APP下载

驴奶和牛奶游离氨基酸成分分析研究

2017-12-06涂裔伟姚路迅

中国乳业 2017年10期
关键词:分析仪游离生理

文/李 宇 赵 兵 蒋 慧 张 静 涂裔伟 姚路迅 贾 薇 张 奇*

〔中驭(北京)生物工程有限公司〕

驴奶和牛奶游离氨基酸成分分析研究

文/李 宇 赵 兵 蒋 慧 张 静 涂裔伟 姚路迅 贾 薇 张 奇*

〔中驭(北京)生物工程有限公司〕

采用氨基酸分析仪对驴奶和牛奶中游离的氨基酸进行分析,比较其中的必需氨基酸和特殊生理功能氨基酸的相对含量。结果表明,驴奶的游离氨基酸中所含有的必需氨基酸、小儿/早产儿必需氨基酸及特殊生理活性氨基酸等均高于牛奶。可见在游离氨基酸相对含量方面,驴奶表现出显著优于牛奶的营养价值和保健功效。

驴奶;牛奶;游离氨基酸;氨基酸分析仪

氨基酸是构成蛋白质的基本单位,赋予蛋白质特定的分子结构形态,使其的分子具有生化活性,其可分为必需氨基酸、非必需氨基酸、特殊生理活性氨基酸。

必需氨基酸是指人体自身不能合成或合成速度不能满足人体需要,需要从食物中摄取的一类氨基酸,成人必需氨基酸包括赖氨酸(Lysine,Lys)、色氨酸(Tryptophane,Trp)、苯丙氨酸(Phenylalanine,Phe)、蛋氨酸(Methionine,Met)、苏氨酸(Threonine,Thr)、异亮氨酸(Isoleucine,Ile)、亮氨酸(Leucine,Leu)、缬氨酸(Valine,Val),小儿生长发育必需氨基酸除上述氨基酸外还包括组氨酸(Histidine,His),早产儿必需氨基酸还包括精氨酸(Arginine,Arg)、胱氨酸(Cystine,Cys)、酪氨酸(Tyrosine,Tyr)和牛磺酸(Taurine,Tau)。

除上述必需氨基酸和人体能够自身合成的10 种常规氨基酸外,研究人员对诸多非常规氨基酸进行研究,发现许多非常规氨基酸具有极强的生理活性,如磷脂酰丝氨酸(Phosphatidylserine,P-Ser)、肌氨酸(Sarcosine,Sar)、β-丙氨酸(beta-Alanine,β-Ala)、瓜氨酸(Citrulline,Cit)、γ-氨基丁酸(γ-aminobutyric acid,GABA)、鸟氨酸(Ornithine,Orn)等。

常规的18 种氨基酸在人体生理活动过程中除参与蛋白合成、脂肪和糖代谢、氮平衡、维生素辅酶合成等功能外,其中的必需氨基酸和特殊生理活性氨基酸还具有营养补充、机体发育、疾病治疗、提升免疫力、调节机体平衡、解毒、抗衰老、抗氧化、缓解机体疲劳等重要功能,详见表1~3。

据报道,驴奶是最接近人乳的天然乳[1],其营养价值高,远优于牛奶与羊奶。《本草纲目》中对驴奶的描述为:“气味甘,冷利,无毒,热频饮之可治气郁,解小儿热毒,不生痘疹”[2]。国外有研究表明,驴奶具有抗氧化,增强人体免疫力的功能[3~4]。

本文对新疆维吾尔自治区喀什地区的驴奶中游离氨基酸组分进行分析,与牛奶相比,发现驴奶的游离氨基酸中必需氨基酸和特殊生理活性氨基酸的含量均显著高于牛奶。可见在氨基酸含量和比例上,驴奶的营养价值和保健功效均显著优于牛奶。

1 材料与方法

1.1 材料

新鲜驴奶采自新疆维吾尔自治区喀什地区,牛奶为商品化市售纯牛奶,混合氨基酸标准工作液购于北京普鹏科技有限公司,其它试剂均为国产分析纯。

1.2 样品处理

取9 mL新鲜驴奶和牛奶,分别加入1 mL三氯乙酸,充分振荡混匀后静置10 min,12 000 r/min离心,收集上清液,4 ℃保存备用。每个样品3 个重复。

1.3 氨基酸成分分析

将上述制备好的样品进行1 000倍稀释后以全自动氨基酸分析仪(L-8800型,日本日立)进行分析测定,色谱柱为4.6 mm×60 mm,色谱条件:柱温为62 ℃,泵流速为0.4 mL/min,进样体积为50 μL,检测器为35 ℃,检测波长为440 nm。混合氨基酸标准工作液以相同体积注入氨基酸分析仪,以外标法通过峰面积计算样品测定液中氨基酸的浓度。

