GC-based analysis of plant stanyl fatty acid esters in enriched foods

Approaches for the capillary gas chromatographic (GC) based analysis of intact plant stanyl esters in enriched foods were developed. Reference compounds were synthesized by enzyme-catalyzed transesterifications. Their identities were confirmed by means of mass spectrometry. Using a medium polar trifluoropropylmethyl polysiloxane stationary phase, long-chain plant stanyl esters could be separated according to their stanol moieties and their fatty acid chains. Thermal degradation during GC analysis was compensated by determining response factors; calibrations were performed for ten individual plant stanyl esters. For the analysis of low-fat products (skimmed milk drinking yogurts), the GC separation was combined with a "fast extraction" under acidic conditions. For fat-based foods (margarines), online coupled LC-GC offered an elegant and efficient way to avoid time-consuming sample preparation steps. The robust and rapid methods allow conclusions on both, the stanol profiles and the fatty acid moieties, and thus provide a basis for the authentication of this type of functional food ingredients.     

Characterization of sphingolipids from sunflower seeds with altered fatty acid composition

Sphingolipids are a group of lipids that are derived from long-chain 1,3-dihydroxy-2-amino bases and that are involved in important processes in plants. Long-chain bases are usually found bound to long-chain fatty acids forming ceramides, the lipophilic moiety of the most common sphingolipid classes found in plant tissues: glucosyl-ceramides and glucosyl inositol phosphoryl-ceramides (GIPCs). The developing sunflower seed kernel is a tissue rich in sphingolipids, although, importantly, its glycerolipid composition can vary if some steps of the fatty acid synthesis are altered. Here, the sphingolipid composition of the seed from different sunflower mutants with altered fatty acid compositions was studied. The long-chain base composition and content were analyzed, and it was found to be similar in all of the mutants studied. The sphingolipid species were also determined by mass spectrometry, and some differences were found in highly saturated sunflower mutants, which contained higher levels of GIPC, ceramides, and hydroxyl-ceramides.     

Changes in fatty acid composition with age and environment in different types of peat profiles in Japan

Fatty acid composition of the crude lipid fraction of peat was investigated using several typical peat profiles in Japan. Fatty acid composition varied with the peat layers accumulated in a peat profile since 32,000 years BP. Deposition of long-range transported volcanic ash tephras also affected the composition remarkably due to the acceleration of decomposition. Fatty acid composition differed among high moor, transitional moor, and low moor peat profiles sampled in several locations in Hokkaido, Japan. The difference in the plants involved in the formation of peat was considered to be a very important factor determining the fatty acid composition. For example, arachidic acid was considered to be a good indicator for the presence of reed, while the percentage of stearic acid tended to be high in the high moor peat profiles dominated by sphagnum. As the fatty acids with a longer chain length were more stable than those with a shorter chain length, the percentage of longer fatty acids (C24 and C26) tended to increase and that of shorter fatty acids (CI4 and CI6) to decrease in the lower peat profiles. The drying of peatland also exerted a similar effect on the fatty acid composition.     

Development of a simplified method for the determination of folates in baker's yeast by HPLC with ultraviolet and fluorescence detection

A simplified HPLC method for rapid determination of folates in yeast with ultraviolet and fluorescence detection without sample purification has been developed. By use of the column Aquasil C(18), specially designed for polar analytes, and gradient elution, it was possible to separate and determine five folate derivatives: tetrahydrofolate, 5-methyltetrahydrofolate, and 5-formyltetrahydrofolate with fluorescence detection, and 10-formylfolic acid and folic acid with ultraviolet detection. The sample preparation required only a small amount of dry yeast (25-50 mg) and included an extraction of folates by heat treatment and deconjugation of folate polyglutamates to monoglutamates with the use of rat serum conjugase. Validation involved investigation of matrix effects, determination of recovery by standard addition method, repeatability, and stability tests. The dominating folate forms in commercial dry baker's yeast were found to be tetrahydrafolate and 5-methyltetrahydrofolate with a total folate content of 2890 microg/100 g (63.4 nmol/g). The simplicity of the method makes it suitable for folate screening studies of different yeast strains.     

