Development of a rapid method for the sequential extraction and subsequent quantification of fatty acids and sugars from avocado mesocarp tissue

Methods devised for oil extraction from avocado (Persea americana Mill.) mesocarp (e.g., Soxhlet) are usually lengthy and require operation at high temperature. Moreover, methods for extracting sugars from avocado tissue (e.g., 80% ethanol, v/v) do not allow for lipids to be easily measured from the same sample. This study describes a new simple method that enabled sequential extraction and subsequent quantification of both fatty acids and sugars from the same avocado mesocarp tissue sample. Freeze-dried mesocarp samples of avocado cv. Hass fruit of different ripening stages were extracted by homogenization with hexane and the oil extracts quantified for fatty acid composition by GC. The resulting filter residues were readily usable for sugar extraction with methanol (62.5%, v/v). For comparison, oil was also extracted using the standard Soxhlet technique and the resulting thimble residue extracted for sugars as before. An additional experiment was carried out whereby filter residues were also extracted using ethanol. Average oil yield using the Soxhlet technique was significantly (P < 0.05) higher than that obtained by homogenization with hexane, although the difference remained very slight, and fatty acid profiles of the oil extracts following both methods were very similar. Oil recovery improved with increasing ripeness of the fruit with minor differences observed in the fatty acid composition during postharvest ripening. After lipid removal, methanolic extraction was superior in recovering sucrose and perseitol as compared to 80% ethanol (v/v), whereas mannoheptulose recovery was not affected by solvent used. The method presented has the benefits of shorter extraction time, lower extraction temperature, and reduced amount of solvent and can be used for sequential extraction of fatty acids and sugars from the same sample.     

Formation of modified fatty acids and oxyphytosterols during refining of low erucic acid rapeseed oil

Formation of trans fatty acids and cyclic fatty acid monomers was investigated during refining of low erucic acid rapeseed oil. The first steps of the refining process, that is, degumming, neutralization, and bleaching, hardly modified the fatty acid profile. In contrast, deodorization produced substantial quantities of trans fatty acids (>5% of total fatty acids) and small amounts of cyclic fatty acid monomers (650 mg of cyclic fatty acid monomers/kg of oil) when severe conditions (5-6 h at 250 degrees C) were used. Alpha-linolenic acid was the main precursor of cyclic fatty acid monomers. The influence of deodorization on the chemical composition of low erucic acid rapeseed oil was studied additionally. Whereas free fatty acids, peroxides, and tocopherols decreased, neither total polar compounds nor oxyphytosterols changed during deodorization. Oxyphytosterols were identified by GC-MS. Three oxyphytosterols not yet observed in oil were tentatively identified as 6beta-hydroxycampestanol, 6beta-hydroxysitostanol, and 6beta-hydroxybrassicastanol. Brassicasterol oxides were the most abundant oxyphytosterols.     

A quantitative method for the determination of cyclopropenoid fatty acids in cottonseed, cottonseed meal, and cottonseed oil (Gossypium hirsutum) by high-performance liquid chromatography

Cyclopropenoid fatty acids (CPFAs), found in cottonseed, have been shown to have detrimental health effects to susceptible livestock. Previous quantitative analytical methods for the determination of CPFAs expressed these acids in terms of their relative abundance with respect to other fatty acids in the oil, necessitating the concurrent analysis of other fatty acids. The proposed analytical method describes the quantitation of three relevant CPFAs for cotton (malvalic acid, sterculic acid, and dihydrosterculic acid) in cottonseed in micrograms per gram fresh weight of sample. The method involves extraction of the oil, saponification, and derivatization of the free fatty acids with 2-bromoacetophenone to give the phenacyl esters. These esters are then separated by dual-column reverse-phase high-performance liquid chromatography and quantitated via external standards. This is the first method to include external calibration standards for CPFAs and, as such, is capable of direct quantification with no further data conversion required. CPFA data generated from the analysis of cottonseed, cottonseed meal, and cottonseed oil produced in the United States in 2002 are presented.     

