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(1)Journal of the Korean Chemical Society 2004, Vol. 48, No. 6 Printed in the Republic of Korea. . . V`çj šÏR :šJbB~ Öz>² ~ö R Öê. J ^ %*. Ó"&v š& wÏ"¦ (2004. 8. 27 7>) Optimum pH of the Reduction of Hydrogen Peroxide at a Tobacco Plant Tissue Based Amperometric Biosensor Kil-Joong Yoon* Division of Applied Sciences, Cheongju University, Cheongju 360-764, Korea (Received August 27, 2004). *Ú: "Öz>², *~», :šJbB, ‚' Öê Keywords: Hydrogen Peroxide, Amperometry, Biosensor, Optimum pH. * V βº ;‚ ÖWš¾ r¢ÒW χ 7öB æW j ¢b* ‚Wj ©º . V¢B β>wj šÏ~  bîj ;ï~¶ B·>º β*, ¯ :š JbBº Ú¶ pH'öB β / K~ ‚š ¢ Ú¾ºæ {ž>Ú¢ ‚ .  þ öBº ږ çj ê²>#ö b«* "Öz>² ;ïj *‚ :šJbB¢ B·~, © ~ *Vz' ßWj V~,  V˚ Ëç> 6ê& Ú "Ö z>² bB¢ BB~¶ žKš zb–, áÚê Ö " j >Nf ‚: ® . š 7öº î‚Ú Michaelis-Menten š†j wÏ~ :šJbB~ pH ~šW ‚ÿê¢ &VŽb‚B, š·W¶W ֞ Î ²Wî~ / >w";š ‘EH+S→EHS→EH+P’ ª j, Ò Úö šÒ~º "Özβ ÿ*β (isozyme) ~ šÒ¢ *Vz'ž O»b‚ {ž† > ®rj C& ~Vê ~& . "Öz>²º ö>¾Ò¾ ²ëö ÒÏ>–, b', çb, ª* ~ R W 5 îB‚ ÒÏ>º 7º‚ ëÏ £š . V ¢B "Öz>²¢ ³~ ;{~² ;ï~º © f &® 7º‚ ¢š .  ¢^öBº  ’ n;‚ ^ßWj <º * bîj d~ 7, V 1-4. 5-7. 8. 8a,9. 10. 11. ´š ŽF~ ®º "Özβ& – 6w*~f n ;‚ ^ ßWj ¾æÞ º Ò j BÒ~, š¢ šÏ‚ *j B·~  ßWj *Vz'ž O »b‚ V~& . ß® *^~ n;Wf ^ *~&

(2) z>º *~ G;j &Ë ~&b–,  ^*~ ’V~ Öê ~šWj &VŽb‚B V– çj šÏ‚ "Öz>² ;ï :šJbB~ ‚' Ö ê¢ ê† > ®î . Vö  Ö"¢ ~ ¶ ‚ .. þ 8\ # £ B·B *~ *Vz' ßWj ÚÚV *‚ B~*{*~êº EPSILON(BAS Model EPSILON cyclic voltammograph, Bioanalytical System, Inc., U.S.. j šÏ~ áîb–, Vöº Ag/AgCl V&* (BAS, MF2052)" Pt –*š Ö>î . šž ~ Î ^G;f Vƒê, KIPP & ZONEN(BD 111, Holand)& ÖB EG&G Model 362(Princeton Applied Research, U.S.A.) ¢;**V¢ ÒÏ~& . *šî χ~ pH {žf R&jÏχ pH 4, 7, 10(Shinyo Pure Chemicals Co., Japan, S.P.C. GR Reagent)j š A.). V654V.