土壤中木质素磺酸盐对土壤溶液化学和磷脂脂肪酸组成的影响

An incubation experiment （Exp. 1） with three soils, two from Australia and one from Norway, was carried out to investigate the fate of dissolved BorreGro （a lignosulfonate, produced by Borregaard LignoTech Company, Norway） at different concentrations （0, 10 and 100 mg C L-1） in soil solutions. A rhizobox experiment （Exp. 2） was also done in a Norwegian clay soil, mixed with four levels of BorreGro-carbon （BG-C） added （0, 2, 20 and 200 mg BG-C kg-1） to test the impact of BorreGro on root growth, rhizosphere chemistry （pH, metals and dissolved organic carbon （DOC）） and the composition of phospholipid fatty acids （PLFAs）. The BorreGro addition increased the concentration of Mn due to the high concentrations in BorreGro. The BorreGro addition to soil had an indirect but significant impact on the rhizosphere chemistry and PLFAs. The lowest amounts of added BorreGro facilitated the DOC excretion at plant roots, and thereby increased the bacterial and fungal biomass, likely as an effect of increased Mn solubility from BorreGro in the root zone.     

Effects of lactoferrin, phytic acid, and EDTA on oxidation in two food emulsions enriched with long-chain polyunsaturated fatty acids

The influence of the addition of metal chelators on oxidative stability was studied in a milk drink and in a mayonnaise system containing highly polyunsaturated lipids. Milk drinks containing 5% (w/w) of specific structured lipid were supplemented with lactoferrin (6-24 muM) and stored at 2 degrees C for up to 9 weeks. Mayonnaise samples with 16% fish oil and 64% rapeseed oil (w/w) were supplemented with either lactoferrin (8-32 muM), phytic acid (16-124 muM), or EDTA (16-64 muM) and were stored at 20 degrees C for up to 4 weeks. The effect of the metal chelators was evaluated by determination of peroxide values, secondary volatile oxidation products, and sensory analysis. Lactoferrin reduced the oxidation when added in concentrations of 12 muM in the milk drink and 8 muM in the mayonnaise, whereas it was a prooxidant at higher concentrations in both systems. In mayonnaise, EDTA was an effective metal chelator even at 16 muM, whereas phytic acid did not exert a distinct protective effect against oxidation. The differences in the equimolar effects of the metal chelators are proposed to be due to differences in their binding constants to iron and their different stabilities toward heat and low pH.     

温度和秸秆质量调节与秸秆分解相关的微生物磷脂脂肪酸（PLFA）组成

Variations in temperature and moisture play an important role in soil organic matter (SOM) decomposition. However, relationships between changes in microbial community composition induced by increasing temperature and SOM decomposition are still unclear. The present study was conducted to investigate the effects of temperature and moisture levels on soil respiration and microbial communities involved in straw decomposition and elucidate the impact of microbial communities on straw mass loss. A 120-d litterbag experiment was conducted using wheat and maize straw at three levels of soil moisture (40%, 70%, and 90% of water-holding capacity) and temperature (15, 25, and 35°C). The microbial communities were then assessed by phospholipid fatty acid (PLFA) analysis. With the exception of fungal PLFAs in maize straw at day 120, the PLFAs indicative of Gram-negative bacteria and fungi decreased with increasing temperatures. Temperature and straw C/N ratio significantly affected the microbial PLFA composition at the early stage, while soil microbial biomass carbon (C) had a stronger effect than straw C/N ratio at the later stage. Soil moisture levels exhibited no significant effect on microbial PLFA composition. Total PLFAs significantly influenced straw mass loss at the early stage of decomposition, but not at the later stage. In addition, the ratio of Gram-negative and Gram-positive bacterial PLFAs was negatively correlated with the straw mass loss. These results indicated that shifts in microbial PLFA composition induced by temperature, straw quality, and microbial C sources could lead to changes in straw decomposition.     

