基于代谢组学筛选表征茶花鸡肌肉中特征风味的水溶性化合物

doi:10.3864/j.issn.0578-1752.2020.08.012

Abstract

Abstract:

【Objective】 The biomarkers in the chest and thigh muscles of Chahua Chicken before and after heat treatment were screened by using metabolomics combined with multivariate statistical methods, the aim of this study was to screen and characterize water-soluble compounds in different parts of Chahua Chicken, so as to provide scientific and theoretical basis for establishing evaluation standards of high-quality local chicken.【Method】Using liquid chromatography-quadrupole electrostatic field orbitrap mass spectrometry (LC-Q-Exactive-MS) and 300-day-old Yunnan native Chahua Chicken, the water-soluble compounds in the chest and thigh muscles before and after heat treatment were tested. Also, the spectrum was identified based on the network database Metlin, the relative content of each substance were calculated by the concentration of DL-o-Chlorophenylalanine, and the multivariate statistical analysis was used to detect substances. The relationship model between metabolite expression and sample groups was established by using orthogonal partial least squares discrimination analysis (OPLS-DA) to predict the sample category. The representative biomarkers was screened by S-plot to determine whether the model had over-fitting by observing the slope of the line of the 200 test plots of the permutations, and to discriminate the influence intensity and interpretation ability of each metabolite expression pattern on the classification and discrimination of each group of samples by the variable important of the projection (VIP). Therefore, the study achieved the purpose of screening markers: combining the calculation for Taste Activity Value (TAV) by relative content results with sensory evaluation results evaluate the characteristic flavor of Chahua Chicken muscles.【Result】The results showed that 106 species and 43 metabolites were detected in the chest and thigh muscles of Chahua Chicken before and after heat treatment in the positive ion mode and the negative ion mode, including amino acids, vitamins, nucleotides, organic acids, and fatty acids. Among them, creatine, carnosine, amino acid, and nucleotide were water-soluble compounds which had differences between before and after heat treatment of Chahua Chicken muscles, which could be used as a biomarker for the characteristics of the tribute. The TAV value results showed that the largest TAV value in the chest muscles was gamma-aminobutyric acid (TAV=23.00), and the followed by Histidine (TAV=1.14). The highest TAV value in the thigh muscles was gamma-aminobutyric acid (TAV=37.50), and the followed by Betaine (TAV=2.00). Through sensory evaluation, it was found that the overall taste of the chest muscles and thigh muscles was mainly sweet and umami.【Conclusion】After metabolomics analysis, the characteristic taste substances were obtained, and through the taste theory and sensory evaluation values, the study determined that the taste of the breast and leg muscles of Chahua chicken was mainly sweet and fresh meat, and the leg muscles were superior to the chest muscles.

Key words: Chahua Chicken, muscles, liquid chromatography-quadrupole electrostatic field orbitrap-mass spectrometry (LC- Q-Exactive-MS), Metabonomics, water-soluble compound

ZHAO WenHua,WANG GuiYing,XUN Wen,YU YuanRui,GE ChangRong,LIAO GuoZhou. Selection of Water-Soluble Compounds by Characteristic Flavor in Chahua Chicken Muscles Based on Metabolomics[J].Scientia Agricultura Sinica, 2020, 53(8): 1627-1642.

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Figures/Tables 11

Table 1

Fig. 1

Table 2

Metabolites of muscles samples in positive ion mode"

