亚麻酸异构体的气相色谱-质谱分析

本文采用气相色谱-质谱法对亚麻酸异构体的出峰顺序和质谱特征进行了分析研究。结果表明:脂肪酸在强极性毛细管柱BPX-70(120 m×0.25 mm×0.25μm)上的出峰顺序具有一定的规律性,所测亚麻酸异构体中,出峰顺序依次为C18:3 9t,12t,15t、C18:39c,12t,15t、C18:3 9t,12c,15t、C18:3 9t,12t,15c、C18:3 9c,12c,15t、C18:3 9c,12t,15c、C18:3 9t,12c,15c和C18:3 9c,12c,15c。根据断裂规律和质谱特征分析,α-亚麻酸甲酯和γ-亚麻酸甲酯的特征离子分别为m/z 79、108(ω离子)、236(α离子)、261、292和m/z 79、150(ω离子)、194(α离子)、261、292。在8种α-亚麻酸顺反异构体中,当12位双键为顺式时,基峰离子为m/z 79,并具有明显的ω离子(m/z 108)和α离子(m/z 236);而当12位双键为反式时,α离子消失,ω离子强度减弱,并表现出离子强度m/z107108,且同时当15位双键为反式时,基峰离子为m/z 95;而当15位双键为顺式时,基峰离子为m/z 67。双键位置和顺反构象对亚麻酸的质谱特征产生显著影响,且12位顺反异构的影响大于9位和15位的顺反异构。     

酸碱对大豆油脂中反式脂肪酸的影响

以常规加热为对照组,通过气相色谱法分析H3PO4处理或NaOH处理对非共轭亚油酸(C18∶2)的热诱导顺/反异构化的分子机制。由加热试验可知,C18∶2-9c,12t和C18∶2-9t,12c为加热大豆油脂中的主要反式异构体。随着加热温度逐渐升高、加热时间的逐渐增强,反式亚油酸含量随之增加。3种油样加热后产生反式脂肪酸的量的排序为:NaOH处理后油样>H3PO4处理后油样>未处理油样。在加热过程中,生成单反式亚油酸的量高于生成双反式亚油酸的量。在温度260℃加热8 h时,经NaOH处理的加热油脂产生的单反式亚油酸(C18∶2-9c,12t)最多,达1.222%。通过Gauss软件对油脂中的双键进行分析,可知常规加热形成C18∶2-9t,12t需要跨越能量分别为235.986,223.846 kJ/mol两个能垒,而当油脂进行H3PO4处理或NaOH处理后产生的反式亚油酸的能垒降低,表明H3PO4处理或NaOH处理均会增加加热油脂反式异构体的几率,其中NaOH处理的影响最大。     

亚麻籽油煎炸薯条过程中不同构型反式α-亚麻酸的含量及分布

为研究不同煎炸温度下不同构型反式α-亚麻酸(TALA)在煎炸薯条和煎炸油中的分布情况,在170、200℃下进行亚麻籽油煎炸薯条实验,比较不同温度的煎炸过程中不同构型的TALA在煎炸油中的含量变化及TALA在煎炸薯条和煎炸油中含量的比值变化情况。结果表明：两个煎炸温度下煎炸油中均检测到6种构型的TALA,即9t,12c,15c-C18∶3、9c,12t,15c-C18∶3、9c,12c,15t-C18∶3、9c,12t,15t-C18∶3、9t,12t,15c-C18∶3和9t,12c,15t-C18∶3,而三反式构型的9t,12t,15t-C18∶3在整个煎炸过程中并未形成,6种构型的TALA均在200℃煎炸亚麻籽油中含量更多;煎炸亚麻籽油中TALA以单反式构型为主,约占TALA总含量的94%;3种单反式α-亚麻酸含量均随煎炸时间的延长和煎炸温度的升高呈显著增加趋势,3种双反式α-亚麻酸的含量从大到小依次为9t,12c,15t-C18∶3、9c,12t,15t-C18∶3、9t,12t,15c-C18∶3,且煎炸温度和煎炸时间对9t,12c,15t-C18∶3含量影响较大,对另外两种的...     