(3) V`çj šÏR :šJ>*~ Öz>² ~ö R Öê. Ï~ ¦;B Digital pH/mV/Temp meter(Suntex SP701, Taiwan)j šÏ~ šÚr . w&(:0.1 mm)f JªJ¢f '' Flukaf SigmaÒ B®šî b–, Vî‚ ÒÏB "Öz>²º Junsei(35% EP)Ò B®j šÏ~& . þ χf ¢>'b‚ ÒÏ> º jÏbî~ ãÖ 7º*‚ pH–;š ®&Ë~æ‚ 0.1 M~ NaCl(Shinyo Pure Chemicals Co., Japan, 99.5%) 7W*šîj šÏ~& . Ò š *šî χ~ pHº cf HCl(Orintal Chem. Co., Korea)" NaOH(Shinyo Pure Chemicals Co., Japan)j '.® &~ –;~ & .. *~ 9; ê²># * ³öB *¶ *bî‚ ÒÏF. Úbî, ferrocene 0.09 gj 10 mL~ CHCl ö Ÿ w & 0.91 g" b‚ ê, 60 CöB š–~ CHCl j B–‚ . áÚê ferrocene Ž w&f J ªJ¢j 55:45(wt/wt)~ jN‚ b~ ê²>#j 3. o. 3. 655. ò . V –çf 90ª šç ^β¾š&‚ 2ê ~–, jºï~ –çj ~ 60 CöB >ªj B– ‚ ê, š >#ö b«* j*® b‚ r, JZ 7/š B>º çã 6 mm~ polyethylene Â2ö j ò *b‚ Òς . o. 5b. Ö # 8 *~ *Vz' ß9 Fig. 1f Öê& pH 7.75ž *šî χ 7öB  V–ç Žïš 10%ž *~ *Vz' ßWj  "º B~*{*~êš . a, b, c, d~ ³B BB ‚ *{ "Ò>îb–, af bº *šîö Vîχš &šææ p~j ãÖ~, Ò cf dº 10 mL * šî χö 0.10 M H O 200 µL& &šê ãÖ~ æzB ÎÛj " ® . r~ OËb‚ *{ " Ò(0.0 mV→ −300.0 mV)~º a~ ãÖ, r** Ã& ö V¢ ~ö*~& 6ê'b‚ Ã&~º ©j  2. 2. Fig. 1. Cyclic voltammograms of the tobacco plant tissue modified carbon paste electrode without(a, b) and with(c, d) 200 µL of 0.10 M H2O2 in 10 mL of 0.10 M NaCl( pH: 7.75, scan rate: 10 mV/sec, tissue %: 10). The current difference between a and c is shown(inset). 2004, Vol. 48, No. 6.

(4) J^7. 656. " ®º–, šº *ö ŽFB V–ç ¶Ú&   &æ zb~ bbž 6j 6n† r, * ê öB ’–& {ž>æ pf *Vz >wš ê Ú¦ öB ¢Ú¾ ®j &ËWš ®b–, *** Ã& ‚ ž‚ Ï**~~ Ã&& Žþ V‚ ©b‚ " > ® . c~ ãÖ r** "Ò , −40 mV(vs. Ag/AgCl) 6öB Vîš Î&>¶ **~~ /ς æz ¢ " ® . š æzº Vî~ Î&ö Vž~ , −40 mV šêö áÚæº *{*~ê~ ;& a f jv~ *~Ã& žö æ;B 6š ìº ©b‚ j Ã&B *~ af c~ Nšº Jç "Öz>²~ ~öö Vž‚ ©š¢ šê ZO† ©š . *~ a f c~ N¢ ~, ©j *{ö &~ ê‚ © š Fig. 1~ cö "Ú^® . *~~ N, ¯ ^*~ º *{ö &~ Z&‚ ©j " > ®º–, šº * **~ æzö &~ çF'b‚ æ~º ‹~ ã Ö¾, – Ã&~º ~ *–ç~ ãÖfº &– 'š . ò VB Ã&B *~& *{ Ã&ö Z&~² ¢;‚ ©f ~ö>º z«š ¢«š –, ©š "Öz>²ö VžŽj ÝAŽ~ " ® . ^*~ G;j *‚ ***º * .~ ã Ö  ÔºÚ −150 mV(vs. Ag/AgCl)‚ ;~&. . Fig. 2º  þöB ^& áÚæº *;'ž ";j " ® . pH 9.37~ 10 mL *šî Ï ‡ 7öB 19%~ V–çj Ž~ ®º *j šÏ~º ã֞–, Vîš ìº çöB **ê 9,12. 6,8b,10. Fig. 2. Typical time course of signal showing the experimental determination of the response current and the saturation time. The measurement was under the following conditions. pH= 9.37, tissue%=19, electrode potential=−150 mV(vs. Ag/AgCl). 30 µL of 0.10 M hydrogen peroxide was added into 10 mL electrolytic solution.. Fig. 3. Effect of the tissue composition on the response current at pH 7.75. 50 µL of 0.10 M H2O2 solution was added.. 'Ï , /Ï® ;WB Ï**~º & Û 15. ã" ê :ûF >&b‚ 6²~ ^G; ö 'Ëj ~æ pº . :ûF~ æz& ìº ç , ¯ *~ ãÖ **ê ;W ê 70.& ã">î j r, 0.10 M H O Vîχ 30 µL& Î&>š ^ *~& W>º–,  *~º &Û 13.¢ ã"~š z>,  ’Vº 180 nAªj " ® . ‚ Þ * ‚šö šÒ† > ®º ‚W¶Ò~ >º *  bîš Ž~º –ç~ Žï æzf ç7' &N š ® . *~ V–ç Žïòj æz* ^* ~~ æz¢ &V‚ Ö"& Fig. 3ö "Úr . * ** −150 mVöB 0.10 M H O χ 50 µL¢ &† r, ^*~º –ç~ Žï æzö &ڂ çFW (y=16.7x−54.2, y: current(nA), x: tissue %, R=0.993)ª j " ® . ãËWö Všš – ^¢ áV *~ *š  ôf –çj ކ ©š º’> ¾, Žï%& Ã&†>ƒ ê²>#~ 6ê& ²ž~ –, *~ > ÒÏb‚ ž‚ * ‚š~ "¶š *~~ Ò*Wö ’² ¶çj "æ‚, G;ö ÒÏF 2êB V–ç~ Žïf 19%‚ ;~& . *~. þÖ"‚¦8 áÚê þ –š, ¯ *** −150 mV f –ç Žï 19%ž *j 0.10 M NaCl χ 10 mL ö *~Ê,  þ pH8öB 0.10 M VîÏ ‡ 30 µL¢ º&~&j r áÚæº *~ i(nA)~ 8. š Table 1ö "Ú^ ® . Ò š 8 j pHö &~ ê‚ ©š Fig. 4ö "Ú^ ® . Fig. 4&  "º ãËWf pH 8.00" pH 10.0 ҚöB &8 š ;W> ®rj r > ® . Wî‚ ’WB Î (potential step). 2. 2. 2. 2. Journal of the Korean Chemical Society.