IUPAC gas chromatographic method for determination of fatty acid composition: collaborative study.

An international collaborative study of IUPAC methods II.D.19 and II.D.25 for preparation and GLC analysis of fatty acid methyl esters was begun in 1976. The IUPAC methodology, applicable to animal and vegetable oils and fats and fatty acids from all sources, contains special instructions for preparation and analysis of methyl esters of fatty acids containing 4 or more carbon atoms (analysis of milk fat). Twenty-three collaborators participated in the analysis of 5 known mixtures, 4 vegetable oils, 1 fish oil, and 2 butterfats. Several blind duplicate samples were included. The experimental data were subjected to statistical analysis to examine intra- and interlaboratory variation. Reproducibility and accuracy data for the higher fatty acid (14:0-22:1) mixtures and fish and vegetable oils were satisfactory and were in good agreement with results from an AOCS Smalley Committee check sample program involving analysis of the same samples. Typical coefficients of variation (%) at various concentrations were 15 (2% level), 8.5 (5% level), 7 (10% level), and 3 (50% level). Low recoveries and poor reproducibility were characteristics of results obtained for butyric acid in the butterfat and related known mixtures. A coefficient of variation of about 19% was found for analysis of butyric acid in butterfat, vs coefficients of variation in the range of 4-13% for similar levels of other components in butterfat and other samples. The IUPAC methodology for GLC analysis of fats and oils other than milk fats has been adopted by the AOAC as official first action to replace the current GLC method, 28.063-28.067.     

Stearidonic acid soybean oil enriched with palmitic acid at the sn-2 position by enzymatic interesterification for use as human milk fat analogues

Stearidonic acid (SDA, C18:4n-3) enriched soybean oil may be added to the diet to increase intake of omega-3 fatty acids (FAs). Human milk fat has ≥60% of palmitic acid (PA), by weight, esterified at the sn-2 position to improve absorption of fat and calcium in infants. Enzymatic interesterification of SDA soybean oil and tripalmitin produced structured lipids (SLs) enriched with PA at the sn-2 position of the triacylglycerol. Reactions were catalyzed by Novozym 435 or Lipozyme TL IM under various conditions of time, temperature, and substrate mole ratio. Response surface methodology was used to design the experiments. Model optimization conditions were predicted to be 1:2 substrate mole ratio at 50 °C for 18 h with 10% (by weight) Lipozyme TL IM resulting in 6.82 ± 1.87% total SDA and 67.19 ± 9.59% PA at sn-2; 1:2 substrate mole ratio at 50 °C for 15.6 h resulting in 8.01 ± 2.41% total SDA and 64.43 ± 13.69% PA at sn-2 with 10% (by weight) Novozym 435 as the biocatalyst. The SLs may be useful as human milk fat analogues for infant formula formulation with health benefits of the omega-3 FAs.     

Effects of temperature and life stage on the fatty acid composition of Collembola

Fatty acid (FA) analysis is used as a promising tool to investigate trophic interactions in soil food webs. The FA profile of neutral lipids in consumers is affected by the diet, and the occurrence and amount of certain FAs can reflect feeding strategies. We investigated the lipid composition of the Collembola Folsomia candida, Heteromurus nitidus and Protaphorura fimata with the fungus Chaetomium globosum as food source. The impact of environmental temperature and life stage was assessed, with special respect to linoleic acid (18:2ω6,9) as a marker FA for fungal feeding. In all Collembola species the ratio of C16/C18 in neutral lipid fatty acids (NLFAs) increased with decreasing temperature. In the NLFAs of F. candida and H. nitidus the Unsaturation Index and the amount of 18:2ω6,9 decreased with temperature, whereas in P. fimata effects were the opposite. The composition of phospholipid fatty acids (PLFAs) differed between species, but was little affected by temperature. The degree of unsaturation in NLFAs increased with the age of Collembola, mainly due to higher amounts of 18:2ω6,9 and a lower proportion of 18:1ω9. The biomarker linoleic acid represented over 20% of FAs in all fungal feeding Collembola. Despite considerable influence of temperature and life stage on its proportion, the amount was always higher than in individuals reared on other diets. This suggests that linoleic acid can serve as marker for fungal feeding independent of such physiological variations in Collembola.     

