Antioxidative Activity and Nitrite Scavenging Ability of Ethanol Extract from Phyllostachys bambusoides

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(1)KOREAN J. FOOD SCI. TECHNOL. Vol. 36, No. 2, pp. 306~310 (2004). ©The Korean Society of Food Science and Technology. ¾Z öêRºÂb~ Öz Î" 5 jîÖ" ²·Ï ªêjÁ¾B1Á9ßz* ö%L ~*^ö £¶öB*, 8.¶\² 1 ·L N~:¶¦. Antioxidative Activity and Nitrite Scavenging Ability of Ethanol Extract from Phyllostachys bambusoides Jin A Lim, Young Soon Na1, and Seung Hwa Baek* Department of Herbal Resources, Professional Graduate School of Oriental Medicine and Institute of Basic Natural Sciences, Wonkwang University 1 Division of Fashion and Beauty, Konyang University Efficacy of antioxidative activity and nitrite scavenging ability of ethanol extract of Phyllostachys bambusoides S. et Z. (P. bambusoides) was investigated. Electron-donating ability of ethanol extract at RC50 was 116.75 µg/mL. After addition of 0.92 mg/mL ethanol extract, autooxidation of pyrogallol decreased to 44% by superoxide dismutase-like activity. In antioxidative activity of ethanol extract against linoleic acid during incubation times of 4 and 6 day at 40oC, TBA values decreased by 74.76 and 58.48% with addition of 50 mg/mL, respectively. Nitrite scavenging ability showed the most remarkable effect at pH 1.2, decreasing to 43% by addition of 0.2 mg/mL. These results suggest that ethanol extract of P. bambusoides can be used in bioactive and functional material. Key words: Phyllostachys bambusoides, antioxidative activity, nitrite scavenging ability. B. . GB®¾ >Ö&® ö BïB Î&>º îÖ"¾ jîÖ"f GB®~ Bï 5 Gï~ n;zö V öò j î¢ Clostridium botulinumö & ;·Ïj ¾æÚ, G~ >W" ÖOWj BF~º ö 7º j (15). . îÖ"f ²zV&ÚöB $º ®~ &Ë 7ö îÖ~ö Î²¾ îÖ" ~ö^ö ~~ jîÖ"b ~ö> j îÖ"f 2/ 5 3/ amine~f >w~ nitrosoaminej W ~º ©b rJ^ ® (16). Nitrosaminef ÚÚöB diazoalkane(CnH2nN2)b æz~ Ö¾ Wî $º ^Ú~ W ªj alkylz b zj FB ~(17), $ jîÖ "f ¶ ëWj < ®V r^ö ¢; ³ê ç ê ³ S .7~ Î&nj Öz* zÞ Î&nÃ (methemoglobinemia)j FB (18). Mirvish (19)f ascorbic acidö ~ nitrosoamine WÛBVËj ~& Millard > wWbö ~ ÛB·Ï Kato (20)ö ~ >î . $ ®7ö phenolic guaiacol, resorcinol ~ phenolê bî. nitroz >wj ;K~² ÛB º Ò >îb (21), jÞ ¢jºÂb(18), ºÂb(22), ªU~(23) Ò Ö«¶f ."ºÂb(24)öB jîÖ" ²·Ï > ® . &¾Zº â"f jÝ~æò *V& Ïî © , Ö Ò¾¢öº 5³ö 10«, 4æ« ª>Ú ®, " 7¦ Îö ¶¢, &' ®«f å&(Sasamorpha purpurascens Nakai var. borealis Nakai), ^&(Phyllostachys reticulata koch). ÚÚöB Özf &NB *çb ^ïö Ò~º æ îf Ö²öB F¾>º superoxide anion radical, hydroxyl radical, singlet oxygen 5 H2O2 ~ WÖ²f Ö~ "Öz bj ò , ~ ³>wö ~~ alcohol~, aldehyde~, ketone~ j Wb ÚÚöB DNA¢ ¶ç* zj FB öò jî¢, ^z, ^ï ª, æOÖz ' Ò' Ë¢ ¢bÎ (1-3). æ.ræ WÖ²f free radical Wj Oæ~V * ÒÏ>º ÖzBº α-tocopherol, BHT, BHA, PG, TBHQ, ascorbic acid rJ^ ®b (4), 7 Öz Î"& ÚÂ BHTf BHAº æöW 5 ëW æ'>Ú(5), *Òº ÒÏï Ï6>º ;. . V¢B " n*W" &Ëç ^B& >æ pº Â Ö zB BBj * ôf & ê> ®b, z{(6), ¦f (7), ¯¾Z(8), rö(9), &v(10), J¶(11), ªU~(12), º ´(13), æ{¾Z(14) öB Öz Î"& > ® .. *Corresponding author : Seung Hwa Baek, Department of Herbal Resources, Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan, Chonbuk 570-749, Korea Tel: 82-63-850-6225 Fax: 82-63-841-4893 E-mail: [email protected] 306.

