超临界CO2萃取红麸杨果实中黄颜木素及其体外抗氧化活性研究

doi:10.3969/j.issn.0253-2417.2020.03.014

摘要/Abstract

摘要：

在超微粉碎机上将红麸杨果实于0~10℃条件下粉碎，并用石油醚（60~90℃）脱脂，然后采用超临界CO₂萃取果粉中黄颜木素。研究了适宜的夹带剂，并通过正交试验优化超临界CO₂萃取工艺条件，考察了所得黄颜木素的体外总抗氧化能力及其对DPPH·、超氧阴离子自由基（O₂^-·）与羟基自由基（·OH）的清除效果。结果显示：红麸杨果实经超微粉碎后粒径可达0.1~1 μm，细胞破壁率达95%以上，适宜的夹带剂为体积分数70%乙醇，萃取最佳条件为：果粉与夹带剂固液比5：3（g：mL）、萃取压力30 MPa、萃取温度55℃、萃取时间2 h、分离釜Ⅰ的温度55℃、压力10 MPa、分离釜Ⅱ的温度35℃、压力5 MPa，此条件下黄颜木素萃取物得率为0.46%，纯度为80.26%。该纯度黄颜木素具有较强的体外抗氧化活性，对DPPH·、O₂^-·和·OH的半数清除浓度（IC₅₀）分别为6.93、59.63和9.05 mg/L；与同浓度的V_C比较，对DPPH·和O₂^-·的清除率接近V_C，而对·OH的清除能力和总抗氧化能力优于V_C。

关键词: 红麸杨果实, 黄颜木素, 超微粉碎, 超临界萃取, HPLC

Abstract:

Rhus punjabensis var. sinica Rehd. fruit was superfinely crushed at 0-10℃ and degreased with petroleum ether (60-90℃), and then the dihydrofisetin was extracted from fruit powder by supercritical CO₂ extraction. The suitable entrainer was chosen, and the extraction conditions were optimized by orthogonal experiment. The total antioxidant capacity of the obtained dihydrofisetin and its scavenging effects on DPPH·, superoxide anion radicals (O₂^-·) and hydroxyl radicals (·OH) were investigated. The results showed that the particle size of R. punjabensis var. sinica fruit powder could be up to 0.1-1 μm after ultrafine-pulverization and cell-wall breaking ratio was over 95%; the suitable entrainer was ethanol with volume fraction of 70%, the optimal conditions for the extraction were solid-liquid ratio 5:3(g:mL), extraction pressure 30 MPa, extraction temperature 55℃, extraction time 2 h, separating axeⅠtemperature 55℃ and pressure 10 MPa, separating axeⅡtemperature 40℃ and pressure 5 MPa. Under these conditions, the yield of dihydrofisetin was 0.46%, the purity was 80.26%. The dihydrofisetin had strong antioxidant activity, and the half-clearance concentration(IC₅₀) of DPPH·, O₂^-· and ·OH were 6.93, 59.63 and 9.05 mg/L, respectively. The radical scavenging abilities on DPPH· and O₂^-· were close to those of V_C. The scavenging capacity of ·OH and total antioxidant capacity were better than those of V_C.

Key words: Rhus punjabensis var. sinica Rehd. fruit, dihydrofisetin, ultrafine-pulverization, supercritical extraction, HPLC

中图分类号:

TQ35

董爱文,李维新,王欢,邹亚珂. 超临界CO₂萃取红麸杨果实中黄颜木素及其体外抗氧化活性研究[J]. 林产化学与工业, 2020, 40(3): 108-114.

Aiwen DONG,Weixin LI,Huan WANG,Yake ZOU. Extraction of Dihydrofisetin from Rhus punjabensis var. sinica Rehd. Fruit by Supercritical CO₂ and Its Antioxidant Activity in vitro[J]. Chemistry and Industry of Forest Products, 2020, 40(3): 108-114.

