乳脂肪甘油三酯测定与分析方法研究进展

doi:10.12160/j.issn.1672-5190.2020.02.013

摘要/Abstract

摘要： 脂类是极其复杂多样的生物分子,仅已报道的乳脂肪甘油三酯（triacylglycerol,TAG）就达400多种,其中,乳脂中含量高于1%的近60种。目前液相色谱—质谱联用技术（liquid chromatography-mass spectrometry,LC-MS）是分离和鉴定TAG分子的主要手段,分离技术有反相高效液相色谱（reversed-phase high-performance liquid chromatography,RP-HPLC）、银离子高效液相色谱（silver ion high-performance liquid chromatography,Ag⁺-HPLC）、二维高效液相色谱（two-dimensional high-performance liquid chromatography,2D-HPLC）和超临界流体色谱（supercritical fluid chromatography,SFC）,分子电离常用电喷雾电离（electrospray ionization,ESI）和大气压化学电离（atmospheric pressure chemical ionization,APCI）。质谱分析可以确定分子量和脂肪酸位置,但分子异构体鉴定仍面临挑战。我国针对牛、山羊、马、骆驼、牦牛和水牛等家畜的TAG缺乏系统研究。TAG谱的建立不仅可以评价乳脂肪的营养特征,指导饲养方式科学改进,也有助于建立物种、饲养方式、产地和加工方式的鉴定模型。对乳脂肪甘油三酯测定与分析方法的研究进展进行综述,以期为乳制品质量控制以及食品完整性和真实性鉴定提供理论依据。

关键词: 甘油三酯, 脂肪酸, 乳脂肪, 液相色谱, 质谱

Abstract: Lipids are extremely complex and diverse biomolecules, and at least more than 400 types of milk fat triacylglycerol （TAG） have been reported. The content of nearly 60 reported types of TAG exceed 1% in milk fat. At present, liquid chromatography-mass spectrometry （LC-MS） is the commonly used method to separate and identify TAG molecules, including reversed-phase high-performance liquid chromatography （RP-HPLC）, silver ion high-performance liquid chromatography （Ag⁺-HPLC）, two-dimensional high-performance liquid chromatography （2D-HPLC）, and supercritical fluid chromatography （SFC）. Moreover, the mainly used molecular ionization methods are electrospray ionization （ESI） and atmospheric pressure chemical ionization （APCI）. Mass spectrometry can determine molecular weight and fatty acid positions, but molecular isomer identification remains a challenge. Systematic researches on TAG for cattle, goats, horses, camels, yaks and buffalos are limited in China. The establishment of the TAG profile can not only evaluate the nutritional characteristics of milk fat and guide the scientific improvement of breeding methods, but also help to establish an identification model of species, breeding methods, origin and processing methods of milk. This article reviews the research progress on determination and analysis methods of milk fat triacylglycerol, in hoping to provide a theoretical basis for dairy product quality control as well as food integrity and authenticity identification.

Key words: triacylglycerol, fatty acids, milk fat, liquid chromatography, mass spectrometry

中图分类号:

TS252.7

刘宇婷, 郭军, 许昀晖, 吴艳. 乳脂肪甘油三酯测定与分析方法研究进展[J]. 畜牧与饲料科学, 2020, 41(2): 56-65.

LIU Yu-ting, GUO Jun, XU Yun-hui, WU Yan. Research Progress on the Determination and Analysis Methods of Milk Fat Triacylglycerol[J]. Animal Husbandry and Feed Science, 2020, 41(2): 56-65.