Diet alters the fatty acid composition of individual phospholipid classes in beef muscle

The objective of this study was to investigate the effect of diet on the distribution of phospholipid classes and fatty acid profiles of individual phospholipid classes in longissimus muscle of beef. An experiment was established to examine the effect of pasture-based versus concentrate diet offered to two different breeds (German Holstein and German Simmental bulls) to enhance the content of beneficial fatty acids in beef and improve the meat quality for the consumer. High-performance thin-layer chromatography was utilized to separate the phospholipid classes. The fatty acid composition of the individual phospholipid classes was determined by gas chromatography. The main phospholipid classes in the muscle were phosphatidylethanolamine and phosphatidylcholine, representing approximately 60% of the total phospholipids, followed by phosphatidylinositol ranging between 11.8 and 14.8%. The results have shown that the fatty acid profiles in the detected seven phospholipid classes can be affected by different feeding systems. Pasture-based feeding resulted in an enrichment of total and individual n-3 fatty acids in all phospholipid classes of muscle lipids of bulls compared with those fed on concentrate. In contrast, pasture-based diet significantly decreased the proportion of total and individual n-6 fatty acids in phospholipid classes, except in the sphingomyelin fraction. The total saturated fatty acid proportions in the phospholipid classes were different and ranged between 4.5% in the cardiolipin fraction and 50.5% in the sphingomyelin fraction of muscle lipids of bulls. Furthermore, the diet effects on the saturated fatty acid proportion in the different phospholipid classes differ widely. The results have shown that the C18:1 trans and CLA profiles in the detected seven phospholipid classes can be affected by different feeding systems.     

分提橄榄油中主要脂肪酸成分傅里叶变换红外光谱法测量（英）

为了测量从橄榄油中分提的高、低熔点油脂的脂肪酸成分,在4 000～600 cm-1 的范围测量了31个含有不同脂肪酸成分植物油的傅里叶变换红外光谱,用于建立偏最小二乘（PLS）回归分析校正模型。在油脂的傅里叶变换红外光谱变量和脂肪酸组成变量之间建立了交叉验证的PLS校正模型。为了校正油酸和亚油酸含量,在4?000～600 cm-1 的频率范围,经平滑,二阶导数,规范化处理的红外光谱获得了最好的交叉验证校正模型和最佳的预测结果。PLS校正模型预测结果表明,与高熔点橄榄油（油酸,72.29%,亚油酸,9.98%）相比,低熔点橄榄油含有较高的油酸含量和亚油酸含量（油酸,77.46%,亚油酸,12.51%）,预测的结果与气相色谱测量的结果有很好的一致性。建立的PLS校正模型预测橄榄油的不饱和脂肪酸含量具有较好的相关性。该研究为分提油脂质量的判别评价提供了便捷的方法。     

Effects of nitric oxide on fatty acid composition in peach fruits during storage

Peach (Prunus persica (L.) Batsch., cv. Feicheng) fruits were fumigated with 0, 5, 10, and 15 microL/L nitric oxide (NO), and the effects of NO on rot index, ion leakage, and the lipid compositions in peach fruits were studied. The results showed that the treatment with 10 microL/L NO significantly increased the contents of the palmitoleic (16:1), oleic (18:1), and linolenic (18:3) acid in phospholipid (PL), while it decreased the contents of linoleic (18:2) acid in PL. The content of monoacylglycerol (MG) in fruit fumigated with 10 microL/L NO were higher than in the control but lower in fruit fumigated with 15 microL/L NO. The treatment with 10 microL/L NO increased the content of MG, decreased the contents of diacylglycerol (DG) and triacylglycerol (TG). However, there was no significant difference in the contents of free fatty acid (FFA). The compositions of MG, DG, TG, and FFA in peach fruit were also changed by the treatment with 10 microL/L NO. It could preserve the content of 18:3, maintain the integrity of membrane, and prevent the softening and rotting of peach fruit. The content of 18:1 was detected in PL and FFA but not in MG, DG, and TG. This might be due to the different compositions of MG, DG, and TG.     