ATR-Fourier transform mid-infrared spectroscopy for determination of trans fatty acids in ground cereal products without oil extraction

Fourier transform mid-infrared (FT-IR) spectroscopy was investigated as a method of analysis for trans fatty acid content of cereal products without the need for prior oil extraction. Spectra were obtained, with an FT-IR spectrometer equipped with an attenuated total reflectance (ATR) device, of ground samples pressed onto the diamond ATR surface, and trans fatty acids were measured by a modification of AOAC Method 996.01. Partial least-squares (PLS) models were developed for the prediction of trans fatty acids in ground samples using several wavenumber selections on the basis of bands related to lipids. The models (n = 79) predicted trans fatty acids in ground samples with standard error of cross-validation (SECV) of 1.10-1.25 (range 0-12.4) % and R2 of 0.85-0.88 and in validation samples (n = 26) with standard error of performance (SEP) of 0.96-1.12 (range 0-12.2) % and r2 of 0.89-0.92, indicating sufficient accuracy for screening. Sample trans fatty acid % was predicted as accurately with the fingerprint region (1500-900 cm(-1)) as with the entire range (4000-650 cm(-1)) indicating, in concert with the regression coefficients, the importance of the isolated trans double bonds at 966 cm(-1) in development of the model. Data is also presented on prediction of trans fatty acids using the spectra of residual oil films on the ATR surface after removing the solid portion of the sample.     

Quantitation of fatty acids, sterols, and tocopherols in turpentine (Pistacia terebinthus Chia) growing wild in Turkey

The chemical composition (fatty acids, tocopherols, and sterols) of the oil from 14 samples of turpentine (Pistacia terebinthus L.) fruits is presented in this study. The oil content of the samples varied in a relatively small range between 38.4 g/100 g and 45.1 g/100 g. The dominating fatty acid of the oil is oleic acid, which accounted for 43.0 to 51.3% of the total fatty acids. The total content of vitamin E active compounds in the oils ranged between 396.8 and 517.7 mg/kg. The predominant isomers were alpha- and gamma-tocopherol, with approximate equal amounts between about 110 and 150 mg/kg. The seed oil of P. terebinthus also contained different tocotrienols, with gamma-tocotrienol as the dominate compound of this group, which amounted to between 79 and 114 mg/kg. The total content of sterols of the oils was determined to be between 1341.3 and 1802.5 mg/kg, with beta-sitosterol as the predominent sterol that accounted for more than 80% of the total amount of sterols. Other sterols in noteworthy amounts were campesterol, Delta5-avenasterol, and stigmasterol, which came to about 3-5% of the total sterols.     

水酶法提取山核桃油脂工艺研究

为优化水酶法提取山核桃油脂工艺,以山核桃为原料,采用水酶法提取油脂,在单因素试验基础上,采用响应面法研究木瓜蛋白酶用量、酶解时间、pH值、酶解温度和料液比对山核桃油提取率的影响;采用气相色谱-质谱联用技术分析提取油脂的脂肪酸组成,并对比了压榨法、溶剂法和水酶法3种方式对油脂理化性质的影响。结果表明,水酶法提取山核桃油脂工艺的最佳条件为:木瓜蛋白酶量0.17%,酶解时间150 min,pH值6.34,酶解温度54.43℃、料液比1∶5,在此条件下山核桃中油脂提取率为81.32%。采用气相色谱-质谱联用技术对提取油脂的脂肪酸组成进行分析,共测出12种脂肪酸,棕榈酸、油酸、亚油酸、亚麻酸是4种主要脂肪酸,其中饱和脂肪酸(SFA)占7.61%,单不饱和脂肪酸(MUFA)占69.10%,多不饱和脂肪酸(PUFA)占23.29%。对比3种提油方式发现,水酶法是一种较为理想的油脂提取方法。本研究结果为水酶法提取山核桃油脂生产提供了理论依据。     

Application of ethyl esters and d3-methyl esters as internal standards for the gas chromatographic quantification of transesterified fatty acid methyl esters in food