(5) 8`çj šÏR :šJbB~ Öz>² ~ö R Öê Table 1. Effect of pH on the response current and the saturation time. 30 µL of 0.10 M hydrogen peroxide was added pH. i(nA). t(sec). pH. i(nA). t(sec). 3.56 4.07 4.68 5.08 5.55 6.10 6.80 7.60. 33 45 60 83 105 120 140 150. 52 48 44 34 26 21 20 18. 8.26 9.07 9.78 10.45 11.19 11.78 12.30. 158 177 153 132 117 84 64. 18 15 18 20 22 26 28. 657. ©j r > ® . V–çj Òς Öz>² bB ~ ‚' Öêö &~  ôf \& šÚ^¢ † ©š¾, * êöBº š 8j ‚' Öê¢ š ê ZO† ©š . Table 1öº Fig. 2öB áÚê ©  ÿ¢‚ O»b‚ G;B *~~

(6) z*š "Ú^ ®, pHö 8ž ©~ æz& Fig. 4ö Žþ ê >Ú® . pHæzö 8ž

(7) z*~ æz ãËWf *~ ’Vf >& ãËWj " ®b–, ©j Ò.š~‚ O;f š . š ~ ‚²8 14.5 secº pH8š 8.93¢ H áÚæº–, š 8f *öB *~~ ’V‚¦8 áÚê 8.517 ’² šæ p . šº pH 8.93~ ·W¶ ³ê& š·W¶WÖ(diprotic acid) ž jžÖ~ ºV‚ ;WB β~ ‚W¶Ò~ «Ú ' \–ö &~ Vî~ ç^·Ïš &Ë ÎN'š >êƒ 'Ëj ~ ®rj ö~ " ® .. Ö V Fig. 4. pH-dependence of the current( ù ) and the saturation time ( ø ). Other conditions are the same as in Fig. 2.. ²~ ‚W¶Òº χ 7ö šÒ~º ·W¶~ îO š &˂ šš ö jž öb‚ \W> Ú ® . æ‚ Ï‡~ pHæzº β ‚W¶Ò~ «Ú' \–, ¯ β~ / Kö ’² 'Ëj † > ® . VB pHæzö 8¢ ‚& *~¢ ¾æÚ º pH8š šÒ~º ©f ·W¶~ îOš β  Wî Vî~ «Ú' ֏j ‚'z* / Kj ‚&z~º pH& šÒŽj Ò~ ® . þ'b ‚ pH8òj ^ª~ ‚'pH¢ {ž† > ìbæ ‚ ºïš‚¦8 ºÂB Özβ¢ šÏ‚ * ~ ãÖ f ?š Öz>²~ ~öŽ>& &ÖÊ ª

