酸化防止剤力価評価を目的としたDPPHおよびABTSラジカ …

1 DPPH ABTS .. ISSN 13441213. , . , . , . , . , . , . , . , . , . / 40 2 . p. 55-63. 2014 3 .. Tsukuba Business-Academia Cooperation Support Center, Agriculture, Forestry and Fisheries Research Council Secretariat (3) VOL..40 2. 014 C.. 5. 5. DPPH. ABTS . *. 1 * 1. * 1. 2. *. * 2 *. 2. 3 *. * 4 *. 19. *1 . *2 . *3 . *4 . C. omp arati veDPPHa n dABTSR a dic alSca ven gingA. cti vityAss ays f orEva lua tin gNatu ralA. nti oxi dan tsasFoodA. ddit ives YAMAUCHI Ryoko*l FUKAMIZU Sayaka . KOHAMA Yukiko* . SHIMAMURA Tomoko KASHIWAGIT akehiro*2 UKEDA Hiroyuki*2 . AKIYA M A H. iro shi MATSUI Toshiro*4 and ISHIKAWA Hiroya . *1 DepartmentofFoodandHealthSciences InternationalCollegeofArtsSciences . Fukuoka Women's Univers it 1. y 1 1 Kasumigaoka Higashi ku Fukuoka 813 8529.

2 *2 Facul ofAgriculture KochiUniversi " B 200 Monobe Nankoku Kochi 783 8502. I. *3 DivisionofFoodAdditives National1nstituteofHealthSciences . 1 18 1 Kamiyoga S eta g aya k . u o 158 8501. *4 Facul ofAgriculture GraduateSchoolofKyushu Unive . 6 10 1 H. ako zak i H. iga si . ku Fukuoka 812 8581. To assesst h eq ualit yo fn atu rala nti oxidantsu seda sf ooda ddi tive s i ti si mpor tantt oe stabli sh method fore val uatinga nt ioxid anta ct ivi nt hiss tudy t he DPPH and ABTS r adi cals cavenging methods were evalua ted. The antio xidan ta ctivit ieso f21r ef erenceantioxi dants incl udingfl av onoids . poly phenols vita mins aminoa ci ds andp eptid e s were determinedbyt h esem ethods .I nb otha ssays a cat echolandap yroga llolm oi etyc ontr ibutedt oa nt ioxida n tacti vit naddit ion a3 hydro xylg roupi n theC.

3 Ringo fthef l avonoi dsandt hegall ica c i de s term oietyoft hec at echins were a lsoi mp or tantfor ac tivi n tioxid anta c tiv itie sd etermin edbyDPPHandABTS were s imi larw i tht hee xc e ptio no f cat echolcompounds f orwhichv alueswere~ molTE/ molh ig herint he DPPHa ssaytha nint he ABTSa ss a y . We c onc ludet hatDPPH r efl ectsr eact ivat iono ft hecatec holm oietyoft hea n ti o xidant s. Ac o rrel ations tud yo f DPPH and ABTS w i tht he FRAP method showed t h att he DPPH method ref lecte df erri c red ucingabil itymorep recis elyt ha nd idt he ABTSa s say. Tec o nclud et hatth e DPPH. assaywouldbes uit ableasas tand ardmethodf ore val uatingn a turalantio xidantsasf oo da dd itives . 2 0. 13;Accepted] a 0 2014).

4 Keywords:a nti oxi dan ts p oly phe nol DPPH. ABTS. ca tec holcompound DPPH ABTS . *1 8. 13 . 852. 9 1 1 1. S Correspondinga uth or E. ma il:i shi kaw a@f wu. ac. jp *2 7. 83 . 850. 2 2. 00. *3 1. 58 . 850. 1 1 18 1. *4 8. 12 . 858. 1 6 10 1. 5. 6 L 0 2. 014 (4). WST 1 3 .. 25 . 8 6 436 2 5 2 7 WST 1 . 365 1) 2) 2 .. DPPH ABTS .. 7 .. l .. DPPH. ABTS SET.. DPPH ABTS .. 3) . Hydrogen . AtomTra nsf er(HAT) S. ing leE. lec tro nTr ans fer . (SET. ) HAT 21 DPPH . ABTS DPPH ABTS. o xygen . ra dic alabsorban ceca pac ity(ORAC) 4) l owd ens ity DPPH ABTS . li pop rot ein (LDL) . 5. ) FRAP . SET 1 .. 1. 1 . dip hen yl . 2 p icr ylh ydr azy l (DPPH) . 2 2. ' az ino bis(3 et hyl ben zot hiaz oli ne 6 . sulf onicaci d 1.. (ABTS) 7) fe rri cr ed ucingant iox idan tp o ten tia l.