Assessment of oil content and fatty acid composition variability in two economically important Hibiscus species

The Hibiscus genus encompasses more than 300 species, but kenaf (Hibiscus cannabinus L.) and roselle (Hibiscus sabdariffa L.) are the two most economically important species within the genus. Seeds from these two Hibiscus species contain a relatively high amount of oil with two unusual fatty acids: dihydrosterculic and vernolic acids. The fatty acid composition in the oil can directly affect oil quality and its utilization. However, the variability in oil content and fatty acid composition for these two species is unclear. For these two species, 329 available accessions were acquired from the USDA germplasm collection. Their oil content and fatty acid composition were determined by nuclear magnetic resonance (NMR) and gas chromatography (GC), respectively. Using NMR and GC analyses, we found that Hibiscus seeds on average contained 18% oil and seed oil was composed of six major fatty acids (each >1%) and seven minor fatty acids (each <1%). Hibiscus cannabinus seeds contained significantly higher amounts of oil (18.14%), palmitic (20.75%), oleic (28.91%), vernolic acids (VA, 4.16%), and significantly lower amounts of stearic (3.96%), linoleic (39.49%), and dihydrosterculic acids (DHSA, 1.08%) than H. sabdariffa seeds (17.35%, 18.52%, 25.16%, 3.52%, 4.31%, 44.72%, and 1.57%, respectively). For edible oils, a higher oleic/linoleic (O/L) ratio and lower level of DHSA are preferred, and for industrial oils a high level of VA is preferred. Our results indicate that seeds from H. cannabinus may be of higher quality than H. sabdariffa seeds for these reasons. Significant variability in oil content and major fatty acids was also detected within both species. The variability in oil content and fatty acid composition revealed from this study will be useful for exploring seed utilization and developing new cultivars in these Hibiscus species.     

Nondestructive determination of oil content and fatty acid composition in perilla seeds by near-infrared spectroscopy

Near-infrared reflectance spectroscopy (NIRS) was used as a rapid and nondestructive method to determine the oil content and fatty acid composition in intact seeds of perilla [Perilla frutescens var. japonica (Hassk.) Hara] germplasms in Korea. A total of 397 samples (about 2 g of intact seeds) were scanned in the reflectance mode of a scanning monochromator, and the reference values for the oil content and fatty acid composition were measured by gravimetric method and gas-liquid chromatography, respectively. Calibration equations for oil and individual fatty acids were developed using modified partial least-squares regression with internal cross validation (n = 297). The equations for oil and oleic and linolenic acid had lower standard errors of cross-validation (SECV), higher R2 (coefficient of determination in calibration), and higher ratio of unexplained variance divided by variance (1-VR) values than those for palmitic, stearic, and linoleic acid. Prediction of an external validation set (n = 100) showed significant correlation between reference values and NIRS estimated values based on the standard error of prediction (SEP), r2 (coefficient of determination in prediction), and the ratio of standard deviation (SD) of reference data to SEP. The models for oil content and major fatty acids, oleic and linolenic acid, had relatively higher values of SD/SEP(C) and r2 (more than 3.0 and 0.9, respectively), thereby characterizing those equations as having good quantitative information, whereas those of palmitic, stearic, and linoleic acid had lower values (below 2.0 and 0.7, respectively), unsuitable for screening purposes. The results indicated that NIRS could be used to rapidly determine oil content and fatty acid composition (oleic and linolenic acid) in perilla seeds in the breeding programs for development of high-quality perilla oil.