编号 No.	代谢物 Metabolite	编号 No.	代谢物 Metabolite	编号 No.	代谢物 Metabolite
1	肌酸 Creatine	501	氨基丁酸 gamma-Aminobutyric acid	1156	黄嘌呤 Xanthine
3	L-鹅肌肽 L-Anserine	517	泛酸 Pantothenic Acid	1213	α-亚麻酸 α-Linolenic Acid
11	肌酐 Creatinine	525	L-异亮氨酸 L-Isoleucine	1236	高香草基酸Homovanillic acid
13	左旋肉碱 L-Carnitine	546	磷酸胆碱Phosphocholine	1259	鸟嘌呤 Guanine
15	甜菜碱 Betaine	553	5'-甲基硫腺苷 5'-Methylthioadenosine	1296	聚氧乙烯辛烷基苯酚醚 9-OxoODE
37	乙酰肉碱 Acetylcarnitine	557	鞘氨醇 Sphingosine	1308	烟酰胺 Niacinamide
44	N-[4-(乙酰基氨基)苯基]-3-氧代丁酰胺 Hydrouracil	561	叔亮氨酸 Butyrylcarnitine	1363	二氢鞘氨醇 Sphinganine
65	亚油酸肉碱 Linoleylcarnitine	563	甘油磷酸胆碱 Glycerophosphocholine	1403	1-肉豆蔻酰-2-羟基-sn-甘油-3-磷酸胆碱LysoPC(14:0)
88	1-甲基组氨酸 1-Methylhistidine	579	L-精氨酸 L-Arginine	1431	丙氨酰 - 脯氨酸 Alanyl-Proline
108	次黄嘌呤 Hypoxanthine	591	1-十六烷酰-sn-甘油-3-磷酸乙醇胺 LysoPE (0:0/16:0)	1446	鸟嘌呤核苷 Guanosine
124	L-氯化棕榈酰肉碱 L-Palmitoylcarnitine	617	二十四碳六烯酸 tetracosahexaenoic acid	1458	苯乙酸 Phenylacetic acid
146	L-苯丙氨酸 L-Phenylalanine	618	L-丝氨酸 L-Serine	1472	硬脂酸酰胺 Stearamide
148	1-十五烷氧基-2-羟基-sn-甘油-3-磷酸胆碱 LysoPC(15:0)	625	N-十四碳酰-L-丝氨酸 Decanoylcarnitine	1513	腺苷酸 Adenosine monophosphate
219	异亮脯氨酸 Isoleucylproline	641	牛磺胆酸 Taurocholic acid	1518	油酸 Oleic Acid
228	L-组氨酸 L-Histidine	676	高肌肽 Homocarnosine	1558	N6-乙酰-L-赖氨酸 N6-Acetyl-L-lysine
233	乙酰胆碱 Acetylcholine	692	植物鞘氨醇 Phytosphingosine	1594	α-维生素E α-Tocopherol
237	N-乙酰-L-组氨酸 N-Acetyl-L-Histidine	694	DL-2-氨基正辛酸 DL-2-Aminooctanoic acid	1607	亚油酸 Linoleic acid
242	肌苷 Inosine	712	苯丙酮酸 Phenylpyruvic acid	1608	维生素 ARetinol
247	胆碱 Choline	733	L-脯氨酰基-L-苯丙氨酸L-prolyl-L-phenylalanine	1611	腺嘌呤Adenine
255	3-(硬脂酰氧基)-4-(三甲基铵基)丁酸酯 DL-Stearoylcarnitine	753	顺-13-二十二烯酰胺 13Z-Docosenamide	1633	油酸酰胺 Oleamide
282	L-谷氨酰胺 L-Glutamine	809	腺苷 Adenosine	1717	1-磷酸鞘氨醇 Sphingosine 1-phosphate
283	1-苯乙胺 1-Phenylethylamine	829	2-苯基乙酰胺 2-Phenylacetamide	1861	尿嘧啶 Uracil
288	谷胱甘肽 Glutathione	835	花生酸 Arachidonic Acid	1877	谷酰氨基-色氨酸 Glutaminyl-Tryptophan
302	十四左旋肉碱 Tetradecanoylcarnitine	897	L-谷氨酸 L-Glutamate	1929	烟酸 Niacin
303	L-色氨酸 L-Tryptophan	922	二十二酰胺 Docosanamide	1944	十八酰胺乙醇 Stearoylethanolamide
332	1-棕榈酰-2-羟基-sn-甘油-3-磷酸胆碱 LysoPC(16:0)	927	组织胺 Histamine	1991	脱氧胞苷 Deoxycytidine
357	β-丙氨酸 β-Alanine	936	肌苷酸 IMP	1994	苹果酸 Malic acid
386	磺熊去氧胆酸 Tauroursodeoxycholic acid	959	L-2-氨乙基丝氨酸磷酸酯 L-2-Aminoethyl seryl phosphate	2099	脱氧腺嘌呤核苷 Deoxyadenosine
414	辛酰基肉碱 L-Octanoylcarnitine	1005	L-苏氨酸 L-Threonine	2103	甘氨酰-L-亮氨酸 Glycyl-L-leucine
427	焦谷氨酸 Pyroglutamic acid	1017	吡哆胺 Pyridoxamine	2104	尿刊酸 Urocanic acid
446	L-酪氨酸 L-Tyrosine	1056	2-哌啶甲酸 DL-pipecolic acid	2138	脯氨酸甜菜碱 Prolinebetaine
459	牛磺酸 Taurine	1067	乙酰双氢硫辛酸酰胺 S-Acetyldihydrolipoamide	2143	尿酸 Uric acid
465	棕榈酸酰胺 Palmitic amide	1078	邻苯二甲酸二丁酯 Alpha-CEHC	2203	亚油酰胺 Linoleamide
467	L-脯氨酸 L-Proline	1092	3Α-羟基-7-氧代-5Β-胆烷酸 Nutriacholic acid	2291	维生素A酸 Retinoic acid
476	磷酸羟基丙酮酸Phosphohydroxypyruvic acid	1114	蛋氨酸亚矾 Methionine sulfoxide	2296	苯甲酸 Benzoic acid
2036	5-L-谷氨酰-L-半胱氨酸 γ-Glu-Cys

