Gas chromatography-chemical ionization-mass spectrometric fatty acid analysis of a commercial supercritical carbon dioxide lipid extract from New Zealand green-lipped mussel (Perna canaliculus)

Supercritical fluid extracts of New Zealand green-lipped mussels (NZGLM) have been suggested to have therapeutic properties related to their oil components. The large number of minor FA in NZGLM extract was characterized by a GC-CIMS/MS method that excels at identification of double-bond positions in FAME. The extract contained five major lipid classes: sterol esters, TAG, FFA, sterols, and polar lipids. The total FA content of the lipid extract was 0.664 g/mL. Fifty-three unsaturated FA (UFA) were fully identified, of which 37 were PUFA, and a further 21 UFA were detected for which concentrations were too low for assignment of double-bond positions. There were 17 saturated FA, with 14∶0, 16∶0, and 18∶0 present in the greatest concentration. The 10 n−3 PUFA detected included 20∶5n−3 and 22∶6n−3, the two main n−3 FA; n−3 PUFA at low concentrations were 18∶3, 18∶4, 20∶3, 20∶4, 21∶5, 22∶5, 24∶6, and 28∶8. There were 43 UFA from the n−4, n−5, n−6, n−7, n−8, n−9, n−10, n−11 families, with 16∶2n−4, 16∶1n−5, 18∶1n−5, 18∶2n−6, 20∶4n−6, 16∶1n−7, 20∶1n−7, 16∶1n−9, 18∶1n−9, and 20∶1n−9 being the most abundant. In general, we estimated that FAME concentrations greater than 0.05% (w/w) were sufficient to assign double-bond positions. In total, 91 FA were detected in an extract of the NZGLM, whereas previous studies of fresh flesh from the NZGLM had reported identification of 42 FA. These data demonstrate a remarkable diversity of NZGLM FA.

Effects of variation in modulator temperature during cryogenic modulation in comprehensive two-dimensional gas chromatography

Many modulation systems in comprehensive 2D GC (GC×GC) are based on cryogenic methods. High trapping temperatures in these systems can result in ineffective trapping of the more volatile compounds, whilst temperatures that are too low can prevent efficient remobilisation of some compounds. To better understand the trapping and release of compounds over a wide range of volatilities, we have investigated a number of different constant temperature modulator settings, and have also examined a constant temperature differential between the cryo-trap and the chromatographic oven. These investigations have led us to modify the temperature regulation capabilities of the longitudinally modulated cryogenic system (LMCS). In contrast to the current system, where the user sets a constant temperature for the cooling chamber, the user now sets the temperature difference between the cryo-trap and the chromatographic oven. In this configuration, the cooling chamber temperature increases during the chromatographic run, tracking the oven temperature ramp. This produces more efficient, volatility-dependent modulation, and increases the range of volatile compounds that can be analysed under optimal trap-and-release conditions within a single analytical run. This system also reduces cryogenic fluid consumption.

Analysis of residues of prometryne and acetochlor in soil-water system by solid-phase extraction and gas chromatography/mass spectrometry

A highly sensitive and simple analytical method was developed for analyzing the binary mixed pesticides of prometryne and acetochlor in soil–water system by gas chromatography/mass spectrometry (GC/MS). The sample solution was first purified by C18 solid-phase extraction column, which was leached by acetone. The leachate was enriched to 1.0 mL by pressure blowing concentrator and then analyzed by GC/MS. The linear calibration curves were showed in the range of 1–15 μg/mL with a correlation coefficient of 0.9991. The average recoveries (n = 5) were between 95.3 and 115.7%, with relative standard deviations ranged from 1.71 and 7.95%. The limits of detection of Prometryne/Acetochlor were up to 0.06 and 0.17 μg/mL, respectively. This method provides a reliable approach to examine and evaluate the residues of prometryne and acetochlor in the soil–water system.

Analysis of mammalian cell proliferation and macromolecule synthesis using deuterated water and gas chromatography-mass spectrometry.

