Extraction and identification of α-glucosidase-inhibitory components from Phellinus baumii

doi:10.3969/j.issn.1004-1524.2022.05.09

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (5): 949-958.DOI: 10.3969/j.issn.1004-1524.2022.05.09

• Animal Science • Previous Articles Next Articles

Extraction and identification of α-glucosidase-inhibitory components from Phellinus baumii

ZHENG Meiyu¹(), WANG Lu¹^,², LIU Zhe¹, ZHANG Wenjuan¹^,², GAO Pu¹^,³, LU Shengmin¹^,^*()

1. Key Laboratory of Fruit and Vegetables Postharvest and Processing Technology Research of Zhejiang Province, Key Laboratory of Postharvest Handling of Fruits, Ministry of Agriculture and Rural Affairs, Institute of Food Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. College of Food and Pharmaceutical Sciences, Zhejiang Ocean University, Zhoushan 316022, Zhejiang, China
3. College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China

Received:2021-01-06 Online:2022-05-25 Published:2022-06-06
Contact: LU Shengmin

Abstract

Abstract:

To screen and extract the α-glucosidase-inhibitory components in Phellinus baumii,and to identify the constituents in the extracts, the four technological parameters were optimized by α-glucosidase-inhibitory rate and total flavonoid content, and the constituents were identified by UHPLC-Q-Exactive Orbitrap MS. The extracts by 60% ethanol exhibited significant inhibitory activity. The optimized extraction by orthogonal test were achieved using 60% of ethanol volume fraction at 80 ℃ for 3 h with a solid-to-liquid ratio of 1:30 for two times, whose flavonoid extraction ratio reached 159.3 mg·g^-1. Forty five components were identified by LC-MS, among which phenolic substances were in the majority. The extracts with high flavonoids content and high α-glucosidase-inhibitory activity was obtained by optimized technology, containing abundant polyphenols.

Key words: Phellinus baumii, α-glucosidase, extract, component identification

CLC Number:

ZHENG Meiyu, WANG Lu, LIU Zhe, ZHANG Wenjuan, GAO Pu, LU Shengmin. Extraction and identification of α-glucosidase-inhibitory components from Phellinus baumii[J]. Acta Agriculturae Zhejiangensis, 2022, 34(5): 949-958.

Figures/Tables 6

References 25

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指标 Index	稀释倍数 Dilution ratio	乙醇体积分数Ethanol volume fraction/%
指标 Index	稀释倍数 Dilution ratio	0	20	40	60	80
α-葡萄糖苷酶	10	93.3±0.3	100±0.6	100±0.1	100±0.3	100±0.8
α-glucosidase	100	8.1±2.1	91.7±3.3	100±0.3	100±0.1	100±0.1
	200	0	0	40.7±2.2	100±1.7	100±1.8
	250				99.4±0.1	53.9±1.9
	400				78.1±2.3	41.5±3.5
	500				45.6±4.4	22.7±5.9
α-淀粉酶α-amylase	0	17.1±3.3	37.9±5.7	49.9±5.2	69.2±4.2	62.4±3.3

指标 Index	稀释倍数 Dilution ratio	乙醇体积分数Ethanol volume fraction/%
指标 Index	稀释倍数 Dilution ratio	0	20	40	60	80
α-葡萄糖苷酶	10	93.3±0.3	100±0.6	100±0.1	100±0.3	100±0.8
α-glucosidase	100	8.1±2.1	91.7±3.3	100±0.3	100±0.1	100±0.1
	200	0	0	40.7±2.2	100±1.7	100±1.8
	250				99.4±0.1	53.9±1.9
	400				78.1±2.3	41.5±3.5
	500				45.6±4.4	22.7±5.9
α-淀粉酶α-amylase	0	17.1±3.3	37.9±5.7	49.9±5.2	69.2±4.2	62.4±3.3

