Chard Root-Tissue Based Biosensor for the Determination of Dopamine

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1 Printed in the Republic of Korea w q w «z *Á½x Á ¼ Á û w w w w w w w w yw ( ) Chard Root-Tissue Based Biosensor for the Determination of Dopamine Hyoshik Kwon*, Hyunjung Kim, Kil-Joong Yoon, and Yongnam Pak School of Science Education, Chungbuk National University, Cheongju , Korea Division of Applied Sciences, Chongju University, , Korea Department of Chemistry Education, Korea National University of Education, Cheong-Won, Chungbuk , Korea (Received January 23, 2007) : Chard Root-Tissue Based Biosensor, Dopamine, Carbon Paste Electrode Keywords: Chard Root-Tissue Based Biosensor, Dopamine, Carbon Paste Electrode 1. y z w w 1962 Clark w š, 1967 Updike w 1 tw. 2 ( ) l z š w w z Mascini w šw, š g y w. 3 z, Wang, ùù, š þ phenol w š, Gunasingham x sw 4 glucose, 5 Rosa ˆ ù j w» 6 w. w Conrath w», 7 Lee dithizone w cyanide û w.» 8». Dopamine w w., dopamine qk (Parkinson s disease) 9, š (muscular immobility), (neural crest tumor) 10 š w š š. 11 Dopamine w tyrosinase, catechol oxidase lactase w z dopamine ùù,,,, e w dopamine ƒ š Tyrosinase, catechol oxidase lactase û p (specificity), û, û x ùkü, 3~ s s z y y yw z ƒ q z j. Tan Chen l 22 q ü w j» w ww w w. w dopamine w 3 p ùkû š šw. 17,18 291

2 292 «z Á½x Á ¼ Á û Biosensor dopamine w wù Sidwell Rechnitz 21 w š ƒ oil, ùù ùù. ùù polyphenol oxidase z current ùkù dihydroxy xk quinone»yw y j dopamine w. Caruso 22 Cara(Dioscorea bulbifera) l w polyphenol oxidase w dopamine w. Pravda 23 š j m v (HPLC)»yw» k tyrosinase š w š k (SCPE) w dopamine w w.» k ph j w. Karina y (zucchini) w dopamine w šw w M šw. 24 ¾ š w dopamine»yw p š w.»» x 20 ml v BAS Model VC-2 Voltammetric cell w. cell teflon,» Ag/AgCl (Model Re-1 BAS) š w Fig. 1. d EG&G Princeton Applied Research Model 264A Sacanning Potentiostat w, Kipp & Zonnen recorder w. j vr(100 µl) Gilson w. Graphite powder Kanto Chemical Co. Inc. t, w KH 2PO 4 K 2HPO 4 Shinyo,» w dopamine Aldrich t w. w mineral oil, ferrocene chloroform Aldrich t w. w z» w ascorbic acid yœ œ z t, glucose yw z ( ) t, potassium tartrate, sodium bicarbonate, Fig. 1. Curremt-time recordings obtained at (a) 9% ferrocene modified carbon paste electrode, (b) 40% chard root tissue modified carbon paste electrode, (c) 40% chard root tissue and 9% ferrocene modified carbon paste electrode on increasing the dopamine substrate concentration. Successive increments of M dopamine. Operating potential, V. ph=6.53, M phosphate buffer. Stirred 300 rpm. glycine œ z (, GR) t, urea Osaka Hayashi Pure Chemical Industries Ltd., Japan t w. w w w w, 4 o C w þ š Áò k s g w v w. ph M KH 2PO M K 2HPO 4 v ww w, ph d w digital ph/ion meter(model DP-215)ƒ. w, w x w. y d 10% 9% ferrocene sww g ferrocene 20 ml CHCl 3 z 0.91 g graphite powder ³ w yw k hot plate g. yw 0.60 g w mineral oil 0.40 g yww k z, g q w g ƒw z, yw k k 6 mm, ¼