Deuterated water (²H₂O), a stable isotopic tracer, provides a convenient and reliable way to label multiple cellular biomass components (macromolecules), thus permitting the calculation of their synthesis rates. Here, we have combined ²H₂O labelling, GC-MS analysis and a novel cell fractionation method to extract multiple biomass components (DNA, protein and lipids) from the one biological sample, thus permitting the simultaneous measurement of DNA (cell proliferation), protein and lipid synthesis rates. We have used this approach to characterize the turnover rates and metabolism of a panel of mammalian cells in vitro (muscle C2C12 and colon cancer cell lines). Our data show that in actively-proliferating cells, biomass synthesis rates are strongly linked to the rate of cell division. Furthermore, in both proliferating and non-proliferating cells, it is the lipid pool that undergoes the most rapid turnover when compared to DNA and protein. Finally, our data in human colon cancer cell lines reveal a marked heterogeneity in the reliance on the de novo lipogenic pathway, with the cells being dependent on both 'self-made' and exogenously-derived fatty acid.

Surface energy of silk fibroin and mechanical properties of silk cocoon composites

Silkworm cocoons are biological composite structures protecting the silkworms against environmental damage and physical attack by natural predators. In particular, some outdoor reared silk cocoons exhibit outstanding mechanical properties that are relevant to the higher level protection required to enhance the survival chance of silkworms while supporting their metabolic activity. The performance of composite materials strongly depends on the adhesion between the fiber reinforcement and matrix, with the surface properties of the fibers playing a key role in determining the level of adhesion achieved. For this reason it is important to study the surface properties of silk fibroin to further understand the composite properties of the cocoons. In this work, both the mechanical properties of the silk cocoons and silk fibroin were studied. The surface topography was examined using scanning probe microscopy (SPM), which revealed distinct longitudinal ridges and striations along the fiber axis of the four silk fiber types. The fibers were found to exhibit heterogeneity in surface energy as evidenced from inverse gas chromatography (IGC) measurements. The combination of excellent mechanical properties and the more energetically heterogeneous surface nature of the wild A. pernyi silk fibroin fibers correlates well with the excellent mechanical properties of the A. pernyi cocoons. This journal is

Relationship between processing, surface energy and bulk properties of ultrafine silk particles

Silk particles of different sizes and shapes were produced by milling and interactions with a series of polar and non-polar gaseous probes were investigated using an inverse gas chromatography technique. The surface energy of all silk materials is mostly determined by long range dispersive interactions such as van der Waals forces. The surface energy increases and surface energy heterogeneity widens after milling. All samples have amphoteric surfaces and the concentration of acidic groups increases after milling while the surfaces remain predominantly basic. We also examined powder compression and flow behaviours using a rheometer. Increase in surface energy, surface area, and static charges in sub-micron air jet milled particles contributed to their aggregation and therefore improved flowability. However they collapse under large pressures and form highly cohesive powder. Alkaline hydrolysis resulted in more crystalline fibres which on milling produced particles with higher density, lower surface energy and improved flowability. The compressibility, bulk density and cohesion of the powders depend on the surface energy as well as on particle size, surface area, aggregation state and the testing conditions, notably the consolidated and unconsolidated states. The study has helped in understanding how surface energy and flowability of particles can be changed via different fabrication approaches.

Structure–property relationships of elementary bamboo fibers

Natural fibers are promising alternatives to synthetic fibers because of their sustainability, low environmental footprint and specific properties desirable for a wide range of technical engineering applications. The industrial implementation of fine grade natural bamboo fibers, including technical (100–200 microns) and elementary fibers (<30 microns) has been of increasing interest in recent times because these fibers offer a unique set of properties including high tensile strength, antibacterial and UV absorption. However to date, very little scientific effort has been devoted to fully understand the inter-correlation between their mechanical, physico-chemical, microstructural and morphological properties. In this paper, we report for the first time the structure–property relationship of elementary bamboo fibers. The impact of the inner microstructural organization of fibers (including the micro-fibrils angle) and physico-chemical factors such as the cellulose content and crystallinity index, on the tensile performance of these fibers is discussed in detail. This work also provides an insight into the application of bamboo fibers as natural and low-cost sorbent material for the removal of Cu2+ metal ions from model industrial wastewater. The metal ion adsorption properties of the fibers are correlated to surface energy analysis obtained from inverse gas chromatography.