毛细管气相色谱法测定精炼和氢化大豆油中的反式脂肪酸

采用CP—Sil88强极性毛细管柱气相色谱法对精炼和氢化大豆油中的顺、反异构和位置异构的脂肪酸进行定性、定量分析。结果表明，精炼和氢化大豆油中主要顺、反式脂肪酸实现了很好地分离，各种位置异构的反式脂肪酸也实现了较好地分离。采用归一化法对各种脂肪酸进行定量分析，结果表明，精炼大豆油中反式脂肪酸为总脂肪酸含量的3．45％，以△9c，12t—C18：2、△9t，12c—C18：2和△9c，12c，15t—C18：3、△9t，12c，15c—C18：3 4种形式为主；氢化大豆油中反式脂肪酸为总脂肪酸含量的38．73％，以△9t—C18：1、△10t—C18：1、△11t—C18：1 3种形式为主的反式十八碳单烯酸（t—C18：1）占反式脂肪酸总量的90．81％。     

亚麻籽油煎炸薯条过程中不同构型反式α-亚麻酸的含量及分布北大核心CSCD

为研究不同煎炸温度下不同构型反式α-亚麻酸(TALA)在煎炸薯条和煎炸油中的分布情况,在170、200℃下进行亚麻籽油煎炸薯条实验,比较不同温度的煎炸过程中不同构型的TALA在煎炸油中的含量变化及TALA在煎炸薯条和煎炸油中含量的比值变化情况。结果表明:两个煎炸温度下煎炸油中均检测到6种构型的TALA,即9t,12c,15c-C18∶3、9c,12t,15c-C18∶3、9c,12c,15t-C18∶3、9c,12t,15t-C18∶3、9t,12t,15c-C18∶3和9t,12c,15t-C18∶3,而三反式构型的9t,12t,15t-C18∶3在整个煎炸过程中并未形成,6种构型的TALA均在200℃煎炸亚麻籽油中含量更多;煎炸亚麻籽油中TALA以单反式构型为主,约占TALA总含量的94%;3种单反式α-亚麻酸含量均随煎炸时间的延长和煎炸温度的升高呈显著增加趋势,3种双反式α-亚麻酸的含量从大到小依次为9t,12c,15t-C18∶3、9c,12t,15t-C18∶3、9t,12t,15c-C18∶3,且煎炸温度和煎炸时间对9t,12c,15t-C18∶3含量影响较大,对另外两种的影响较小;煎炸油中检测到的6种TALA在200℃煎炸薯条中均有检出,而在170℃煎炸薯条中仅检出了5种,未检出9t,12t,15c-C18∶3;薯条中各构型TALA的含量通常低于煎炸油的,170℃煎炸时,大部分TALA在薯条和煎炸油中的含量比值随煎炸时间延长逐渐变小,而200℃煎炸时,大部分TALA含量比值在整个煎炸过程中差异不显著。     

载气类型对气相色谱法分离顺-反脂肪酸异构体的影响

采用气相色谱-火焰离子化检测器,CP-Sil88毛细管柱为分析柱,考察在不同载气（氦气和氮气）和不同加热温度（等温180 ℃和梯度升温）条件下对不同食品基质中C18:1、C18:2和C18:3顺-反脂肪酸异构体分离度和定量的影响。结果显示,以氦气为载气,在起酥油、黄油、饼干和蛋糕4 种食品基质中,C18:1 13t,14t(6-8c)和C18:1 9c的分离度大于1.0,可清晰分辨并准确定量不同浓度C18:1的4t～13t,14t异构体,不会低估C18:1反式脂肪酸异构体总量。在植物油中,载气类型对C18:2顺反异构体的分离度和含量无影响;相比氮气,氦气在C18:3顺反异构体分离上具有一定优越性,但总量无显著差异。氦气作为同时测定食品中的反式脂肪酸、饱和脂肪酸、单不饱和脂肪酸和多不饱和脂肪酸的载气,定量准确且样品适用性广。     

