Method Development for the Sample Preparation and Quantitative Analysis of Synthetic Colors in Foods

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(1)KOREAN J. FOOD SCI. TECHNOL. Vol. 36, No. 6, pp. 893~899 (2004). ©The Korean Society of Food Science and Technology. ® % Ïæï²~ òê *¾ÒO» {ã 5 ï ªC ;9Á

(2) >Á;ß]1,* ®~£®n*Ó ®Î&>", 1ã\v ®«". Method Development for the Sample Preparation and Quantitative Analysis of Synthetic Colors in Foods Sung-Kwan Park, Tal-Su Lee, and Seung-Kook Park1,* Food Additives Division, Korea Food and Drug Administration Department of Food Science and Biotechnology, Kyung Hee University. 1. Method for sample preparation and quantitative analysis of 19 permitted and non-permitted synthetic colors in foods was developed based on reversed-phase ion-pairing high performance liquid chromatography. For color extraction of samples, deionized water was added, and pH was appropriately adjusted with 1% ammonia water. Any undissolved matters were extracted with 50% ethanol or 70% methanol. Lipid in snacks was first removed using n-hexane with centrifugation, water was added to extract colors, followed by clean-up and concentration using Sep-Pak C18 cartridge. Recovery efficiencies at known concentrations of 19 standard food colors spiked into foods were in 90.3-97.9% range for soft drink, 79.2-101.9% for candy, 84.1-103.4% for jelly, 86.4-100.8% for chewing gum, 83.5-103.4% for ice cream, and 78.5-95.6% for snack. Key words: synthetic food colors, sample preparation, ion-pairing HPLC. B. . f ëWj ï&~ n*~ «ÃB © . ÖÒ¾¢ öBº Ïï² 'ï 2^, 'ï 3^, 'ï 40^, 'ï 102^, ï 4^, ï 5^, ï 3^, Óï 1^, Óï 2^~ 9« 5 rª. 7«('ï 3^, 'ï 102^ B) ®ö ÒÏ > ®ê ®Î&b*ö îÏ>Ú ®b, $ Â ®, ~, f~~, ~, Zæ >~ ®ö ÒÏj .æ~º ; ÒÏV&b J;>Ú ® (3). ® 7 Ïæï²~ ªC»b ®*öº ®ö Ò ÏB æï²ö &~ æîÆ¢b 5 ;[îÆ ¢bö ~ ;W»ò >>Ú ®b¾ O» f ®ö Ò~º bî Rf 8j æzÒ > ®, ö V¢ ªC&ç®" B> &ï²* Nö ~ ªC ~ ;&ê 5 ;{ê& ÎÚî > ®b, $ ;ïj > ìº 6 ® . HPLC¢ Ï æï² ªCö & (1,4-12)ê ®b¾ ò¦V ï²¢ ºÂ~º *¾Ò ";" Obîj B~V * ;B"; ¢¦ &ç® 5 ï²ö ;>Ú · ®ö 'Ï~V ¦~¾ º& ' & jº~ Ôf ²>N ~ ^B ®*¦Ò V& öB ;þ»b 'Ï~Vöº 6 ® . V¢B æï²~ ÒÏV& *> ¦¢ {~¾ >« ® 7~ ®îÏ æï²¢ ;{~² ¦Â~ ;ï'b ª C &Ë ªC»~ BB jº ; . &®~ >Â«ö V ï² ÒÏï Bö &j~ ¢ Û æï²~ Sï Ò, *ê ï&ö jº " ' "¢ {~V * ; (13,14)f ÖÒ¾¢öB ®. " ~ æz &®~ >º& Ã&ö V¢ ®Î&b~ ÒÏ Ã&> ®b ö V¢ Î&b~ S n*Wö & &ê ¸jæ ®º º^ . Oïòº Â®j &, ~º ";ö B > ®º æïOæf »Ãê, &~Ëç ~ Ï'b ÒÏ> ®b¾(1,2) ® * Òò¢ ³V * *zÏ'b ÒÏ>¾ jºç b ÎÏ~º ãÖ f ®*»öB êÏ® ÒÏj B $º .æ~ ® (3). Ïæï²º &'W Ú¾B Oïò ô Ï~ ® . Cêæ 7ö FB ÊFÒ(benzene ring)¾ ¾* î2 Ò(naphthalene ring)¦V W bîB *Ò îÏ > ®º ©f Îv >ÏW~ ÖWæï²ö ³~, z çb jê(azo type), ºvê(xanthene type), ÞÒ¾öz êê(triphenylmethan type)f :ê(sulfonated indigo type) ª~> ® (1,2). ¢>'b ®öB ¢>' ï²~ Ò Ïïf 5-100 ppm ;ê, ¢¦ ï²º FW ¢¦ ®Ú ÒÏ .æ>Vê ®b¾ *Ò Ïï² ÒÏ î&B © *Corresponding author: Seung-Kook Park, Department of Food Science and Biotechnology, Kyung Hee University, Yongin-Si, 449-701, Korea Tel: 82-31-201-2655 Fax: 82-31-204-8116 E-mail: [email protected] 893.