Table 2

Table 3

Metabolites of muscles samples in negative ion mode"

编号 No.	代谢物 Metabolite	编号 No.	代谢物 Metabolite
356	尿苷 Uridine	1102	L-谷氨酰基5-磷酸 L-Glutamyl 5-phosphate
499	十一烷二酸 Undecanedioic acid	1083	吲哚乳酸 Indolelactic acid
1762	血栓素 B3TXB3	1793	谷氨酰二肽 Glutamylphenylalanine
258	苏糖酸 Threonic acid	1019	谷氨酰丙氨酸 Glutamylalanine
1454	四氢皮质醇 Tetrahydrocortisol	318	γ-谷氨酰谷氨酸 Gamma Glutamylglutamic acid
533	四氢皮质甾酮 Tetrahydrocorticosterone	1778	二十烷二酸 Eicosanedioic acid
1323	硬脂肪酸 Stearic acid	1343	脱氧尿苷一磷酸 dUMP
875	鞘氨醇1-磷酸 Sphingosine 1-phosphate	1814	十二烷二酸 Dodecanedioic acid
139	景天庚酮糖 Sedoheptulose	198	D-葡萄糖-6-磷酸二钠 D-Glucose 6-phosphate
276	脯氨酰羟脯氨酸 Prolylhydroxyproline	482	脱氧胆酸 Deoxycholic acid
1658	苯乙酮酸 Phenylglyoxylic acid	1609	D-生物素 D-Biotin
1756	苯丁酰谷氨酰胺 Phenylbutyrylglutamine	1811	皮质醇 Cortisol
721	第三类前列腺素 PGE3	1514	顺-11-二十碳烯酸 cis-gondoic acid
1495	前列腺素 PGE2	1366	顺式-11-二十碳烯酸 cis-9-palmitoleic acid
1435	前列腺素 E1PGE1	454	胆酸 Cholic acid
1661	棕榈酸 Palmitic acid	1758	肾上腺酸 Adrenic Acid
1375	十八烷二酸 Octadecanedioic acid	769	(5Z,7E,11Z,14Z)-9-羟基-5,7,11,14-二十碳四烯酸 9-HETE
1705	肉豆蔻油酸 Myristoleic acid	534	（8R）-8-氢过氧基亚油酸 8R-HpODE
1821	1-羟基-2-肉豆蔻酰-sn-甘油-3-磷酸乙醇胺 LysoPE(0:0/14:0)	680	L-4-羟基色氨酸 4-Hydroxy-L-threonine
1109	L-脯氨酰-L-苯丙氨酸 L-prolyl-L-phenylalanine	1859	3-吲哚丙酸3-Indolepropionic acid
467	1-（1Z十六碳烯酰基）-sn-甘油-3-磷酸 LPA(P-16:0e/0:0)	1177	3-吲哚丁酸 3-Indolebutyric acid
1106	2棕榈酰甘油-3-磷酸 LPA(0:0/16:0)

Table 3

Fig. 2

Fig. 3

Table 4

Fig. 4

Table 5

Table 6

Taste substance and TAV value in muscles of Chahua chicken"