1.4 统计分析

用SPSS 17.0软件进行统计分析,结果以平均值±标准差表示,方差齐性检验。采用单因素方差分析(ANOVA),组间两两比较(LSD),检验水准P=0.05,以P<0.05为差异显著,P <0.01为差异极显著。

2 结果与分析

2.1 必需氨基酸比较分析

采用氨基酸分析仪对驴奶和牛奶中游离氨基酸进行检测,并对必需氨基酸进行比较(表1),发现驴奶的游离氨基酸中,8 种必需氨基酸的含量均高于牛奶,差异均极显著(P<0.01),其中驴奶中游离的Thr和Phe比牛奶高20 倍以上;此外,驴奶中含有人体所必需的Trp,而在本研究选用的牛奶中未检测出该氨基酸。

2.2 小儿/早产儿必需氨基酸分析

小儿或早产儿除成人所必需的氨基酸外,还需补充Tau、Tyr、His、Arg和Cys等氨基酸,通过对牛奶与驴奶中游离氨基酸含量进行分析(表2),发现驴奶中均含这5 种游离氨基酸,其中Cys含量高于牛奶,但不具备显著性差异,其它4 种氨基酸的相对含量均高于牛奶,且差异均极显著(P<0.01),其中重要的His在驴奶中的相对含量高于牛奶20 倍以上。

表1 8 种必需氨基酸含量检测结果(n=3)

表2 小儿与早产儿生长发育所必须的氨基酸含量检测结果(n=3)

表3 具有生理活性的氨基酸含量检测结果(n=3)

2.3 其它重要生理活性氨基酸

鲜乳中除了常规的18 种氨基酸外,还含有一些非常规的特殊氨基酸,其中部分氨基酸具有十分重要的生理活性,对驴奶和牛奶的游离氨基酸进行分析(表3),发现驴奶和牛奶游离氨基酸中均含有生理活性氨基酸。具备显著生理功能的氨基酸在驴奶中的相对含量均高于牛奶,其中肌氨酸(Sar)表现为显著性差异(P<0.05),其它氨基酸表现出极显著差异(P<0.01),如具备提高免疫力、抗氧化、降低血压、抵抗焦虑等功能的GABA,在驴奶中的游离含量为牛奶的13 倍以上,此外提升免疫力的Cit,在驴奶中的游离含量为牛奶的26 倍。

3 结论

本文对驴奶和牛奶的游离氨基酸中的必需氨基酸、特殊生理活性氨基酸进行比较分析研究,发现驴奶游离氨基酸中的必需氨基酸和特殊生理活性氨基酸的含量均高于牛奶。由此可见,在游离氨基酸相对含量方面,驴奶的营养价值及保健功效均高于牛奶,更适合用于营养补充及疾病的补充治疗,具有极高的应用和推广价值。

[1] Guo H Y,Pang K,Zhang X Y,et al.Composition,physiochemical properties,nitrogen fraction distribution,and amino acid profile of donkey milk[J]. Journal of Dairy Science,2007,90(4):1635-1643.

[2] 李时珍. 本草纲目(金陵版排印本)兽部第五十卷[M]. 北京:人民卫生出版社,2006.

[3] Chiozzi R Z,Capriotti A L,Cavaliere C,et al. Purification and identification of endogenous antioxidant and ACE-inhibitory peptides from donkey milk by multidimensional liquid chromatography and nanoHPLC-high resolution mass spectrometry[J]. Analyticalamp; Bioanalytical Chemistry,2016,408(20):5657-5666.

[4] Amati L,Marzulli G,Martulli M,et al.Donkey and goat milk intake and modulation of the human aged immune response[J].Current Pharmaceutical Design,2010,16(7):864-869.

[5] Zhang Q,Chen X,Eicher S D,et al. Effect of threonine on secretory immune system using a chicken intestinal ex vivo model with lipopolysaccharide challenge[J]. Poultry Science,2017,96(9):3043-3051.

[6] Khedr N F,Khedr E G. Branched chain amino acids supplementation modulates TGF-β1/smad signaling pathway and interleukins in CCl4-induced liver fibrosis[J].Fundamental amp; Clinical Pharmacology,2017,31(5):534-545.

[7] Hes M,Gliszczyńska-SwigIo A,Gramza-MichaIowska A. The effect of antioxidants on quantitative changes of lysine and methionine in linoleic acid emulsions at different pH conditions[J].Acta Scientiarum Polonorum, Technologia Alimentaria,2017,16(1):53-67.

[8] Campos-Ferraz P L,Bozza T,Nicastro H,et al. Distinct effects of leucine or a mixture of the branched-chain amino acids(leucine,isoleucine,and valine)supplementation on resistance to fatigue,and muscle and liver-glycogen degradation,in trained rats[J]. Nutrition,2013,29(11-12):1388-1394.

