绵马贯众多酚的分离纯化及其抗氧化活性研究

doi:10.3969/j.issn.0253-2417.2019.02.012

摘要/Abstract

摘要：

采用70%乙醇提取绵马贯众多酚，并通过吸附实验考察了7种大孔树脂对绵马贯众多酚粗提物吸附率和解吸率的影响，发现HPD100树脂对绵马贯众多酚的纯化效果较好；进一步分析了样品质量浓度、上样速度、解吸乙醇体积分数、洗脱速度及样品pH值对HPD100树脂纯化绵马贯众多酚的影响，得出最佳纯化工艺条件为：样品溶液质量浓度为2.00 g/L，pH值为5，上样速度为1 mL/min，解吸乙醇体积分数为70%及洗脱速度为2 mL/min。该工艺条件下，绵马贯众多酚纯度由纯化前的27.64%提高到纯化后的54.00%(得率为6.46%)，纯化效果显著。纯化后绵马贯众多酚抗氧化活性显著增加，对DPPH自由基(DPPH·)及羟基自由基(·OH)的IC₅₀值分别为0.01和0.49 g/L(纯化前为0.05和1.79 g/L)，0.04 g/L时对DPPH·的清除率为90.82%，与Vc相当；质量浓度为0.05 g/L时，纯化后绵马贯众多酚对铁离子还原能力为0.47，明显高于纯化前的0.25；且在酸性条件下DPPH·清除能力以及铁离子还原能力更高，在碱性条件下·OH清除能力更高。

关键词: 绵马贯众, 多酚, 大孔树脂纯化, 抗氧化活性

Abstract:

The polyphenols were extracted from Cyrtomium fortune with 70% ethanol, and then purified with 7 macroporous resins. The results showed that HPD100 resin had the best purification effect based on the adsorption rate and desorption rate. The effect of sample mass concentration, loading speed of sample, volume fraction of desorption ethanol, desorption speed and sample pH value on the purification of HPD100 resin was further analyzed, and the optimum purification conditions were obtained as follows: mass concentration of sample solution 2.00 g/L, pH value 5, loading speed of sample 1 mL/min, volume fraction of desorption ethanol 70% and desorption speed 2 mL/min. Under these conditions, the content of polyphenols increased from 27.64% to 54.00% (the yield of polyphenols was 6.46%). The purified polyphenols showed higher antioxidant activity. For the purified polyphenols, the IC₅₀of DPPH radical (DPPH·) and hydroxyl radical (·OH) were 0.01 and 0.49 g/L, respectively; but for the crude polyphenols, the IC₅₀ of DPPH· and ·OH were 0.05 and 1.79 g/L, respectively. At 0.04 g/L, the DPPH· scavenging activity of the purified polyphenols was 90.82%, almost similar to that of Vc. At 0.05 g/L, ferric ion reducing power of the purified polyphenols was 0.47, but the crude polyphenols was only 0.25. Furthermore, the purified polyphenols exhibited better DPPH· scavenging activity and ferric ion reduction ability under acidic conditions and the hydroxyl radical scavenging activity under alkaline condition.

Key words: Cyrtomium fortune, polyphenols, purification with macroporous resin, antioxidant activity

中图分类号:

TQ35

张慧芳,王叶,户维超,袁澍,袁明. 绵马贯众多酚的分离纯化及其抗氧化活性研究[J]. 林产化学与工业, 2019, 39(2): 81-88.

Huifang ZHANG,Ye WANG,Weichao HU,Shu YUAN,Ming YUAN. Purification and Antioxidation Activity of Polyphenol from Cyrtomium fortune[J]. Chemistry and Industry of Forest Products, 2019, 39(2): 81-88.