试验号 Test number	乙醇体积分数 Ethanol volume fraction/%	温度 Temperature/℃	提取时间 Extracting time/h	料液比 Solid-liquid ratio	黄酮提取率 Flavonoids extraction ratio/(mg·g^-1)	抑制率 Inhibitory ratio/%
1	40	60	1	1:10	13.9±0.7	48.4±1.2
2	40	70	2	1:20	40.0±1.2	90.9±2.2
3	40	80	3	1:30	70.0±3.2	89.7±2.4
4	60	60	2	1:30	46.7±4.5	90.9±1.6
5	60	70	3	1:10	24.5±1.6	80.5±0.7
6	60	80	1	1:20	46.9±1.4	89.2±1.2
7	80	60	3	1:20	26.5±0.7	95.5±0.5
8	80	70	1	1:30	29.6±0.5	85.0±1.1
9	80	80	2	1:10	19.6±0.2	39.8±2.1
K1j	123.9	87.0	90.4	57.9
K2j	118.1	94.1	106.3	113.4
K3j	75.6	136.5	120.9	146.3
R	48.3	49.5	30.5	88.3
K1'j	229.0	234.8	222.6	168.7
K2'j	260.6	256.4	221.5	275.6
K3'j	220.4	218.7	265.8	265.6
R'	40.2	37.7	44.2	96.9

试验号 Test number	乙醇体积分数 Ethanol volume fraction/%	温度 Temperature/℃	提取时间 Extracting time/h	料液比 Solid-liquid ratio	黄酮提取率 Flavonoids extraction ratio/(mg·g^-1)	抑制率 Inhibitory ratio/%
1	40	60	1	1:10	13.9±0.7	48.4±1.2
2	40	70	2	1:20	40.0±1.2	90.9±2.2
3	40	80	3	1:30	70.0±3.2	89.7±2.4
4	60	60	2	1:30	46.7±4.5	90.9±1.6
5	60	70	3	1:10	24.5±1.6	80.5±0.7
6	60	80	1	1:20	46.9±1.4	89.2±1.2
7	80	60	3	1:20	26.5±0.7	95.5±0.5
8	80	70	1	1:30	29.6±0.5	85.0±1.1
9	80	80	2	1:10	19.6±0.2	39.8±2.1
K1j	123.9	87.0	90.4	57.9
K2j	118.1	94.1	106.3	113.4
K3j	75.6	136.5	120.9	146.3
R	48.3	49.5	30.5	88.3
K1'j	229.0	234.8	222.6	168.7
K2'j	260.6	256.4	221.5	275.6
K3'j	220.4	218.7	265.8	265.6
R'	40.2	37.7	44.2	96.9

指标 Index	稀释倍数 Dilution ratio	第1次 The first	第2次 The second	第3次 The third
提取率Extraction ratio/(mg·g^-1)		86.4±0.9	72.9±2.4	17.5±1.8
抑制率Inhibitory ratio/%	100	99.1±0.5	99.5±0.1	99.9±0.2
	200	93.1±1.9	95.4±0.8	55.5±2.5
	400	49.0±3.8	55.5±2.8	57.5±3.2

Extraction and identification of α-glucosidase-inhibitory components from Phellinus baumii

