基于代谢组学的‘云抗10号’晒青茶加工过程代谢物变化

doi:10.3864/j.issn.0578-1752.2020.02.010

Abstract

Abstract:

【Objective】The ultrahigh phase liquid chromatography/mass spectrometry (LC-MS) combined technique of metabolomics was used to explore the changes of metabolites in the processing of sun-dried green tea of Camellia sinenis var. assamica cv. Yunkang 10, and to find the iconic metabolites affecting the formation of sun-dried green tea quality. Further study on the change path of these substances would lay a foundation for understanding the formation mechanism of sun-dried green tea quality. 【Method】In the process of making Yunkang 10 sun-dried green tea, 3 samples of fresh leaves, rolled leaves and sun-dried leaves were taken respectively. After the samples were pretreated, the metabolites in the three groups of samples were detected by LC-MS and identified by mass spectrometry database. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used to analyze the detection data of three groups of samples. Metabolites with significant differences were screened out by PLS-DA method. 【Result】LC-MS analysis method for the endogenous metabolites of Yunkang 10 fresh leaves, rolled leaves and sun-dried leaves was established, and commercial mass spectrometry database was used for rapid identification of the detected metabolites. The metabolome data were imported into SIMCA-P software for principal component analysis and partial least squares discriminant analysis, and the metabolome data could be used to distinguish the three groups of samples. By LC/MS technique on Yunkang 10 sun-dried green tea and its processing tea, in combination with multivariate statistical analysis, there were 701 kinds of metabolites were significant differences among the fresh leaves, rolling leaves and sun-dried leaves, and 116 kinds metabolites between fresh leaves and rolling leaves, 158 kinds of metabolites between sun-dried leaves and fresh leaves, and 48 kinds of metabolites between sun-dried leaves and rolling leaves were found. By searching KEGG database to analyze metabolites, these metabolites were mainly related to amino acid metabolism, polyphenol metabolism and other energy metabolism pathways. 【Conclusion】LC-MS technique could be used to distinguish fresh leaf group, rolled leaf group and sun-dried leaf group of Yunkang 10, which proved that metabolomics technology could reveal the chemical changes of metabolites in sun-dried green tea to some extent. The key metabolites were found in the study could provide a theoretical basis for evaluating the quality of sun-dried green tea, and lay a theoretical foundation for exploring the formation of "sunburn taste" of sun-dried green tea and the formation mechanism of sun-dried green tea quality.

Key words: liquid chromatography/mass spectrometry, sun-dried green tea, Yunkang 10, metabolomics, metabolites

DAI YuQiao,Lü CaiYou,HE LuNan,YI Chao,LIU XueYan,HUANG Wen,CHEN JiaMin. Metabolic Changes in the Processing of Yunkang 10 Sun-Dried Green Tea based on Metabolomics[J].Scientia Agricultura Sinica, 2020, 53(2): 357-370.

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Figures/Tables 14

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Table 1

Fig. 5

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Table 2

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Table 4

Table 5

Metabolites of amino acids and their derivatives between fresh leaves, rolling leaves and sun-dried leaves"