(8) ¢ ;W† ©š¢º &; ~ö &ÖÊ Ž>‚  Ò.š~~ O;, 9. i = (A ⁄ ( w* sqrt ( pi ⁄ 2 )) )* exp ( –2* ( pH – 8.517) ⁄ w). 2. –39.17. j áî . VB Af w~ 8f '' 1715f 6.90 š–, Ž>~ ‚&8f pH8š 8.517¢ H áÚæº 2004, Vol. 48, No. 6. Öz-~ö, ¯ *¶~ šÿš >>>º β>wf *Vz'ž O»b‚ >w~ ³ê¾ >w z î ¾ö ‚ ;¢ áj > ® . V–çj

(9) Ž‚ *j B·~, š‚¦8 áÚê Öz>²~ pH ~šW ~ö*~f, ^*~&

(10) z>º *~ æ z ãËWj ªC~ V Öz>² ªšÎ²~ ‚' Öê¢ êÂ~& . v O»b‚ áÚê ‚' Ö êº ’² šæ p~b–, '' 8.517 8.93šî . š ¢^f 2004-2006jê Ó"&L F\ –Wj(ßêB)ö ~~ \>îb–, šö pš 6Òãî .. žÏ^ò 1. Ruan, C.; Yang, R.; Chen, X.; Deng, J. Electroanal. Chem. 1998, 455, 121. 2. Sergeyeva, T. A.; Lavric, N. V.; Rachkov, A. E.; Kazantseva, Z. I.; Piletsky, S. A.; El’skaya, A. V. Anal. Chim. Acta, 1999, 391, 289. 3. Bongiovanni, C.; Ferri, T.; Poscia, A.; Varalli, M.; Santucci, R.; Desideri, A. Bioelectrochem. 2001, 54, 17. 4. Liu, S. Q.; Ju, H. X. Anal. Biochem. 2002, 307, 110. 5. (a) Kwon, H. S.; Kim, K. K.; Lee, C. G. J. Kor. Chem. Soc. 1996, 40, 4, 278. (b) Yoon, K. J.; Pyun, S. Y.; Kwon, H. S. J. Kor. Chem. Soc. 1997, 41, 7, 343..

(11) 658 6. Yoon, K. J.; Kim, K. J.; Kwon, H. S. J. Kor. Chem. Soc. 1999, 43, 3, 271. 7. Lee, B. G.; Yoon, K. J.; Kwon, H. S. Anal. Sci. Tech. 2000, 13(3), 315. 8. (a) Yoon, K. J. Bull. Korean Chem. Soc. 2004, 25, 7, 991. (b) Yoon, K. j.; Lee, B. G.; Kwon, H. S. Anal. Sci. Tech. 2000, 13(1), 41. (c) Lee, B. G.; Kwon, H. S.; Kim, K. E.; Yoon, K. J. Anal. Sci. Tech. 2001, 14(3), 286. 9. Yoon, K. J. Anal. Sci. Tech. 2003, 16(6), 504. 10. (a) Kirchner, J. R. Encyclopedia of Chemical Technol-. J^% ogy; Wiley-Interscience; New York, U.S.A., 1981; Vol. 13, p12. (b) Irving Sax, N. Hawley’s Condensed Chemical Dictionary; Van Nostrand Reinhold: New York, U.S.A., 1987; p 618. 11. (a) Lin, M. S.; Tham, S. Y.; Rechnitz, G. A. Electroanalysis 1990, 2, 511. (b) Fang, Y.; Cai, R.; Deng, J. Electroanalysis 1992, 4, 819. (c) Oungpipat, W.; Alexander, P. W.; Southwell-Keely, P. Anal. Chim. Acta, 1995, 309, 35. 12. Yoon, K. J.; Kwon, H. S. Anal. Sci. Tech. 2004, accepted.. Journal of the Korean Chemical Society.

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