(2) &¾Z öêRºÂb~ Öz Î" 5 jîÖ" ²·Ï. 5 ~&(Sasa coreana Nakai) . &¾Z´f ÿ~~òöB ##¢~ Úâ, bt£, 7³, .{ ö "*º»b ÒÏ>Ú z, 5 ê ·Ï" zÎ"ê ®º © b rJ^ ® (25). " &¾ZºÂb~ Kö & ô f & > ®b¾(26,27) ÒWö & º j ; . ö öBº &¾Z *V~ VËW ®²Ò $º ®Î&bB~ &ËWj ¦Æ~¶ &¾Z *V¢ öêR ºÂ~ ÖzÎ"f jîÖ" ²Ëj Ò~& .. Òò 5 O» Òò þö ÒÏ &¾Zº 3jÖ {&¾Z(Phyllostachys bambusoides S. et Z.)B *Î ·öB 2003j 3úö *V¢ j~ r ê ÒÏ~& . α,α'-diphenyl-β-picrylhydrazyl (DPPH), pyrogallol, linoleic acid, trichloroacetic acid(TCA), 2thiobarbituric acid(TBA), sodium nitrite, sulfanilic acid Ò α-naphthylaminef Sigma(USA)öB «~&b, Væ £f ªCÏ /ç~ £j ÒÏ~& . ò~ B B &¾Z *V¢ ªê ê, ªö 780 gö öêR 4 L ¢ Î&~ NöB 24*ÿn 3² v>ºÂ~& . . r, öêR ºÂf "æ(Whatman, No. 2) "~ 6{ ³» ê ê ÿÖ~&b, áÚê &¾Z*V öêR ºÂb(35.59 g, 4.56%)f þò ÒÏ>î . *¶Ë G; *¶Ë(electron donating ability)f Blosis O»(28)ö ~ DPPH free radical ²»ö ~ G;>î . ¯, òº z êRö &j~ 0.3 mM DPPH Ïj Î&~ Nö B 30ª* O~ ê 517 nmöB 7ê¢ G;~& . *¶ Ëf ò¢ Î&~æ pf &~ 7ê¢ 50% 6² Êº jº ò~ ·j RC50b ~ ¾æÚî . Superoxide dismutase(SOD) FÒW G; Marklundf Marklund~ O»(29)ö V¢ ' ³êê ò(0.2 mL)ö pH 8.5 ; Tris-HCl buffer(50 mM tris[hydroxymethyl]amino-methane containing 10 mM EDTA, 3 mL)f 7.2 mM pyrogallol(0.2 mL)¢ &~ 25oCöB 10ª* O~~&b , 1 N HCl(1 mL) >wj ;æÎ ê 420 nmöB 7ê ¢ G;~& . SOD FÒWf 100−[(òÎ&~ 7ê/Z Î&~ 7ê)Ü100]b ¾æÚî . TBA& G; VîÏf 0.1 M phosphate buffer(pH 7.0)f ethanolj 4 : 1 b Ï ö linoleic acid¢ 0.03 M >ê Î&~& . VîÏ(20 mL)ö 0.1 M phosphate buffer(pH 7.0, 19.2 mL)f ' ³êê ò(0.8 mL)¢ Î& ê 40oC NVöB êû~B ã'b b(2.0 mL)j ~ ªC~& . * b(2.0 mL)ö 35% TCA(1.0 mL)f 0.75% TBA(2.0 mL)¢ & r 30. ÿn êû* 95oC >»çöB 40ª ÿn >w8 . >wj NöB ï'* acetic acid(1.0 