5 (FRAP) 8) f t ran s ( ) .. ( ) ( ) . (ESR. ) ( . 9. ) ( ) . ) 2 (4 . io d o phe n y l) . 3 ( ) ( ) . (4 n i trop henyl ) 5 (2 4 d isul foph e nyl) 2H L ( ) . te trazol onoso diu msa lt(WST 1. ) 10) ( ) . L .. (+/ ) ( ). ( ) . 90% . 95% .. (:t) 6 hydroxy 2 5 78. tetr ame thylchr omane 3 2 . car box yli ca cid ( ) (SIGMA) . DPPH ABTS 1 1 . diphen yl 2 pic ryl hydra zyl (5) . J DPPH ABTS 5. 7. (DPPH) ( ) 2 . 2' . azi no . bis(. 3 e thy 1be n (. Ac) ABTS. z oth iaz o1in e 6 su1. phon ica s id) (ABTS) (Roche . D. iag nost ics) ( DPPH (%) .. ) 2 4 . 6 T. ri(. 2 pyr idyl ) 1 3 . 5 t ria zin e (TPTZ) (1) TEAC. (. mo 1TE. / . ( ) ( ) ( 0. 1) (2) . ) 5.. FRAP.. U 2001 FRAP DUDONNE . ( ) 1 0t 0 A 300t A FRAP . 2.. ( ( pH. ):10m MTPTZ :20mM.

6 5m g FeCb =10: 1 1) 3m R 3 7'. C 30 . ( ) M. ill iQ ( 593nm . ) ( (. x) . ) (110W 15~30 (. y) . ) .. 3.. DPPH (DPPH ) TEAC ( mo1TE/ mol) . DPPH 1110 DPPH.. 2. 2. 00t A. 100m MTris DPPH ABTS 2. 1 . HC1 (. pH7..4)8. 00t A. O..20m MDPPH (. Tab 1e1) TEAC . 1m {! 1. 0 3. 0 Tab1e2 . 517nm 6 . (A. s) ( . ( A. c) 3 ) 1 ..2. O. 0m MDPPH ( ) 3 4 (( ) . (. EC) ( ) (EGC) ( ). (. Ac) (ECG) ( ) . (A As) (. 1) (EGCG)) 1. 1 . (%) 1. 0 (. tra ns . L . (%)= (. Ac . As)/AcX 1. 00 .. (. H1) H. L (+/ ) m . ) . ( ) I. C5. 0 . (. mol /mR. ) (. 2) B . (TEAC) DPPH . (. mo 1TE/. 0. 1) >.. TEAC ( 0 ) = ( IC. 1TE/ mo1 5. a B . ( o l/m R))/ IC 0. 5 ( ol /mR. ) (. 2). B 2. ' 4. ' . 4.. ABTS (ABTS ) mo1TE/. ABTS 10 ABTS mo1 . workings olu tio n 7mM ABTS 5m {.}}

7 ! 140mM DPPH. B . 8 8tA . 12~16 734nm . 0..7: 2 C 3 . ABTS C 3 3 . workingso lut ion 1m{! 1. 00t A . 1. 0 3 0. 'C 4 0..96 mo1TE/ mo1 . 734nm ( A. s) C 3 O..8~ 1. 0 mo1. TE/ mo1 . 5. 8 VOL..40 2. 014 (6). T. abe1 R. l efe ren cecompounds Sample S. ubs tit uen t S. tru ctu res F. lav ono ls Kaempfe rol 3..4 ' OH. Querc etin 3 3' 4' OH. Myric etin 3 3' 4' 5' OH. Morin 3..2 '..4 ' OH. Q. uerc eti n3 . glu cos ide 3 rahamnose 3. ' 4' OH. Ru tin 3 ruti n o se 3. ' 4' OH. F. lav ons L. ute oli n 3'.4' OH. O. D: F. lav an . 3 o ls E. ( ) pi cate chin(EC) 3. 3' 4' OH. E. ( ) pi gal loca tec hin (EGC) 3. 3 ' 4. ' OH. E. ( ) pi cate chingal lat e (ECG) 3. ga lli ca c id 3. ' 4' OH. E. ( ) pi gal loca tec hingall ate (EGCG) 3. ga lli ca c id 3.}

