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Corresponding Author(s)

陈复生(1963—), 男, 河南工业大学教授, 博士。E-mail: fushengc@haut.edu.cn

Abstract

Lysinoalanine (LAL) is a cross-linked amino acid formed through the addition of lysine and dehydroalanine. High-protein foods are prone to forming LAL during food processing (such as alkaline treatment, heat treatment). LAL may induce nephrotoxicity, affect human digestive function, and can also reduce the content of essential amino acids in food, reducing the nutritional value of food. Currently, LAL quantification primarily relies on liquid chromatography (LC) and liquid chromatography-mass spectrometry (LC-MS) methods. This study reviews and discusses the detection methods of LAL from three aspects: pretreatment, derivatization, and instrumental detection, aiming to provide insights for developing high-sensitivity, more accurate, and stable LAL detection methods.

Publication Date

4-3-2026

First Page

66

Last Page

73

DOI

10.13652/j.spjx.1003.5788.2025.80236

References

[1] MCKERCHAR H J,CLERENS S,DOBSON R C J,et al.Protein-protein crosslinking in food:proteomic characterisation methods,consequences and applications [J].Trends in Food Science & Technology,2019,86:217-229.
[2] RENZONE G,ARENA S,SCALONI A.Cross-linking reactions in food proteins and proteomic approaches for their detection [J].Mass Spectrometry Reviews,2022,41(5):861-898.
[3] ROMBOUTS I,LAMBRECHT M A,CARPENTIER S C,et al.Identification of lanthionine and lysinoalanine in heat-treated wheat gliadin and bovine serum albumin using tandem mass spectrometry with higher-energy collisional dissociation [J].Amino Acids,2016,48(4):959-971.
[4] AKILLIOĞLU H G,CHATTERTON D E W,LUND M N.Maillard reaction products and amino acid cross-links in liquid infant formula:effects of UHT treatment and storage [J].Food Chemistry,2022,396:133687.
[5] 刘旭辉.减压法对皮蛋 LAL 产生的影响及蛋清凝胶机理的研究[D].天津:天津科技大学,2019:30-31.LIU X.Effect of vacuum decompression technology on LAL production of preserved eggs and research on mechanism of egg white gel [D].Tianjin:Tianjin University of Science and Technology,2019:30-31.
[6] HOU F R,DING W H,QU W J,et al.Alkali solution extraction of rice residue protein isolates:influence of alkali concentration on protein functional,structural properties and lysinoalanine formation [J].Food Chemistry,2017,218:207-215.
[7] HELLWIG M,MATTHES R,PETO A,et al.N-ε-fructosyllysine and N-ε-carboxymethyllysine,but not lysinoalanine,are available for absorption after simulated gastrointestinal digestion [J].Amino Acids,2014,46(2):289-299.
[8] BOSCHIN G,D'AGOSTINA A,RINALDI A,et al.Lysinoalanine content of formulas for enteral nutrition [J].Journal of Dairy Science,2003,86(7):2 283-2 287.
[9] FRIEDMAN M.Chemistry,biochemistry,nutrition,and microbiology of lysinoalanine,lanthionine,and histidinoalanine in food and other proteins [J].Journal of Agricultural and Food Chemistry,1999,47(4):1 295-1 319.
[10] FRIEDMAN M,LEVIN C E.Nutritional and medicinal aspects of D-amino acids [J].Amino Acids,2012,42(5):1 553-1 582.
[11] 罗序英.禽蛋碱加工过程中赖丙氨酸的形成与控制研究 [D].南昌:南昌大学,2014:19-25.LUO X.Study on lysinoalanine formation and control in alkali-pickled eggs [D].Nanchang:Nanchang University,2014:19-25.