Photocatalytic reduction of carbon dioxide into gaseous hydrocarbon using TiO2 pellets

It has been shown that CO₂ could be transformed into hydrocarbons when it is in contact with water vapour and catalysts under UV irradiation. This paper presents an experimental set-up to study the process employing a new approach of heterogeneous photocatalysis using pellet form of catalyst instead of immobilized catalysts on solid substrates. In the experiment, CO₂ mixed with water vapour in saturation state was discharged into a quartz reactor containing porous TiO₂ pellets and illuminated by various UV lamps of different wavelengths for 48 h continuously. The gaseous products extracted were identified using gas chromatography. The results confirmed that CO₂ could be reformed in the presence of water vapour and TiO₂ pellets into CH₄ under continuous UV irradiation at room conditions. It showed that when UVC (253.7 nm) light was used, total yield of methane was approximately 200 ppm which was a fairly good reduction yield as compared to those obtained from the processes using immobilized catalysts through thin-film technique and anchoring method. CO and H₂ were also detected. Switching from UVC to UVA (365 nm) resulted in significant decrease in the product yields. The pellet form of catalyst has been found to be attractive for use in further research on photocatalytic reduction of CO₂.

Study of polycyclic aromatic hydrocarbons emissions from burning of local wood species in Australia

Four kinds of Australian local wood species were burned in a domestic wood heater installed in a laboratory. The selected wood species include pine, red gum, yellow box and sugar gum, that are the most popular domestic fuel wood in Australia. Particulate matter emissions from burning of each load of wood were sampled isokinetically on filter media from the flue by standard stack emission sampling train. The particle laden filters then went for Gas Chromatography/ Mass Spectrometer (GC/MS) analysis to determine polycyclic aromatic hydrocarbons (PAHs) concentrations. The sampling was conducted under two different burning conditions – air inlet of the combustion chamber fully open and half open. Approximately 15 types of PAHs were detected. Emission factors were derived as microgram of PAHs /kg of wood burned. Total particulate emission factors were also obtained from gravimetric measurement before and after the sampling. PAH emission profiles for four species were generated from the results. Comparisons of emission factors have been conducted among different species of wood, as well as under different burning conditions, ie. fast burning and slowing burning. According to the derived emission factors, pine displayed the highest level of PAHs among the four species, followed by red gum and yellow box, whereas sugar gum showed the lowest level of PAHs. Emission factors were compared between each type of wood under two different burning conditions, the slow burning condition, which was air inlet half open, clearly showed higher PAH levels compared to the fast burning condition. Total PAH fractions on particulate matter were calculated and compared among wood types under two burning conditions. During the fast burning condition, red gum and pine have the higher percentage of PAH to total particulate matter emission than sugar gum and yellow box. When changed to slow burning, the PAH fraction on particulate matter are all increased with sugar gum having the largest increase.

A whole body, in vivo, fatty A\acid balance method to quantify PUFA metabolism (desaturation, elongation and beta-oxidation)

Currently there are several contrasting methods utilized for estimating elongation and desaturation of fatty acids and their general metabolism. The majority of these methods involve an ex vivo approach, requiring expensive and sophisticated equipment, likely to result in considerable variation in enzyme activity between and within species. In the present paper we introduce a further development of the whole-body fatty acid balance method for the estimation of the elongation and desaturation of fatty acids. This method though receiving considerable attention because of its simplicity and reliability has yet to be presented in detail. Theoretically, the whole-body fatty acid balance method can potentially be applied to any organism and requires little more than a gas chromatography unit for fatty acid analysis and elementary calculations. As such in this paper we attempt to spell out in detail the theoretical basis and the methods of application drawing specific examples. Using the present method it is possible to measure the fate of individual fatty acids towards desaturation, elongation and oxidation and calculate the elongase, Δ-6 desaturase and Δ-5 desaturase activities.

Low dose supplementation with two different marine oils does not reduce pro-inflammatory eicosanoids and cytokines in vivo

