代谢组学在农产品营养品质检测分析中的应用

doi:10.3864/j.issn.0578-1752.2019.18.009

中国农业科学 ›› 2019, Vol. 52 ›› Issue (18): 3163-3176.doi: 10.3864/j.issn.0578-1752.2019.18.009

代谢组学在农产品营养品质检测分析中的应用

许彦阳¹,姚桂晓^1,³,刘平香¹,赵洁¹,王昕璐¹,孙君茂²(),钱永忠¹()

^1. 中国农业科学院农业质量标准与检测技术研究所/农业农村部农产品质量安全重点实验室,北京 100081
^2. 农业农村部食物与营养发展研究所, 北京 100081
^3. 西安理工大学印刷包装与数字媒体学院,西安 710048

收稿日期:2019-02-22 接受日期:2019-04-23 出版日期:2019-09-16 发布日期:2019-09-23
通讯作者: 孙君茂,钱永忠
作者简介:许彦阳,Tel：010-82106539;E-mail：xuyanyang@caas.cn。
基金资助:
国家自然科学基金(31701519);科技部食品安全关键技术研发专项(2017YFC1600705)

Review on the Application of Metabolomic Approaches to Investigate and Analysis the Nutrition and Quality of Agro-Products

XU YanYang¹,YAO GuiXiao^1,³,LIU PingXiang¹,ZHAO Jie¹,WANG XinLu¹,SUN JunMao²(),QIAN YongZhong¹()

^1. Institute of Quality Standards and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-Product Quality and Safety, Ministry of Agriculture and Rural Affairs, Beijing 100081
^2. Institute of Food and Nutrition Development, Ministry of Agriculture and Rural Affairs, Beijing 100081
^3. Xi’an University of Technology, Xi’an 710048;

Received:2019-02-22 Accepted:2019-04-23 Online:2019-09-16 Published:2019-09-23
Contact: JunMao SUN,YongZhong QIAN

摘要/Abstract

摘要：

科学检测和分析农产品营养品质对提升优质农产品的营养水平具有重要作用。由于农产品中营养成分组成复杂,已有的分析方法只能针对已知营养素的浓度、功能等进行分析,无法对农产品中存在的大量其他功能性活性物质进行分析鉴定。代谢组学技术通过高通量化学分析技术对生物样品中的小分子代谢产物进行定性和定量分析,在具有特殊营养功能的小分子物质分析中具有突出优势。代谢组学技术的引入,为农产品营养成分表征及差异性分析,产地溯源及真伪鉴别,生长储藏过程中营养成分变化规律揭示,营养功能成分作用机制研究等提供了新方法,也为膳食结构的优化调整提供了新策略。本文对代谢组学研究方法的新进展,包括样品制备、代谢物分析鉴定以及数据分析等进行了综述,总结了代谢组学技术在农产品营养成分表征及差异性分析、产地溯源及真伪鉴别、生长储藏过程中营养成分变化规律以及营养功能成分作用机制等方面的应用,旨在为我国农业高质量发展提供思路。在样品制备方法方面,首先通过快速改变样品所处的环境条件,如添加强酸、强碱或液氮冷冻等技术终止新陈代谢相关酶的活性。然后针对代谢物的极性,选择不同的提取溶剂,从而获得较高的提取率。在样品分析方法方面,核磁共振、色谱质谱和毛细管电泳质谱等技术得到了广泛地应用。其中,色谱质谱联用技术将色谱的有效分离和质谱的准确定量相结合,已经成为代谢组学中使用最广泛的分析技术。在数据处理及分析方面,无监督分析中的主成分分析和有监督分析中的正交偏最小二乘法–判别分析是目前应用最多的数据分析方法。通过通路分析软件的富集分析和拓扑分析,可以明确与代谢物差异相关性最高的代谢通路,对差异代谢产物的机制进行分析和解释。在农产品营养成分分析应用方面,通过对农产品中初生代谢产物及次生代谢产物进行全面表征,形成农产品独特的代谢指纹图谱,从而实现营养成分的差异性分析;通过非特定目标物的检测和无监督分析方法,实现不同产地农产品的组间差异鉴别和差异化合物筛选;通过农产品生长过程中关键成分的消长规律及合成机理分析,指导最佳收获期;通过体液代谢谱和生物标志物的检测,系统研究营养功能成分与生物体代谢之间的交互作用,为营养功能成分作用机制研究及膳食指导提供有价值的信息。