图/表 4

图1

表1

表2

图2

参考文献 19

1	中国科学院中国植物志编辑委员会. 中国植物志[M]. 北京: 科学出版社, 1999: 67- 69.
	Editorial Board of 《Flora Republicae Popularis Sinicac》, CAS . Flora Republicae Popularis Sinicae[M]. Beijing: China Science Publishing & Media Ltd., 1999: 67- 69.
2	江苏新医学院. 中药大辞典[M]. 上海: 上海人民出版社, 1997: 1817- 1819.
	Jiangsu New Medical College . Chinese Medicine Dictionary[M]. Shanghai: Shanghai People's Publishing House, 1997: 1817- 1819.
3	ZHANG M , ZHANG J P , JI H T , et al. Effect of six flavolloids on proliferation of hepatic stellate cells in vitro[J]. Acta Pharmacologica Sinica, 2000, 21 (3): 253- 256.
4	董爱文, 姚姝凤, 赵虹桥, 等. 酶辅助提取协同大孔树脂纯化红麸杨果实黄颜木素[J]. 林产化学与工业, 2016, 36 (6): 100- 106. doi: 10.3969/j.issn.0253-2417.2016.06.016
	DONG A W , YAO S F , ZHAO H Q , et al. Enzyme-assisted extraction and macroporous resin purification of fustin from Rhus punjabensis var. Sinica Rehd.Fruit[J]. Chemistry and Industry of Forest Products, 2016, 36 (6): 100- 106. doi: 10.3969/j.issn.0253-2417.2016.06.016
5	赵军.盐肤木抗肿瘤活性成分的研究[D].天津:天津大学, 2006.
	ZHAO J.Studies on the antitumor constituents of Rhus chinensis[D]. Tianjin: Tianjin University, 2006.
6	曹冉冉, 高嘉屿, 刘华清, 等. 皂角刺中二氢黄酮醇类化合物及其细胞毒活性研究[J]. 中草药, 2016, 47 (5): 707- 711.
	CAO R R , GAO J Y , LIU H Q , et al. Flavanonol compounds from thorns of Gleditsie Spina and their cytotoxicity[J]. Chinese Traditional and Herbal Drugs, 2016, 47 (5): 707- 711.
7	张珉, 张俊平, 王杰松, 等. 黄颜木素对HSC-T6细胞增殖和胶原合成的影响[J]. 第二军医大学学报, 1999, 20 (5): 304- 305. doi: 10.3321/j.issn:0258-879X.1999.05.011
	ZHANG M , ZHANG J P , WANG J S , et al. Effects of fisetin on proliferation and collagen synthesis of HSC-T6 cells[J]. Academic Journal of Second Military Medical University, 1999, 20 (5): 304- 305. doi: 10.3321/j.issn:0258-879X.1999.05.011
8	陈虹霞, 王成章. 漆树黄酮类化合物研究进展[J]. 天然产物研究与开发, 2013, 25 (12): 1752- 1758.
	CHEN H X , WANG C Z . Progress in flavonoid compounds from Rhus verniciflua Stokes[J]. Natural Product Research and Development, 2013, 25 (12): 1752- 1758.
9	郭丽冰, 孙丽丽, 邓琪, 等. 降香中黄酮类化学成分研究(Ⅱ)[J]. 中药材, 2010, 33 (6): 915- 917.
	GUO L B , SUN L L , DENG Q , et al. Studies on the flavonoids from lignum Dalbergiae odoriferae(Ⅱ)[J]. Journal of Chinese Medicinal Materials, 2010, 33 (6): 915- 917.
10	赵军, 崔承彬, 蔡兵, 等. 国产盐肤木属植物的研究进展[J]. 解放军医学学报, 2006, 22 (1): 48- 51.
	ZHAO J , CUI C B , CAI B , et al. The research progress of domestic plants of the genus poison sumac[J]. The People's Liberation Army Medical Journals, 2006, 22 (1): 48- 51.
11	陈海霞, 肖顺汉. 皂角刺抗肿瘤作用有效成分研究进展[J]. 四川生理科学杂志, 2010, 32 (1): 35- 37. doi: 10.3969/j.issn.1671-3885.2010.01.016
	CHEN H X , XIAO S H . Study progress on antitumor active ingredients from Spina gleditsiae[J]. Sichuan Physiological Science Magazine, 2010, 32 (1): 35- 37. doi: 10.3969/j.issn.1671-3885.2010.01.016
12	张蓓蓓. 植物黄酮类化合物的研究[J]. 科技视界, 2018, (23): 155- 157.
	ZHANG B B . Study on plant flavonoids[J]. Science & Technology Vision, 2018, (23): 155- 157.
13	万新焕, 陈新梅, 马山, 等. 黄酮类化合物提取新方法的应用[J]. 中草药, 2019, 50 (15): 3691- 3699. doi: 10.7501/j.issn.0253-2670.2019.15.027
	WAN X H , CHEN X M , MA S , et al. Applications of new methods in extraction of flavonoids from Chinese materia medica[J]. Chinese Traditional and Herbal Drugs, 2019, 50 (15): 3691- 3699. doi: 10.7501/j.issn.0253-2670.2019.15.027
14	金建忠, 哈成勇. 超临界CO₂萃取落叶松木材挥发油的工艺研究[J]. 林产化学与工业, 2009, 29 (5): 91- 94. doi: 10.3321/j.issn:0253-2417.2009.05.017
	JIN J Z , HA C Y . Study on extraction of volatile oil from larch wood by supercritical carbon dioxide[J]. Chemistry and Industry of Forest Products, 2009, 29 (5): 91- 94. doi: 10.3321/j.issn:0253-2417.2009.05.017
15	容凡丁. 超临界萃取分离精细化工技术的应用探讨[J]. 生物化工, 2018, 4 (6): 154- 158. doi: 10.3969/j.issn.2096-0387.2018.06.045
	RONG F D . Application of supercritical fluid extraction for fine chemical separation technology[J]. Biological Chemical Engineering, 2018, 4 (6): 154- 158. doi: 10.3969/j.issn.2096-0387.2018.06.045
16	董爱文.紫花地丁中芹菜素的提取与分离纯化技术研究[D].长沙:湖南农业大学, 2008.
	DONG A W.Study on extraction separation and purification of apigenin from Viola yedoensis Makino[D]. Changsha: Hunan Agricultural University, 2008.
17	钟方丽, 王文姣, 王晓林, 等. 刺玫果总黄酮的双水相萃取工艺及其抗氧化能力[J]. 林产化学与工业, 2016, 36 (4): 64- 72. doi: 10.3969/j.issn.0253-2417.2016.04.009
	ZHONG F L , WANG W J , WANG X L , et al. Aqueous two-phase extraction of total flavonoids in fruit of Rosa davurica Pall.and its antioxidant ability[J]. Chemistry and Industry of Forest Products, 2016, 36 (4): 64- 72. doi: 10.3969/j.issn.0253-2417.2016.04.009
18	李雪, 王宏伟, 郑东然, 等. 杜香叶多糖的提取及抗氧化和抗肿瘤活性研究[J]. 林产化学与工业, 2017, 37 (5): 133- 138. doi: 10.3969/j.issn.0253-2417.2017.05.018
	LI X , WANG H W , ZHENG D R , et al. Extraction and antioxidant, antiproliferative activities of polysaccharide from Ledum palustre L.leaves[J]. Chemistry and Industry of Forest Products, 2017, 37 (5): 133- 138. doi: 10.3969/j.issn.0253-2417.2017.05.018
19	曹会凯.山楂中黄酮类化合物的提取及成分分析[D].秦皇岛:河北科技师范学院, 2013.
	CAO H K.Extraction and component analysis of flavonoids from Crataegus pinnatifida Bge[D]. Qinghuangdao: Hebei Normal University of Science and Technology, 2013.