3 Fig. 2. Variation in current with ph. Other conditions were same as in Fig mm mx ó 1 mm g Cu w w Fig. 2. d ƒƒ, 10 ml swwš cell M dopamine 20 µl ƒw z ƒ z yj» w d w. š ƒ d 300 rpm. v ferrocene w w š, w 4 o C þ w x d w. š w f. w q w 293, š z peroxidase dopamine dopaminequinone y jš, y peroxidase»(potentiostat) w dopamine y. y d w. wr, w z w š,» w z w ùkù. ƒ y w»(stem), (leaf), (root) ƒ ƒw. p 3.4 yƒ j ùkû,» ƒ 7 ƒw. dopamine x w. Dopamine ph y Fig. 1 (ph=6.53) 10 ml M dopamine 20 µlƒ 10z ƒ 40% 9% ferrocene sww (c), 40% sw w (b), 9% ferrocene sww (a) (-0.15V vs. Ag/Agcl) y x. Fig. 1(a) œ dopamine ƒ ùkü. sww (a), sww (b) (a) w ùkû (c) w w. dopamine ferrocene z w ùküš. ferrocene sww (c)» y w ùküš, ü k ùküš. Fig. 2 ph ùkü x. Tu 25 ph w w š š w. 32 o C, ph=6.0~8.0 peak currentƒ ùkû. Chen Tan 26 ƒ ph, ph=6.5, ph=5.56, ùù ph=5.56, (mushroom) ph=7.4 ùkþ š šw. x V

4 294 «z Á½x Á ¼ Á û Fig. 3. Dependence of the biocatalytic current on the operating potential. Other conditions were same as in Fig. 1. Fig. 4. The effect of the tissue composition on the response the carbon paste electrode. Other conditions were same as in Fig. 1. (0.050 M KH 2PO 4: M K 2HPO 4=8 : 2) ph =6.53 ƒ š, ù û wš. x ƒ j ùkü ph=6.53 kw. w y Fig. 3 w š. ph= M M dopamine 20 µl ùkù d w V l V¾ 0.05 V y k V y ƒ j z w. x ƒ V kw. w y y w y j w š. k w yƒ z w y w». k x w 9% ferrocene œm w xw. 15% 60% w Fig. 4 ùkû 40% w w ƒ ùküš. Fig. 4 15% 40%¾ ƒ ƒw w ƒw t ƒ ƒw 27, Gunasingham (» 40%) ƒ w w ƒ wš. 28 Mediator(ferrocene) w y y Fig. 6 ferrocene w y y š. ph= M 40%, d V (vs. Ag/AgCl), dopamine M š w. Fig. 1 w

5 w q w 295 w w ferrocene w w ùküš., ferrocene w j» w e. ferrocene w 3% 13%¾ 2% y j d w. x 9% w w ƒ ùküš. Oungpipat ferrocene ƒw ferrocene w œw» ƒ ƒw š wš 29, Senda ƒ ƒ, yƒ k j ƒ w, ƒ w š w. 30 Fig. 5. Optimization of mediator concentration in the carbon paste electrode. Other conditions were same as in Fig. 1. w w Table 1 w w r» w, w x w e š ƒ ƒ kw w. Sotomayor 31 ascorbic acid, uric acid, acetaminophen dopamine w ascorbic acid wƒ ùkû, uric acid, acetaminophen y j» w š šw. x ¾ x w, M dopamine ƒw z» M w. ascorbic acid, glucose, glycine, potassium tartrate, sodium bicarbonate, urea dopamine yj» Table 1. Interference studies of some important dopamine donors in the presence of dopamine Compound * S X,dopamine/S dopamine Ascorbic acid 1 Glucose 1 Glycine 1 Potassium tartrate 1 Sodium bicarbonate 1 Urea 1 Fig. 6. Carlibration curve for dopamine. Other conditions were same as in Fig. 1. *Each injection changed in [interference] = M and [dopamine]= M **S X,dopamine: Steady-state sensitivity of interference in the presence of dopamine S dopamine : Steady-state sensitivity of dopamine S X,dopamine/S dopamine : Relative sensitivity

6 296 «z Á½x Á ¼ Á û Fig. 7. Calibration curve for dopamine in range of M ~ M of dopamine. Other conditions were same as in Fig. 1. w w wš. dopamine w w w e š w w. Dopamine š Fig. 6 dopamine x w š ùkü. Dopamine ƒw w ƒw. w, Fig. 7 dopamine, ~ w y ùkü. - ùkû. wr, Fig. 8 w ƒ ùkü. Michaelis-Menten» ([S]) y g z ([ν]) dwš Lineweaver-Burk w, z w. 29 Fig. 8 w z ùküš. w dopamine r Fig. 8. Lineweaver-Burk plot of the same data as in Fig. 9. Other conditions were same as in Fig. 1. wš yw d w. q ƒw y 95%ƒ ¾, t 95% 12 s, dopamine w M. k š k, dopamine w x w. ph=6.53(0.01 M ) ƒ ùkþš, V y ƒ š ùkü. w, 40% 9% ferrocene sww ƒ ùkü. q ƒw y 95%ƒ ¾, t 95% 12s, dopamine w M. w dopamine ƒw w ƒw š, w ùkû.

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