关键词: 代谢组学, 农产品, 营养, 品质

Abstract:

Scientific evaluation of the nutrition and quality of agricultural products is essential for improving the nutrition level of agro-products. Because of the complex composition of nutrients in agro-products, the existing analytical methods can only analyze the concentration and function of known nutrients but cannot analyze and identify a large number of unknown functional substances. On the basis of high-throughput chemical analyses, metabolomics can qualitatively and quantitatively analyze endogenous and exogenous metabolites of biological samples. Therefore, metabolomics has outstanding advantages in the analysis of small molecular substances with special nutritional functions in agricultural products; it has advantages like providing new methods for the characterization and differential analysis of nutrient components, traceability and authenticity of identification, variation analysis of functional substances during growth and storage, and the effect mechanisms of functional components. It also provides new strategies for structural optimization of dietary requirements. In this paper, the recent advances in metabolomics research, including sample preparation, metabolite analysis, data processing, differential metabolite identification, and metabolic pathway analysis were reviewed. This work summed up the application of metabolomics in the characterization and difference analysis of metabolites, traceability and authenticity identification of origin, metabolite variation in the process of storage, and the evaluation of nutritional functions to provide theoretical bases and practical references for high-quality agricultural development in China. In the field of sample preparation, the activity of metabolism-related enzymes is first terminated by rapidly changing the environmental conditions, such as adding strong acid (alkali) or freezing in liquid nitrogen. Different extraction solvents are selected based on the polarities of the metabolites to obtain a higher extraction rate. In the field of sample analysis methods, technologies, such as nuclear magnetic resonance spectroscopy, chromatography mass spectrometry and capillary electrophoresis-mass spectrometry, have been widely used. Among them, the combination of chromatography and mass spectrometry has become the most commonly used analytical technique in metabolomics. In the field of data processing and analysis, principal component analysis and orthogonal partial least squares-discriminant analysis are the most common data analysis techniques. Through enrichment and topological analysis, the metabolic pathway with the highest correlation to differential metabolites can be identified, and the reason of differential metabolites can be explained and analyzed. In the field of evaluation of nutrition and quality of agricultural products, through the comprehensive characterization of primary metabolites and secondary metabolites in agricultural products, unique fingerprints of agricultural products are established and used for differential analysis, whereas through non-specific target analysis and unsupervised analysis methods, differences between groups and relating metabolites can be identified. Via concentration analysis of key components in the growth process of agricultural products, the best harvest periods can be provided. Interaction studies between functional components and metabolism of organisms based on the detection of humoral metabolism and biomarkers can provide valuable information for dietary guidance.

Key words: metabolomics, agro-product, nutrition, quality

许彦阳,姚桂晓,刘平香,赵洁,王昕璐,孙君茂,钱永忠. 代谢组学在农产品营养品质检测分析中的应用[J]. 中国农业科学, 2019, 52(18): 3163-3176.

XU YanYang,YAO GuiXiao,LIU PingXiang,ZHAO Jie,WANG XinLu,SUN JunMao,QIAN YongZhong. Review on the Application of Metabolomic Approaches to Investigate and Analysis the Nutrition and Quality of Agro-Products[J]. Scientia Agricultura Sinica, 2019, 52(18): 3163-3176.