[9] Chen C F,Tai Y K. A rare haemoglobin variant(Hb Phnom Penh)manifesting as a falsely high haemoglobin A1c value on ion-exchange chromatography[J]. Singapore Medical Journal,2014,55(8):126-128.

[10] Yang W,He Y,Liu S,et al. Integrative transcriptomic analysis of NAFLD animal model reveals dysregulated genes and pathways in metabolism[J]. Gene,2016,595(1):99-108.

[11] Cynober L,Bier D M,Kadowaki M,et al.Proposals for uUpper limits of safe intake for arginine and tryptophan in young adults and an upper limit of safe intake for leucine in the elderly[J]. Journal of Nutrition,2016,146(12):2652-2654.

[12] Petermann M,González C,Hirsch S,et al. Leucine and glucose turnover in chronic alcoholics during early abstinence and after an ethanol load[J]. Alcoholism,Clinical and Experimental Research,1993,17(6):1295-1300.

[13] Strasser B,Sperner-Unterweger B,Fuchs D,et al. Mechanisms of inflammationassociated depression:immune influences on tryptophan and phenylalanine metabolisms[J]. Current Topics Behavioral Neurosciences,2017,31:95-115.

[14] Canto de Souza L,Provensi G,Vullo D,et al. Carbonic anhydrase activation enhances object recognition memory in mice through phosphorylation of the extracellular signal-regulated kinase in the cortex and the hippocampus[J].Neuropharmacology,2017,118:148-156.

[15] Tripathi A K,Kumari T,Tandon A,et al. Selective phenylalanine to proline substitution for improved antimicrobial and anticancer activities of peptides designed on phenylalanine heptad repeat[J]. Acta Biomaterialia,2017,57:170-186.

[16] Boulet L,Faure P,Flore P,et al.Simultaneous determination of tryptophan and 8 metabolites in human plasma by liquid chromatography/tandem mass spectrometry[J]. Journal of Chromatography.B,B i o m e d i c a l S c i e n c e s a n d Applications,2017,1054:36-43.

[17] Lv L,Ge W,Liu Y,et al. Lysine-specific demethylase 1 inhibitor rescues the osteogenic ability of mesenchymal stem cells under osteoporotic conditions by modulating H3K4 methylation[J]. Bone Research,2016,4:16037.

[18] Korompokis K,Östman E,Dougkas A.The impact of liquid preloads varying in macronutrient content on postprandial kinetics of amino acids relative to appetite in healthy adults[J]. Appetite,2016,107:511-520.

[19] 陈仁. 营养保健食品[M]. 北京:中国轻工业出版社,2001.

[20] 高勇. 食品组分对铁的化学状态和其生物利用率的影响及其机制研究:[硕士学位论文][D].北京:中国疾病预防控制中心,2013:7-75.

[21] 赵华伟. 组氨酸对局灶性脑缺血引起的长期脑损伤的保护作用:[硕士学位论文][D].州:浙江大学医学院,2010:1-25.

[22] Li X W,Li F,Liu J,et al. Effect of antepartum taurine supplementation in regulating the activity of Rho family factors and promoting the proliferation of neural stem cells in neonatal rats with fetal growth restriction[J]. Zhongguo Dang Dai Er Ke Za Zhi,2016,18(11):1158-1165.

[23] Walczewska M,Ciszek-Lenda M,Surmiak M,et al. Impact of taurine on innate and adaptive immunity as the result of HOCl neutralization[J]. Advances in Experimental Medicine and Biology,2015,803:109-120.

[24] El Idrissi A,Okeke E,Yan X,et al. Taurine regulation of blood pressure and vasoactivity[J].Advances in Experimental Medicine and Biology,2013,775:407-425.

[25] El-Batal A I,El-Sayyad G S,El-Ghamry A,et al. Melanin-gamma rays assistants for bismuth oxide nanoparticles synthesis at room temperature for enhancing antimicrobial,and photocatalytic activity[J]. Journal of Photochemistry and Photobiology B-biology,2017,173:120-139.

[26] Jiang H,Guo S,Xiao D,et al. Arginine deiminase expressed in vivo,driven by human telomerase reverse transcriptase promoter,displays high hepatoma targeting and oncolytic efficiency[J]. Oncotarget,2017,8(23):37694-37704.

[27] Mondanelli G,Ugel S,Grohmann U,et al. The immune regulation in cancer by the amino acid metabolizing enzymes ARG and IDO[J]. Current Opinion in Pharmacology,2017,35:30-39.