序号 No.	保留时间 Retention time (min)	物质 Metabolites	YK10-1	YK10-2	YK10-3	类型 Type
1	0.79	天门冬氨酸二葡糖苷 Aspartic acid di-O-glucoside	567000±31000	2093333.33±218250.62	2993333.33±685298.00	上调 Up
2	1.24	L-亮氨酸 Aspartic acid di-O-glucoside	192000±27874.72	1002666.67±33842.77	990333.33±37287.17	上调 Up
3	1.14	L-(-)-酪氨酸L-(-)-Tyrosine	867000±70149.84	3750000±193132.078	4370000±167032.93	上调 Up
4	1.23	L-异亮氨酸L-Isoleucine	103766.67±17417.33	499333.33±21501.94	500000±15394.80	up
5	1.97	L-苯丙氨酸L-Phenylalanine	528000±39949.97	2470000±115325.63	2420000±115325.63	上调 Up
6	0.8	2-氨基己二酸 (L-高谷氨酸) 2-Aminoadipic acid (L-Homoglutamic acid)	330000±11357.82	1270000±155241.75	1420000±36055.51	上调 Up
7	0.84	缬氨酸Dl-Norvaline	8706666.67±428524.60	35466666.67±3720663.02	34933333.33±1871719.35	上调 Up
8	0.84	L-缬氨酸L-Valine	1953333.33±132035.35	7136666.67±828573.07	7423333.33±460470.77	上调 Up
9	0.71	L-酵母氨酸L-Saccharopine	181000±16522.71	773333.33±61174.61	667333.33±122964.76	上调 Up
10	0.68	L-(+)-赖氨酸L-(+)-Lysine	8870000±478852.80	31666666.67±642910.05	31300000±3122499.00	上调 Up
11	0.81	DL-高半胱氨酸DL-homocysteine	139333.33±11015.14	461333.33±47077.95	479333.33±25696.95	上调 Up
12	1.21	DL-多巴 3,4-Dihydroxy-DL-phenylalanine	4706.67±2740.55	14033.33±3362.04	15800±4253.23	上调 Up
13	0.65	蛋氨酸亚砜Methionine sulfoxide	152333.33±25658.01	639000±11135.53	507000±37643.06	上调 Up
14	0.72	L-谷氨酸 O-己糖苷 L-Glutamic acid O-glucoside	43066.67±3027.10	62033.33±13041.60	140333.33±5131.60	上调 Up
15	0.73	L-天冬酰胺L-Asparagine	126666.67±16258.33	420333.33±73493.76	407666.67±48675.80	上调 Up
16	1.24	S-甲基谷胱甘肽 S-(methyl) glutathione	6613.33±2116.63	8910±2098.36	19900±2128.38	上调 Up
17	0.74	同型丝氨酸L-Homoserine	12933.33±2967.04	26400±3751.00	38800±5350.70	上调 Up
18	0.68	L-组氨酸L-Histidine	1153333.33±76376.26	3736666.67±265015.72	3280000±360555.13	上调 Up
19	3.52	L-苯丙氨酸-L-苯丙氨酸Phe-Phe	42533.33±7850.05	144333.33±	109433.33±25162.74	上调 Up
20	0.77	L-(-)-胱氨酸L-(-)-Cystine	30866.67±11184.07	24200±7937.88	69866.67±14216.31	上调 Up
21	2.05	N′-甲酰基犬尿氨酸 N′-Formylkynurenine	705333.33±111540.73	612333.33±25890.80	1593333.33±56862.41	上调 Up
22	0.81	高胱氨酸L-Homocystine	1593333.33±3646.00	51466.67±17333.88	61366.67±9022.38	上调 Up
23	1.93	N-甘氨酰-L-亮氨酸 N-Glycyl-L-leucine	32666.67±2668.96	78633.33±5641.22	67333.33±6493.33	上调 Up
24	1.14	谷胱甘肽还原型 Glutathione reduced form	135733.33±92073.96	447000±303605.01	34066.67±12702.10	下调 Down
25	0.74	2-氨基异丁酸 2-Aminoisobutyric acid	64700000±4853864.44	23033333.33±1331665.62	25333333.33±2683902.63	下调 Down
26	2.43	N-乙酰基蛋氨酸 N-Acetylmethionine	147233.33±46476.48	111666.67±9712.53	66400±2066.40	下调 Down
27	0.99	L-茶氨酸L-Theanine	3820000±137356	4600000±558748.60	4490000±331763	上调 Up
28	0.77	γ-氨基丁酸γ-aminobutyric acid	1903333.33±295014.13	5510000±508625.60	5783333.33±508625.60	下调 Down

Table 5

Table 6

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样品 Tea sample	R²X	R²Y	Q²
YK10-1_vs_YK10-2	0.514	0.996	0.923
YK10-1_vs_YK10-3	0.547	0.997	0.937
YK10-2_vs_YK10-3	0.377	0.986	0.778

序号 No.	代谢物种类 Species of metabolites	总数 Amount	上调数量 Increase the number	下调数量 Reduce the number
1	脂质类Lipids	44	37	7
2	氨基酸及其衍生物Amino acid and its derivatives	27	23	4
3	黄酮类Flavone	25	19	6
4	核苷酸及其衍生物Nucleotide and its derivates	18	18	0
5	苯甲酸及其衍生物Benzoic acid and its derivatives	6	5	1
6	羟基肉桂酰衍生物Hydroxycinnamoyl derivatives	11	8	3
7	有机酸及其衍生物Organic acids	15	10	5
8	维生素Vitamins	4	2	2
9	儿茶素及其衍生物Catechin and its derivatives	1	1	0
10	花青素Anthocyanins	2	0	2
11	生物碱Alkaloids	1	1	0
12	胆碱类Cholines	3	2	1
13	酚胺Phenolamides	3	3	0
14	糖类Carbohydrates	2	2	0
15	萜类Terpenoids	1	0	1
16	吡啶Pyridine derivatives	1	1	0
17	奎宁酸及其衍生物Quinate and its derivatives	2	1	1
18	香豆素及其衍生物Coumarins	3	3	0
19	色胺及其衍生物Tryptamine derivatives	1	0	1