In view of the reported potential anti-inflammatory activity of the New Zealand green lipped mussel (NZGLM), we aimed to compare the effect of low dose marine oil supplementation, from mussels and fish, in reducing blood markers of inflammation. Thirty apparently healthy males and females were recruited from the general public in Melbourne, Australia to participate in a double blind, randomised, parallel intervention study. Subjects were consuming approximately 73 mg of omega-3 long chain polyunsaturated fatty acids (n-3 LCPUFA) daily in their background diet prior to the commencement of the intervention. Subjects were randomly assigned to consume either 2 mL/day of the NZGLM oil preparation (mixed with olive oil and dl-alpha-tocopherol) or fish oil preparation (also mixed with olive oil and dl-alpha-tocopherol) for six weeks. Two mL of the oils contained 241 mg and 181 mg of n-3 LCPUFA, respectively. Neutrophil phospholipid fatty acids, serum thromboxane B2 (TXB2), stimulated monocyte production of prostaglandin E2 (PGE2), interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNFalpha) were measured. During the intervention, the total intakes of n-3 LCPUFA from the background diet and the supplements were 199 mg/d and 173 mg/day for the NZGLM and FO groups, respectively. Following six weeks of supplementation, both groups showed a small, but significant increase in neutrophil phospholipid proportion of eicosapentaenoic acid. The NZGLM group also showed a significant increase in docosahexaenoic acid levels. There were no significant changes with time or treatment for TXB2, PGE2, IL-1 beta or TNFalpha. This study showed that low dose supplementation with n-3 LCPUFA from two different marine oil preparations showed no difference in inflammatory markers in this group of healthy individuals. Further studies are warranted including dose response trials and studies in populations with inflammatory conditions.

Photodegradation, interaction with iron oxides and bioavailability of dissolved organic matter from forested floodplain sources

Photochemical degradation of dissolved organic matter (DOM) can influence food webs by altering the availability of carbon to microbial communities, and may be particularly important following periods of high DOM input (e.g. flooding of forested floodplains). Iron oxides can facilitate these reactions, but their influence on subsequent organic products is poorly understood. Degradation experiments with billabong (= oxbow lake) water and river red gum (Eucalyptus camaldulensis) leaf leachate were conducted to assess the importance of these reactions in floodplain systems. Photochemical degradation of DOM in sunlight-irradiated quartz tubes (with and without amorphous iron oxide) was studied using gas chromatography and UV-visible spectroscopy. Photochemical reactions generated gaseous products and small organic acids. Bioavailability of billabong DOM increased following irradiation, whereas that of leaf leachate was not significantly altered. Fluorescence excitation-emission spectra suggested that the humic component of billabong organic matter was particularly susceptible to degradation, and the source of DOM influenced the changes observed. The addition of amorphous iron oxide increased rates of photochemical degradation of leachate and billabong DOM. The importance of photochemical reactions to aquatic systems will depend on the source of the DOM and its starting bioavailability, whereas inputs of freshly formed iron oxides will accelerate the processes.

Associations between fatty acids in colostrum and breast milk and risk of allergic disease

Background Exposure to n-3 polyunsaturated fatty acids (PUFA) in early life is hypothesized to offer protection against atopic disease. However, there is controversy in this area, and we have previously observed that high levels of n-3 fatty acid (FA) in colostrum are associated with increased risk of allergic sensitization.
Objective The aim of the study was to assess the relationship between FA profile in breast milk and risk of childhood atopic disease.
Methods A high-risk birth cohort was recruited, and a total of 224 mothers provided a sample of colostrum (n = 194) and/or 3-month expressed breast milk (n = 118). FA concentrations were determined by gas chromatography. Presence of eczema, asthma and rhinitis were prospectively documented up to 7 years of age.
Results High levels of n-3 22:5 FA (docosapentaenoic acid, DPA) in colostrum were associated with increased risk of infantile atopic eczema [odds ratio (OR) = 1.66 per 1 standard deviation increase, 95% confidence interval (CI) = 1.11–2.48], while total n-3 concentration in breast milk was associated with increased risk of non-atopic eczema (OR = 1.60, 95% CI = 1.03–2.50). Higher levels of total n-6 FA in colostrum were associated with increased risk of childhood rhinitis (OR = 1.59, 95% CI = 1.12–2.25). There was no evidence of associations between FA profile and risk of asthma.
Conclusion In this cohort of high-risk children, a number of modest associations were observed between FA concentrations in colostrum and breast milk and allergic disease outcomes. Further research in this area with larger sample sizes is needed.

