分心木水提物的抗氧化性分析及组分鉴定

doi:10.13304/j.nykjdb.2023.0863

摘要/Abstract

摘要：

为研究核桃分心木（diaphragma juglandis fructus， DJF）水提物最佳提取工艺及抗氧化性，选取液料比、超声时间及超声温度为影响因素，以1，1-二苯基-2-三硝基苯肼（1，1-diphenyl-2-trinitrophenylhydrazine， DPPH）自由基清除率为响应值，进行单因素试验，采用响应面法优化超声辅助提取DJF水提物的工艺条件，并对分心木水提物进行组分分析及抗氧化性研究。结果表明，通过Box-Benhnken模型确定DJF水提物的最佳工艺为液料比92 mL∶1 g、超声温度58 ℃、超声时间19 min，所得DPPH自由基清除率为68.95%；通过高效液相色谱法（high performance liquid chromatography，HPLC）对DJF水提物进行组分分析发现，DJF水提物中多酚含量为0.28 mg·mL^-1，多糖含量为0.33 mg·mL^-1，主要活性成分含量为槲皮素245 μg·mg^-1、没食子酸18 μg·mg^-1、半乳糖98 μg·mg^-1；DJF水提物抗氧化能力试验表明，DJF水提物2，2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸（2，2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid，ABTS）自由基清除率及DPPH自由基清除率总体低于维生素C（vitamin C， Vc），总还原力大于Vc，表明其具有较好的抗氧化能力，是一种潜在的抗氧化食品优质原料。以上研究结果为DJF活性成分提取提供了数据支撑，为进一步探究DJF功能成分提供了理论依据。

关键词: 核桃分心木, 水提物, 响应面, 抗氧化, 液相色谱

Abstract:

In order to study the optimal extraction process and oxidation resistance of the aqueous extract of diaphragma juglandis fructus （DJF）， the liquid-material ratio， ultrasonic time and ultrasonic temperature were selected as the influencing factors， and the free radical clearance rate of 1，1-diphenyl-2-trinitrophenylhydrazine （DPPH） was used as the response value. The response surface method was used to optimize the process conditions of ultrasonic-assisted extraction of DJF water extracts， and the component analysis and oxidation resistance of the water extracts from distracted wood were studied. The results showed that the optimal process for determining the aqueous extract of DJF by the Box-Benhnken model was as follows： liquid-material ratio of 92 mL∶?1 g， ultrasonic temperature of 58 °C， ultrasonic time of 19 min， and the obtained DPPH free radical clearance rate was 68.95%. High performance liquid chromatography （HPLC） showed that the content of polyphenols in the DJF water extracts was 0.28 mg·mL^-1， and the contents of polysaccharides were 0.33 mg·mL^-1， and the main active components were quercetin 245 μg·mg^-1， gallic acid 18 μg·mg^-1 and galactose 98 μg·mg^-1. The results of DJF water extract antioxidant capacity test showed that the free radical clearance rate of DPPH and free radical clearance rate of 2，2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid （ABTS） of DJF water extract were generally lower than those of vitamin C （Vc）， and the total reducing power was greater than that of Vc， indicating that it had better antioxidant capacity and it was a potential antioxidant food quality raw material. Above results provided data support for the extraction of DJF active ingredients， and provided a theoretical basis for further exploration of DJF functional components.

Key words: diaphragma juglandis fructus, water extracts, response surface, antioxidant, liquid chromat-ography

中图分类号:

S184

苗春霖, 许欢欢, 贾紫毅, 何爱民, 吉洋洋, 牟德华, 高山. 分心木水提物的抗氧化性分析及组分鉴定[J]. 中国农业科技导报, 2025, 27(5): 182-192.

Chunlin MIAO, Huanhuan XU, Ziyi JIA, Aimin HE, Yangyang JI, Dehua MOU, Shan GAO. Oxidation Resistance Analysis and Component Identification of Distracted Wood Water Extracts[J]. Journal of Agricultural Science and Technology, 2025, 27(5): 182-192.