呈味 Taste	名称 Name	阈值^[19,20,21] Threshold (mmoL∙L^-1)	原料胸肉 Raw breast muscle		熟胸肉 Cooked breast muscle		原料腿肉 Raw thigh muscle		熟腿肉 Cooked thigh muscle
呈味 Taste	名称 Name	阈值^[19,20,21] Threshold (mmoL∙L^-1)	含量 (mmoL∙L^-1)	TAV	含量 (mmoL∙L^-1)	TAV	含量 (mmoL∙L^-1)	TAV	含量 (mmoL∙L^-1)	TAV
酸涩味Sour	苹果酸Malic acid	3.70	0.03±0.00a	0.01	0.02±0.00b	0.01	0.02±0.00b	0.01	0.02±0.00b	0.01
酸涩味Sour	γ-氨基丁酸 γ-Aminobutyric acid	0.02	0.24±0.05c	12.00	0.46±0.06b	23.00	0.70±0.10a	35.00	0.75±0.13a	37.50
	其他氨基酸OAA		0.27±0.05c		0.48±0.07b		0.72±0.10a		0.77±0.13a
苦味Bitter	色氨酸 L-Tryptophan	5.00	1.50±0.30a	0.30	1.18±0.19a	0.24	1.17±0.15a	0.23	0.32±0.03b	0.06
苦味Bitter	次黄嘌呤Hypoxanthine	9.00	1.21±0.14b	0.13	5.91±2.08a	0.66	2.47±0.02b	0.27	2.45±0.62b	0.27
苦味Bitter	异亮氨酸L-Isoleucine	11.00	0.52±0.15b	0.05	0.56±0.13b	0.05	0.80±0.22a	0.07	0.24±0.02c	0.02
苦味Bitter	酪氨酸L-Tyrosine	5.00	0.89±0.05	0.18	0.69±0.10	0.14	0.71±0.10	0.14	0.94±0.27	0.19
苦味Bitter	精氨酸L-Arginine	2.87	0.38±0.11a	0.13	0.21±0.03b	0.07	0.43±0.06a	0.15	0.39±0.11a	0.14
苦味Bitter	组氨酸L-Histidine	3.22	3.03±0.02ab	0.94	3.69±0.86a	1.14	2.46±0.62bc	0.76	1.82±0.54c	0.56
苦味Bitter	苯丙氨酸L-Phenylalanine	3.03	0.32±0.07a	0.11	0.43±0.08a	0.14	0.36±0.10a	0.12	0.11±0.06b	0.04
	苦味氨基酸BFA		7.89±0.43b		12.14±2.00a		8.77±0.88b		6.27±1.25b
甜味Sweet	脯氨酸L-Proline	26.00	0.67±0.18b	0.03	0.82±0.12b	0.03	0.81±0.11b	0.03	1.54±0.50a	0.06
甜味Sweet	丝氨酸L-Serine	30.00	0.08±0.01b	0.00	0.02±0.00c	0.00	0.66±0.06a	0.02	0.08±0.02b	0.00
甜味Sweet	苏氨酸L-Threonine	40.00	0.12±0.03	0.00	0.11±0.03	0.00	0.11±0.01	0.00	0.12±0.05	0.00
甜味Sweet	D-葡萄糖6-磷酸 D-Glucose 6-phosphate	48.00	9.21±2.29a	0.19	9.42±1.96a	0.20	1.65±0.19b	0.03	1.59±0.52b	0.03
甜味Sweet	甜菜碱Betaine	20.01	4.58±1.02c	0.23	5.90±0.89c	0.29	14.03±3.66b	0.70	39.98±6.38a	2.00
	甜味氨基酸SFA		14.74±2.1b		16.36±1.37b		17.34±3.81b		43.30±7.22a
鲜味Umami	腺甘酸 Adenosine monophosphate	4.00	0.01±0.00	0.00	0.01±0.00	0.00	0.01±0.00	0.00	0.01±0.00	0.00
鲜味Umami	谷氨酸盐L-Glutamate	3.40	0.03±0.01b	0.01	0.07±0.01a	0.02	0.08±0.02a	0.02	0.04±0.02b	0.01
鲜味Umami	肌苷酸IMP	0.68	0.11±0.02ab	0.16	0.07±0.02b	0.10	0.12±0.03a	0.18	0.08±0.03ab	0.12
	鲜味氨基酸UFA		0.14±0.02b		0.15±0.02b		0.22±0.05a		0.13±0.05b