银离子固相萃取法测定反刍动物反式脂肪酸异构体含量

利用氨丙基硅胶柱从反刍动物脂质中分离甘油三酯和磷脂,以氢化大豆油作为对照。采用银离子固相萃取柱分离甘油三酯和磷脂中反式脂肪酸异构体,气相色谱仪测定反式脂肪酸含量。结果显示:在3种反刍动物脂质中检测出trans-16:1,trans-18:1和trans18:2 3类反式脂肪酸,其中总脂质和甘油三酯总反式脂肪酸含量最高的是羊肉脂质,分别占总脂肪酸的7.64%和7.38%;磷脂中总反式脂肪酸最高的是牛肉脂质,占总脂肪酸的5.89%。银离子固相萃取柱能够从反刍动物脂质中分离出9种trans18:1异构体,其中含量最高的是11t18:1,分别为总脂质、甘油三酯和磷脂trans18:1的30.33%~39.03%,38.08%~45.09%和27.10%~44.47%。通过PLS-DA分析3种反刍动物反式脂肪酸异构体之间的差异,羊肉脂质中甘油三酯中的9t18:1和磷脂中的11t18:1含量均显著高于牛奶脂质和牛肉脂质(P<0.05);牛肉脂质中磷脂中的9t18:1和15t18:1含量均显著高于牛奶脂质和羊肉脂质(P<0.05);牛奶脂质磷脂中的12t18:1和16t18:1的含量均显著高于牛肉脂质和羊肉脂质(P<0.05)。反刍动物磷脂中的反式脂肪酸异构体有较大差异。     

电化学氢化制备低反式脂肪酸大豆油脂的分析

采用Agilent112-88A7 HP-88毛细管色谱柱的气相色谱和Avatar 370傅里叶转换红外光谱相结合的方法,对质子转移膜式电化学氢化制备的低反式脂肪酸的大豆油脂进行分析,其中气相色谱法中各脂肪酸的检出限为3.2~11.4μg/mL,相对标准偏差为0.30%~1.45%,试验表明两种方法对反式脂肪酸的定量结果一致;经过6 h的电化学氢化反应,当氢化大豆油脂的碘价由原料油的130.04 g I2/100 g下降到88.86g I2/100 g时,亚麻酸9c,12c,15cC18∶3质量分数由6.84%下降到0.28%,硬脂酸C18∶0质量分数由4.36%上升到10.44%,产品中反式脂肪酸质量分数为8.62%,主要是7.15%的9tC18∶1,0.74%的9t,12tC18∶2和0.36%的9t,12t,15tC18∶3。     

人造奶油中反式脂肪酸含量分析

采用氢氧化钠-甲醇对样品中的脂肪酸和反式脂肪酸进行甲酯化处理,气相色谱仪测定,以十一碳酸甘油三酯为内标进行定量。分析测定了国内5家知名焙烤企业抽检的41种不同用途的人造奶油中反式脂肪酸含量,通过分析发现:不同用途的人造奶油脂肪酸含量和组成有所差异,而反式脂肪酸的组成基本一致:主要是C18:1 6t、C18:19t、C18:1 11t和C18:2 9c12t、C18:2 9t12c。反式脂肪酸含量范围为0.271%反式脂肪酸含量(占总脂肪酸)8.245%。其中反式脂肪酸含量(占总脂肪酸)1%的共有35种,占总抽检数的85.4%;含量3%的有两种,占总抽检数的4.88%。本次研究较客观地反映了目前国内主流市场上人造奶油中反式脂肪酸含量情况,为国家风险管理部门进一步加强政策引导和宣传力度,引导消费者培养和强化消费前阅读营养标签的习惯提供数据依据。     

中国居民膳食中原料食物的各种反式脂肪酸的调研

通过本实验室建立的Ag+-TLC/GC连用技术测定我国日常膳食组成的原料食物中反式脂肪酸(TFAs)含量。测定结果表明我国食物中普遍存在反式脂肪酸,9t C18:1、10t-C18:1、11t C18:1和9t12t C18:2是其存在的主要形式。反刍动物产品中存在较高的9t C16:1、t C18:1和9t12t C18:2等各种反式脂肪酸(仅羊肉中未检出9t12t C18:2),且均以t C18:1为主,其中奶粉、奶油、鲜牛奶和酸奶等奶制品中含量最高的反式脂肪酸异构体均为11t C18:1。羊肉中9t C18:1、10t C18:1和11t C18:1的含量比较均衡,其它反刍动物产品中12t C18:1、13/14t C18:1、15t C18:1、16t C18:1的总量达到总反式脂肪酸含量的(25.9±1.8)%。坚果类食品中,TFA主要为t C18:1和9t12t C18:2,且9t12t C18:2含量约为总TFAs的10%。其它原料食物,如水果、蔬菜、畜禽肉、鱼虾及腌菜中TFAs含量极低。     

Biohydrogenation, duodenal flow, and intestinal digestibility of trans fatty acids and conjugated linoleic acids in response to dietary forage:concentrate ratio and linseed oil in dairy cows