Ethyl esters (FAEE) and trideuterium-labeled methyl esters (d3-FAME) of fatty acids were prepared and investigated regarding their suitability as internal standards (IS) for the determination of fatty acids as methyl esters (FAME). On CP-Sil 88, ethyl esters of odd-numbered fatty acids eluted approximately 0.5 min after the respective FAME, and only coelutions with minor FAME were observed. Depending on the problem, one or even many FAEE can be added as IS for the quantification of FAME by both GC-FID and GC-MS. By contrast, d3-FAME coeluted with FAME on the polar GC column, and the use of the former as IS requires application of GC-MS. In the SIM mode, m/z 77 and 90 are suggested for d3-methyl esters of saturated fatty acids, whereas m/z 88 and 101 are recommended for ethyl esters of saturated fatty acids. These m/z values give either no or very low response for FAME and can thus be used for the analysis of FAME in food by GC-MS in the SIM mode. Fatty acids in sunflower oil and mozzarella cheese were quantified using five saturated FAEE as IS. Gravimetric studies showed that the transesterification procedure could be carried out without of loss of fatty acids. GC-EI/MS full scan analysis was suitable for the quantitative determination of all unsaturated fatty acids in both food samples, whereas GC-EI/MS in the SIM mode was particularly valuable for quantifying minor fatty acids. The novel GC-EI/MS/SIM method using fatty acid ethyl esters as internal standards can be used to quantify individual fatty acids only, that is, without determination of all fatty acids (the common 100% method), although this is present. This was demonstrated by the exclusive quantification of selected fatty acids including methyl-branched fatty acids, erucic acid (18:1n-9trans), and polyunsaturated fatty acids in cod liver oil and goat's milk fat.     

Characterization of medium-chain triacylglycerol (MCT)-enriched seed oil from Cinnamomum camphora (Lauraceae) and its oxidative stability

Medium-chain triacylglycerol (MCT)-enriched oil was extracted by supercritical fluid extraction of carbon dioxide (SFE-CO(2)) from Cinnamomum camphora seeds. The SFE-CO(2) process was optimized using the Box-Behnken design (BBD). The maximum oil yield (42.82%) was obtained under the optimal SFE-CO(2) conditions: extraction pressure, 21.16 MPa; extraction temperature, 45.67 °C; and extraction time, 2.38 h. Subsequently, the physicochemical characteristics, fatty acid composition, triacylglycerol (TAG) composition, tocopherol content, and DSC profile as well as oxidative stabilities of C. camphora seed oil (CCSO) were studied. Results showed that CCSO contained two major medium-chain fatty acids, capric acid (53.27%) and lauric acid (39.93%). The predominant TAG species in CCSO was LaCC/CLaC (ECN 32, 79.29%). Meanwhile, it can be found that CCSO had much higher oxidative stabilities than coconut oil due to the higher content of tocopherols in CCSO (α-tocopherol, 8.67 ± 0.51 mg/100 g; γ-tocopherol, 22.6 ± 1.02 mg/100 g; δ-tocopherol, 8.38 ± 0.47 mg/100 g). Conclusively, CCSO with such a high level of MCTs and high oxidative stabilities could be potentially applied in special food for specific persons such as weak patients and overweight persons because oils enriched in MCTs can be rapidly absorbed into body to provide energy without fat accumulation.     

Evaluation of fatty acid extraction methods for Thraustochytrium sp. ONC-T18

Various extraction methods were assessed in their capacity to extract fatty acids from a dried biomass of Thraustochytrium sp. ONC-T18. Direct saponification using KOH in ethanol or in hexane:ethanol was one of the most efficient techniques to extract lipids (697 mg g(-1)). The highest amount of fatty acids (714 mg g(-1)) was extracted using a miniaturized Bligh and Dyer extraction technique. The use of ultrasonics to break down cell walls while extracting with solvents (methanol:chloroform) also offered high extraction yields of fatty acids (609 mg g(-1)). Moreover, when the transesterification mixture used for a direct transesterification method was doubled, the extraction of fatty acids increased approximately 77% (from 392 to 696 mg g(-1)). This work showed that Thraustochytrium sp. ONC-T18 has the ability to produce over 700 mg g(-1) of lipids, including more than 165 mg g(-1) of docosahexaenoic acid, which makes this microorganism a potential candidate for the commercial production of polyunsaturated fatty acids. Finally, other lipids, such as myristic, palmitic, palmitoleic, and oleic acids, were also produced and recovered in significant amounts (54, 196, 123, and 81 mg g(-1)), respectively.     