图/表 12

参考文献 30

1	LIU R X, ZHAO Z Y, DAI S J,et al.. Identification and quantification of bioactive compounds in Diaphragma juglandis fructus by UHPLC-Q-orbitrap HRMS and UHPLC-MS/MS [J]. J. Agric. Food Chem.,2019,67(13):3811-3825.
2	CHEN Y, SUN X Q, FANG L T,et al.. Optimization of ultrasound-assisted extraction of polyphenols from Ilex latifolia using response surface methodology and evaluation of their antioxidant activity [J/OL]. Molecules, 2022,27(13):3999 [2023-10-23]. .
3	MENG Q R, WANG Y Q, CHEN F,et al..Polysaccharides from Diaphragma juglandis fructus:extraction optimization,antitumor,and immune-enhancement effects [J].Int.J.Biol.Macromol.,2018,115:835-845.
4	洪茜茜,叶永丽,张银志,等.核桃分心木化学成分及功能活性研究进展[J].食品研究与开发,2021,42(7):194-202.
	HONG Q Q, YE Y L, ZHANG Y Z, et al.. Research progress on chemical constituents and functional activities of Diaphragma juglandis fructus [J]. Food Res.Dev.,2021,42(7):194-202.
5	WEI N, WANG X S, WU Y Y, et al.. Comparative study on anti-inflammatory effect of polysaccharides from vinegar-baked Radix bupleuri using different methods [J].ACS Omega,2023,8(32):29253-29261.
6	HUANG X, LI S, DING R,et al.. Antitumor effects of polysaccharides from medicinal lower plants:a review [J/OL].Int. J. Biol. Macromol., 2023,252:126313 [2023-10-23]..
7	YAMAGUCHI T, YASUI K, FUJII S,et al.. Lentilactobacillus hilgardii H-50 strongly inhibits lipopolysaccharide-induced inflammatory responses in mouse splenocytes via its specific surface layer proteins [J/OL].J.Appl.Microbiol.,2023,134(3):lxad021 [2023-10-23]..
8	蒋红芝,覃微,李海冬.核桃分心木单宁提取及其对羊毛的染色工艺[J].毛纺科技,2022,50(7):31-36.
	JIANG H Z, QIN W, LI H D. Study on extraction technology of tannin and dyeing technology of wool from walnut-wood [J].Wool Text. J.,2022,50(7):31-36.
9	WANG S, WU H, ZHANG X,et al.. Preparation of nano-selenium from chestnut polysaccharide and characterization of its antioxidant activity [J/OL].Front. Nutr.,2022,9:1054601[2023-10-23]. .
10	刘昕茹. 荞麦麸皮多酚的强化提取及其复合物的性质研究[D]. 天津:天津科技大学, 2022.
	LIU X R. Study on the intensified extraction of polyphenols from buckwheat bran and properties of their complex [D]. Tianjin: Tianjin University of Science and Technology, 2022.
11	熊芳芳,马佳琪,李敏.猴头菇菌渣中多糖提取工艺的优化及其体外抗氧化活性研究[J].饲料研究,2023,46(18):92-96.
	XIONG F F, MA J Q, LI M.Study on optimization of extraction process of polysaccharide from Hericium erinaceus residues and in vitro antioxidant activity [J]. Feed Res.,2023,46(18):92-96.
12	陈肖,曾维艳,王翔,等.草果中不同多酚类体外抗氧化活性研究[J].食品与发酵科技,2023,59(3):43-46, 75.
	CHEN X, ZENG W Y, WANG X, et al.. Study on in vitro antioxidant activities of polyphenols from Amomum tsaoko [J].Food Ferment. Sci. Technol., 2023,59(3):43-46, 75.
13	辛二娜. 黄粉虫抗氧化多糖的制备、纯化、分离及组成和活性研究[D]. 晋中:山西农业大学, 2022.
	XIN E N. Studies on the preparation,purification,isolation,composition and activity of the antioxidant Tenebrio molitor polysaccharide [D]. Jinzhong:Shanxi Agricultural University, 2022.
14	MIAO J N, SHI W, ZHANG J Q,et al.. Response surface methodology for the fermentation of polysaccharides from Auricularia auricula using Trichoderma viride and their antioxidant activities [J].Int.J.Biol.Macromol.,2020,155:393-402.
15	冯李院,侯洪波,邢江艳,等.响应面微波辅助酸法优化提取南瓜皮果胶工艺研究[J].包装与食品机械,2023,41(4):39-45.
	FENG L Y, HOU H B, XING J Y, et al.. Study on optimized extraction of pectin from pumpkin peel by response surface combined with microwave-assisted acid method [J].Packag.Food Mach., 2023,41(4):39-45.
16	张红娇. 紫苏多糖的分离纯化、结构表征及应用研究[D]. 太原:中北大学, 2022.