参考文献

[1] MANUEL,BUCHGRABER,FRANZ,et al.Triacylglycerol profiling by using chromatographic techniques[J].European Journal of Lipid Science and Technology,2004,106(9):621-648.
[2] INNIS S M .Dietary triacylglycerol structure and its role in infant nutrition[J].Advances in Nutrition,2011,2(3):275-283.
[3] JENSEN R G .The composition of bovine milk lipids:January 1995 to December 2000[J].Journal of Dairy Science,2002,85(2):295-350.
[4] IUPAC,IUB.IUPAC-IUB Commission on Biochemical Nomenclature (CBN)[J].European Journal of Biochemistry,1967,2(2):127-131.
[5] 高希西. 乳脂肪甘油三酯分析及黄油分馏物组成与物化特性研究[D].沈阳:沈阳农业大学,2016.
[6] GERMAN J B,DILLARD C J .Composition,structure and absorption of milk lipids:a source of energy,fat-soluble nutrients and bioactive molecules[J].Critical Reviews in Food Science and Nutrition,2006,46(1):57-92.
[7] DIMICK P S,REDDY S Y,ZIEGLER G R .Chemical and thermal characteristics of milk-fat fractions isolated by a melt crystallization[J].Journal of the American Oil Chemists′ Society,1996,73(12):1647-1652.
[8] NARINE S S,MARANGONI A G.Relating structure of fat crystal networks to mechanical properties:A review[J].Food Research International,1999,32(4):227-248.
[9] SMIDDY M A,HUPPERTZ T,RUTH S M V. Triacylglycerol and melting profiles of milk fat from several species[J].International Dairy Journal,2012,24(2):64-69.
[10] AGANGA A A,AMARTEIFIO J O,NKILE N.Effect of stage of lactation on nutrient composition of Tswana sheep and goat′s milk[J].Journal of Food Composition and Analysis,2002,15(5):533-543.
[11] 张鑫. 有机与非有机原料乳及UHT牛乳的脂肪酸特征研究[D].呼和浩特:内蒙古农业大学,2018.
[12] 杨月欣,葛可佑.中国营养科学全书(上册)[M].2版. 北京:人民卫生出版社,2019.
[13] DAMODARAN S,FENNEMA O R,PARKIN K L.食品化学[M].江波,杨瑞金,钟芳,等,译. 4版. 北京:中国轻工业出版社,2013.
[14] LIU Z,EZERNIEKS V,WANG J,et al.Heat stress in dairy cattle alters lipid composition of milk[J].Scientific Reports,2017,7(1):961.
[15] ZHOU Q,GAO B,ZHANG X,et al.Chemical profiling of triacylglycerols and diacylglycerols in cow milk fat by ultra-performance convergence chromatography combined with a quadrupole time-of-flight mass spectrometry[J].Food Chemistry,2014,143:199-204.
[16] GRESTI J,BUGAUT M,MANIONGUI C,et al.Composition of molecular species of triacylglycerols in bovine milk fat[J].Journal of Dairy Science,1993,76(7):1850-1869.
[17] MOTTRAM H R,EVERSHED R P.Elucidation of the composition of bovine milk fat triacylglycerols using high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry[J].Journal of Chromatography A,2001,926(2):239-253.
[18] BECCARIA M,SULLINI G,CACCIOLA F,et al.High performance characterization of triacylglycerols in milk and milk-related samples by liquid chromatography and mass spectrometry[J].Journal of Chromatography A,2014,1360:172-187.
[19] 涂安琪,杜振霞.超临界流体色谱—四极杆飞行时间质谱快速分析牛奶与羊奶中的甘油三酯组分[J].质谱学报,2017,38(2):217-226.
[20] FOLCH J.A simple method for the isolation and purification of total lipides from animal tissues[J].The Journal of Biological Chemistry,1957,226(1):497-509.
[21] BLIGH E G,DYER W J.A rapid method of total lipid extraction and purification[J].Canadian Journal of Biochemistry and Physiology,1959,37(8):911-917.
[22] BRUCKDORFER K R,EDWARDS P A,GREEN C .Properties of aqueous dispersions of phospholipid and cholesterol[J].FEBS Journal,1968,4(4):506-511.
[23] LIU Z,ROCHFORT S,COCKS B G.Optimization of a single phase method for lipid extraction from milk[J].Journal of Chromatography A,2016,1458:145-149.
