Screening of Antioxidant Sites of Dendrobium aurantiacum Rchb. f. var. denneanum (Kerr.) Z. H. Tsi. Leaves Based on Grey Relational Degree Analysis

doi:10.3969/j.issn.1000-2561.2020.08.026

Abstract

Abstract:

The leaves of Dendrobium aurantiacum Rchb. var. f. denneanum (Kerr.) Z. H. Tsi. were coarse-extracted with different concentrations of ethanol. And then with different polar solvents, the petroleum ether layer, dichloromethane layer, ethyl acetate layer, n-butanol layer and water layer were obtained from 60% ethanol crude extract, whose antioxidant activity was higher. Those layer obtained were assayed with the ultra performance liquid chromatography (UPLC) and UV-spectrophotometric method. With DPPH antioxidant activity as an index, the grey correlation degree correlation analysis was used to screen the main antioxidant activity. The highest amount of flavonoids, 24.89%, was in the n-butyl alcohol layer, and the highest amount of polysaccharide, 0.23%, layer was in the water layer. Each extraction layer had antioxidant activity, and the free radical clearance ability in order from strong to weak was water layer, n-butanol layer, ethyl acetate layer, dichloromethane layer, petroleum ether layer. Besides, the n-butanol layer and water layer were selected as the main antioxidant active parts. And the antioxidant activity was the result of the synergistic action of flavonoids and polysaccharides, which could provide some guidance for further extraction of antioxidant active compounds and development of natural antioxidants.

Key words: leaves of Dendrobium aurantiacum Rchb. var. f. denneanum (Kerr.) Z. H. Tsi., gray relational degree, antioxidant active site, total flavonoids, polysaccharide

CLC Number:

S567
R284

RUAN Peihua,DENG Hong,FU Yongmei,ZHANG Shu,GUO Cai’e,HUANG Shubin,LI Zhichao. Screening of Antioxidant Sites of Dendrobium aurantiacum Rchb. f. var. denneanum (Kerr.) Z. H. Tsi. Leaves Based on Grey Relational Degree Analysis[J]. Chinese Journal of Tropical Crops, 2020, 41(8): 1700-1707.