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参考文献 23

1	崔月曦, 刘合刚. 贯众的研究进展[J]. 中国现代中药, 2014, 16 (12): 1043- 1048.
	CUI Y X , LIU H G . The advance of research on Cytomium fortune[J]. Modern Chinese Medicine, 2014, 16 (12): 1043- 1048.
2	林红梅, 韩忠明, 吴劲松, 等. 粗茎鳞毛蕨种群形态形状及其生态可塑性的研究[J]. 吉林农业大学学报, 2006, 28 (4): 398- 401, 406. doi: 10.3969/j.issn.1000-5684.2006.04.012
	LIN H M , HAN Z M , WU J S , et al. Study on morphologic characters of Dryopteris crassirhizoma population and its ecological plasticity[J]. Journal of Jilin Agricultural University, 2006, 28 (4): 398- 401, 406. doi: 10.3969/j.issn.1000-5684.2006.04.012
3	高增平, 李国富, 陆蕴如. 绵马贯众抗疟活性筛选[J]. 北京中医药大学学报, 2002, 25 (4): 45- 46. doi: 10.3321/j.issn:1006-2157.2002.04.015
	GAO Z P , LI G F , LU Y R . Screening of the antimalarial parts of the extract of Guanzhong[J]. Journal of Beijing University of Traditional Chinese Medicine, 2002, 25 (4): 45- 46. doi: 10.3321/j.issn:1006-2157.2002.04.015
4	刘钊.绵马贯众抗感染活性成分研究[D].北京:北京协和医学院, 2010.
	LIU Z. Studies on anti-infective constituents of Dryopteris crassirhizoma Nakai[D]. Beijing: Peking Union Medical College, 2010.
5	赵艳红, 李建科, 李国荣. 石榴皮多酚纯化及其抗氧化活性表征[J]. 食品科学, 2010, 31 (11): 31- 37.
	ZHAO Y H , LI J K , LI G R . Purification with macroporous adsorbent resins and in vitro antioxidant evaluation of pomegranate peel polyphenols[J]. Food Science, 2010, 31 (11): 31- 37.
6	赵晋彤, 贾莹莹, 李苏苏, 等. 绵马贯众总多酚超声提取工艺的优化及其抗氧化活性[J]. 中成药, 2018, 40 (3): 729- 732. doi: 10.3969/j.issn.1001-1528.2018.03.047
	ZHAO J T , JIA Y Y , LI S S , et al. Optimization of ultrasonic extraction process and its antioxidant activity for the total polyphenols of Cyrtomium fortune[J]. Chinese traditional patent medicine, 2018, 40 (3): 729- 732. doi: 10.3969/j.issn.1001-1528.2018.03.047
7	曹广尚, 王信, 杨培民. 阴离子交换树脂分离纯化绵马贯众总多酚的工艺研究[J]. 中草药, 2014, 45 (9): 1265- 1269.
	CHAO G S , WANG X , YANG P M . Separation and purification of total poly phenol from Dryopteris crassirhizoma with anion exchange resin[J]. Chinese Traditional and Herbal drugs, 2014, 45 (9): 1265- 1269.
8	GAO Z P , YU Z F , YUE T L , et al. Optimization of polyphenols removal from kiwifruit juice using a macroporous resin[J]. Journal of the Science of Food and Agriculture, 2017, 97 (8): 2498- 2509. doi: 10.1002/jsfa.2017.97.issue-8
9	甘芝霖, 倪元颖, 郭悦, 等. 大孔树脂分离纯化玫瑰果多酚及其抗氧化性[J]. 农业工程学报, 2015, 31 (24): 298- 306. doi: 10.11975/j.issn.1002-6819.2015.24.045
	GAN Z L , NI Y Y , GUO Y , et al. Separation and purification of rosehip polyphenols by macroporous resin and its antioxidant activity[J]. Transactions of the Chinese Society of Agricultural Engineering, 2015, 31 (24): 298- 306. doi: 10.11975/j.issn.1002-6819.2015.24.045
10	KOFFI E N , LE GUERNEVÉ C , LOZANO P R , et al. Polyphenol extraction and characterization of Justicia secunda Vahl leaves for traditional medicinal uses[J]. Industrial Crops and Products, 2013, 49, 682- 689. doi: 10.1016/j.indcrop.2013.06.001
11	薄采颖, 郑光耀, 高丽萍, 等. 亚临界水提取协同大孔树脂纯化杨树芽总黄酮[J]. 林产化学与工业, 2014, 34 (4): 107- 112.
	BO C Y , ZHENG G Y , GAO L P , et al. Extraction and purification of flavonoids from poplar buds with subcritical water extraction coordinated with macroporous resin adsorption[J]. Chemistry and Industry of Forest Products, 2014, 34 (4): 107- 112.
12	LIN L Z , ZHAO H F , DONG Y , et al. Macroporous resin purification behavior of phenolics and rosmarinic acid from Rabdosia serra(Maxim.) Hara leaf[J]. Food chemistry, 2012, 130 (2): 417- 424. doi: 10.1016/j.foodchem.2011.07.069
13	ZHU K X , YAO S W , ZHANG Y J , et al. Effects of in vitro saliva, gastric and intestinal digestion on the chemical properties, antioxidant activity of polysaccharide from Artocarpus heterophyllus Lam. (Jackfruit) pulp[J]. Food Hydrocolloids, 2018, 87, 925- 959.
14	高凝轩, 李斌, 杜姗姗, 等. 黑果腺肋花楸多酚稳定性的研究[J]. 食品科学, 2016, 37 (23): 20- 24. doi: 10.7506/spkx1002-6630-201623004
	GAO N X , LI B , DU S S , et al. Stabllity of polyphenols from Aronia melanocarpa fruits[J]. Food Science, 2016, 37 (23): 20- 24. doi: 10.7506/spkx1002-6630-201623004
15	田强, 张梦娜, 赵琳, 等. 厚朴叶不同溶剂提取物抗氧化活性比较分析[J]. 食品研究与开发, 2018, 39 (16): 37- 42. doi: 10.3969/j.issn.1005-6521.2018.16.007
	TIAN Q , ZHANG M N , ZHAO L , et al. Comparative evaluation of antioxidant activities of different solvent extracts from Magnolia offcinalis leaves[J]. Food Research and Development, 2018, 39 (16): 37- 42. doi: 10.3969/j.issn.1005-6521.2018.16.007
16	唐琦, 胡广林, 刘金芳, 等. 体外模拟消化对萌发红小豆的抗氧化活性的分析[J]. 食品科技, 2018, 43 (1): 67- 76.
	TANG Q , HU G L , LIU J F , et al. Determination of antioxidant activity of germinated red beans by stimulated gastrointestinal digestion[J]. Food Science and Technology, 2018, 43 (1): 67- 76.
17	BURAN T J , SANDHU A K , LI Z , et al. Adsorption/desorption characteristics and separation of anthocyanins and polyphenols from blueberries using macroporous adsorbent resins[J]. Journal of Food Engineering, 2014, 128, 167- 173. doi: 10.1016/j.jfoodeng.2013.12.029
18	张星, 董欣, 刘振春, 等. 大孔树脂纯化黑豆异黄酮的工艺优化[J]. 西北农业科技大学学报, 2018, 46 (1): 136- 145, 154.
	ZHANG X , DONG X , LIU Z C , et al. Optimization of purification process of black beans disoflavones with macroporous adsorption resin[J]. Journal of Northwest A & F University, 2018, 46 (1): 136- 145, 154.
19	朱洁, 王红宝, 孙佳君, 等. 梨幼果多酚的纯化及其抗氧化性[J]. 食品科学, 2017, 38 (5): 14- 20.
	ZHU J , WANG H B , SUN J J , et al. Purifixation and antioxidant activity of polyphenols from young pear fruits[J]. Food Science, 2017, 38 (5): 14- 20.
20	ZHAO Z Y , DONG L L , WU Y L , et al. Preliminary separation and purification of rutin and quercetin from Euonymus alatus(Thunb.) Siebold extracts by macroporous resins[J]. Food and Bioproducts Processing, 2011, 89 (4): 266- 272. doi: 10.1016/j.fbp.2010.11.001
21	蔡彬新, 周逢芳, 杨惠婷. 山竹果皮总黄酮富集工艺研究[J]. 中国农学通报, 2017, 33 (32): 34- 38.
	CAI B X , ZHOU F F , YANG H T . Purification condition of flavonoids from Garcinia mangostana pericarp[J]. Chinese Agricultural Science Bulletin, 2017, 33 (32): 34- 38.
22	YANG Z Z , TANG H T , SHAO Q , et al. Enrichment and purification of the bioactive flavonoids from flower of Abelmoschus manihot (L.) medic using macroporous resins[J]. Molecules, 2018, 23, 2449. doi: 10.3390/molecules23102449
23	张一鸣, 吴跃中, 杨士花, 等. 云南黑大麦多酚优化提取纯及化工艺[J]. 食品科技, 2018, 43 (5): 206- 213.
	ZHANG Y M , WU Y Z , YANG S H , et al. Optimization of extraction and purification of polyphenols from Yunnan black hulless barley[J]. Food Science and Technology, 2018, 43 (5): 206- 213.