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图/表 2

表1

代谢组学在农产品营养品质检测分析中的应用"

应用 Application	农产品种类 Agro-product	检测技术 Detection technology	数据处理方法 Data analysis	生物标志物 Biomarker	参考文献 Reference
营养成分表征及差异性分析 Characterization and difference analysis of nutritional components	番茄 Tomato	LC-MS/MS	PCA	氨基酸(Amino acid)、6-甲基-5-庚烯-2-酮(6- Methyl-5-hepten-2-one)、香叶基丙酮(Geranyl acetone)等	[67]
	洋葱 Onion	LC/ ESI-QTOF-MS	ANOVA, HC, PCA	低聚果糖(Fructooligosaccharides)、氨基酸(Proteinogenic amino acids)、多肽(Peptides)、S-半胱氨酸(S-substituted cysteine conjugates)、黄酮(Flavonoids)及皂苷类(Flavonoids)	[68]
	葡萄 Grapevine berry	UHPLC-ESI-QTOF-MS	PCA, OPLS-DA	花色苷(Anthocyanins)、芪类化合物(Stilbenoids)、原花青素(Procyanidin)等	[69]
	蟹 Crab	¹H-NMR	PCA	谷氨酸(Glutamate)、丙氨酸(Alanine)、甘氨酸(Glycine)、龙虾碱(Lobsterine)、乳酸(Lactic acid)、甜菜碱(Betaine)和牛磺酸(Taurine)	[70]
	番茄 Tomato	HS/SPME/GC-MS	ANOVA, MCA, PCA, HCA	邻甲基苯乙酮(O-methylacetophenone)、苯甲酮(Benzophenone)等	[71]
	芜菁 Brassica crops	GC-MS	PCA, PLS regression analysis, ANOVA	L-谷氨酰胺(L-glutamine)、L-天冬酰胺(L- asparagine)、棉子糖(Raffinose)、麦芽糖(Maltose)、苹果酸(Malic acid)和异硫氰酸烯丙酯(Allyl isothiocyanate)	[72]
	蔓越莓 Cranberry	UPLC-TOF-MS	PCA, PLS-DA	花青素(Procyanidin)	[73]
	树莓 Raspberries	UPLC-TOF-MS	Pattern analysis	花青素(Procyanidin)	[74]
	马铃薯 Potatoes	HPLC,GC-TOF-MS	ANOVA, PCA	花青素(Procyanidin)	[75]
	番茄 Tomato	¹H-NMR,LC-FID, GC-FID	PCA, SOM	甘露糖(Mannose)、胆碱(Choline)、油酸(Oleic)、硬脂酸(Stearic)、亚油酸(Linoleic)、亚麻酸(Linolenic)、棕榈酸(Palmitic)、淀粉(Amylum)	[76]
	玉米 Maize	UPLC-MS/MS,GC-MS	PCA, PLS-DA	二氢山萘酚(Dihydrokaempferol)和柚皮素(Naringenin)	[78]
	番茄 Tomato	¹H-NMR	PCA, PLS, ANOVA	谷氨酸(Glutamic acid)、果糖(Fructose)、缬氨酸(Valine)、山奈酚(Kaempferol)、桂皮素苷(Cinnamrin)、γ-氨基丁酸(Gamma-aminobutyric acid)、柠檬酸(Citric acid)、蔗糖(Sucrose)、苯丙氨酸(Phenylalanine)和葫芦巴碱(Trigonelline)	[79]
产地溯源及真伪鉴别 Origin traceability and authenticity identification	大白菜 Cabbage	¹H-NMR	PCA	氨基丁酸(Aminobutyric acid)、天冬酰胺(Asparagine)、亮氨酸(Leucine)、异亮氨酸(Isoleucine)、O-磷酸胆碱(O-phosphocholine)、乙酸苯酯(Phenylacetate)、苯丙氨酸(Phenylalanine)、琥珀酸盐(Succinate)、蔗糖(Sucrose)、酪氨酸(Tyrosine)、缬氨酸(Valine)	[81]
	枸杞 Lycium barbarum	LC-QTOF-MS	PLS-DA, HCA	槲皮素(Quercetin)、山奈酚糖苷(Kaempferol glycosides)、二咖啡酰奎宁酸(Dicaffeoylquinic acid)和酚酸(Phenolic acids)	[82]
	大蒜 Garlic	DART-HRMS, HPLC-ESI-HRMS, DI-ESI-HRMS	OPLS-DA	蒜氨酸(Alliin)、PC (16:0/18:2) (Phosphatidylcholine)和精氨酸(Arginine)	[83]
	大蒜 Garlic	HRMAS-NMR	PLS-DA	烯丙基含硫化合物(Allyl-organosulphurs)、大蒜素(Allicin)	[84]
	榛子 Hazelnut	UPLC-QTOF-MS	PCA-LDA	磷脂酰胆碱(Phosphatidylcholines)、磷脂酰乙醇胺(Phosphatidylethanolamines)、甘油二酯(Diacylglycerols)、三酰基甘油(Triacylglycerols)和γ-生育酚(γ-tocopherol)	[90]