Effect of diet and dietary fatty acids on the transformation and incorporation of C18 fatty acids in double-muscled Belgian Blue young bulls

Three groups of double-muscled Belgian Blue young bulls were fed during different stages of production diets differing in the proportions of linolenic and linoleic acid by including linseed in the concentrate or giving grass silage as main linolenic acid suppliers. Samples of rumen and abomasal contents and of the longissimus thoracis, subcutaneous fat, and liver were taken to analyze the fatty acid pattern with emphasis on the individual trans (t) C18:1 fatty acids and cis-9,trans-11 conjugated linoleic acid (c9t11CLA). Trans C18:1 isomers represented up to 20 g/100 g of total fatty acids in rumen and abomasal contents, whereas the accumulation of c9t11CLA was limited. Total trans C18:1 content in subcutaneous fat and intramuscular fat of the longissimus thoracis comprised 8.4 and 5.2 g/100 g of total fatty acids, respectively, with t11C18:1 being the most abundant one. Compared to rumen contents, subcutaneous and intramuscular fat were enriched in c9t11CLA and contained fewer tC18:1 isomers, resulting in a higher c9t11CLA/t11C18:1 ratio (0.04, 0.22, and 0.22, respectively). This result suggests that the endogenous synthesis of c9t11CLA in adipose tissue by the Delta(9)-desaturase was more important than its ruminal production.     

Measurement of conjugated linoleic acid (CLA) in CLA-rich potato chips by ATR-FTIR spectroscopy

A conjugated linoleic acid (CLA)-rich soy oil has been produced by photoisomerization of soy oil linoleic acid. Nutritional studies have shown that CLA possesses health benefits in terms of reducing certain heart disease and diabetes risk factors. Potato chips are snacks that are readily produced in the CLA-rich soy oil containing CLA levels similar to those of the oil used for frying. The objective of this study was to develop an FTIR method to rapidly determine the CLA content of oil in potato chips. Photoirradiated soy oil samples with ～25% total CLA were mixed with control soy oil, and 100 soy oil samples with total CLA levels ranging from 0.89 to 24.4% were made. Potato chips were fried using each of these 300 g CLA rich soy oil mixtures at 175 °C for approximately 3 min. Duplicate GC-FID fatty acid analyses were conducted on oil extracted from each batch of potato chips. The chip samples were ground and then scanned using ATR-FTIR spectroscopy with the aid of a high-pressure clamp, and duplicate spectra of each sample were averaged to obtain an average spectrum. Calibration models were developed using PLS regression analysis. These correlated the CLA isomer concentrations of potato chips obtained by GC-FID fatty acid analysis with their corresponding FTIR spectral features. The calibration models were fully cross validated and tested using samples that were not used in the calibration sample set. Calibrations for total CLA, trans,trans CLA, trans-10,cis-12 CLA, trans-9,cis-11 CLA, cis-10,trans-12 CLA, and cis-9,trans-11 CLA had coefficients of determinations (R2v) between 0.91 and 0.96 and corresponding root-mean-square error of prediction (RMSEP) ranging from 0.005 to 1.44. The ATR-FTIR technique showed potential as a method for the determination of the CLA levels in unknown potato chip samples.     

Effects of heat and ultraviolet radiation on the oxidative stability of pine nut oil supplemented with carnosic acid

The effects of carnosic acid (CA) of different concentrations (0.05, 0.1, and 0.2 mg/g) and two common antioxidants (butylated hydroxytoluene and α-tocopherol) on oxidative stability in pine nut oil at different accelerated conditions (heating and ultraviolet radiation) were compared. The investigation focused on the increase in peroxide and conjugated diene values, as well as free fatty acid and thiobarbituric acid-reactive substances. The changes in trans fatty acid and aldehyde compound contents were investigated by Fourier transform infrared spectroscopy, while the changes in pinolenic acid content were monitored by gas chromatography-mass spectrometry. The results show that CA was more effective in restraining pine nut oil oxidation under heating, UV-A and UV-B radiation, in which a dose-response relationship was observed. The antioxidant activity of CA was stronger than that of α-tocopherol and butylated hydroxytoluene. Pine nut oil supplemented with 0.2 mg/g CA exhibited favorable antioxidant effects and is preferable for effectively avoiding oxidation.     

