脂肪酸的色谱保留时间规律与质谱特征研究及其在食品分析中的应用

用石油醚提取食品中的脂肪,经甲酯化反应后,采用HP-88(100m×0.25mm,0.33μm)弹性石英毛细管柱分离脂肪酸甲酯的同系物及异构体,GC/MS法测定。研究了不同链长脂肪酸的同系物及异构体的气相色谱出峰顺序,得到其保留时间规律;研究了不同脂肪酸的质谱断裂规律,选择3个特征离子来鉴定脂肪酸成分。建立了3个特征离子确定脂肪酸碳数及双键数目,色谱保留时间规律确定脂肪酸顺反异构体及双键位置异构体的方法。本法无需标准品即可快速测定脂肪酸同系物及异构体的含量,适用于脂肪酸组成的研究;及油脂、食品中脂肪酸,特别是反式脂肪酸的测定。     

气相色谱-质谱联用法分析全氟酰氟

采用气相色谱-质谱联用技术(GC/MS)对电化学氟化法生产的全氟环己烷酰氟产品中主要产物全氟酰氟进行了检测。在60℃下,采用甲醇对全氟环己烷酰氟产品进行甲酯化处理。考察了不同长度,极性及膜厚的毛细管色谱柱的分离效果。以KB-1MS毛细管色谱柱(90 m×0.25 mm×1.0μm)为分离柱,采用GC/MS法对全氟酰氟组成进行了定性与定量分析;结合有机质谱学裂解规律,分别对环状全氟羧酸甲酯、饱和直链全氟羧酸甲酯和单不饱和脂肪酸甲酯的裂解方式和质谱特征进行了分析归纳。通过质谱数据库检索、标准品对照及已知全氟化合物的质谱信息分析,共鉴定出5种全氟酰氟,其中包括两种异构体;测得全氟环己烷酰氟约占总全氟酰氟含量的65%。     

气相色谱/质谱法分析孔石莼中的脂肪酸

建立了孔石莼脂肪酸的气相色谱/质谱(GC/MS)测定方法。使用Folch法提取了孔石莼中的总脂,经过2 mol/L HCl-甲醇溶液的甲酯化处理后,采用GC/MS法对其脂肪酸组成进行了分离分析,同时结合有机质谱学规律,分别对饱和脂肪酸甲酯、单不饱和脂肪酸甲酯和多不饱和脂肪酸甲酯的裂解规律和质谱特征进行了分析归纳。通过质谱数据库检索和标准品对照,鉴定出孔石莼中的24种脂肪酸,其中9,12,15-十八碳三烯酸、4,7,10,13-十六碳四烯酸和6,9,2,15-十八碳四烯酸3种主要多不饱和脂肪酸占总脂肪酸含量的45.14%。通过对孔石莼中脂肪酸的分析,表明特征离子在脂肪酸甲酯尤其是多不饱和脂肪酸甲酯的定性方面具有很好的应用价值。     

超声提取-毛细管气相色谱法全分析倒刺鲃组织中的脂肪酸

采用Folch液超声提取倒刺鲃组织样品中的脂肪,脂质提取液经氢氧化钾-三氟化硼甲酯化后进行气相色谱分析。采用100∶1分流比进样,220～240 ℃程序升温操作,BPX70石英毛细管柱(60 m×0.25 mm)分离,火焰离子化检测器检测。利用标准品对照并结合线性程序升温等当碳链长(ECLT)值对照的方法对脂肪酸组分包括存在双键异位的不饱和脂肪酸进行定性。测定了野生和箱养倒刺鲃肌肉与脂肪中17种脂肪酸的含量。结果表明箱养倒刺鲃的脂肪酸营养结构与野生倒刺鲃无明显区别,不饱和脂肪酸占大多数,单不饱和脂肪酸含量高于多不饱和脂肪酸,n-6族不饱和脂肪酸含量明显高于n-3族。     

全二维气相色谱-四极杆质谱法检测植物油脂中脂肪酸

建立了植物油脂中31种脂肪酸成分的全二维气相色谱-四极杆质谱(GC×GC-qMS)分析方法。样品经甲酯化衍生后,以DB-1柱(30 m×0.25 mm×0.25 μm)作为一维柱、DB-Wax柱(3.2 m×0.1 mm×0.1 μm)作为二维柱组成柱系统进行分离,在调制周期为3.5 s、四极杆质量扫描范围为m/z 40～350的条件下,植物油脂中31种脂肪酸成分可以在50 min内得到准确和灵敏的检测。将本方法应用于实际样品的分析,灵敏度较传统的气相色谱-质谱法提高了100倍以上,一些植物油中微量的脂肪酸成分也因此被检出。该研究不仅为植物油中脂肪酸成分的分析提供了新的技术手段,同时对于确保食用植物油的质量安全、消除食用植物油的掺假伪劣等均有重要意义。     