[12] LUO X Y,TU Y G,ZHAO Y,et al.Effects of sulfhydryl compounds,carbohydrates,organic acids,and sodium sulfite on the formation of lysinoalanine in preserved egg [J].Journal of Food Science,2014,79(8):T1 621-T1 628.
[13] ZHAO Y,LUO X Y,LI J K,et al.Formation of lysinoalanine in egg white under alkali treatment [J].Poultry Science,2016,95(3):660-667.
[14] MONTILLA A,GÓMEZ-RUIZ J Á,OLANO A,et al.A GC-FID method for analysis of Lysinoalanine [J].Molecular Nutrition & Food Research,2007,51(4):415-422.
[15] ZHAO Y,LUO X Y,LI J K,et al.Effect of basic alkali-pickling conditions on the production of lysinoalanine in preserved eggs [J].Poultry Science,2015,94(9):2 272-2 279.
[16] BOSCH L,SANZ M L,MONTILLA A,et al.Simultaneous analysis of lysine,N-ɛ-carboxymethyllysine and lysinoalanine from proteins [J].Journal of Chromatography B,2007,860(1):69-77.
[17] PELLEGRINO L,RESMINI P,DE NONI I,et al.Sensitive determination of lysinoalanine for distinguishing natural from imitation mozzarella cheese 1[J].Journal of Dairy Science,1996,79(5):725-734.
[18] CATTANEO S,MASOTTI F,PELLEGRINO L.Effects of overprocessing on heat damage of UHT milk [J].European Food Research and Technology,2008,226(5):1 099-1 106.
[19] CATTANEO S,MASOTTI F,PELLEGRINO L.Liquid infant formulas:technological tools for limiting heat damage [J].Journal of Agricultural and Food Chemistry,2009,57(22):10 689-10 694.
[20] LIU D R,ZHAO C H,GUO M R.Sodium tripolyphosphate inhibits the formation of lysinoalanine in heat-treated whey protein [J].Journal of Food Processing and Preservation,2017,41(6):e13501.
[21] PELLEGRINO L,CATTANEO S,MASOTTI F,et al.Detection of milk powder and caseinates in Halloumi cheese[J].Journal of Dairy Science,2010,93(8):3 453-3 460.
[22] 张兆丽.碱提米渣分离蛋白过程中赖丙氨酸的生成及超声波技术控制研究 [D].镇江:江苏大学,2020:36-37.ZHANG Z.Study on lysinoalanine generation and inhibition of ultrasound technology in alkali extraction of rice residue protein isolates [D].Zhenjiang:Jiangsu University,2020:36-37.
[23] XU H N,PAN J Y,HAO J,et al.Inhibition of cross-linked lysinoalanine formation in pH 12.5-shifted silkworm pupa protein,and functionality thereof:effect of ultrasonication and glycation [J].International Journal of Biological Macromolecules,2024,256:128120.
[24] XU H N,PAN J Y,MA C F,et al.Effect of carbonyl-amino condensation,non-covalent cross-linking and conformational changes induced by ultrasound-assisted Maillard reaction on lysinoalanine formation in silkworm pupa protein [J].International Journal of Biological Macromolecules,2024,263:130480.
[25] XU H N,PAN J Y,DABBOUR M,et al.Synergistic effects of pH shift and heat treatment on solubility,physicochemical and structural properties,and lysinoalanine formation in silkworm pupa protein isolates [J].Food Research International,2023,165:112554.
[26] ZHANG Z L,WANG Y,DAI C H,et al.Alkali extraction of rice residue protein isolates:effects of alkali treatment conditions on lysinoalanine formation and structural characterization of lysinoalanine-containing protein [J].Food Chemistry,2018,261:176-183.
[27] LI Y H,ZHANG Z L,REN W B,et al.Inhibition effect of ultrasound on the formation of lysinoalanine in rapeseed protein isolates during pH shift treatment [J].Journal of Agricultural and Food Chemistry,2021,69(30):8 536-8 545.