5种提取方法对甲鱼油品质的影响

为探究酶解法、淡碱水解法、溶剂法、超声辅助溶剂法及蒸煮法这5种提取方法对甲鱼油品质的作用效果,以甲鱼脂肪为原料,采用气相色谱仪(GC)和顶空固相微萃取-气质联用仪(HS-SPME-GC-MS)对甲鱼油中脂肪酸组成及挥发性成分变化进行测定分析,并结合感官评价及理化指标分析。结果表明,5种提取方法对甲鱼油的感官与理化特性、脂肪酸组成及挥发性成分存在显著性差异(P<0.05),其中,超声辅助溶剂法的甲鱼油提取率最高,达78.51%,该方法提取的甲鱼油酸价(0.97 mg KOH·g^-1)、过氧化值(2.16 m Eq·kg^-1)均优于其他方法,并含有丰富的油酸、EPA和DHA。此外,超声辅助溶剂法的甲鱼油壬醛、己醛、2-壬酮、2-十一酮及1-戊烯-3-醇主体特征风味化合物的相对含量均较低,鱼腥味较淡,色泽透亮。综上表明,超声辅助溶剂法提取的甲鱼油品质优于其他提取方法。本研究可为今后优化甲鱼油提取方法提供理论依据。     

Comparison of values derived from selected flours by fat acidity methods of the American Association of Cereal Chemists and the International Organization for Standardization

Fat acidity values were determined on 8 commercially available flours by the American Association of Cereal Chemists method (AACC-02-01A) and by the International Organization for Standardization method (ISO-7305). Comparisons of means obtained by these 2 methods indicated that the ISO method produced results that were 1.4-2.5 times greater (P less than 0.001) than those for the AACC method. These data are consistent with a previous report which indicated that acidic materials (phosphates and amino acids) other than fatty acids are extracted by 95% alcohol (ISO method), whereas fatty acids alone are extracted by petroleum ether (AACC method). Fat acidity values obtained by AACC methodology employing Soxhlet and continuous extraction were also compared. No difference in fat acidity means existed with respect to extraction technique except for an aged whole wheat flour sample which produced a higher value (P = 0.05) by continuous extraction.     

Dietary lipid source modulates in vivo fatty acid metabolism in the freshwater fish, Murray cod (Maccullochella peelii peelii)

The aim of the present investigation was to quantify the fate of C18 and long chain polyunsaturated dietary fatty acids in the freshwater fish, Murray cod, using the in vivo, whole-body fatty acid balance method. Juvenile Murray cod were fed one of five iso-nitrogenous, iso-energetic, semipurified experimental diets in which the dietary fish oil (FO) was replaced (0, 25, 50, 75, and 100%) with a blended vegetable oil (VO), specifically formulated to match the major fatty acid classes [saturated fatty acids, monounsaturated fatty acids, n-3 polyunsaturated fatty acids (PUFA), and n-6 PUFA] of cod liver oil (FO). However, the PUFA fraction of the VO was dominated by C18 fatty acids, while C20/22 fatty acids were prevalent in the FO PUFA fraction. Generally, there was a clear reflection of the dietary fatty acid composition across each of the five treatments in the carcass, fillet, and liver. Lipid metabolism was affected by the modification of the dietary lipid source. The desaturation and elongation of C18 PUFAs increased with vegetable oil substitution, supported by the occurrence of longer and higher desaturated homologous fatty acids. However, increased elongase and desaturase activity is unlikely to fulfill the gap observed in fatty acid composition resulting from decreased highly unsaturated fatty acids intake.     