(3) ®"²æ B 36 ² B 6 ^ (2004). 894. ö ÒÏ îÏB 8«(rª. 15«)~ æï² ¢ &çb ;WÁ;ï ªCO»j ¦Æ~& ;{ê 5 Ò *W ¸f ³ÚîÆ¾b(HPLC)¢ F~ ÿ ªC»ö & ¢ >¯ : ® . öBº ÖÒ¾ ¢öB îÏB 9«" öBº ÒÏ îÏ>Ú ®b¾ Ö Ò¾¢öBº îÏ>æ ®æ pf ï² 5 >«®7 ¦ÂÒ f& ®º ®îÏï² 10« C 19«~ æï²¢ &çb ~ ® 7~ ò*¾ÒO»j ¦Æ~ {ã~ N 3 £j Ï ³ÚîÆ¾b O»(13,14)j Ï ~ · ®~ ï²ö & Ò*W" ²>N j { b ® 7 æï²~ ïj ÎN' ³~² ª C~¶ ~& . $ ¢ Æ& ÖÒ¾¢öB FÛ> ® º Ïæï² Ò"~ ¢~ ¦ê Ò~& .. Òò 5 O» Òç® 7öB FÛ> ®º B® 7 Óïrò, Òû, JÒ, *¯ ò, jÊâ, "~ C 6« 120®Ïj F;~ ªC& çb ~& . £ Methanol" acetonitrilef MerckÒ(Darmstadt, Germany)~ HPLC/j, tetrabutylammonium bromide(TBA-Br)º Junsei ChemicalÒ(Tokyo, Japan)~ B®j ÒÏ~&, £f WakoÒ(Osaka, Japan)~ ß/£j ÒÏ~& . þö ÒÏ ï² f TCIÒ(Tokyo, Japan)~ B®b Table 1ö ;Ò~&. . ;BÏ ÞÒæº Sep-Pak C18(Waters, Milford, MA., USA)j zêR 10 mL 5 0.1% TBA-Br >Ï 10 mL¢ ÒÏ ~ Nf ^;Î ©j ÒÏ~& . Ï~ B 'ïêÛ(R2, R3, R40, R102, R104, R105, R106, Azo, R2G)" ïêÛ(Y4, Y5, ORII, QY), Óï ïêÛ(B1, B2, G3, GS, PBV, BBN) ï² 5 bï² &Ï~ 100 ppm Ïj ò ê ¢ C~ 0-50 µg/mL~ ³ê º*öB ¦ ïFj ~& . HPLC ªC ªCö ÒÏ HPLCº WatersÒ(Milford, MA., USA)~ M510 solvent delivery system" 717 autosampler, 486 UV detecter WB ©bB ªÒÏ "f Symmetry C18(3.9 mm i.d.Ü150 mm, 5 µm, Waters, Milford, MA., USA)j ÒÏ ~& . ÿçbº 0.01 M TBA-Brj F 0.025 M ammonium acetatef acetonitrile, methanolj 65 : 25 : 10" 40 : 50 : 10b b v &æ W~ Ï ¢ ÒÏ~ gradient mode ªC~& . ÿç~ F³j 1.0 mL/minb ~ ò¢ 20 µL "«~&b ¦ÂV~ 2Ë" ÿç~ gradient f Table 2ö ;Ò~& . ò % ï²~ ºÂ Óïròf Òû, JÒ, *¯ò~ ï²º Ã~> ºÂ~& , jÊâ" "~º Ö $º CFör¢ ÒÏ~. Table 1. Synthetic