[28] PAN J Y,XU H N,DABBOUR M,et al.Changes in physicochemical,structural and functional properties,and lysinoalanine formation during the unfolding and refolding of pH-shifted black soldier fly larvae albumin [J].International Journal of Biological Macromolecules,2024,272(Pt 1):132801.
[29] AL-SAADI J M S,EASA A M,DEETH H C.Effect of lactose on cross-linking of milk proteins during heat treatments [J].International Journal of Dairy Technology,2013,66(1):1-6.
[30] ZHANG Z L,WANG Y,JIANG H,et al.Effect of dual-frequency ultrasound on the formation of lysinoalanine and structural characterization of rice dreg protein isolates [J].Ultrasonics Sonochemistry,2020,67:105124.
[31] WANG Y,ZHANG Z L,LI Y H,et al.Lysinoalanine formation and conformational characteristics of rice dreg protein isolates by multi-frequency countercurrent S-type sonochemical action[J].Ultrasonics Sonochemistry,2020,69:105257.
[32] AYIM I,MA H L,ALENYOREGE E A,et al.Effect of alkali concentration on functionality,lysinoalanine formation,and structural characteristics of tea residue proteins [J].Journal of Food Process Engineering,2018,41(8):e12877.
[33] FENAILLE F,PARISOD V,VISANI P,et al.Modifications of milk constituents during processing:a preliminary benchmarking study [J].International Dairy Journal,2006,16(7):728-739.
[34] CALABRESE M G,MAMONE G,CAIRA S,et al.Quantitation of lysinoalanine in dairy products by liquid chromatography –mass spectrometry with selective ion monitoring [J].Food Chemistry,2009,116(3):799-805.
[35] NIELSEN S D,LE T T,KNUDSEN L J,et al.Development and application of a multiple reaction monitoring mass spectrometry method for absolute quantification of lysinoalanine and lanthionine in dairy products [J].International Dairy Journal,2020,105:104693.
[36] NIELSEN S D,KNUDSEN L J,BÆKGAARD L T,et al.Influence of lactose on the maillard reaction and dehydroalanine-mediated protein cross-linking in casein and whey [J].Foods,2022,11(7):897.
[37] AGUILERA-TORO M,POULSEN N A,AKKERMAN M,et al.Development in maillard reaction and dehydroalanine pathway markers during storage of UHT milk representing differences in casein micelle size and sedimentation [J].Foods,2022,11(10):1 525.
[38] CARDOSO H B,WIERENGA P A,GRUPPEN H,et al.Maillard induced aggregation of individual milk proteins and interactions involved [J].Food Chemistry,2019,276:652-661.
[39] VAN ROOIJEN C,BOSCH G,VAN DER POEL A F B,et al.Quantitation of maillard reaction products in commercially available pet foods [J].Journal of Agricultural and Food Chemistry,2014,62(35):8 883-8 891.
[40] WANG Q L,JIN Y,XIONG Y L.Heating-aided pH shifting modifies hemp seed protein structure,cross-linking,and emulsifying properties [J].Journal of Agricultural and Food Chemistry,2018,66(41):10 827-10 834.
[41] KOYAMA S,NEMOTO Y,ICHIKAWA M,et al.Effects of suppressing protein structural changes on the excellent gelling properties of dried egg white via dry-heat treatment [J].Food Science and Technology Research,2021,27(2):293-300.
[42] KOYAMA S,OKA D,TSUJII Y,et al.Effects of lanthionine and lysinoalanine on heat-induced gelation of egg white [J].Food Science and Technology Research,2020,26(6):789-795.
[43] AKILLIOĞLU H G,LUND M N.Quantification of advanced glycation end products and amino acid cross-links in foods by high-resolution mass spectrometry:applicability of acid hydrolysis [J].Food Chemistry,2022,366:130601.