GC-MS双内标法测定植物油中脂肪酸组成

建立了植物油中脂肪酸组成及含量的气相色谱-质谱联用双内标定量的检测方法。样品经甲脂化衍生后,通过HP-88毛细管柱(100 m×0.25 mm×0.20μm)分离,在选择离子监测(SIM)模式的条件下,植物油中46种脂肪酸成分在39 min以内得到了较好的分离。利用保留时间和特征离子定性,根据定量离子的峰面积,采用内标法定量。该方法简便、快速、准确、可靠。同时对亚麻籽油中棕榈酸、硬脂酸、油酸、亚油酸和α-亚麻酸进行6次测量,结果的回收率为99.1%~101.6%,变异系数均小于4.00%。将本方法应用于市售样品分析,结果证明这些食用植物油完全符合相关的国家标准。     

超临界CO_2萃取-气相色谱质谱法测定斑点叉尾鮰肌肉中的脂肪酸

采用超临界CO2萃取工艺、气相色谱-电子电离质谱(GC-EI-MS)法,建立了斑点叉尾鮰肌肉中脂肪酸的定性与定量分析方法。在CO2流速为30 L/h、温度45℃、压力25 MPa、时间100 min条件下进行超临界CO2萃取,所得油脂皂化10 min,采用三氟化硼法进行甲酯化衍生20 min,经HP-Innowax毛细管柱分离,电子电离质谱检测,以全扫描模式定性分析,选择离子扫描模式定量分析,饱和脂肪酸、单不饱和脂肪酸、双不饱和脂肪酸、多不饱和脂肪酸的定量离子分别为m/z74,55,67和79。14种脂肪酸甲酯的检出限和定量限分别在2.2～20μg/L和7.4～60μg/L之间;平均回收率为90.0%～111.2%,相对标准偏差为2.0%～5.9%。本方法灵敏度高、重复性好,可用于测定斑点叉尾鮰肌肉样品的脂肪酸。     

气相色谱-质谱对三种硅藻脂肪酸组成的测试规律性分析

借助气相色谱-质谱(GC-MS)联用技术比较分析了角毛藻(Chaetoceros muelleri)、新月菱形藻(Nitzshia closterium)和三角褐指藻(Phaeodactylum tricornutum)等3种硅藻在保留时间和质谱裂解方面的异同性规律.结果表明,强极性柱中各脂肪酸的分离顺序为低碳原子数到高碳原子数,碳原子数相同的组分分离顺序按低不饱和度到高不饱和度,弱极性柱中色谱出峰顺序也为低碳原子数到高碳原子数,与强极性柱一致,但碳原子数相同的组分分离顺序则从低饱和度到高饱和度,与强极性柱恰好相反;不同脂肪酸的质谱裂解表明,饱和脂肪酸甲酯的基峰为m/z 74,单烯脂肪酸甲酯的基峰为m/z 55,双烯脂肪酸甲酯的基峰为m/z 67,而三烯以上脂肪酸甲酯的基峰则为m/z 79,本文认为基峰这一特点可用来确定不饱和度,以助于快速准确的对脂肪酸甲酯进行定性分析;本文还比较了TMSH和NaOH-CH3OH两种甲酯化方法的区别.在此基础上用面积归一化法测定了硅藻脂肪酸的含量,发现3种硅藻都含有饱和脂肪酸、单不饱和脂肪酸和多不饱和脂肪酸,其中ω-3 PUFAs主要为EPA.     