food colors used as standards for HPLC analysis Permitted colors1) Food Red No. 2 (Amaranth, R2) Food Red No. 3 (Erythrosine, R3) Food Red No. 40 (Allura Red, R40) Food Red No. 102 (Ponceau 4R, R102) Food Blue No. 1 (Brilliant Blue FCF, B1) Food Blue No. 2 (Indigocarmine, B2) Food Green No. 3 (Fast green FCF, G3) Food Yellow No. 4 (Tartrazine, Y4) Food Yellow No. 5 (Sunset Yellow FCF, Y5) Non-permitted colors2) Food Red No. 104 (Phloxine, 104) Food Red No. 105 (Rose Bengal, R105) Food Red No. 106 (Acid Red, R106) Orange II (ORII) Azo Rubine (Azo) Red 2G (R2G) Brilliant Black BN (BBN) Patent Blue V (PBV) Green S (GS) Quinoline Yellow (QY) 1) Aluminum lake of permitted food colors except for Food Red No. 3 and 102 were also used. 2) PBV and QY are the products of Warner Jenkinson in Great Britain.. æOj B r zêR, öêR, ²*6R, zÎîj>, NaOH Ï j ÒÏ~ ºÂÎNj jv~& . ºÂ ê ö ªÒ~ ;bj B~ pH¢ 5-6b ;~ Sep-Pak C18b ;B~& (15). ®b¦V ï²¢ ºÂrö ºÂ ÎNj ¸V *~ ºÂNê¾ ºÂÏ , îz ¾Ò, . r2 ¾Ò &æ O» 7 &Ë ºÂN ¸f O» j F~& . ò~ ;B Sep-Pak C18 ÞÒæ¢ Ò zêR" b ^; ê acetic acidf 1% zÎîj> Ïb Sep-Pak C18 ÞÒæ¢ Û" Êº ï²ºÂ~ pH¢ 5-6b ;~ 0.1% TBA-Brj &~ Sep-Pak C18ö ï²¢ FV . b 10 mL ^;. r 0.1% "Ö-zêR 10 mLf 1% zÎîjW zêR £ 2 mL ÏÂ~& . ® % æï²~ ²>N ¦Æ 6> ®º Óïrò, Òû, *¯ò, "~, jÊâ, JÒ 7 ï²¢ F~æ pº ©j Î®b F~ ' ï²~ ²>Nj Ò~& . ' òö bï² &Ïj Î&~ ò 7 ï 10 µg/g >ê ê òê *¾ Ò O»j Ï~ ' Î®b¦V ï²¢ ºÂ r Sep-Pak C18 ÞÒæ¢ Û"B ;B~& . ÏÂf b 2 mL¢ & r ÃB ³»~ 0.5% zÎîj> 1 mL >V ¢ ^¿~ bj &~ ;{® 10 mL ê HPLC¢ Ò Ï~ ;ïªCj ~ ²>Nj 3² > G;~& ..