8 ' 4. ' 5' OH. O. the rs . t ran s F. eru licA. cid H. or lx ;c H E. lla gicAcid H.. P. yro cat ech ol C. aff eicAcid Or . ;. H. L. As cor bicAcid .. :: : L. Cy ste ine H . N. G. all icA. cid HO. S. esa mol . G. lut ath ion e HOOC1J .. ' .. / COOH. HO. +/ ) . ( To cop her ol C 3 1 .. C 3 . 3 . ( ~4. 86 mol TE/ mol) . DPPH B . B C . 3 C 3 . i nvi vo ECG EGCG (CAA ) molTE/ mol 7. B ~1. 74 molTE/ mol C. (7) . J DPPH ABTS 5. 9. Table2 A. nti oxi dan tac tiv ityo fcompoundsa sdeterminedbyDPPHandABTS. DPPH ABTS. Sample (. umlTE/ mol o ) (. mo lTE/ l Imol ). F. lav ono ls Kaempferol' 1..O. 1: !:0..01 1..00. :! Quercetinb 3..1. 8: !:0. 03 2. 01 0..09. Myricetin' 3..0. 9: !:0. 11 3. 11: ! Morin' 0..9. 7: !:0. 01 0..8. 6:! Querc eti n3.

9 G1u cos ideb 2..2. 2: !:0. 11 0..96 0..04. Rut inb . 240. ::0. ! 11 0..98: !:0..04. F. lavon L. ute oli nb 2..2. 2: !:0..07 1..0. 4: !:0..26. F. lav an . 3 o ls ( ) . Epic atech in (EC)b 2. 73: !:0..08 1..5. 7:! ( ) . Epi gall ocate chin (EGC) 3. 12 0..05 2..98 0..06. ( ) . Epi cat ech ing all ate (ECG)b 4..86: !:0..21 3. 1l :! ( ) . Epi gal loc ate chi nga lla te (EGCG)' 4..83. :!:0..18 4..67 0..08. O. the rs tr ans F. erulic Acid' 0..71: !: 0. 04 0..64 0..06. Pyro cate c holb 2..7. 3:! 1..08. :! Ca eic Acidb 2..46 0..01 1..08. :!:0..04. Ga lli c Acid' 2..52. :! 2. 49. :!:0..20. Sesamol' 0..7. 8: ! 2 2..03. :! El lagi c Acidb 3..84. :!: 0 2..74. :! L . Ascorb icAcid' 0..87. :!: 0 1..0. 6: ! 4. L . Cyste ine' 0..5. 5: !

10 1 1..3. 7: ! 7. Gl utathi on e' 0..7. 0: !:0..00 0..80. :!: 4. (+/ ) Tocopherol' 0..9. 0: !:0..05 0..9. 4: ! 1. D. ataa ree xpr ess eda sth emean : ! SD (. n=3. ) . : DPPH<ABTS. b :DPPH= ABTS. ':DPPH>ABTS. 3 .. ABTS DPPH .. DPPH . EGCG ABTS .. ' ABTS . 2 mol TE/ > EGC>EC EGCG>ECG . mol TEAC. > . 3 .. 2 1 molTE/ mol . molTE/ mol . DPPH ABTS.. 30 . L DPPH ABTS . (+/. ) . 1 molTE/ mol . 6. 0 VOL..40 2. 014 (8). y=O. ( .901x r= ) . DPPH ABTSr: . TEAC. molTE/ mol 2. 1 DPPH . ABTS (TEAC ) DPPH ABTS . 0..77. 9 DPPH. ABTS . ( ) Table2 . DPPH ABTS molTE/. umol . DPPH ABTS . (DPPH>ABTS) DPPH ABTS . (DPPH ABTS) DPPH DPPH . ABTS (DPPH< . ABTS) 3 2. 1 . Table2 . DPPH . DPPH ABTS (. Fig .1) 16).1 DPPH . DPPH . DPPH>ABTS . ABTS DPPH.