Figures/Tables 8

References 22

[1]	吴英详, 余雪芳, 詹兴堆, 等. 千层金叶片醇提物抗氧化活性的研究[J]. 热带作物学报, 2019,40(7):1367-1372.
[2]	邱珊莲, 林宝妹, 张少平, 等. 嘉宝果嫩叶提取物不同极性部位抗氧化及抑制α-葡萄糖苷酶活性研究[J]. 热带作物学报, 2019,40(7):1373-1378.
[3]	Zehiroglu C Ozturk Sarikaya S B. The importance of antioxidants and place in today’s scientific and technological studies[J]. Journal of Food Science and Technology, 2019,56(11):4757-4774. URL PMID
[4]	四川省食品药品监督管理局. 四川省中药材标准: 2010年版[M]. 成都: 四川科学技术出版社, 2011: 414.
[5]	徐悦, 刘宏程, 李鲜. 石斛化学成分、指纹图谱及药理活性研究进展[J]. 中国中医药信息杂志, 2019,26(2):129-132.
[6]	Tang H, Zhao T, Sheng Y, et al. Dendrobium officinale Kimura et Migo: a review on its ethnopharmacology, phytochemistry, and industrialization[J]. Evidence-Based Complementary and Alternative Medicine, 2017:7436259. DOI URL PMID
[7]	周桂芬, 吕圭源. 铁皮石斛不同部位黄酮碳苷类成分及清除DPPH自由基能力比较研究[J]. 中国中药杂志, 2012,37(11):1536-1540.
[8]	李艳萍, 李海燕, 纪晓婉, 等. 金钗石斛叶中总黄酮的提取分离及体外抗阿尔茨海默病活性研究[J]. 中国药房, 2018,29(3):330-333.
[9]	Zhang Y, Zhang L, Liu J, et al. Dendrobium officinale leaves as a new antioxidant source[J]. Journal of Functional Foods, 2017,37:400-415.
[10]	罗颖懿, 李运容, 雷胄熙, 等. 铁皮石斛共性黄酮类成分新西兰牡荆苷Ⅱ的体外抗氧化与诱导HepG2细胞凋亡的作用[J]. 中国实验方剂学杂志, 2019,25(1):43-50.
[11]	高海立, 郁吉锋, 黄路瑶, 等. 铁皮石斛叶总黄酮的大孔树脂纯化工艺及抗氧化活性[J]. 浙江理工大学学报(自然科学版), 2019,41(3):380-386.
[12]	李艳萍, 李海燕, 纪晓婉, 等. 金钗石斛叶中总黄酮的提取分离及体外抗阿尔茨海默病活性研究[J]. 中国药房, 2018,29(3):330-333.
[13]	黄丹丹, 陈欢欢, 黎梅桂, 等. 金钗石斛黄酮苷的含量测定及抗氧化活性研究[J]. 中药新药与临床药理, 2017,28(1):73-77.
[14]	李燕, 赵天明, 黄丽荣, 等. 响应面法优化响铃草总黄酮提取工艺及体外抗氧化活性研究[J]. 食品研究与开发, 2019,49(17):79-84.
[15]	李芳, 魏云, 陈艳杰. 铁皮石斛茎、叶、花中黄酮含量及其体外抗氧化活性研究[J]. 中医学报, 2019,34(5):1020-1023.
[16]	黄萍. 夹江叠鞘石斛入选国家石斛林木种质资源库[EB/ OL].[ 2018-05-02]. http://local.newssc.org/system/20180502/002411999. htm.
[17]	国家药典委员会. 中华人民共和国药典: 2015年版一部[M]. 北京: 中国医药科技出版社, 2015.
[18]	汪洋, 彭潇, 何燕玲, 等. 基于灰色关联分析探索艾纳香非挥发性部位的抗炎活性谱效关系[J]. 中国实验方剂学杂志, 2019,25(4):180-186.
[19]	唐静月, 颜美秋, 齐芳芳, 等. 铁皮石斛花总黄酮提取工艺优化及体外抗氧化活性研究[J]. 浙江中医药大学学报, 2017,41(3):235-242.
[20]	高海立, 郁吉锋, 黄路瑶, 等. 铁皮石斛叶总黄酮的大孔树脂纯化工艺及抗氧化活性[J]. 浙江理工大学学报(自然科学版) 2019,41(3):380-386.
[21]	Singh C B, Devi M C, Thokchom D S, et al. Phytochemical screening, estimation of total phenols, total flavonoids and determination of antioxidant activity in the methanol extract of Dendrobium denudans D. Don stems[J]. Journal of Pharmacognosy and Phytochemistry, 2015,4(4):6-11.
[22]	陈佳江, 郭力, 许莉, 等. 叠鞘石斛茎叶花HPLC- DAD-ELSD对比研究[J]. 时珍国医国药, 2013,24(3):574-575.

提取液 Extracting solution	芹菜素-6,8-二-c-葡萄苷含量 Apigenin-6,8-2-c-glucoside content/%	异夏佛塔苷含量 Isaphoroside content/%	芦丁含量 Rutin content/%	总黄酮含量 Total flavonoid content/%	多糖含量 Polysaccharide content/%	IC₅₀
水	0.100	0.052	0.086	1.00	2.44	1.348
20%乙醇	0.098	0.058	0.110	1.25	0.77	0.402
40%乙醇	0.120	0.080	0.180	2.20	0.73	0.355
60%乙醇	0.120	0.080	0.180	2.39	0.16	0.344
80%乙醇	0.130	0.045	0.150	2.00	-	0.452
95%乙醇	0.015	-	-	0.50	-	1.664

提取液 Extracting solution	芹菜素-6,8-二-c-葡萄苷含量 Apigenin-6,8-2-c-glucoside content/%	异夏佛塔苷含量 Isaphoroside content/%	芦丁含量 Rutin content/%	总黄酮含量 Total flavonoid content/%	多糖含量 Polysaccharide content/%	IC₅₀
水	0.100	0.052	0.086	1.00	2.44	1.348
20%乙醇	0.098	0.058	0.110	1.25	0.77	0.402
40%乙醇	0.120	0.080	0.180	2.20	0.73	0.355
60%乙醇	0.120	0.080	0.180	2.39	0.16	0.344
80%乙醇	0.130	0.045	0.150	2.00	-	0.452
95%乙醇	0.015	-	-	0.50	-	1.664