(4) ® 7 æï²~ ò*¾ÒO» {ã 5 Fï ªC. 895. Table 2. HPLC conditions for the analysis of 19 synthetic food colors Color type. Operating conditions. Yellow. Column Eluent Detector wavelength Flow rate Injection vol.. Symmetry C18 (3.9 mm i.d.Ü150 mm, 5 µm) A1) : B2) = 100 : 0 0 : 100/8 min, curve #8, 0 : 100/15 min UV 420 nm 1.0 mL/min 20 µL. Red. Column Eluent Detector wavelength Flow rate Injection vol.. Symmetry C18 (3.9 mm i.d.Ü150 mm, 5 µm) A : B = 75 : 25 0 : 100/8 min, curve #9, 0 : 100/18 min UV 520 nm 1.0 mL/min 20 µL. Blue & Green. Column Eluent Detector wavelength Flow rate Injection vol.. Symmetry C18 (3.9 mm i.d.Ü150 mm, 5 µm) A : B = 100 : 0 40 : 60/7 min, curve #7, 40 : 60/15 min UV 620 nm 1.0 mL/min 20 µL. Mixed. Column. Symmetry C18 (3.9 mm i.d.Ü150 mm, 5 µm) A : B = 100 : 0/2 min, 100 : 0 60 : 40/24 min, curve #6, 60 : 40 0 : 100/26 min, curve #6, 0 : 100/35 min UV 254 nm 1.0 mL/min 20 µL. Eluent Detector wavelength Flow rate Injection vol. 1). A: 0.025 M ammonium acetate (containing 0.01 M TBA-Br):acetonitrile:methanol = 65 : 25 : 10. B: 0.025 M ammonium acetate (containing 0.01 M TBA-Br):acetonitrile:methanol = 40 : 50 : 10.. 2). Ö 5 V ¦ïF ·W ï²&Ïj 0, 2, 5, 10, 20 5 50 µg/mL~ ³ê B ~ 20 µL¢ HPLCö "«~ áf îÆÎöB ¦ ïFj ·W®j r Îv ·^ çFW(r = 0.9999)j ¾æÚ î (Fig. 1). ÿ¢ ³êöB~ ¦ÂVö & >wf 'ïï² ~ ãÖ R3f R106 ï²'ïêÛ ï² ö j~ ¸ ~, ïï²~ ãÖº Y4, QY, ORII, Y5~ BB, Óï 5 ïï²~ ãÖº G3& &Ë ¸~, BBN &Ë Ô~ . $ êÛê Ïï²~ ¦Âêº 'ïêÛ~ 0.05 µg/g, ï êÛ 0.03 µg/g, Óï 5 ïêÛ 0.01 µg/gî . ò %~ ï²ÏÂ; Óïrò, Òû, JÒ, *¯ò, jÊâ~, "~ 6« ~ òö & ï²~ ÏÂj r" ?f O»ö ~~. ~& . Óïròf Òû, JÒ~º ò £ 5-10 µgj ~ b ï²¢ ºÂ~&b r bö ¾ º ò~ ã Ööº NöB, bö ¾ æ pº òº 50-60oC >»çö B 10-30ª* &N~ v>, ÏÊ¾ îz $º .r 2 ¾Ò~& . Òû" JÒ~º ¾² ¦>ÚB ïï~& ò& rzRj F ãÖöº ÃB³»~ rzRj B~ & . öªÒ(3,500 rpm, 15 min)~ çûj ~ bö ®j*~² Ï>º ãÖº *bö 50-80% öêRj &~ v>~ 2-3* ÿn NöB ºÂ~&b, öêRê ºÂ ®j* ãÖº 1 N "Ö 10 mL¢ ÒÏ~ ºÂ~ & . *¯òf ò¢ £ 5-10 g ~ Óïrò¾ Òû, J Ò~f ÿ¢~² ºÂ~> b $º 1% zÎîj> ºÂ~& .. jÊâ~º ò £ 5-10 gj ~ îz ê b ï²¢ ºÂ~& . v>~B NöB 2-3* $º 50-60oC >»çöB 10-30ª* &N~ ò¢ ÏV . bö ¾ Ï>æ pj ãÖöº b" öêR~ 1 : 1 b $º 1% zÎîj:50% öêR Ïb ºÂ~& . öªÒ~ çû j ~ *bö ï²& OB ãÖº 1 N "Ö 10 mL ºÂ";j >~& . "~º æO ï ¸jB b ï²¢ ºÂ~V& Ú[ òö ÒO ¢ÚÆ > ®bæ(16) ºÂÏ ¢ Ò ~ pH¢ æzBB ºÂj ¦Æ~& . zÎîj>f >Öz¾Þj ÒÏ~ pH¢ r¢ÒWb ;~&, & N^òj ^ öêR, zêR, ²*6R j b" b ~ ºÂÏ ÒÏ~& (16-23). >Öz¾Þb pH¢ ;®j ;B ê ¢¦ï²& æï> ÚÒ& Vº * ç B~ r¢Ò £bº zÎîj>¢ F~& . zêR º öêR ï²ö & z n;'¢º (24) fº Ò þöBº zêR ºÂÏ &Ë ·^~& b" zêR~ bjNf 30 : 70-40 : 60 &Ë '~&. . $ ò~ æOB¢ * ª, :öör, CFör 5 Öj ÒÏ Ö" ªf j7 ¸j ò& ç[~ b[ö Ò~ Ï ~ B& ¦~&b, HPLCö "« ªÒ*çj ¢bÊV r^ö(25) j* ê ò¢ zêRö ÒÏB¢ ~º ~ "; º&'b B~& . :öörº R3& Ï [b ²ï ¯>º ^B& ®ÚB, æOB& ·^~ ï²~ ¯ ìº CF örf Ö 7öB ÏÞÒWj J Öj F~ þö ÒÏ~& . V¢B ò 5-10 gj ~ î z ê æOj F òº 50 mL Öb 2-3² ºÂ~.