Table 6

Fig. 5

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编号 No.	胸肌Breast muscle		编号 No.	腿肌Thigh muscle
编号 No.	Var ID (Primary)	VIP pred	编号 No.	Var ID (Primary)	VIP pred
1	肌酸 Creatine	7.01967	1	肌酸 Creatine	6.56079
2	L-鹅肌肽 L-Anserine	4.57094	2	肌酐 Creatinine	3.23688
3	甜菜碱 Betaine	2.84563	3	左旋肉碱 L-Carnitine	3.22422
4	1-甲基组氨酸1-Methylhistidine	1.83296	4	甜菜碱 Betaine	3.20225
5	N-[4-(乙酰基氨基)苯基]-3-氧代丁酰胺Hydrouracil	1.64535	5	L-鹅肌肽 L-Anserine	3.00584
6	1-棕榈酰-2-羟基-sn-甘油-3-磷酸胆碱 LysoPC(16:0)	1.48292	6	3-(硬脂酰氧基)-4-(三甲基铵基)丁酸酯DL-Stearoylcarnitine	1.81764
7	肌苷 Inosine	1.39204	7	L-谷氨酰胺 L-Glutamine	1.63342
8	L-色氨酸 L-Tryptophan	1.37077	8	次黄嘌呤 Hypoxanthine	1.52174
9	5'-甲基硫腺苷5'-Methylthioadenosine	1.23907	9	N-乙酰-L-组氨酸N-Acetyl-L-Histidine	1.44579
10	N-乙酰-L-组氨酸N-Acetyl-L-Histidine	1.22247	10	L-组氨酸 L-Histidine	1.31488
11	异亮脯氨酸 Isoleucylproline	1.20120	11	乙酰肉碱Acetylcarnitine	1.22599
12	肌酐 Creatinine	1.11686	12	1-棕榈酰-2-羟基-sn-甘油-3-磷酸胆碱LysoPC(16:0)	1.14697

编号 No.	胸肌Breast muscle		编号 No.	腿肌Thigh muscle
编号 No.	Var ID (Primary)	VIP pred	编号 No.	Var ID (Primary)	VIP pred
1	苏糖酸 Threonic acid	2.77134	1	L-4-羟基色氨酸4-Hydroxy-L-threonine	2.76987
2	景天庚酮糖 Sedoheptulose	2.69728	2	L-谷氨酰5-磷酸酯 L-Glutamyl 5-phosphate	2.14000
3	L-4-羟基色氨酸4-Hydroxy-L-threonine	2.38874	3	景天庚酮糖 Sedoheptulose	2.04383
4	尿苷 Uridine	2.21336	4	苏糖酸 Threonic acid	1.94052
5	D-葡萄糖-6-磷酸二钠D-Glucose 6-phosphate	1.51438	5	3-吲哚丁酸 3-Indolebutyric acid	1.58979
6	γ-谷氨酰谷氨酸 Gamma Glutamylglutamic acid	1.31314	6	D-葡萄糖 6-磷酸二钠D-Glucose 6-phosphate	1.57744
7	1-（1Z十六碳烯酰基）-sn-甘油-3-磷酸LPA(P-16:0e/0:0)	1.30657	7	前列腺素 PGE2	1.3055
8	吲哚乳酸 Indolelactic acid	1.12537	8	脯氨酰羟脯氨酸Prolylhydroxyproline	1.27561
9	脯氨酰羟脯氨酸Prolylhydroxyproline	1.08437	9	γ-谷氨酰谷氨酸 Gamma Glutamylglutamic acid	1.19071
10	十一烷二酸 Undecanedioic acid	1.01416	10	1-（1Z十六碳烯酰基）-sn-甘油-3-磷酸LPA(P-16:0e/0:0)	1.01964
			11	谷氨酰丙氨酸 Glutamylalanine	1.00139

Selection of Water-Soluble Compounds by Characteristic Flavor in Chahua Chicken Muscles Based on Metabolomics

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时间 Time (min)	流速 Flow rate (mL∙min^-1)	A (%)	B (%)
0	0.3	100	0
0	0.3	100	0
1.5	0.3	80	20
9.5	0.3	0	100
14.5	0.3	0	100
14.6	0.3	100	0
18	0.3	100	0

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