Tailoring of Atlantic salmon (Salmo salar L.) flesh lipid composition and sensory quality by replacing fish oil with a vegetable oil blend

Atlantic salmon (Salmo salar L.) juveniles were fed either 100% fish oil (FO), 75% vegetable oil (VO), or 100% VO throughout their life cycle to harvest weight followed by a finishing diet period when all groups were fed 100% FO. The two experimental VO diets were tested at two different locations (Scotland and Norway) against the same control diet (100% FO). The VO blend was composed of rapeseed oil, palm oil, and linseed oil using capelin oil as a control for fatty acid class compositions. Flesh fatty acid profiles were measured regularly throughout the experiment, with the times of sampling determined by changes in pellet size/lipid content and fish life stage. Growth and mortality rates were not significantly affected by dietary fatty acid compositions throughout the life cycle, except during the seawater winter period in Norway when both growth and protein utilization were increased in salmon fed 100% VO compared to 100% FO. Flesh fatty acid composition was highly influenced by that of the diet, and after the finishing diet period the weekly intake recommendations of very long chain n-3 polyunsaturated fatty acid (VLCn-3 PUFA) for human health were 80 and 56% satisfied by a 200 g meal of 75% VO and 100% VO flesh, respectively. No effect on flesh astaxanthin levels was observed in relation to changing dietary oil sources. Sensory evaluation showed only minor differences between salmon flesh from the dietary groups, although prior to the finishing diet period, flesh from 100% VO had less rancid and marine characteristics and was preferred over flesh from the other dietary groups by a trained taste panel. After the finishing diet period, the levels of typical vegetable oil fatty acids in flesh were reduced, whereas those of VLCn-3 PUFA increased to levels comparable with a 100% FO fed salmon. No differences in any of the sensory characteristics were observed between dietary groups. By blending VOs to provide balanced levels of dietary fatty acids, up to 100% of the fish oil can be replaced by the VO blend without compromising growth or flesh quality. At the same time, 75% of the dietary fish oil can be replaced without compromising flesh VLCn-3 PUFA content, thereby providing a beneficial nutritional profile for human consumption.     

Turkish Tombul hazelnut (Corylus avellana L.). 2. Lipid characteristics and oxidative stability

The quality of crude hazelnut oil extracted from Tombul (Round) hazelnut, grown in the Giresun province of Turkey, was determined by measuring lipid classes, fatty acids, and fat soluble bioactives (tocopherols and phytosterols). Oxygen uptake, peroxide value, thiobarbituric acid reactive substances, and alpha-tocopherol levels of stripped and crude hazelnut oils in bulk and oil-in-water (o/w) emulsion systems were also evaluated as indices of lipid oxidation over a 21 day storage period at 60 degrees C in the dark. The total lipid content of Tombul hazelnut was 61.2%, of which 98.8% were nonpolar and 1.2% polar constituents. Triacylglycerols were the major nonpolar lipid class and contributed nearly 100% to the total amount. Phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol were the most abundant polar lipids, respectively. Sixteen fatty acids were identified, among which oleic acid contributed 82.7% to the total, followed by linoleic, palmitic, and stearic acids. Unsaturated fatty acids accounted for 92.2% of the total fatty acids present. Among oil soluble bioactives, alpha-tocopherol (38.2 mg/100 g) and beta-sitosterol (105.5 mg/100 g) were predominant in hazelnut oil and comprised 88 and 93% of the total tocopherols and phytosterols present, respectively. The results also showed that both stripped and crude hazelnut oils were more stable in terms of lipid oxidation in the bulk oil as compared to those in an o/w emulsion.     