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序号 No.	t_R	质荷比 m/z	误差 Error (×10^-6)	分子式 Formula	裂解碎片 Pyrolysis fragments MS/MS (+/-)^*	鉴定结果 Identified results	类别 Category
1	2.33	197	-2.27	C₉H₁₀O₅	MS²[197]: 109, 151, 153, 101(+)	丁香酸^[23] Syringic acid	酚类 Phenols
2	2.67	153	2.88	C₇H₆O₄	MS²[153]: 109(+)	原儿茶酸^[18] Protocatechuic acid	酚类 Phenols
3	3.40	139 137	-0.58 -7.57	C₇H₆O₃	MS²[139]: 111, 93, 139, 120(+) MS²[137]: 108(+)	原儿茶醛^[18] Protocatechualdehyde	酚类 Phenols
4	3.73	177	-4.02	C₉H₆O₄	MS²[177]:133, 105(+)	7,8-二羟基香豆素 7,8-dyhydroxy-couma-rin	酚类 Phenols
5	3.79	181 179	-0.16 -4.42	C₉H₈O₄	MS²[181]: 163, 121, 93, 135(+) MS²[179]: 135(-)	咖啡酸^[18] Caffeic acid	酚类 Phenols
6	3.83	191	-2.68	C₁₀H₈O₄	MS²[191]: 147, 121, 165(-)	东莨菪亭^[18] Scopoletin	酚类 Phenols
7	3.85	163	-5.01	C₉H₈O₃	MS²[163]: 119(-)	间羟基肉桂酸 M-hydroxycinnamic acid	酚类 Phenols
8	4.14	287	0.66	C₂₀H₁₂O₈	MS²[287]: 269, 203, 272, 137, 241(-)	二氢山柰酚^[18] Dihydrokaempferol	黄酮 Flavonoids
9	4.22	289	1.28	C₁₃H₆O₈	MS²[289]: 217, 133, 189, 161, 177(-)	PhelligridinJ^[22]	酚类 Phenols
10	4.32	193	-0.65	C₁₀H₈O₄	MS²[193]: 119, 147, 153(+) MS²[191]: 147(-)	7,8-二羟基-4-甲基香豆素 7,8-dihydroxy-4-methyl-coumarin	酚类 Phenols
11	4.51	163	-4.77	C₉H₈O₃	MS²[163]: 119(+)	对香豆酸 p-coumaric acid	酚类 Phenols
12	4.75	179 177	-1.74 -4.50	C₁₀H₁₀O₃	MS²[179]: 143, 115, 161, 105, 133(+) MS²[177]:135, 134(-)	3,4-二羟基苯亚甲基丙酮 3,4-dihydroxy benzylidene acetone	酚类 Phenols
13	5.16	421	0.95	C₂₃H₁₈O₈	MS²[421]: 217, 159, 174, 241(+)	Interfungin B^[18]	酚类 Phenols
14	5.53	465	-1.44	C₂₅H₂₀O₉	MS²[465]: 163, 147, 135, 175, 379, 137(+)	Davallialactone^[18]	酚类 phenols
15	5.84	317 315	-1.23 1.25	C₁₆H₁₂O₇	MS²[317]: 163, 247, 135, 145, 201(+) MS²[315]: 159, 243, 111, 299(-)	鼠李素^[18] Rhamnetin	黄酮 Flavonoids
16	5.91	977 975	-1.40 1.76	C₅₂H₃₂O₂₀	MS²[977]: 203, 245, 175(+) MS²[975]: 243, 199, 175, 403(-)	Phelligridimer A^[18]	酚类 Phenols
17	5.99	465	-1.37	C₂₅H₂₀O₉	MS²[465]: 163, 135, 145, 203, 147(+)	Davallialactone^[18]	酚类 Phenols
18	6.06	381	-1.72	C₂₀H₁₂O₈	MS²[381]: 163, 203, 135, 147, 205(+)	Phelligridin D^[23]	酚类 Phenols
19	6.16	975	-0.44	C₅₂H₃₂O₂₀	MS²[975]: 243, 199, 175, 201, 215(+)	Phelligridimer A^[18]	酚类 Phenols
20	6.50	469 467	-1.90 1.35	C₂₄H₂₀O₁₀	MS²[469]: 163, 135, 183, 145(+) MS²[467]: 159, 135, 175, 201, 243(-)	茶叶花宁 B^[25] Apocynin B	酚类 Phenols
21	6.65	285	2.17	C₁₅H₁₀O₆	MS²[285]: 205, 223, 267(+)	山奈素^[18] Kaempferide	黄酮 Flavonoids
22	6.72	251	-1.71	C₁₃H₁₄O₅	MS²[251]: 163, 135, 145(+)	橘霉素 Notalin	酚类 Phenols