mL)f chloroform(2.0 mL)j &~ êûÎ ê, 3,000 rpmö. 307. B 5ª ÿn öªÒ~ çû~ 7ê¢ 532 nmöB G ;~ ¢ TBA8b ~& (30). jîÖ" ²Ë G; jîÖ" ²Ë(nitrite scavenging ability)f Kato (20)~ O »b G;~& . ¯, 1 mM NaNO2Ï(2 mL)ö ò(1 mL) ¢ &~ 0.1 N HCl(pH 1.2), 0.2 M Ö jÏ(pH 3.0, 6.0)b '' pH 1.2, 3.0, 6.0b ; r >wÏ~ ¦ b¢ 10 mL ~& . Ïj 37oCöB 1* >wÊ ' >w(1 mL)j ~ 2% .ÖÏ(2 mL)" 30% .ÖÏ b Ï Griess reagent(1% sulfanilic acid: 1% naphthylamine = 1 : 1, 0.4 mL)¢ & r b~ NöB 15ª* O~ ê 520 nmöB 7ê¢ G;~& . &º Griess reagent & Ã~>¢ &~ G;~&b, jîÖ" ²Ë f 100−[(òÎ&~ 7ê/ZÎ&~ 7ê)Ü100]b ¾æÚî . Ûê¾Ò Ûê¾Òº ''~ òö & Student's t-test¢ ÒÏ~ meanÛS.D. êÖ~&b, p-value¢ ~ F~Wj ¦Ã~ & .. Ö" 5 V ¾Z*8 öêRºÂb~ *¶Ë ¢> bÚ~ Öz ·Ïö &Bº ôf & ®º Ïb 21«j ¦*ê DPPH¢ Ï~ FÒ¢:¢ ² Î"¢ G; Ö" ^´, 9´ 5 ^*´öB ß'b ¸f Î"& &V>î (31). {&¾Z*V öêRºÂb~ DPPH free radical ²Ëj ò¢ Î&~æ pf &~ 7ê¢ 50% 6²Êº jº òï(RC50)b G; Ö " 116.75 µg/mLb ¾æÒ (Fig. 1). " f (32)f {& ¾Z¢ >ºÂ~ bb~ ³ê& 0.1%& >² B ê *¶·Ïj Ò Ö", #K" *Vº '' 86%, 51.7% ~ ¸f *¶Ëj &b, ´ºÂbf ·Ï £®. ~ ®Ú, öB~ {&¾Z *V öêRºÂ b(RC50 = 116.75 µg/mL)f >ºÂb(ß1 mg/mL) *¶ Ë Ö>~ º ©j "º Ö" 'B . Ò &bî ÒÏB β-carotene" BHA~ RC50 '' 18.02 µg/ mL, 18.48 µg/mL(data not shown)b ¾æ¾ {&¾Z *V ö êR ºÂbf Î"' *¶Ëj º ©b &V>î . ¾Z*8 öêRºÂb~ superoxide dismutase(SOD) F ÒW Öz Î²7~ ~¾ superoxide dimutase(SOD)º ^ö Ú ~ö Ö²«(superoxide)j "Öz>² *~Êº > wj / ~º Î² . SODº 30 KDa ç~ ª¶ïj & ê Wî bî ÚÚö £² >>æ á~ Ú VÂ >(33,34), " pHö ®n;~ (35). V¢B SODf FÒ Wj &æB SOD~ 6j j > ®º SOD FÒ W bîj dº & B® ê¯> ® . V¢B {&¾ Z*V öêR ºÂb~ superoxide(O2−) ÖzÛB·Ïj rj V * superoxidef >w~ .æbîj ¾æÚº pyrogallol ¶ÿÖz>wj G;~ SOD FÒWj G;~& (29). Fig. 2(a)º {&¾Z*V öêR ºÂb~ SOD FÒWj G;.