[28] Santos J C,De Araújo O R,Valentim I B,et al. Choline and cystine deficient diets in animal models with hepatocellular injury:evaluation of oxidative stress and expression of RAGE,TNF-α,and IL-1β[J]. Oxidative Medicine and Cellular Longevity,2015:121925.

[29] Zhang Y Y,Yang L Q,Guo L M. Effect of phosphatidylserine on memory in patients and rats with Alzheimer's disease[J].Genetics and Molecular Research,2015,14(3):9325-9333.

[30] Rae C,Digney A L,McEwan S R,et al.Oral creatine monohydrate supplementation improves brain performance: a doubleblind, placebo-controlled, cross-over trial[J]. Proceedings Biological Sciences,2003,270(1529):2147-2150.

[31] Rapovy S M,Zhao J F,Bricker R L,et al. Differential Requirements for L-Citrulline and L-Arginine during Antimycobacterial Macrophage Activity[J]. Journal of Immunology,2015,195(7):3293-3300.

[32] Breuillard C,Curis E,Le Plénier S,et al. Nitric oxide production by peritoneal macrophages from aged rats:A short term and direct modulation by citrulline[J].Biochimie,2017,133:66-73.

[33] Pandurangan M,Enkhtaivan G,Mistry B,et al. β-Alanine intercede metabolic recovery for amelioration of human cervical and renal tumors[J]. Amino Acids,2017,49(8):1373-1380.

[34] Brencher L,Verhaegh R,Kirsch M.Attenuation of intestinal is chemia reperfusion injury by β-alanine: a potentially glycine-receptor mediated effect[J]. Journal of Surgical Research,2017,211:233-241.

[35] Belviranli M,Okudan N,Revan S,et al.Repeated supramaximal exercise-induced oxidative stress:effect of β-alanine plus creatine supplementation[J]. Asian Journal of Sports Medicine,2016,7(1):e26843.

[36] Glenn J M,Gray M,Stewart R W,et al.Effects of 28-day beta-alanine supplementation on isokinetic exercise performance and body composition in female masters athletes[J]. Journal of Strength and Conditioning Research,2016,30(1):200-207.

[37] Cheng J,Zheng N,Sun X,et al. Feeding rumen-protected gamma-aminobutyric acid enhances the immune response and antioxidant status of heat-stressed lactating dairy cows[J]. Journal of Thermal Biology,2016,60:103-108.

[38] Yoshimura M,Toyoshi T,Sano A,et al.Antihypertensive effect of a gammaaminobutyric acid rich tomato cultivar‘DG03-9’ in spontaneously hypertensive rats[J]. Journal of Agricultural and Food Chemistry,2010,58:615-619.

[39] Rosa S G,Quines C B,Stangherlin E C,et al. Diphenyl diselenide ameliorates monosodium glutamate induced anxietylike behavior in rats by modulating hippocampal BDNF-Akt pathway and uptake of GABA and serotonin[J].Physiology amp; Behavior,2016,155:1-8.

[40] Ho Y Y,Nakato J,Mizushige T,et al.L-Ornithine stimulates growth hormone release in a manner dependent on the ghrelin system[J]. Food amp; Function,2017,8(6):2110-2114.

Analysis of Amino Acids in Milk of Donkey

LI Yu,ZHAO Bing,JIANG Hui,ZHANG Jing,TU Yi-wei,YAO Lu-xun,JIA Wei,ZHANG Qi*
〔 Zhong Yu(Beijing)Biological Engineering Co.,Ltd 〕

An amino acid analyzer was used to analyze the free amino acids in the milk of donkey and milk,comparing the relative contents of essential amino acids and special physiological functions of amino acid. The results showed that the essential amino,children / premature infants essential amino acids and specif i c physiological active amino acids in the free amino acids of donkey milk were higher than that of milk,in terms of the relative contents of free amino acid,donkey milk showed signif i cantly better nutritional and health benef i ts than milk.

donkey milk;milk;amino acid;amino acid analyzer

国家高新技术研究发展计划(863 计划)(2014AA022209)]

李宇(1993-),学士,助理工程师,从事临床营养研究。

★通信作者:张奇(1979-),博士,副教授,从事临床营养、功能食品和药物开发等领域研究。

2017-07-25)

猜你喜欢

分析仪游离生理
游离股前外侧穿支皮瓣修复足踝部软组织缺损
Sekisui CP3000与STA Compact Max凝血分析仪检测项目比对分析
Sievers分析仪(苏伊士)
一种用于深空探测的Chirp变换频谱分析仪设计与实现
莫须有、蜿蜒、夜游离
陶珊珊作品
大脑如何知觉音乐的形式美?——来自电生理的证据
肉鸡铬需要量及其营养生理作用
铜绿微囊藻对锌、镉胁迫的生理响应
妈妈们产后的生理烦恼