提取液 Extracting solution	芹菜素-6,8-二-c-葡萄苷含量 Apigenin-6,8-2-c-glucoside content/%	异夏佛塔苷含量 Isaphoroside content/%	芦丁含量 Rutin content/%	总黄酮含量 Total flavonoid content/%	多糖含量 Polysaccharide content/%	IC₅₀	提取率 Yield/%
石油醚层	0.05	0.03	-	0.99	-	0.700	1.49
二氯甲烷层	0.04	-	-	1.20	-	0.203	1.72
乙酸乙酯层	-	0.02	0.20	13.61	-	0.049	2.43
正丁醇层	0.90	0.86	2.50	24.89	0.08	<0.050	4.59
水层	0.32	0.19	0.26	3.34	0.23	<0.050	14.66

提取液 Extracting solution	芹菜素-6,8-二-c-葡萄苷含量 Apigenin-6,8-2-c-glucoside content/%	异夏佛塔苷含量 Isaphoroside content/%	芦丁含量 Rutin content/%	总黄酮含量 Total flavonoid content/%	多糖含量 Polysaccharide content/%	IC₅₀	提取率 Yield/%
石油醚层	0.05	0.03	-	0.99	-	0.700	1.49
二氯甲烷层	0.04	-	-	1.20	-	0.203	1.72
乙酸乙酯层	-	0.02	0.20	13.61	-	0.049	2.43
正丁醇层	0.90	0.86	2.50	24.89	0.08	<0.050	4.59
水层	0.32	0.19	0.26	3.34	0.23	<0.050	14.66

峰号 Peak No.	石油醚层 Petroleum ether layer	二氯甲烷层 Dichloromethane layer	乙酸乙酯层 Ethyl acetate layer	正丁醇层 N-butyl alcohol layer	水层 water layer
1	20 288	-	19 142	451 026	1 829 283
2	-	-	19 468	304 048	83 766
3	39 050	6 627	322 155	1 626 220	3 990 622
4	64 016	57 638	-	3 836 059	4 892 452
5	36 220	34 246	-	2 480 093	2 843 122
6	68 579	29 756	-	3 235 331	6 050 317
7	3 576	-	34 757	340 894	456 412
8	3 061	-	64 601	727 199	777 343
9	8 090	-	-	262 153	1 072 841
10	26 352	-	34 138	3 049 601	2 126 672
11	-	-	20 746	610 472	316 038
12	-	-	10 025	341 189	354 702
13	-	-	7 820	1 297 389	778 742
14	-	-	9 159	864 973	461 043
15	38 055	-	25 398	3 017 511	1 139 313
16	-	-	-	1 098 217	330 867
17	-	-	23 310	762 339	-
18	-	-	195 945	4 585 479	1 533 071
峰号 Peak No.	石油醚层 Petroleum ether layer	二氯甲烷层 Dichloromethane layer	乙酸乙酯层 Ethyl acetate layer	正丁醇层 N-butyl alcohol layer	水层 water layer
19	-	-	160 648	1 376 165	271 900
20	33 409	-	797 481	907 727	-
21	-	-	48 201	1 178 442	317 954
22	-	-	16 331	573 850	196 917
23	-	-	21 421	289 492	-
24	339 682	-	3 135 440	31 547 334	4 102 194
25	99 090	-	4 732 584	4 529 962	458 601
26	-	20 020	23 051	1 236 235	700 938
27	13 500	109 631	50 931	2 074 721	897 543
28	4 772	16 306	1 889 280	772 590	175 063
29	162 035	582 503	7 898 656	-	109 971
30	-	434 974	388 982	-	-
总黄酮峰面积	875 471	1 234 063	19 911 060	68 785 578	29 462 186
总黄酮含量/%	0.99	1.20	13.61	24.89	3.34