气相色谱同位素比质谱测定不同产地鱼油中5种不饱和脂肪酸碳同位素比值

以IAEA-600咖啡因(δ13C-27.771‰)作为溯源标准,建立了气相色谱同位素质谱技术测定鱼油中功能因子亚油酸(LIA)、亚麻酸(LNA)、花生四烯酸(ARA)、二十碳五烯酸(EPA)、二十二碳六烯酸(DHA)等5种不饱和脂肪酸碳稳定同位素比值δ13C的分析方法。鱼样品先经HCl水解,乙醚液液萃取脂肪,提取的脂肪在2 mol/L KOH-甲醇溶液中反应生成脂肪酸甲酯,采用强极性毛细管气相色谱柱(Sil-88 100 m×0.25 mm×0.2μm)分离,稳定同位素比质谱测定。方法经日内、日间和人员比对验证,表明测定结果稳定,标准偏差小于0.82%。收集了不同产地的241个淡水鱼和深海鱼,对于提取的天然鱼油进行5种不饱和脂肪酸同位素比值分析,测得天然鱼油中5种不饱和脂肪酸同位素比值(δ13C)在-32.87‰~27.07‰之间,经计算,鱼油中不饱和脂肪酸δ13C值与硬脂酸δ13C值之比在-5.79~1.88之间,同时测得了相同的产地、品种鱼油中不饱和脂肪酸δ13C的分布范围,构建了天然鱼油的同位素指纹特征数据库,用于鉴定鱼油真伪。将不同浓度玉米油添加至天然金枪鱼油中进行测定,证明δ13C值的变化与掺入C4植物油量呈良好的线性关系,方法可根据δ13C值的变化鉴别掺假的鱼油。     

气相色谱-质谱法测定化妆品中的三氯叔丁醇

建立了测定化妆品中三氯叔丁醇的气相色谱-质谱分析方法.膏霜、水剂、散粉、香波、唇膏等不同类型的化妆品样品加入50％甲醇-无水乙醇或无水乙醇超声提取后,样品提取液高速离心处理,取上清液经无水硫酸钠脱水,进行气相色谱-质谱定性及定量分析.选用DB-1701(30 m×0.25 mm×0.25μm)石英毛细管柱,程序升温,流...     

Analysis of Polyunsaturated Fatty Acids Using High Performance Liquid Chromatography – Atmospheric Pressure Chemical Ionization Mass Spectrometry

Analysis of polyunsaturated fatty acids using high performance liquid chromatography–atmospheric pressure chemical ionization mass spectrometry is described. The standard fatty acid methyl esters from 16 to 22 carbons were analyzed by LC‐MS with APCI. The effect of orifice voltage and total carbon atoms versus number of double bonds in each homologue on the mass spectra is discussed. The correction coefficients for homologues from saturated fatty acids to hexaenoic acid are also mentioned.     

Gas chromatography-mass spectrometry coupled with pseudo-Sadtler retention indices, for the identification of components in the essential oil of Curcuma longa L.

Gas chromatography-mass spectrometry was applied to the cyclohexane extract of Curcuma longa L. The chromatographic conditions generated retention indices very close i.e., greater than 99.9%, to those reported for structures in the Sadtler Standard Gas Chromatography Retention Index Library. In addition to the extensively reported sesquiterpene ketones, this essential oil extract contained a series of saturated and unsaturated fatty acids. Wiley mass spectra library matching for the free fatty acids, their trimethylsilyl esters and methyl esters narrowed their identity down to a few candidates. Combining this information with the retention indices of the fatty acid methyl esters in the Sadtler library allowed the identification of some of the double bond positions.     

Prediction of equivalent chain lengths from two-dimensional fatty acid retention indices

A recently introduced two-dimensional fatty acid retention index system (2D-FARI) was used as basis for prediction of equivalent chain lengths (ECL) of fatty acid methyl esters (FAME) on a BPX-70 stationary phase. Models for the relationship between 2D-FARI data and ECL values of a calibration sample with 30 common fatty acids were established by a simple multivariate regression. The models were thereafter applied on 2D-FARI data for other FAMEs and used to predict the ECLs for these compounds. The 2D-FARI values for the fatty acids in the calibration sample are given by definition. Thus, the only information necessary to calculate the ECL value for a compound run under identical conditions as the calibration sample is the 2D-FARI values for the compound, which can be acquired from literature data. The method was validated with test sets analysed with different temperature and flow programs. ECLs of various marine FAME and trans isomers of Eicosapentaenoic and Docosahexaenoic acid were predicted with root mean squared error of prediction from 0.002 to 0.012 ECL units.     

Proton affinities of saturated aliphatic methyl esters

The kinetic method was used to determine the proton affinities of methyl esters of several saturated fatty acids. Decompositions of the proton-bound dimers of the methyl esters, AHB⁺, were observed under different conditions with two instruments. The proton affinities (PAs) of the methyl esters increase continually with increasing carbon number in the acid. Equilibrium and initial rate experiments were performed with a Fourier transform ion cyclotron resonance mass spectrometer on the methyl ester of the C₂₂ saturated acid (methyl behenate). These experiments give values for PA (methyl behenate) that are perhaps slightly lower than those obtained with the kinetic method. The PAs of the methyl esters of the fatty acids could be correlated with the equation: PA (ester) = (40.0 ± 2.5)*log(n) + (784.7 ± 3.9) kJ/mol or PA (ester) = (864 ± 2) − (479 ± 41)/n, wheren = number of atoms in the molecule. Proton affinities of smaller sets of 1-alkylamines and 1-alkanols can be fit to similar equations.     