(3) 308. ®"²æ B 36 ² B 2 ^ (2004). Fig. 1. Electron donating ability of ethanol extract from P. bambusoides. The values represent the meanÛstandard deviations for triplicate experiments. *Significantly different from the control values (p<0.05).. ©b 0.23, 0.46 Ò 0.92 mg/mL Î& '' 17.96%, 30.41% Ò 43.88%~ Wê¢ ¾æîb, jvbî ascorbic acidº 0.184 mg/mLöB 76.06%~ SOD FÒWj ªb(Fig. 2(b)) {&¾Z *V öêR ºÂb~ Ö> ÖzK &V>î . $ f (36) G; _ÒÚf _& &¾Z*V öêR ºÂb~ SOD FÒW ¸ ² ¾æ¾º ©j &V > ®î .. ¾Z*8 öêRºÂb~ TBA&ö ~º 'Ë {&¾Z *V öêRºÂb~ æî"Özf &N Öz Î"¢ ¦Æ~V *~ linoleic acidö ºÂbj 12.5, 25, 50 µg/mL ³ê '' Î&~ 40oCöB 6¢ÿn TBA8~ æ z¢ 7ê(532 nm) G;~& (Fig. 3). Ö", &º 4¢" 6¢ ê 0.531, 0.554 TBA8 G;>î ºÂb 50 µg/mLj Î&®jröº 0.134, 0.23 TBA8 ¾æ¾ & ö j ºÂb Î&~ TBA>~& '' 74.76%, 58.48% 6²>î (Fig. 3(a)). $ &bî 25 µg/mL BHA¢ Î& ~ TBA8j G; Ö", &ö j 4¢" 6¢ ê ' '~ TBA 8 76.04%, 80.21% 6²j r > ®î (Fig. 3(b)). f (37)f {&´ bºÂb~ in vitroöB "Özb ; W &Nj TBA»b G; Ö", ³ê ~' &¢ &b 40 mg/mL~ ³êöB 50.9% æî"Öz W Û BÎ"¢ ¾æÞ ~& . *~ Ö" {&¾Z* V öêRºÂbf æî"Özf &NB Öz Î"& Ö> ©b &V>î . ¾Z*8 öêRºÂb~ jîÖ" ²Ë jîÖ¾Þ Ïö {&¾Z*V öêRºÂb 0.2 mg/mL j Î&~ pH j '' pH 1.2, pH 3.0, pH 6.0b ;~ jîÖ"ö & ²Nj G;~& . pH º* º ÚÚ *öB~ pH æz¢ J~&b, ' pH ~. Fig. 2. SOD-like activity of ethanol extract from P. bambusoides (a) and ascorbic acid (b). The values represent the meanÛstandard deviations for triplicate experiments.. Fig. 3. TBA values in linoleic acid substrate containing ethanol extract from P. bambusoides (a) and BHA (b) during storage at 40oC. The values represent the meanÛstandard deviations for triplicate experiments. *Significantly different from the control values (p<0.05)..

(4) &¾Z öêRºÂb~ Öz Î" 5 jîÖ" ²·Ï. 309. 6Ò~ º BK21 Òëæöjö ~ >¯>îb, {&¾Z ¢ B" (")W®ö 6Òãî .. ^. Fig. 4. Nitrite scavenging ability of ethanol extract from P. bambusoides. The values represent the meanÛstandard deviations for triplicate experiments.. (pH 1.2, pH 3.0, pH 6.0)öB jîÖ" ²Ëf '' 43.02%, 35.46%, 9.86%b ¾æ¾ pH ~' jîÖ" ²Ëj & (Fig. 4). f FÒ Ö" Kang (38)f '« phenolW zb~ ³ê¢ 0.1-6 mM >Ïb B ê, jîÖ" ² Nj pHöB G; Ö" pH 1.2öB &Ë ¸² ¾æÒb¾, pH 6.0öBº 10% ;ê &¦ª ç >î ~& . jîÖ"f GB®ö Î&>Ú BïB 5 B Ï> ®b¾, ®7ö Ò~º amine~f >w~ Bzbî nitrosaminej W~º ";f pH& Ôf öB £² ¢Ú¾º ©b rJ^ ® (39). îÞz(nitrosation)ö 'Ëj "º nitriteº nitrous acid(HNO2)¢ ;W~V *B ÖWz> HNO2º H2NO2+b protonz>Ú F' b amidef >w~ nitrosamide¢ ;W . ÖWz (acidification)"; r^ö îÞz>wf " ÚÚ ÖW* (acidic stomach)öB B (40,41). Ö"ö ~~, jî Ö" ²Ë Ú~ *Ú pH " jÝ pH 1.2öB & Ë Ö> ©b G;>Ú {&¾Z *V öêR ºÂbf ÚÚöBê Î"' jîÖ" ²·Ïj Û nitrosamine Wj ÛB ©b 'B .. º. £. º {&¾Z *V~ ÒW 5 VËWj ¦Æ~V *B {&¾Z *V¢ öêR ºÂ~, ºÂb~ Öz Î"f jîÖ" ²Ëj G;~& . Ö", {&¾Z * V öêR ºÂb~ *¶Ë(RC50)f 116.75 µg/mL ¾æ Ò SOD FÒWf ºÂb(0.92 mg/mL)j Î&~&j r 43.88% &Ë ¸² &V>îb, linoleic acidö & Öz Kf ºÂb(50 µg/mL)j Î&~ TBA8j G; Ö" V· * 4¢" 6¢ ã" ê '' 74.76%, 54.48% 6²Nj ª b Î"' Öz Î"¢ ¾æÚî . jîÖ" ²Ë f ºÂb(0.2 mg/mL)j Î&~&j r pH 1.2 öB 43.02% &Ë ¸² ¾æÒb pH& Ã&> 6²~& . ç~ Ö"¢ «~ {&¾Z *V~ öêR ºÂbf Öz Î"f jîÖ" ²Ë ®º bîj F ©b $ B . ºê, {&¾Z~ Öz Î"f jîÖ" ²Ëj ¾ æÚº öbîö & & ê¯>Ú¢ © .. ò. 1. Fridorich I. The biological activity of oxygen radicals. Science 201: 875-881 (1978) 2. Gardner DR, Fridorich I. Superoxide sensitivity of Escherichia coli 6-phospho gluconate dehydratase. J. Biol. Chem. 266: 14781483 (1991) 3. Imlay IA, Linn S. DNA damage and oxygen radical toxicity. Science 240: 1302-1309 (1986) 4. Dziezak JD. Antioxidants. Food Technol. 