(5) 896. ®"²æ B 36 ² B 6 ^ (2004). âö ÊÖê ï²& FB ãÖöº pH¢ 11-12 ; ~ Sep-Pak C18 ÞÒæ¢ Û"Vb ºÂB ï² 7 R3º ÖWÏ 7öB *>æ ;B~ ÃB³» ê Ã B³» >Vö 0.1 N NaOH¢ & R3¢ Ï~ áf r HPLCÏ þÏb ~& . Model ®öB~ Î& ²>N ¦Æ Ïæï²¢ F~æ pf Óïrò, º:, *¯ò, " ~, jÊâ, JÒ 6«~ ªC&çòö & '' 10 µg/g >ê ï²bj Î& ê ²>Nj 3² > G ;~& . Table 3" ? ²>Nf Óïrò 90.3-97.9%, Òû 79.2-101.9%, JÒ 84.1-103.4%, *¯ò 86.4-100.8%, jÊ â 83.5-103.4%, "~ 78.5-95.6% Îv ·^ Ö"¢ á î . "~¾ jÊâf ®ö j ²>N Ô ~º Sep-Pak C18 ÞÒæ ;B~º ";öB ï²~ Ff ÏÂ ·^~æ á®V r^ ©b ÒòB . ß® ÊÖê ï² R3º Wîö O>Ú j*® ÏÂ>æ p , ÖW~öBº 7F~ 'Ë >î (24,26). $ B2~ ãÖº NöB zÎîj >Ïö 30ª* O~® j r 85%& Îj® 100oCöB 5ª òö ~ ª>º (27) ï²ö j n;W ÎÚææ ªC"; 7 Nê ¾ pH .ö F~B þöBº *¾Ò~ : ªC ãÖ ¸f ²>Nj áj > ®î .. Fig. 1. Calibration curves for food colors by HPLC. A: Yellow color types, B: Red color types, C: Blue and green color types.. æOj B r b" zêR~ 30 : 70 bj &~. NöB 2-3* $º >»çöB 10-30ª* &N~ ºÂ~ & . öªÒ ê *ö º~º ï²º 1 N "Ö 10 mL¢ & ºÂj >~&b ºÂö B zêR" Öf ÃB³»~ B~& . ºÂB ï² f Sep-Pak C18b ;B~& . r jÊ. FÛ ® %~ Ïï² ;ï 7öB FÛ> ®º B® 7 Óïrò, Òû, JÒ, *¯ ò, jÊâ, "~~ C 6« 120®Ïj ªC &ç®b þöB 'z>Ú J;B *¾Ò 5 ªCO»j ' Ï~ êÛê $º ÿªCö ~ ï²~ Fïj Ö "º Table 4-6" ? . ÖÒ¾¢öB îÏ>æ pº ï²º ¦ Â>æ p~ &çB®~ Ò"º Îv ¢~~º ©b ¾æÒ . ïêÛ~ ï² 7 ¦ÂB ©f Y5f Y4 v «~î Y4º JÒf "~, Ò ÒûöB ô ¦Â >îb, Y5º Òû" "~öB ô ¦Â>î (Table 4). ÖÒ¾¢öBº ®Î&b* 7 Ïæï²~ ÒÏV&ö B ÒÏï B¢ ~ ®æº pb¾ EU(F#)~ ÒÏï V&ö V Y4f Y5º jrOËòròf "¶~, "~ ö '' 100, 300, 150 mg/kgb &îÏïj J;~& Codex(B®Ï*ö²)öBº JÒö 200 mg/kg, "~ 5 jÊÊö 100 mg/kgb &îÏïj J;~ ® (28). þöB ªC ò 7 JÒ ò~ ®Ï Y4& 270.21 µg/gb Codex~ V&" jv r &îÏïj . ."