) formation in both the gastric and intestinal steps, whereas levels of lipid hydroperoxides remained nearly constant. The presence of digestive compounds was decisive for the TBARS development because TBARS did not change when the cod liver oil was subjected only to the temperature and pH gradient of the GI model. Preformed oxidation products in the cod liver oil resulted in further elevated TBARS levels during the digestion. Addition of hemoglobin (11.5 μM) to emulsified cod liver oil dramatically increased TBARS and lipid hydroperoxide levels during GI digestion, whereas 1 mg α-tocopherol/g oil did not show any protection against oxidation. Specific concern thus needs to be taken in the design of foods containing LC n-3 PUFA to preserve these lipids and avoid harmful oxidation, both before and after consumption.

Quantification of total fatty acids in microalgae: comparison of extraction and transesterification methods.

Determination of microalgaes’ fatty acid content is often done with chloroform and methanol according to the Bligh and Dyer extraction, though faster methods exist. A number of comparisons between the Bligh and Dyer and faster methods have resulted in contradicting data, possibly due to differences in algae used and the different versions of the Bligh and Dyer method applied. Here, various forms of direct-transesterification (D-TE) and two-step transesterification (2-TE), including three versions developed in our lab, are compared with the original Bligh and Dyer (Can J Biochem Physiol 37: 911–917, 1959) extraction and two modifications thereof (Lee et al. J AOAC Int 79:487–492, 1996, and our own acidified version) on microalgae with different cell walls: Isochrysis galbana, Nannochloropsis oculata, and Phaeodactylum tricornutum. In total, fatty acid extracts from 11 methods were separated and quantified by gas chromatography with mass spectrometry. Results show that, for N. oculata and P. tricornutum, methods based on chloroform–methanol underestimated the fatty acid content compared with the 2-TE and D-TE methods, which gave similar results. Moreover, D-TE methods are faster than chloroform–methanol methods and use chemicals that are less toxic. Of the D-TE methods, the ones using hydrochloric acid and sulfuric acid recovered the most fatty acids, while boron trifluoride recovered slightly less. The main qualitative difference between the fatty acids recovered was that the chloroform–methanol methods recovered less saturated fatty acids in P. tricornutum.

Imaging of Lipids in Microalgae with Coherent Anti-Stokes Raman Scattering Microscopy

Lipids are accumulated as giant droplets alongside coalescing emerging droplets under excessive lipid storage, in contrast to the multiple micron-sized droplets formed at normal conditions. Microalgae have great prospects as a sustainable resource of lipids for refinement into nutraceuticals and biodiesel, which increases the need for detailed insights into their intracellular lipid synthesis/storage mechanisms. As an alternative strategy to solvent- and label-based lipid quantification techniques, we introduce time-gated coherent anti-Stokes Raman scattering (CARS) microscopy for monitoring lipid contents in living algae, despite strong autofluorescence from the chloroplasts, at approximately picogram and subcellular levels by probing inherent molecular vibrations. Intracellular lipid droplet synthesis was followed in Phaeodactylum tricornutum algae grown under (1) light/nutrient-replete (control [Ctrl]), (2) light-limited (LL), and (3) nitrogen-starved (NS) conditions. Good correlation (r2 = 0.924) was found between lipid volume data yielded by CARS microscopy and total fatty acid content obtained from gas chromatography-mass spectrometry analysis. In Ctrl and LL cells, micron-sized lipid droplets were found to increase in number throughout the growth phases, particularly in the stationary phase. During more excessive lipid accumulation, as observed in NS cells, promising commercial harvest as biofuels and nutritional lipids, several micron-sized droplets were present already initially during cultivation, which then fused into a single giant droplet toward stationary phase alongside with new droplets emerging. CARS microspectroscopy further indicated lower lipid fluidity in NS cells than in Ctrl and LL cells, potentially due to higher fatty acid saturation. This agreed with the fatty acid profiles gathered by gas chromatography-mass spectrometry. CARS microscopy could thus provide quantitative and semiqualitative data at the single-cell level along with important insights into lipid-accumulating mechanisms, here revealing two different modes for normal and excessive lipid accumulation.

Glazing herring (Clupea harengus) fillets with herring muscle press juice: effect on lipid oxidation development during frozen storage

In this study, fish muscle press juice, PJ, was evaluated as an antioxidative liquid for glazing of fish fillets prior to frozen storage. PJ has in previous studies efficiently protected fish mince lipids against haemoglobin- mediated oxidation. Frozen herring fillet were glazed with water, PJ or 50 mM phosphate buffer of the same pH as herring PJ (6.5). Over 52 weeks, peroxide value (PV), TBA-reactive substances (TBARS), rancid odour and colour were monitored. From 26 to 52 weeks, water-glazed fillets were significantly (P < 0.05) more oxidised compared to fillets glazed with PJ or buffer, most clearly shown by PV and TBARS data. The protective effects of both PJ and buffer compared to water could indicate a role, for example, of the physiological ionic strength. The finding that PV became elevated in PJ which was frozen stored alone in a thin layer indicates that PJ stripped from, for example, trace lipids and haemoglobin would provide better protection.