树脂 resin	极性 polarity	粒径/mm particle size	孔径/nm pore size	比表面积/(m²·g^-1) specific surface area
S-8	强极性strong polarity	0.30~1.25	28~30	100~120
X-5	非极性nonpolar	0.32~1.25	290~300	500~600
DM301	中极性moderate polarity	0.30~1.25	13~17	330~380
D101	非极性nonpolar	0.30~1.25	9~10	500~550
CAD-40	中极性moderate polarity	0.30~1.25	7~8	450~500
AB-8	非极性nonpolar	0.30~1.25	130~140	450~520
HPD100	非极性nonpolar	0.30~1.25	8.50~9.00	650~700

树脂 resin	吸附量/(mg·g^-1) adsorption capacity	吸附率/% adsorption ratio	解吸量/(mg·g^-1) desorption capacity	解吸率/% desorption ratio
S-8	18.86±0.09	91.06	2.31±0.05	12.27
X-5	18.83±0.10	90.90	16.33±0.69	86.71
DM301	18.23±0.32	88.12	13.53±0.53	74.13
D101	18.85±0.08	91.03	17.45±0.41	92.56
CAD-40	18.68±0.26	90.20	16.68±0.83	89.28
AB-8	18.55±0.23	89.55	17.14±0.64	92.42
HPD100	18.72±0.38	90.35	17.92±1.20	95.76

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