表1

续表1

代谢组学在农产品营养品质检测分析中的应用"

应用 Application	农产品种类 Agro-product	检测技术 Detection technology	数据处理方法 Data analysis	生物标志物 Biomarker	参考文献 Reference
	榛子 Hazelnut	LC-ESI-QqQ-MS	PCA-LDA, ANOVA	PE(18:2/18:2) (1,2-dilinoleoyl-sn glycero-3- phosphoethanolamine)、PC(18:2/18:2) (1,2- dilinoleoyl-sn-glycero-3-phosphocholine)、DG (18:2/18:2) (1,3-100 dilinoleoyl-rac-glycerol)	[91]
生长储藏过程中营养成分变化规律 Changes of nutritional components during growth and storage	草莓 Strawberry	GC-MS,HPLC-MS	PCA, PLS-DA	游离氨基酸(Free amino acids)	[92]
	葡萄 Grapevine berry	GC-MS	HCA, PCA, PLS-DA, ANOVA	糖类(Sugars)和氨基酸(Amino acids)	[93]
	番茄 Tomato	GC-MS	PLS-DA, ANOVA	甘露糖(Mannose)、柠檬酸(Citramalic)、葡萄糖酸(Gluconic)和酮-1-古洛糖酸(Keto-l-gulonic acids)等	[94]
	鸡蛋 Chicken eggs	HPLC-QTOF-MS	PCA	胆碱(Choline)	[48]
	大豆 Soybean	¹H-NMR	PCA, PLS-DA	脯氨酸(Proline)、赖氨酸(Lysine)和硫(Sulfur)	[96]
营养功能成分作用机制及对代谢的影响 Functional mechanism of nutritional components and their effects on metabolism	生姜 Ginger	HPLC-QTOF-MS	ANOVA	D-葡糖醛酸-6 (D-glucurono-6)、3-内酯(3- lactone)、甘油-3-磷酸(Glycerol-3-phosphate)、丙酮酸(Pyruvic acid)、石胆酸(Lithocholic acid)、2-吡啶甲酸(2-pyrocatechuic acid)和前列腺素E1 (Prostaglandin El)等	[100]
	枸杞 Lycium barbarum	GC-TOF-MS	PCA, OPLS-DA	丙氨酸(Alanine)、胸腺嘧啶脱氧核苷酸(Thymidine deoxynucleotide)	[101]
	绿茶 Green tea	LC-MS	PCA, OPLS-DA, PLS regression analysis	多酚(Polyphenol)	[102]
	苦瓜 Bitter gourd	NMR	PCA, PLS-DA	柠檬酸(Citramalic)、琥珀酸(Succinate)、肌氨酸(Sarcosine)、丙酮酸(Pyruvic acid)等	[107]
	白茶 Origanum dictamnus tea	¹H-NMR	PCA, OPLS-DA	马尿酸(Hippurate)和肌酐(Creatinine)	[108]
	饮食模式 Dietary modulation	¹H-NMR	OPLS-DA	肉碱(Carnitine)、乙酰肉碱(Acetylcarnitine)、三甲胺-N-氧化物(Trimethylamine-N-oxid)、对-羟基苯乙酸酯(p-hydroxyphenylacetate)	[109]

续表1

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代谢组学在农产品营养品质检测分析中的应用

Review on the Application of Metabolomic Approaches to Investigate and Analysis the Nutrition and Quality of Agro-Products

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