Dichloromethane as a solvent for lipid extraction and assessment of lipid classes and fatty acids from samples of different natures

The usefulness of the solvent mixture dichloromethane/methanol for lipid extraction and the determination of lipid classes and fatty acids in samples of different natures was conducted. Two different extraction methods were compared, one containing chloroform/methanol, another containing dichloromethane/methanol. Total lipid extraction showed some minor differences but no variation in the lipid classes. Regarding the fatty acid profile, in Echium virescens seeds, 17 major fatty acids could be identified and quantified, and all were equally extracted when either solvent system was employed. In Echium acanthocarpum hairy roots, 17 major fatty acids were quantified, showing some statistical differences for one cell line in favor of chloroform. The data obtained from the liquid nutrient medium were also comparable. The cod roe sample showed 31 major fatty acids, showing no statistical differences between the two solvent systems. Contrarily, the CH 2Cl 2 method was able to extract 31 main fatty acids found in European seabass dorsal muscle more efficiently than the CHCl 3 method. The results indicate that, for lipid extraction and fatty acid assessment, dichloromethane/methanol can readily replace the commonly employed chloroform/methanol, thus avoiding the major health, security, and regulatory problems associated with the use of chloroform.     

Study on Different Emulsifiers to Retain Fatty Fraction During Extrusion of Fatty Flours

Doughs made from wheat and almond flours, water, and five types of emulsifiers commonly used in confectionary and bakery products (soy lecithin, sucrose esters, mono‐glycerides, mono‐ and di‐glycerides of fatty acids, and diacetyl tartaric acid esters of monoglycerides [DATEM]) were studied. To evaluate the additive ability to retain the fatty fraction during the extrusion process, electrical conductivity was measured and fat loss (%) that occurs during extrusion processing was determined. The electrical conductivity measurements showed that the lower and better concentration of soy lecithin and mixed mono‐ and di‐glycerides of fatty acids to obtain an oil‐water emulsion was 0.2 g/100 g of dough, while for sucrose esters and mono‐glycerides of fatty acids, it was 0.7 g/100 g of dough. No efficacy for DATEM was observed. The fat loss results showed that sucrose esters were the most suitable emulsifiers for retaining the fatty fraction during extrusion processing, even at a very low amounts (2 g/kg of dough).     

放牧与围栏内蒙古针茅草原土壤微生物生物量碳、氮变化及微生物群落结构PLFA分析

以内蒙古贝加尔针茅草原、大针茅草原和克氏针茅草原为研究对象,采用氯仿熏蒸法和磷脂脂肪酸（PLFA）分析方法研究了放牧与围栏条件下内蒙古针茅草原土壤微生物生物量和群落结构特征的变化情况。研究表明放牧与围栏草地土壤微生物生物量和群落结构差异显著。氯仿熏蒸法分析结果表明内蒙古针茅草原土壤微生物生物量碳的含量介于166.6-703.5mg·kg^-1之间,微生物生物量氮含量介于30.34-92.15mg·kg^-1之间,其中贝加尔针茅草原土壤微生物生物量碳、氮最高,大针茅草原次之,克氏针茅草原则最低。放牧条件下,贝加尔针茅草原、大针茅草原土壤微生物生物量碳、氮显著低于围栏草地,克氏针茅草原则无显著变化。PLFA分析结果显示,内蒙古针茅草原土壤微生物群落PLFAs种类、含量丰富,共检测出28种PLFA生物标记磷脂脂肪酸,并且以直链饱和脂肪酸和支链饱和脂肪酸为主,相对含量占总量的2/3左右,其中贝加尔针茅草原土壤微生物含量最丰富,其围栏样地土壤的PLFA含量达到27.3nmol·g-1,大针茅草原和克氏针茅草原依次降低。围栏条件下,各类型草原土壤细菌脂肪酸与总PLFA含量均显著高于放牧草地,真菌脂肪酸含量则因草原类型不同各有差异;放牧导致各类型草原革兰氏阳性细菌PLFAs/革兰氏阴性细菌PLFAs（GPPLFAs/GNPLFAs）比值显著降低,而除了克氏针茅草原,细菌PLFAs/真菌PLFAs比值则显著升高。PLFAs主成分分析表明,放牧和围栏处理对内蒙古针茅草原土壤微生物群落结构产生影响,且围栏处理的影响程度大于放牧处理。经相关分析表明,氯仿熏蒸法和PLFA分析方法之间有很好的一致性,且土壤微生物PLFAs与土壤有机质、全磷、硝态氮显著相关。