序号 No	A 果粉与夹带剂固液比(g:mL) solid-liquid ratio of bloom and entrainer	B 萃取压力/MPa pressure	C 萃取温度/℃ temperature	D 萃取时间/h time	得率/% yield
1	5:2	25	35	1	0.304
2	5:2	30	45	2	0.485
3	5:2	40	55	3	0.428
4	5:3	25	45	3	0.436
5	5:3	30	55	1	0.587
6	5:3	40	35	2	0.559
7	5:4	25	55	2	0.486
8	5:4	30	35	3	0.508
9	5:4	40	45	1	0.315
k₁	0.406	0.510	0.457	0.402
k₂	0.527	0.527	0.412	0.510
k₃	0.436	0.434	0.500	0.457
R	0.121	0.093	0.088	0.108

方差来源 sources of variation	平方和 ss	自由度 df	均方 mean square	F值 F value	P值 P value
A	0.024	2	0.012	2.000	>0.05
B	0.023	2	0.012	1.917	>0.05
C	0.012	2	0.006	1.000	>0.05
D	0.017	2	0.009	1.417	>0.05
误差error	0.010	2	0.005	F_0.05(2, 2)=19

[1]	陈丹丹, 郭雪峰, 赵蕾, 邵思越. HPLC法同时检测刚竹属竹叶中4种黄酮碳苷的含量[J]. 林产化学与工业, 2019, 39(5): 108-114.
[2]	董爱文, 姚姝凤, 赵虹桥, 成江, 伍陵. 酶辅助提取协同大孔树脂纯化红麸杨果实黄颜木素[J]. 林产化学与工业, 2016, 36(6): 100-106.
[3]	刘普, 李小方, 高嘉屿, 尹卫平, 邓瑞雪. 流苏花精油的超临界CO₂萃取及抗菌活性研究[J]. 林产化学与工业, 2015, 35(6): 126-132.
[4]	黎海龙, 李成国, 秦光辉, 刘健, 甘礼惠, 谢茹胜, 龙敏南. HPLC结合MALDI-TOF-MS分析木聚糖水解产物[J]. 林产化学与工业, 2015, 35(5): 10-14.
[5]	何源峰;王成章;陆榕. HPLC-MS法表征坝漆酚类化合物的结构[J]. 林产化学与工业, 2012, 32(6): 47-51.
[6]	冯国东;周永红;郭小昕;胡立红;. 微波液化木材动力学研究及产物分析[J]. 林产化学与工业, 2010, 30(3): 66-72.
[7]	孙新超;杨波;许源;杨健;赵榆林;王晶;王翠敏. 紫苏籽的超临界CO₂萃取及β-环糊精包合一体化技术研究[J]. 林产化学与工业, 2010, 30(3): 73-77.
[8]	李正;杭悦宇;周义锋. 中国主要产区何首乌以二苯乙烯苷含量为指标的种质评价[J]. 林产化学与工业, 2003, 23(04): 37-41.
[9]	钱学仁;李坚. 木材亚-超临界动态萃取过程解析[J]. 林产化学与工业, 2000, 20(3): 6-12.

超临界CO₂萃取红麸杨果实中黄颜木素及其体外抗氧化活性研究

Extraction of Dihydrofisetin from Rhus punjabensis var. sinica Rehd. Fruit by Supercritical CO₂ and Its Antioxidant Activity in vitro

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 4

参考文献 19

相关文章 9

编辑推荐

Metrics

本文评价

关于本刊

投稿指南

在线期刊

相关单位

联系我们