Study on Purification Technology and Antioxidant Activity of Total Flavonoid from Eriobotryae Folium

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1 Interleukin-1beta causes pulmonary inflammation, emphysema, and airway remodeling in the adult murine lung [J]. Am J Respir Cell Mol Biol, 2005, 32(4): [10] FAN C H, LI S Y, LI M, et al. The clinical significane of IFN-γ, IL-32, IL-1β in chronic obstructive pulmonary disease [J]. Chin Foreign Med Treat(), 2011(19): [11] ZHANG W, GU M M. Correlation of blood stasis and chronic obstructive pulmonary disease [J]. Guide J Tradit Chin Med Pharm( ), 2012, 18(7): * ( ) ABTS D101 D101 1 g 1.5 g 10% 40% 90% 80 ml h 1 (12 BV h 1 ) 40% D101 40% ABTS D101 R285.5 A (2014) Study on Purification Technology and Antioxidant Activity of Total Flavonoid from Eriobotryae Folium LÜ Han, TENG Jiehui, CHEN Jian, MA Li, REN Bingru, LI Weilin * (Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing , China) ABSTRACT: OBJECTIVE To optimize the separate and purify technology of total flavonoid from Eriobotryae Folium, and to evaluate the antioxidant activity of total flavonoid. METHODS Investigate the adsorption capacity and purification activity of resin, polyamide, silica gel. ABTS free radical-scavenging assay was used to test the antioxidant activity. RESULTS Resin D101 has the best capacity to adsorb and purify total flavonoids, the optimal procedure was: 1 g D101 resin, extract solution (1.5 g powder of Eriobotryae Folium), water, 10%, 40% and 90% ethanol gradient elute, adsorbing rate 80 ml h 1 (12 BV h 1 ), 40% fraction was eluted and purified again by resin D101 with same process, then the 40% fraction was concentrated and dried to obtain total flavonoid. Total flavonoid showed a certain ABTS free radical-scavenging activity. CONCLUSION The repeated adsorption using resin D101 is the ideal method to separate and purify total flavonoid from Eriobotryae Folium.The total flavonoid of Eriobotryae Folium has a certain antioxidant activity. KEY WORDS: Eriobotryae Folium; total flavonoids; macroporous adsorbing resin; antioxidant activity Eriobotrya japonica (Thunb.) Lindl. [1] [2] [3-5] [6] [7] ( )-(BM ) Tel: (025) xiaohan1814@163.com * Tel: (025) lwlcnbg@mail.cnbg.net 40 Chin J Mod Appl Pharm, 2014 January, Vol.31 No

2 1 1.1 MP120-2 ( ) TU-1810 ( )NKA NKA-9 HPD-600 HPD-417 HPD-100 HPD-450 HPD-722 AB-8 DM130 D101 (30~60 )(100~200 ) ( %) 1.2 Eriobotrya Japonica(Thumb.) Lindl mg 70% 25 ml mg ml ml 0.3 ml 5% NaNO 2 6 min 0.3 ml 10% Al(NO 3 ) 3 6 min 4.0 ml 4% NaOH 70% 10 ml 10~15 min 510 nm (x) (y) y= x r= (n=3) ml 10 ml 0.3 ml 5% NaNO 2 6 min 0.3 ml 10% Al(NO 3 ) 3 6 min 4.0 ml 4% NaOH 70% 10 ml 10~15 min 510 nm g 70% 2 1 h 2 24 h r min 1 10 min 200 ml mg ml % 24 h 95% (1 3) D101 NKA HPD-100 HPD-722 AB-8 HPD-450 DM130 NKA-9 HPD-600 HPD g 5.00 ml ( )=( )/( )= ( )/ % 2.2 ( )=( )/(%)= /100% D101 5 g ( 3.3 ml 6.5 ml) 15 ml 40 ml h 1 2 ml D101 5 g 5 ml ml h 1 ( BV h 1 BV ) Chin J Mod Appl Pharm, 2014 January, Vol.31 No.1 41