23	6.80	453 451	-1.15 1.14	C₂₄H₂₀O₉	MS²[453]: 163, 379, 147, 175, 137, 351(+) MS²[451]: 243, 159, 247, 175, 199(+)	Phellibaumin E^[18]	酚类 Phenols
24	6.98	453 451	-0.79 0.74	C₂₄H₂₀O₉	MS²[453]: 163, 379, 147, 175, 137, 351(+) MS²[451]: 159, 243, 247, 135, 175, 219(-)	Phellibaumin E^[18]	酚类 Phenols
25	7.22	465	-0.99	C₂₅H₂₀O₉	MS²[465]: 163, 379, 175, 135, 137(+)	Davallialactone^[18]	酚类 Phenols
26	7.44	461	1.18	C₂₅H₁₈O₉	MS²[461]: 159, 241, 257, 333, 213(-)	Inoscavin A^[18]	酚类 Phenols
27	7.62	421	0.66	C₂₃H₁₈O₈	MS²[421]: 159, 217, 175, 115269.1756 (60), 174(-)	Interfungin B^[18]	酚类 Phenols
28	7.71	593	0.24	C₃₂H₁₈O₁₂	MS²[593]: 311, 135, 159, 265, 431(-)	Phelligrindin G^[23]	酚类 Phenols
29	7.88	381	-0.90	C₂₀H₁₂O₈	MS²[381]: 163, 135, 145, 147, 219(+)	桑色素 D^[18] Morin D	黄酮 Flavonoids
30	7.96	379	-0.96	C₂₂H₁₈O₆	MS²[379]: 261, 169, 197, 141(+)	桑黄黄酮A^[20] Kuwanon A	黄酮 Flavonoids
31	8.00	461	0.64	C₂₅H₁₈O₉	MS²[461]: 159, 377, 257, 241, 214(+)	Inoscavin A^[18]	酚类 Phenols
32	8.07	147	-0.86	C₉H₆O₂	MS²[147]: 91, 119, 105(+)	香豆素^[24] coumarin	酚类 Phenols
33	8.45	381	0.07	C₂₀H₁₂O₈	MS²[381]: 163, 135, 145, 219(+)	桑色素 D^[19] Morin D	黄酮 Flavonoids
34	8.89	377	0.75	C₂₁H₁₄O₇	MS²[377]: 241, 159, 145, 333, 305, 197(-)	Phellifuropyranone A^[18]	黄酮 Flavonoids
35	9.31	421 419	-0.86 0.95	C₂₃H₁₆O₈	MS²[421]: 357, 163, 137, 385, 329(+) MS²[419]:283, 257, 241, 332, 223(-)	Inoscavin C^[22]	黄酮 Flavonoids
36	11.11	345	0.12	C₁₈H₁₆O₇	MS²[345]: 315, 297, 197, 169(+)	3,3',7-三甲氧基-4',5-二羟基黄酮 3,3',7-trimethoxy-4', 5-dihydroxy flavonoid	黄酮 Flavonoids
37	11.25	345	-1.01	C₁₈H₁₆O₇	MS²[345]: 91, 315,201, 154, 105, 99, 55, 67, 79(+)	3, 3', 7-三甲氧基-4',5-二羟基黄酮 3,3'7-trimethoxy-4',5-dihydroxy flavonoid	黄酮 Flavonoids
38	13.21	363	-6.68	C₂₁H₃₂O₅	MS²[363]: 295, 135, 179, 195, 277(-)	Phellinignincisterol A^[25]	甾体 Sterides
39	13.23	291	1.35	C₁₈H₂₈O₃	MS²[291]: 113, 139), 121, 185, 125, 247, 197(-)	Phellilins C^[24]	倍半萜 Sesquiter- penes
40	13.35	291	1.04	C₁₈H₂₈O₃	MS²[291]: 185, 121, 125, 197(-)	Phellilins C^[24]	倍半萜 Sesquiter- penes
41	13.87	315	-1.75	C₂₀H₂₆O₃	MS²[315]: 161, 269, 297, 147, 121(+)	咖啡豆醇 Kahweol	三萜类 Triterpenes
42	14.15	315	-1.46	C₂₀H₂₆O₃	MS²[315]: 161, 269, 147, 297, 121(+)	咖啡豆醇 Kahweol	三萜类 triterpenes
43	15.65	299	0.12	C₂₀H₂₈O₂	MS²[299]: 184, 185, 183, 119, 120, 185(-)	脱氢枞酸 Dehydroabietic acid	二萜 Diterpenes
44	15.72	455	-2.50	C₃₀H₄₈O₃	MS²[455]: 139, 339, 411, 379(-)	桦木酸 Betulinic acid	三萜类 Triterpenes
45	16.28	455	-0.28	C₃₀H₄₈O₃	MS²[455]: 339, 367, 411(-)	熊果酸 Ursolic acid	三萜类 Triterpenes

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