序号 No.	保留时间 Retention time (min)	代谢物 Metabolites	YK10-1	YK10-2	YK10-3	类型 Type
1	2.53	咖啡酰原儿茶酸 Protocatechuic acid O-glucoside	1050333.33±217647.27	6006666.67±575528.74	9486666.67±5827661.05	上调 Up
2	3.05	原儿茶醛Protocatechuic aldehyde	55100±7100	72833.33±3499.05	101300±7291.78	上调 Up
3	4.02	4-甲基儿茶酚4-Methylcatechol	15433.33±1767.30	24200±3143.25	24633.33±1703.92	上调 Up
4	2.38	二没食子儿茶素 Gallocatechin-gallocatechin	529666.67±85242.79	590000±43347.43	582000±71021.12	上调 Up
5	3.49	没食子儿茶素-儿茶素 Gallocatechin-catechin	6836.67±1506.40	4870±1384.59	2560±2470.49	下调 Down
6	2.3	原儿茶酸Protocatechuic acid	194000±15394.80	196666.67±26764.40	123900±41179.24	下调 Down
7	3.51	三儿茶素Catechin-catechin-catechin	3713333.33±385010.82	3863333.33±346458.27	3073333.33±213853.53	下调 Down
8	3.45	表儿茶素表阿夫儿茶精 Epicatechin-epiafzelechin	281000±22869.19	281666.67±54811.80	253000±26000	下调 Down
9	2.99	儿茶素Catechin	3960000±240208.24	3890000±208086.52	3650000±715821.21	下调 Down
10	2.44	没食子儿茶素(+)-Gallocatechin (GC)	1576666.67±63508.53	1376666.67±56862.41	1470000±78102.50	下调 Down
11	2.53	表没食子酸儿茶素 Epigallocatechin (EGC)	1500000±80000	1313333.33±50332.23	1400000±52915.03	下调 Down
12	3.18	表儿茶素L-Epicatechin	3306666.67±117189.31	3316666.67±138684.29	3146666.67±565803.26	下调 Down
13	3.28	表没食子酸儿茶素没食子酸酯Epigallate catechin gallate (EGCG)	3886666.67±210792.16	4140000±186815.42	3766666.67±213853.53	下调 Down
14	3.83	表儿茶素没食子酸 Epicatechin gallate (ECG)	15666666.67±808290.38	15900000±529150.26	15300000±608276.25	下调 Down

序号 No.	保留时间 Retention time (min)	物质 Metabolites	YK10-1	YK10-2	YK10-3	类型 Type
1	6.73	白杨素Chrysin	0±0.00	8880±782.37	14883.33±5566.49	上调 Up
2	330.2	丁香亭Syringetin	4490±1406.84	17366.67±782.37	33666.67±12483.72	上调 Up
3	5.08	槲皮素Quercetin	3046666.67±336501.61	10096666.67±505008.25	13400000±1907878.40	上调 Up
4	5.08	桑色素水合物Morin	2883333.33±361155.55	9653333.33±609289.20	12400000±1708800.75	上调 Up
5	4.58	二氢山奈酚Aromadedrin (Dihydrokaempferol)	978666.67±132673.79	3026666.67±206478.41	3600000±170880.07	上调 Up
6	5.69	山奈酚Kaempferol	3753333.33±480555.23	10603333.33±1347973.79	12543333.33±3894051.02	上调 Up
7	5.73	金圣草(黄)素Chrysoeriol	369666.67±37581.02	939666.67±42770.71	1196666.67±90737.72	上调 Up
8	5.81	异鼠李素Isorhamnetin	12453.33±3177.19	14000000±1708800.75	37733.33±5131.60	上调 Up
9	5.86	3,7-二氧-甲基槲皮素 Di-O-methylquercetin	136666.67±30022.21	249666.67±19655.36	353000±17435.60	上调 Up
10	6.98	金合欢素Acacetin	12046.67±2802.59	240000±20297.78	29700±2600	上调 Up
11	5.62	芹菜素Apigenin	504666.67±72507.47	829333.33±64002.60	1234666.67±390800.89	上调 Up
12	7.23	毡毛美洲茶素Velutin	1983.33±221.89	5923.33±2653.72	4543.33±320.36	上调 Up
13	7.17	华良姜素Kumatakenin	13866.67±1193.04	46366.67±27164.38	30333.33±1858.31	上调 Up
14	3.54	异鼠李素-3-O-新橙皮糖苷 Isorhamnetin 3-O-neohesperidoside	519666.67±198807.28	303333.33±51964.73	216333.33±62043.00	下调 Down
15	4.02	芹菜素 7-O-新橘皮糖苷 (野漆树苷) Apigenin 7-O-neohesperidoside (Rhoifolin)	15966666.67±3807011.08	10786666.67±920072.46	6953333.33±1276727.59	下调 Down

序号 No.	保留时间 Retention time (min)	物质 Metabolite	YK10-1	YK10-2	YK10-3	类型 Type
1	1.16	哌啶Piperidine	5443333.33±494098.51	19800000±1873499.4	18500000±781024.97	上调 Up
2	0.79	葫芦巴碱Trigonelline	164666.67±54993.94	318000±125391.39	308666.67±112669.13	上调 Up
3	0.78	甜菜碱Betaine	13733333.33±929157.32	17566666.67±1833939.29	24300000±12931743.89	上调 Up
4	2.64	茶碱Theophylline	6455066.67±578935.03	7286366.67±194360.4	6545566.67±1001309.43	上调 Up
5	1.45	大麦芽碱Hordenine	417333.33±33246.55	472333.33±66905.41	391000±34655.45	下调 Down
6	3.14	咖啡碱Caffeine	6883000±232758.27	6826466.67±97485.61	6241100±380824.22	下调 Down
7	4.64	异喹啉Isoquinoline	112366.67±24056.25	119466.67±28075.85	88566.67±3442.87	下调 Down
8	2.38	可可碱Theobromine	7260000±401497.2	6323333.33±230289.67	5460000±326955.65	下调 Down

Metabolic Changes in the Processing of Yunkang 10 Sun-Dried Green Tea based on Metabolomics

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