	ZHANG H J. Isolation, purification, structural characterization and application of perilla polysaccharides [D]. Taiyuan:North University of China, 2022.
17	张韬,陈启航,韦海秋,等.加拿大一枝黄花多糖提取工艺优化及抗氧化活性研究[J].浙江海洋大学学报(自然科学版),2022,41(3):272-278.
	ZHANG T, CHEN Q H, WEI H Q, et al.. Study on extraction process and antioxidant activity of Solidago canadensis L. [J]. J.Zhejiang Ocean. Univ. (Nat.Sci.), 2022,41(3):272-278.
18	GAO J Y, HU D Y, SHEN Y, et al.. Optimization of ultrasonic-assisted polysaccharide extraction from Hyperici Perforati Herba using response surface methodology and assessment of its antioxidant activity [J]. Int. J. Biol. Macromol., 2023,225:255-265.
19	ZHENG J W, ZHANG X X, HERRERA-BALANDRANO D D, et al.. Extraction optimization of Arctium lappa L. polysaccharides by box-behnken response surface design and their antioxidant capacity [J/OL]. Starch-Stärke, 2022,74(9-10):2100305 [2023-10-23]. .
20	LIU J, BAI J, SHAO C,et al.. Optimization of ultrasound-assisted aqueous two-phase extraction of polysaccharides from seabuckthorn fruits using response methodology,physicochemical characterization and bioactivities [J]. J. Sci. Food Agric.,2023,103(6):3168-3183.
21	李湘,文瑶.响应面法优化荞麦花黄酮提取工艺研究[J].云南化工,2023,50(8):38-41.
	LI X, WEN Y.Response surface methodology to optimize the extraction process of buckwheat flower flavonoids [J].Yunnan Chem. Technol.,2023,50(8):38-41.
22	CHEN X Q, JIA X D, YANG S, et al.. Optimization of ultrasonic-assisted extraction of flavonoids,polysaccharides,and eleutherosides from Acanthopanax senticosus using response surface methodology in development of health wine [J/OL]. LWT, 2022,165:113725 [2023-10-23]. .
23	BU X Y, XU Y Q, ZHAO M M, et al.. Simultaneous extraction of polysaccharides and polyphenols from blackcurrant fruits:comparison between response surface methodology and artificial neural networks [J/OL]. Ind. Crops Prod., 2021,170:113682 [2023-10-23]. .
24	ZHANG H, LI H, ZHANG Z, et al.. Optimization of ultrasound-assisted extraction of polysaccharides from perilla seed meal by response surface methodology:characterization and in vitro antioxidant activities [J]. J. Food Sci., 2021,86(2):306-318.
25	刘佳, 周亚梅, 张潇丹, 等. 响应面法优化牛膝多糖的微波法提取工艺研究[J]. 农产品加工, 2023(7): 27-31.
	LIU J, ZHOU Y M, ZHANG X D, et al.. Optmization of ultrasound-assisted extraction of tricholomamatsutake mycelium polysaccharide using response surface methology [J]. Agric. Prod. Process., 2023(7): 27-31.
26	FENG Y N, ZHANG X F. Polysaccharide extracted from Huperzia serrata using response surface methodology and its biological activity [J]. Int. J. Biol. Macromol.,2020,157:267-275.
27	侯登勇,张建,何颖,等.核桃分心木水提物抗疲劳作用研究[J].中国食物与营养,2021,27(9):73-77.
	HOU D Y, ZHANG J, HE Y,et al.. Anti-fatigue activity study of aqueous extract from diaphragma juglandis fructus in walnut [J].Food Nutr. China,2021,27(9):73-77.
28	吴修利,徐雷,谢英,等.超声波辅助提取油莎豆粕水溶性多糖及其抗氧化性[J].食品研究与开发,2022,43(16):81-87.
	WU X L, XU L, XIE Y,et al..Ultrasonic-assisted extraction and antioxidant activity of water-soluble polysaccharides from Cyperus esculentus meals [J]. Food Res. Dev.,2022,43(16):81-87.
29	ZHANG X X, NI Z J, ZHANG F, et al.. Physicochemical and antioxidant properties of Lycium barbarum seed dreg polysaccharides prepared by continuous extraction [J/OL].Food Chem., 2022,14:100282 [2023-10-23]. .
30	MEDLEJ M K, CHERRI B, NASSER G, et al.. Optimization of polysaccharides extraction from a wild species of Ornithogalum combining ultrasound and maceration and their anti-oxidant properties [J]. Int. J. Biol. Macromol., 2020,161:958-968.