Duodenal flows of hydrogenation intermediates in response to changes in dietary forage:concentrate ratio (F:C) and linseed oil were evaluated using 4 lactating Holstein cows fed a low (65:35 forage to concentrate) or high (35:65) concentrate diet without (LC, HC) added oil or with linseed oil (LCO, HCO) at 3% of DM. A 4 x 4 Latin square design was implemented for 5 wk. Lower hydrogenation of 18:2n-6 and 18:3n-3 was observed with HC, but it increased with LCO or HCO. Duodenal flow of total conjugated linoleic acids (CLA) increased by 1.40 (LCO) to 3.01 (HCO) g/d with linseed oil. This response was associated with greater flows of cis9,trans11- (+0.21 to +0.55 g/d), trans11,cis13- (+0.33 to +0.36), trans11,trans13- (+1.01 to +1.15 g/d), and trans,trans-CLA (+0.12 to +0.72 g/d). Trans10,cis12-CLA flow averaged 0.08 g/d and was not affected by F:C or oil. trans11,cis15-18:2 flow increased by 8.5 (LCO) to 62 (HCO) g/d in response to linseed oil. Total trans-18:1 flow was 37 g/d in cows fed LC and increased to 81 g/d with HC. Feeding oil increased total trans-18:1 to the greatest extent with HCO. Flow of trans10-18:1 was lower with LC than with HC (1.46 vs. 20 g/d). Linseed oil increased trans11-18:1 flow by 40 (LCO) to 113 g/d (HCO). Feeding LCO and HCO also increased flows of trans6+7+8-, trans13+14-, trans15-, and trans16-18:1. Apparent intestinal digestibility of trans-18:1 isomers was largely unaffected by concentrate level and ranged between 67 and 95%. Linseed oil increased digestibility of nearly all isomers by 3 to 16 percentage units. Digestibility of cis9,trans11-CLA was greater in cows fed HC (55%) compared with cows fed LC (32%) and was not affected by linseed oil. Data suggest that high concentrate diets enhanced ruminal outflow of trans10-18:1. We provide initial in vivo evidence that supplemental 18:3n-3 is hydrogenated to trans11,cis15-18:2, trans11-18:1, trans13+14-18:1, trans15-18:1, trans6+7+8-18:1, and trans16-18:1 primarily.     

11种品牌玉米油脂肪酸及异构体的主成分分析

对市售的11种品牌玉米油开展了脂肪酸(包括脂肪酸及其异构体)组成与含量的调查研究。采用气相色谱技术,结合主成分分析法,分析了不同品牌玉米油中脂肪酸的差异性和相关性。结果表明,11种品牌玉米油中共鉴定出19种脂肪酸,包括7种饱和脂肪酸,5种不饱和脂肪酸,6种反式脂肪酸(TFAs)和1种共轭亚油酸(t,t-CLAs),平均含量分别为14.18×10~(-2) g/g,80.32×10~(-2) g/g,1.47×10~(-2) g/g和0.23×10~(-2) g/g;其中,亚油酸(C18:2-9c12c)、油酸(C18:1-9c)和棕榈酸(C16:0)依次是玉米油中含量最多的3种脂肪酸;不同品牌玉米油中脂肪酸含量存在显著性差异(p0.05)。主成分分析揭示了不同品牌玉米油在脂肪酸含量上的相似性、差异性及脂肪酸之间相关程度,TFAs和CLAs正相关程度高。本研究为评价市售玉米油脂肪酸品质特征提供了较为全面的数据支撑。     

The kinetics of thermally induced isomerisation of trilinolenin,an improved gas chromatography study

The isomerisation reaction kinetics of heated trilinolenin was examined to better understand the mechanisms and controlling steps of the reaction. Trilinolenin was placed in glass ampoules and sealed before heating to 200, 220, 230 and 240 °C for regular time intervals. The contents were analysed using a newly established gas chromatography (GC) method that had been successfully applied to the separation of trans linolenic acid (TLNA) isomers. The kinetic studies show that the consumption of LNA is a second‐order reaction while the formations of trans LNA (TLNA) isomers are zero‐order reactions. The rate of the isomerisation reactions increases steadily with temperature and time, and the isomerisation mechanisms involved in the formation of TLNA isomers during heating are verified. These results suggest that controlling the formation of single TLNA isomers may be a key way to reduce TLNA isomers during heating of edible oils.     