Capillary electrochromatographic enantioseparations using a packed capillary with a 3 microm OD-type chiral packing

A technique for separating methyl esters of monounsaturated fatty acids by argentation chromatography using silver nitrate-impregnated TLC plates is described. Monounsaturated fatty acid methyl esters are separated from polyunsaturated and saturated fatty acid methyl esters and the monounsaturated fatty methyl esters are resolved according to chain length. cis isomers are well resolved from the corresponding trans isomers. R(F) values for individual monounsaturated fatty acids are very reproducible. The potential of the technique in metabolic studies is demonstrated in the chain elongation of [14C]-18:1(n-9) and delta-9 desaturation of [14C]-18:0 by human skin fibroblasts. Recoveries of individual [14C]-fatty acids for scintillation counting exceed 94%.     

Equivalent chain lengths of all C4-C23 saturated monomethyl branched fatty acid methyl esters on methylsilicone OV-1 stationary phase

Isomer mixtures of monomethyl branched saturated C7-C23 fatty acid methyl esters (FAME) were prepared by performing a methylene insertion reaction to the straight chain FAME and this study model was completed by using commercially available standards of C4-C7 FAME. The equivalent chain lengths (ECL) of all 220 C4-C23 monomethyl branched FAME on OV-1 stationary phase were measured, achieving an average repeatability of ±0.0004 ECL units. The monomethyl branched FAME was identified by GC on the basis of regularity of the fractional chain lengths (FCL) dependence on the number of carbon atoms (C(z)) of individual homologous series of methyl 2-, 3-, …, 21-FAME. The prediction of retention of the first homologues, having the new position of methyl group beginning at higher carbon atoms number, and analogously for the second, third, fourth, and other members of the homologous series, allowed the dependence FCL=f(C(z)) for the first and subsequent members of beginning homologous of monomethyl derivatives of FAME. The identification was confirmed by mass spectrometry. All of the methyl isomers of FAME, which could not be completely separated by gas chromatography due to having a methyl group in surroundings of the middle of the carbon chain, were resolved by mass spectrometry using deconvolution in a SIM-mode. Measured gas chromatographic and mass spectrometric data were applied for identification of the monomethyl branched saturated FAME in tongue coating.     

Analytical Characterization of Fatty Acids Composition of Datura alba Seed Oil by Gas Chromatography Mass Spectrometry

Methyl ester derivatives of fatty acids were analyzed for the determination of the constituents of Datura alba seed oil. Gas chromatography coupled to mass spectrometer was used for these analyses. Results delivered that there were saturated as well as unsaturated fatty acids in Datura alba seed oil. Total of 15 different fatty acid components were identified and quantified. Methyl linoleate was found in highest concentration (16.22%) among the identified analytes of interest. In addition methyl esters of Palmitic acid (6.59%), Oleic acid (5.41%) and Stearic acid (1.35%) were found. Concentrations of rest of the detected fatty acids were less than 1%. From the literature it appears that no such work has been performed for the determination of fatty acids in Datura alba seed oil.     

Identification of fatty acids in gas chromatography by application of different temperature and pressure programs on a single capillary column

A method for the identification of fatty acid methyl esters (FAME) based on the analysis of shifts in equivalent chain lengths (ECL) is described. The method is based on two-dimensional retention data achieved on one capillary column. Various temperature and pressure programs are applied on the same cyanopropyl column and the shifts in the ECL values are analysed by multivariate methods. The chain length, number of double bonds, and the double bond positions can be determined with high accuracy. The same procedure is suitable for determination of the number of trans and cis double bonds in trans fatty acids, and for detection of artefacts in fatty acid methyl ester chromatograms.     

南海Spirasterella中脂肪酸酯组分的鉴定

用色谱—质谱联用技术分析了中国南海海绵Spirasterella aurivilli Lindgren中脂肪酸酯组分。鉴定了29种脂肪酸酯,其中20种饱和脂肪酸乙酯,8种不饱和脂肪酸乙酯及1种饱和脂肪酸甲酯。并发现该海绵中含有自然界少见的10种奇数碳脂肪酸乙酯。