化合物 Compound	时间 Time/min	乙腈 Acetonitrile/%
多酚 Polyphenol	0~2	5~15
	2~5	15~20
	5~20	20
	20~24	20~50
	24~26	50~100
	26~28	100~50
	28~30	50~20
	30~40	20~5
多糖 Polysaccharide	0~28	17
	28~40	17~30
	40~45	30
	45	30~17
	45~50	17

化合物 Compound	时间 Time/min	乙腈 Acetonitrile/%
多酚 Polyphenol	0~2	5~15
	2~5	15~20
	5~20	20
	20~24	20~50
	24~26	50~100
	26~28	100~50
	28~30	50~20
	30~40	20~5
多糖 Polysaccharide	0~28	17
	28~40	17~30
	40~45	30
	45	30~17
	45~50	17

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型 Model	679.00	9	75.56	42.76	<0.000 1
A：超声时间 Ultrasound time	18.07	1	18.07	10.23	0.015 1^*
B：超声温度 Ultrasonic temperature	82.27	1	82.27	46.56	0.000 2^*
C：液料比 Liquid-to-materialratio	25.16	1	25.16	14.24	0.007 0^**
AB	39.33	1	39.33	22.26	0.002 2
AC	0.06	1	0.06	0.03	0.864 0
BC	4.08	1	4.08	2.31	0.172 5
A²	71.36	1	71.36	40.39	0.000 4
B²	293.59	1	293.59	166.16	<0.000 1
C²	98.77	1	98.77	55.90	0.000 1
残差Residuals	12.37	7	1.77
失拟误差Misfit error	9.50	3	3.17	4.41	0.092 9
纯误差Pure error	2.87	4	0.72
总和Sum	692.37	16

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型 Model	679.00	9	75.56	42.76	<0.000 1
A：超声时间 Ultrasound time	18.07	1	18.07	10.23	0.015 1^*
B：超声温度 Ultrasonic temperature	82.27	1	82.27	46.56	0.000 2^*
C：液料比 Liquid-to-materialratio	25.16	1	25.16	14.24	0.007 0^**
AB	39.33	1	39.33	22.26	0.002 2
AC	0.06	1	0.06	0.03	0.864 0
BC	4.08	1	4.08	2.31	0.172 5
A²	71.36	1	71.36	40.39	0.000 4
B²	293.59	1	293.59	166.16	<0.000 1
C²	98.77	1	98.77	55.90	0.000 1
残差Residuals	12.37	7	1.77
失拟误差Misfit error	9.50	3	3.17	4.41	0.092 9
纯误差Pure error	2.87	4	0.72
总和Sum	692.37	16

标品 Standard product	线性回归方程 Linear regression equation	决定系数 R²
没食子酸Gallic acid	y=3 676 602.5x+12 429 215.1	0.999 2
绿原酸Chlorogenic acid	y=9 780 671.5x+25 070.5	0.999 5
儿茶素Catechins	y=18 976 942.0x-1 578 523.2	0.999 8
芦丁Rutin	y=8 195 692.0x-242 787.6	0.999 3
槲皮素Quercetin	y=5 324 594.0x+1 967 979.0	0.999 3
甘露糖Mannose	y=19 806.7+14 563 801.1x	0.999 9
半乳糖Galactose	y=-7 354.5+11 882 050.3x	0.999 8
葡萄糖Glucose	y=48 476.0+17 936 410.2x	0.999 9
阿拉伯糖Arabinose	y=203 770.4+23 019 451.9x	0.999 8