3 2.5.3 D g 10 ml 80 ml h g D ml 20% 40% 95% ml h 1 40% g 10 ml D g 10% 15% 20% 30% 40% 50% 90% 300 ml h 1 BAW[(4 1 1)] 3% AlCl nm 20%~40% 60 1 h 70% (30~60 )10 g 10% 20% 30% 40% 50% 60% 90% 300 ml h 1 20%~50% 60 1 h 70% (100~200 )0.5 g (100~200 )1.0 g 95% 70% 300 ml h 1 95% 60 1 h 70% D g 10% 40% 90% 600 ml h 1 40% 60 1 h 70% 2.7 ABTS DMSO 10 mg ml 1 DMSO ABTS 7 mmol L 1 ABTS 2.45 mmol L 1 ( ) 12~16 h ABTS nm µl 190 µl ABTS 200 µl 10 s 10 min nm (A i ) 10 µl DMSO (A 0 ) 190 µl ABTS (A j ) 4 C A = A A 0 ( i j) A 0 100% IC 50 [8] NKA NKA-9 D D101 2>4.0 ml 5 ml 1 g 1.0 ml ( 1.5 g 42 Chin J Mod Appl Pharm, 2014 January, Vol.31 No

4 ) 1 Tab 1 The dynamic adsorption effect of macroporous resins % D NKA HPD HPD AB HPD DM NKA HPD HPD Tab 2 The investigation of sample volumn ml ml mg ml D101 3>80 ml h 1 D101 ( ml mg ml 1 ) 80 ml h 1 (12 BV h -1 ) 3 Tab 3 The investigation of adsorption velocity /ml h 1 mg ml D % 30% BAW[(4 1 1)] AlCl nm 20% 30% 40% 10% 50% 90% 20%~40% 10% 40% 90% 40% 4 Tab 4 The investigation of alcohol concentration g % 10% % % % % % % D101 1 g 1.5 g 10% 40% 90% 80 ml h 1 (12 BV h 1 ) 40% D101 D101 20% 30% 40% 10% 50% 90% 20%~40% % 30% 40% 50% 10% 60% 90% 20%~50% % 70% 95% D Chin J Mod Appl Pharm, 2014 January, Vol.31 No.1 43

5 5 3 Tab 5 The yield and content of total flavonoid prepared by three different purify methods g % % D % 3.4 ABTS ABTS IC 50 ( )µg ml 1 IC 50 C[( )µg ml 1 ] 4 D101 1 g 5.0 ml ( 1.5 g ) 10% 40% 95% 12 BV h 1 40%40% D101 40% % REFERENCES [1] LI T, XU R Q, LINW J, et al. Establishment and optimization of ISSR reaction system for Eriobotryae Folium from Fujian [J]. Chin J Mod Appl Pharm(), 2012, 29(4): [2] STATE ADMINISTRATION OF TRADITIONAL CHINESE MEDICINE, CHINESE MATERIA MEDICA EDITORIAL BOARD. Chinese Herbal Medicine() [M]. Vol. Shanghai: Shanghai Science and Technology Press, 1999: 140. [3] SHA N, LIANG J Y. Development of Eriobotrya japonica(thumb.) Lindl. [J]. Strait Pharm J(), 2006, 18(1): [4] LU H, LI W L, PEI Y P, et al. Detection of flavonoids in Eriobotrya japonica(thunb.) Lindl. by HPLC-MS n [J]. Res Pract Chin Med( ), 2009, 22(6): [5] LUO M H, LU H, LI W L. Determination of total flavonol in leaves of Eriobotrya japonica by HPLC [J]. Lishizhen Med Mater Med Res(), 2011, 22(3): [6] HUANG A L. Progress on the pharmacologic action of flavonoid [J]. Anhui Agricult Sci Bull(), 2007, 3(10): [7] LIU Y, ZHU X T. Optimization of purification process for total flavonoids from leaves of Eriobotrya japonica by macroporous resin [J]. Chin J Exp Tradit Med Form( ), 2012, 18(11): [8] RER P N. Antioxidant activity applying an improved abts radical cation decolorization assay [J]. Free Radic Biol Med, 1999, 26(9/10): ( ) ELISA Balb/c 4 3E02 4B01 5D03 4C04 ELISA 4 IgG2b IgG2b IgG1 IgG1 ELISA Y= X r= ~1 000 ng ml ng ml % RSD 0.68%(n=6) ELISA ELISA R962 B (2014) ( )(2011Y0035) (JA12171) Tel: yyyfj@163.com 44 Chin J Mod Appl Pharm, 2014 January, Vol.31 No