Lipid composition of cashmere goat fibres

This study examined the differences in the chemical composition, particularly fatty acids, of the lipid extracted from the fibre of bucks, does and castrated goats. The study provides a more detailed understanding of the chemical composition of buck fibre lipid and how it varies throughout the year, and also details the effect of body region and nutrition on the production and chemical composition of lipid from buck fibre. Lipid was extracted with either petroleum ether (non-polar) or chloroform/methanol azeotrope (polar) and analysed by gas chromatography and gas chromatography-mass spectrometry. The more polar solvent system extracted larger amounts of lipid and more of each individual fatty acid. The following buck specific ethyl branched fatty acids were identified: 2-ethylhexanoic, 4-ethylhexanoic, 2-ethyloctanoic, 4-ethyloctanoic, 6-ethyloctanoic, 2-ethyldecanoic, 4-ethyldecanoic, 2-ethyldodecanoic, 6-ethyldodecanoic, 4-ethyldodecanoic, 2-ethyltetradecanoic, 6-ethyltetradecanoic, 4-ethyltetradecanoic, 2-ethylhexadecanoic and 4-ethyloctadecanoic acids. Of these buck specific fatty acids only 4-ethylhexanoic (T), 4-ethyloctanoic, 4-ethyldecanoic, 4-ethyldodecanoic, 6-ethyldodecanoic (T), 4-ethyltetradecanoic, 2-ethylhexadecanoic (T) and 4-ethylhexadecanoic acids have been previously identified or tentatively identified (T) in buck fibre extracts. This shows that the chemical composition of buck fibre lipid is more complex than previously reported, and that it may be more difficult than previously thought to artificially duplicate the odour of the buck. Buck fibre samples had lower average concentrations of 2-methylpropanoic, 2-methylbutanoic, iso-pentadecanoic, anteiso-pentadecanoic, iso-hexadecanoic, anteiso-heptadecanoic, iso-octadecanoic and anteiso-nonadecanoic acids as compared with fibre samples from does, spayed does, or wethers that were castrated at one month of age. The reduced concentrations of these fatty acids in buck fibre extracts were likely to be due to the synthesis of ethyl branched derivatives of iso and anteiso fatty acids. Buck fibre samples had higher concentrations of benzoic acid as compared with fibre samples from does, spayed does, or wethers that were castrated at one month of age. The significance of these results is that non buck specific fatty acids may also make a contribution to the odour of bucks. When fibre samples were collected at various times throughout the year, it was found that the bucks had increased amounts of lipid and ethyl branched fatty acids in fibre samples shorn from March to September, as compared with fibre samples shorn in November and January. The increase in the amount of lipid and ethyl branched fatty acids corresponded with both the rutting period of the buck and the period when the buck odour was increased. This suggests that ethyl branched fatty acids could be pheromones. The variation in lipid content and fatty acid composition was also examined between fibre samples collected from different body regions of the buck during April, as alterations in sebaceous gland activity around the neck during rutting have been reported. It was found that the average amount of lipid in the neck region of the bucks was not statistically higher than the average amounts in the midside and hind regions. However, the ethyl branched fatty acid concentrations were statistically higher in the fibre from around the neck as compared with the fibre from the other body regions, which is consistent with the odour of the buck being most pronounced around the head and neck region. The lipid content and composition of fibre samples from bucks fed high and low quality diets (lucerne and pangola grass, respectively) was examined to determine the effect of nutrition on buck specific components. The high quality diet increased the amount of lipid and ethyl branched fatty acids in fibre samples collected in April from the neck, midside and hind regions, as compared with fibre samples from the corresponding body regions from bucks fed the low quality diet. Thus it may be possible for the pheromone levels of bucks to be increased by simply providing them with good nutrition. The lipid content and ethyl branched fatty acid concentrations of fibre samples increased earlier in the year for the lucerne fed bucks as compared with the pangola grass fed bucks. The lucerne fed bucks had increased concentrations of ethyl branched fatty acids in fibre samples shorn during December to June (6 months) whereas the pangola grass fed bucks had increased concentrations of ethyl branched fatty acids in fibre samples shorn during April to August (4 months). These observations show that good nutrition can result in both the earlier production of ethyl branched fatty acids and an extended period when ethyl branched fatty acids are produced. This suggests that nutrition can be used to manipulate pheromone levels in the buck. The period when the ethyl branched fatty acids were increased corresponded with the period when the plasma luteinizing hormone (LH) and testosterone concentrations, odour and sebaceous gland volume of the bucks were increased, which supports the assumption that ethyl branched fatty acids are involved in odour production and act as pheromones.