[44] 聂雪梅,王菡,许秀丽,等.Q-Orbitrap 高分辨质谱法测定牛奶 中 赖 丙 氨 酸 及 其 含 量 随 温 度 变 化 的 规 律 [J].食 品 科 学,2020,41(4):280-286.NIE X M,WANG H,XU X L,et al.Determination of lysinoalanine in milk products by Q-Orbitrap high resolution mass spectrometry and variation in its content with temperature|[J].Food Science,2020,41(4):280-286.
[45] CHEN Z F,LEINISCH F,GRECO I,et al.Characterisation and quantification of protein oxidative modifications and amino acid racemisation in powdered infant milk formula [J].Free Radical Research,2019,53(1):68-81.
[46] DELATOUR T,HEGELE J,PARISOD V,et al.Analysis of advanced glycation endproducts in dairy products by isotope dilution liquid chromatography-electrospray tandem mass spectrometry.The particular case of carboxymethyllysine [J].Journal of Chromatography A,2009,1 216(12):2 371-2 381.
[47] 陈铭,吴帅,黄轶群,等.山奈、白芷和香叶对猪肉加热过程中 晚 期 糖 化 终 末 产 物 含 量 的 影 响 [J].食 品 与 机 械,2019,35(4):1-6,13.CHEN M,WU S,HUANG Y Q,et al.Effects of galangal,angelica root and bay leaves on the formation of advanced glycation end-products in pork during heating [J].Food & Machinery,2019,35(4):1-6,13.
[48] SCHEIJEN J L J M,CLEVERS E,ENGELEN L,et al.Analysis of advanced glycation endproducts in selected food items by ultra-performance liquid chromatography tandem mass spectrometry:presentation of a dietary AGE database [J].Food Chemistry,2016,190:1 145-1 150.
[49] 蔡雯雯,张根生,山永凯,等.枸杞果汁中多酚类化合物鉴定[J].食品与机械,2024,40(12):129-136.CAI W W,ZHANG G S,SHAN Y K,et al.Identification of polyphenolic compounds in wolfberry juice [J].Food & Machinery,2024,40(12):129-136.
[50] 袁江月,何佳,孙军杰,等.UPLC-MS/MS 分析紫薯酒发酵前后 花 色 苷 种 类 和 含 量 变 化 [J].食 品 与 机 械,2021,37(6):56-61,68.YUAN J Y,HE J,SUN J J,et al.Analysis of anthocyanins in purple sweet potato wine before and after fermentation by UPLC-MS/MS [J].Food & Machinery,2021,37(6):56-61,68.
[51] 齐麟,张长波,刘仲齐.邻苯二甲醛柱前衍生化时间对反相高 效 液 相 色 谱 法 测 定 稻 米 氨 基 酸 含 量 的 影 响 [J].食 品 安 全质量检测学报,2022,13(2):449-456.QI L,ZHANG C B,LIU Z Q.Effects of pre-column derivatization time of O-phthalaldehyde on determination of amino acids content in rice by reversed-phase high performance liquid chromatography [J].Journal of Food Safety & Quality,2022,13(2):449-456.
[52] 卢 兰 香,郑 红,武 传 香,等.AQC 柱 前 衍 生 /UPLC-MS/MS 测定蜂蜜中的内源性氨基酸和牛磺酸成分 [J].分析测试学报,2022,41(12):1 738-1 745.LU L X,ZHENG H,WU C X,et al.Determination of endogenous amino acids and taurine in honey by ultra-performance liquid chromatography-tandem mass spectrometry with AQC pre-column derivation [J].Journal of Instrumental Analysis,2022,41(12):1 738-1 745.
[53] 黄莹,黄晶,杨晓,等.PITC 柱前衍生—高效液相色谱法测大米 肽 中 的 16种 氨 基 酸 含 量 [J].食 品 工 业 科 技,2023,44(24):279-285.HUANG Y,HUANG J,YANG X,et al.Determination of 16 amino acids in rice peptides by PITC pre-column derivatization high performance liquid chromatography [J].Science and Technology of Food Industry,2023,44(24):279-285.
[54] BADOUD R,PRATZ G.Simple and rapid quantitative determination of lysinoalanine and protein hydrolysate amino acids by high-performance liquid chromatography after derivatization with dansyl chloride [J].Chromatographia,1984,19(1):155-164.
[55] ZHOU S K,XIA Y M,HAO L H,et al.Optimization of liquid chromatography and mass spectrometry parameters based on LC-QQQ:a case study on lysinoalanine [J].Journal of Chromatography B,2025,1 251:124427.

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