[24] LI M,YANG L,BAI Y,et al.Analytical methods in lipidomics and their applications[J].Analytical Chemistry,2014,86(1):161-175.
[25] KÓFELER H C,FAULAND A,RECHBERGER G N,et al. Mass spectrometry based lipidomics:An overview of technological platforms[J].Metabolites,2012,2(4):19-38.
[26] LAAKSO P,MANNINEN P.Identification of milk fat triacylglycerols by capillary supercritical fluid chromatography-atmospheric pressure chemical lonization mass spectrometry[J].Lipids,1997,32(12):1285-1295.
[27] DANIELA G,CLAUDIO M,VALERIA G,et al.HPLC-APCI analysis of triacylglycerols in milk fat from different sources[J].European Journal of Lipid Science and Technology,2011,113(2):197-207.
[28] LÍSA M,CÍFKOVÁ E,HOLCAPEK M. Lipidomic profiling of biological tissues using off-line two-dimensional high-performance liquid chromatography-mass spectrometry[J].Journal of Chromatography A,2011,1218(31):5146-5156.
[29] LÍSA M,HOLCAPEK M. Characterization of triacylglycerol enantiomers using chiral HPLC/APCI-MS and synthesis of enantiomeric triacylglycerols[J]. Analytical Chemistry,2013,85(3):1852-1859.
[30] YAMADA T,UCHIKATA T,SAKAMOTO S,et al.Supercritical fluid chromatography/Orbitrap mass spectrometry based lipidomics platform coupled with automated lipid identification software for accurate lipid profiling[J].Journal of Chromatography A,2013,1301:237-242.
[31] VERMA A,AMBATIPUDI K.Challenges and opportunities of bovine milk analysis by mass spectrometry[J].Clinical Proteomics,2016,13(1):8.
[32] MU H,HØY C E. The digestion of dietary triacylglycerols[J].Progress in Lipid Research, 2004,43(2):105-133.
[33] BOHN T,CARRIERE F,DAY L,et al.Correlation between in vitro and in vivo data on food digestion. What can we predict with static in vitro digestion models?[J].Critical Reviews in Food Science and Nutrition,2017,58(13):2239-2261.
[34] MICHALSKI M C.Specific molecular and colloidal structures of milk fat affecting lipolysis,absorption and postprandial lipemia[J].European Journal of Lipid Science and Technology,2009,111(5):413-431.
[35] LIU Z Q,SIMONE R,BEN C.Milk lipidomics:What we know and what we don′t[J].Progress in Lipid Research,2018,71:70-85.
[36] MU H,PORSGAARD T.The metabolism of structured triacylglycerols[J].Progress in Lipid Research,2005,44(6):430-448.
[37] SANDRA P,DERMAUX A,FERRAZ V,et al.Analysis of triglycerides by capillary electrochromatography[J]. Journal of Microcolumn Separations,1997,9(5):409-419.
[38] SEGALL S D,ARTZ W E,RASLAN D S,et al.Analysis of triacylglycerol isomers in Malaysian cocoa butter using HPLC-mass spectrometry[J].Food Research International,2005,38(2):167-174.
[39] ANDRIKOPOULOS,NIKOLAOS K .Chromatographic and spectroscopic methods in the analysis of triacylglycerol species and regiospecific isomers of oils and fats[J].Critical Reviews in Food Science and Nutrition,2002,42(5):473-505.
[40] TEN-DOMÉNECH,BELTRÁN-ITURAT E,HERRERO-MARTÍNEZ J M,et al. Triacylglycerol analysis in human milk and other mammalian species:Small-scale sample preparation,characterization,and statistical classification using HPLC-ELSD profiles[J]. Journal of Agricultural and Food Chemistry,2015,63(24):5761-5770.
[41] MOTTRAM H R,EVERSHED R P.Elucidation of the composition of bovine milk fat triacylglycerols using high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry[J]. Journal of Chromatography A,2001,926(2):239-253.