Conjugated linoleic acid in ewe milk fat

Ewe milk fat from five different herds was studied to determine the content of conjugated linoleic acid (CLA) isomers. Research was carried out by combining gas chromatography-mass spectrometry (GC-MS) of fatty acid methyl esters (FAME) and 4,4-dimethyloxazolyne derivatives (DMOX) with silver ion-high performance liquid chromatography (Ag+-HPLC). Reconstructed mass spectral profiles of CLA characteristic ions from DMOX were used to identify positional isomers and Ag+-HPLC to quantify them. Total CLA content varied from 0.57 to 0.97 g/100 g of total fatty acids. FAME and DMOX were separated into a complex mixture of minor isomers and major rumenic acid (9-cis 11-trans C18:2) by GC-MS using a 100-m polar capillary column. Rumenic acid would represent more than 75% of total CLA. 11-trans 13-trans, 11-13 cis/trans plus trans/cis and 7-9 cis/trans plus trans/cis were the main CLA isomers after rumenic acid. Minor amounts of 8-10 and 10-12 C18:2 isomers were also found. Although most of the isomers were present in each herd's milk, differences in content were observed for some CLA species.     

Effects of antioxidants on heat-induced trans fatty acid formation in triolein and trilinolein

To elucidate the relation between heat-induced cis/trans isomerisation and thermal oxidative degradation of double bonds in unsaturated lipids, we investigated the effects of several edible antioxidants on these two molecular structural changes of double bonds in triolein (cis-9, 18:1) and trilinolein (cis-9, cis-12, 18:2), which were heated at 180 °C. trans Isomerisation and oxidative degradation of each cis double bond in the triacylglycerols during heating were evaluated on the basis of the increase in the amount of trans isomers and decrease in the amount of cis isomers by gas chromatography analysis. The synchronous suppression in trans isomerisation and cis deterioration of double bonds in these triacylglycerols were found by addition of antioxidants, whose inhibitory effects were associated with their kinds and concentrations. When triolein was heated in the presence of antioxidant, suppression of heat-induced trans isomerisation was directly proportional to that of cis deterioration. The results of our analysis suggested that the appropriate addition of antioxidants to edible oils during processing and cooking would facilitate the control of not only thermal oxidative degradation but also heat-induced trans isomerisation of double bonds in unsaturated lipids.     

Conjugated alpha-linolenic acid isomers in bovine milk and muscle

Conjugated linolenic acids (CLnA) are octadecatrienoic fatty acid isomers with at least 2 conjugated double bonds. Various CLnA isomers occur naturally, and some isomers could be formed by ruminants from dietary α-linolenic acid. Ruminant biohydrogenation of polyunsaturated fatty acids gives rise to the formation of numerous metabolites having conjugated or nonconjugated structures. The objectives of this study were to identify and characterize CLnA isomers in milk fat and muscle lipid extracts from cattle fed a high-forage diet. The analysis of total fatty acid methyl esters revealed levels of total CLnA of 0.39% in a single milk lipid extract and 0.34% in a single muscle lipid extract. Fatty acid methyl esters were fractionated by argentation thin-layer chromatography. A fraction containing dienoic fatty acids as well as CLnA isomers was isolated and analyzed. The double bond positions of CLnA isomers (cis-9, trans-11, cis-15 and cis-9, trans-13, cis-15 18:3) were confirmed by mass spectrometry of their 4,4-dimethyloxazoline derivatives. Mass spectra of the cis-9, trans-13, cis-15 18:3 isomer was characterized by an intense ion at m/z 236 attributable to the formation of 2 stabilized allylic radical fragments, whereas this intense ion corresponding to the stabilized radical fragments was located at m/z 262 for the cis-9, trans-11, cis-15 18:3 isomer. The gap of 12 amu between m/z 250 and 262 confirmed the occurrence of a double bond in position Δ¹³. Configuration of the double bonds of standards having similar mass spectra and gas-liquid chromatographic retention times was confirmed by ¹H nuclear magnetic resonance. We also showed that both CLnA isomers were found in the muscle lipid extract, whereas only the cis-9, trans-11, cis-15 18:3 isomer was identified in the milk lipid extract. This study appears to be the first to identify 2 CLnA isomers in bovine muscle lipid extract.