40: 94-102 (1986) 5. Branen AL. Toxicological and biochemistry of butylated hydroxyanisole and butylated hydroxytoluene. J. Am. Oil Chem. Soc. 52: 59-63 (1975) 6. Mang YS, Park HK. Antioxidant activity of ethanol extract from Dodok (Codonopsis lanceolata). Korean J. Food Sci. Technol. 23: 311-316 (1991) 7. Ahn CY, Hyun KH, Park KH. Investigation of antioxidative substance in black sesame seed. Korean J. Food Sci. Technol. 24: 31-36 (1992) 8. Choi U, Shin DH, Chang YS, Shin JI. Antioxidant activity of ethanol extract from Rhus javanica Linne on edible oil. Korean J. Food Sci. Technol. 24: 320-325 (1992) 9. Woo N, Ahn MS, Lee KY. Antioxidant effect of Aloe (Aloe arborescences) extracts on linoleic acid and soybean oil. Korean J. Soc. Food Sci. 11: 536-541 (1995) 10. Bae EA, Moon GS. A study on the antioxidative activities of Korean soybeans. J. Korean Soc. Food Sci. Nutr. 26: 203-208 (1997) 11. Jung GT, Ju IO, Choi JS, Hong JS. The antioxidative, antimicrobial and nitrate scavenging effects of Schizandra Chinesis RUPRECHT (Omija) seed. Korean J. Food Sci. Technol. 32: 928935 (2000) 12. Jeong EJ. Antioxidative and nitrite scavenging effects of solvent extracts from Gyrophora esculenta. Korean J. Food Nutr. 11: 426430 (1998) 13. Kim SM, Cho YS, Sung SK. Effect of ethanol extracts in Pinus densiflora, Lithospermum erthrorhizon on lipid oxidation of oil emulsion. J. Korean Soc. Food Sci. Nutr. 28: 984-989 (1999) 14. Cha JY, Kim HJ, Chung CH, Cho YS. Antioxidative activities and contents of polyphenolic compound of Cudrania tricuspidata. J. Korean Soc. Food Sci. Nutr. 28: 1310-1315 (1999) 15. Christiansen LN, Tompkin RB, Shaparis AB, Kueper TV, Johnston RW, Kautter DA, Kolari OJ. Effect of sodium nitrite on toxin production by Clostridium botulinum in bacon. Appl. Microbiol. 27: 733-7 (1974) 16. Macrae R, Robinson RK, Sadler MJ. Encyclopedia of Food Science Food Technology and Nutrition. Academic Press, New York, NY, USA. pp. 3240-3249 (1993) 17. Bartsh H, Ohshima H, Pignatell B. Inhibition of endogenous nitrosation: Mechanism and implications in human cancer prevention. Mut. Res. 202: 307-324 (1988) 18. Kim DS, Ahn BW, Yeum DM, Lee DH, Kim SB, Park YH. Degradation of carcinogenic nitrosamine formation factor by natural food components. 1. Nitrite-scavenging effects of vegetable extracts. Bull. Korean Fish. Soc. 20: 463-468 (1987) 19. Mirvish SS. Kinetics of dimethylamine nitrosation in relation to nitrosamine carcinogenesis. J. Natl. Cancer Inst. 44: 633-639 (1970) 20. Kato H, Lee IE, Chyuen NV, Kim SB, Hayase F. Inhibitory of nitrosamine formation by nondialyzable melanoidins. Agric. Biol. Chem. 51(1): 1333-1338 (1987) 21. Cooney RV, Ross PD. N-Nitrosation and N-nitration of morpholine by nitrogen dioxide in aqueous solution: Effects of vanillin and related phenols. J. Agric. Food Chem. 35: 789-796 (1987) 22. Kim SB, Ahn BW, Yeum DM, Lee DH, Park YH, Kim DS. Deg-.

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