[42] 韦伟,屠海云,王红青,等.Silver-ion HPLC测定1,3-二油酸-2-棕榈酸三酰甘油的含量[J].中国粮油学报,2014,29(1):105-109.
[43] MACDOUGALL K M,MCNICHOL J,MCGINN P J,et al.Triacylglycerol profiling of microalgae strains for biofuel feedstock by liquid chromatography-high-resolution mass spectrometry[J].Analytical and Bioanalytical Chemistry,2011,401(8):2609-2616.
[44] HSU F F,TURK J.Electrospray ionization multiple-stage linear ion-trap mass spectrometry for structural elucidation of triacylglycerols:assignment of fatty acyl groups on the glycerol backbone and location of double bonds[J].Journal of American Society for Mass Spectromtry,2010,21(4):657-669.
[45] KALLIO H,YLI-JOKIPIIK,KURVINEN J P,et al.Regioisomerism of triacylglycerols in lard,tallow,yolk,chicken skin,palm oil,palm olein, palm stearin, and a transesterified blend of palm stearin and coconut oil analyzed by tandem mass spectrometry[J].Journal of Agricultural and Food Chemistry,2001,49(7):3363-3369.
[46] NIKOLOVA-DAMYANOVA B.Retention of lipids in silver ion high-performance liquid chromatography:Facts and assumptions[J].Journal of Chromatography A,2009,1216(10):1815-1824.
[47] RUIZ-GUTIÉRREZ V,BARRON L J R. Methods for the analysis of triacylglycerols[J].Journal of Chromatography B:Biomedical Sciences and Applications,1995,671(1/2):133-168.
[48] NIKOLOVA-DAMYANOVA B,CHRISTIE W W,HERSLÓF B. Mechanistic aspects of fatty acid retention in silver ion chromatography[J].Journal of Chromatography A,1996,749(1/2):47-54.
[49] DUGO P,KUMM T,CHIOFALO B,et al.Separation of triacylglycerols in a complex lipidic matrix by using comprehensive two-dimensional liquid chromatography coupled with atmosheric pressure chemical ionization mass spectrometric detection[J].Journal of Separation Science,2015,29(8):1146-1154.
[50] 朱文星,杨锐,徐永威,等.超高效合相色谱串联四级杆飞行时间质谱定性分析中/长链结构甘油三酯组成[J].中国药学杂志,2016,51(15):1324-1329.
[51] LEE J W,UCHIKATA T,MATSUBARA A,et al.Application of supercritical fluid chromatography/mass spectrometry to lipid profiling of soybean[J].Journal of Bioscience and Bioengineering,2012,113(2):262-268.
[52] GAO B,LUO Y,LU W,et al.Triacylglycerol compositions of sunflower,corn and soybean oils examined with supercritical CO2 ultra-performance convergence chromatography combined with quadrupole time-of-flight mass spectrometry[J].Food Chemistry,2017,218:569-574.
[53] GRAND-GUILLAUME P A,VEUTHEY J L,GUILLARME D. Comparison of ultra-high performance supercritical fluid chromatography and ultra-high performance liquid chromatography for the analysis of pharmaceutical compounds[J].Journal of Chromatography A,2012,1266:158-167.
[54] LEE J W,NAGAI T,GOTOH N,et al.Profiling of regioisomeric triacylglycerols in edible oils by supercritical fluid chromatography/tandem mass spectrometry[J].Journal of Chromatography B,2014,966:193-199.
[55] GUIOCHON G,TARAFDER A.Fundamental challenges and opportunities for preparative supercritical fluid chromatography[J].Journal of Chromatography A,2011,1218(8):1037-1114.
[56] KALÍKOVÁ K,ŠLECHTOVÁ T,VOZKA J,et al. Supercritical fluid chromatography as a tool for enantioselective separation:A review[J].Analytica Chimica Acta,2014,821:1-33.
[57] 张星河,齐策,陶冠军,等.超高效合相色谱串联四级杆飞行时间质谱快速分析植物油的甘油三酯组成[J].中国油脂,2018,43(11):134-139.
[58] BIRD S S,MARUR V R,SNIATYNSKI M J,et al.Serum lipidomics profiling using LC-MS and high-energy collisional dissociation fragmentation:Focus on triglyceride detection and characterization[J].Analytical Chemistry,2011,83(17):6648-6657.
[59] FULAND A,HARALD K,MARTIN T,et al.A comprehensive method for lipid profiling by liquid chromatography-ion cyclotron resonance mass spectrometry[J].Journal of Lipid Research,2011,52(12):2314-2322.
[60] LI Q,ZHAO Y,ZHU D,et al.Lipidomics profiling of goat milk,soymilk and bovine milk by UPLC-Q-exactive orbitrap mass spectrometry[J].Food Chemistry,2017,224(1):302-309.
[61] ZHOU X R,CALLAHAN D L,SHRESTHA P,et al.Lipidomic analysis of Arabidopsis seed genetically engineered to contain DHA[J].Frontiers in Plant Science,2014,5:419.
[62] COSSIGNANI L,POLLINI L,BLASI F,et al.Authentication of milk by direct and indirect analysis of triacylglycerol molecular species[J].Journal of Dairy Science,2019,102(7):5871-5882.
[63] YAMADA T,UCHIKATA T,SAKAMOTO S,et al.Development of a lipid profiling system using reverse-phase liquid chromatography coupled to high-resolution mass spectrometry with rapid polarity switching and an automated lipid identification software[J].Journal of Chromatography A,2013,1292:211-218.
[64] STRAARUP E M,LAURITZEN L,FAERK J,et al.The stereospecific triacylglycerol structures and fatty acid profiles of human milk and infant formulas[J].Journal of Pediatric Gastroenterology and Nutrition,2006,42(3):293-299.
[65] 韩瑞丽. 牛乳中甘油三酯sn-2位脂肪酸组成的测定方法研究[D].青岛:青岛农业大学,2009.
[66] MARTIN J C,BOUGNOUX P,ANTOINE J M,et al.Triacylglycerol structure of human colostrum and mature milk[J].Lipids,1993,28(7):637-643.
[67] VYSSOTSKI M,BLOOR S J,LAGUTIN K,et al.Efficient separation and analysis of triacylglycerols:Quantitation of β palmitate(OPO)in oils and infant formulas[J].Journal of Agricultural and Food Chemistry,2015,63(26):5982-5992.
[68] 徐同成,王文亮,程安玮,等.甘油三酯中脂肪酸分布位置检测方法研究进展[J].粮油加工,2010(11):37-40.
[69] CVACKA J,KRAFKOVÁ E,JIROS P,et al.Computer-assisted interpretation of atmospheric pressure chemical ionization mass spectra of triacylglycerols[J].Rapid Communications in Mass Spectrometry,2006,20(23):3586-3594.
[70] MOTTRAM H R,WOODBURY S E,EVERSHED R P.Identification of triacylglycerol positional isomers present in vegetable oils by high performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry[J].Rapid Communications in Mass Spectrometry,1997,11(12):1240-1252.
[71] MOTTRAM H R,CROSSMAN Z M,EVERSHED R P.Regiospecific characterisation of the triacylglycerols in animal fats using high performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry[J].Analyst,2001,126(7):1018-1024.
[72] HOLCAPEK M,DVORÁKOVÁ H,LÍSA M,et al. Regioisomeric analysis of triacylglycerols using silver-ion liquid chromatography-atmospheric pressure chemical ionization mass spectrometry:Comparison of five different mass analyzers[J].Journal of Chromatography A,2010,1217(52):8186-8194.
[73] HOLCAPEK M,JNDERA P,ZDERADICKA P,et al.Characterization of triacylglycerol and diacylglycerol composition of plant oils using high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry[J].Journal of Chromatography A,2003,1010(2):195-215.
[74] PALMQUIST D L.Milk Fat:Origin of Fatty Acids and Influence of Nutritional Factors Thereon[M]//FOX P F,McSWEENEY P L H. 3rd ed. Advanced Dairy Chemistry Volume 2 Lipids. US:Springer,2007.
[75] PITAS R E,SAMPUGNA J,JENSEN R G .Triglyceride structure of cows′ milk fat. Ⅰ. Preliminary observations on the fatty acid composition of positions 1,2,and 3[J].Journal of Dairy Science,1967,50(8):1332-1336.