~ ÒÏ~& . 'ïêÛ~ ï² 7öBº R2, R40 5 R3& ¦Â>î ß® R2f R40f 6«~~ ® ÎvöB ¦ Â>î . R3º Òû" JÒ, *¯ò, "~öBò ¦Â>î. (Table 5). 'ïêÛ ï² ö & Codex~ ÒÏï V&ö V R2f R3º JÒöº 200 mg/kg, "~ 5 jÊÊ öº R2& 50, R3& 100 mg/kgb &îÏïj ;~ ®. . $ EUöBº R40~ ãÖ jrWOËr~ö 100, "¶ ~ö 300, "~ö 150, ÊÛö 200 mg/kgb &îÏïj J; ¹ ® (28). 6«~~ ® 7 EU $º CODEX~ V&j ."~ ÒÏ ãÖ¢ ÚÚ R40f Ò ûöB vB~ ò& '' 143.92, 301.16 mg/kgb ."~& "~öBê B~ ò& 357.85 mg/kgb EU~ & îÏï . ¸~ . $ R2º JÒò 7 Bf j.

(6) ® 7 æï²~ ò*¾ÒO» {ã 5 Fï ªC. 897. Table 3. Recovery efficiencies (%) for 19 food colors spiked to the several foods followed by HPLC analysis Recovery efficiencies (%)1). Sample Food color B2 QY GS Y5 Y4 R40 R2 G3 R2G B1 R102 R106 Azo BBN PB ORII R3 R104 R105. Soft drink. Candy. Jelly. Chewing gum. Ice cream. Cereals. 95.3Û0.93 97.7Û1.01 97.2Û0.93 97.2Û1.01 96.7Û0.79 97.9Û1.12 96.5Û0.95 97.8Û1.06 92.0Û1.81 96.1Û1.49 97.0Û1.03 97.0Û0.95 96.5Û1.54 97.6Û0.96 96.4Û1.58 96.5Û1.14 95.1Û2.03 95.5Û0.50 90.3Û1.82. 79.2Û0.68 101.9Û0.47 96.6Û1.10 93.2Û2.30 100.3Û1.75 99.1Û1.42 100.4Û1.52 94.9Û1.31 96.2Û1.21 98.4Û1.60 92.9Û1.55 99.7Û1.53 95.9Û1.10 90.8Û0.78 95.0Û0.80 91.3Û2.42 95.9Û1.89 96.4Û0.90 91.7Û2.11. 103.4Û1.64 98.4Û1.09 93.1Û1.65 97.1Û0.76 101.2Û1.01 96.9Û1.14 96.7Û1.11 94.9Û1.13 97.8Û1.10 100.8Û2.32 95.9Û1.15 96.3Û1.13 92.3Û2.11 97.9Û1.20 95.6Û1.20 95.4Û0.99 97.5Û1.32 97.2Û1.55 84.1Û0.43. 99.1Û0.51 98.6Û0.42 93.5Û0.38 97.7Û0.41 95.1Û0.62 96.2Û0.28 97.0Û0.36 96.0Û0.44 98.9Û0.89 100.8Û0.65 97.0Û0.31 97.3Û0.52 91.2Û0.16 98.8Û0.57 96.4Û0.57 95.4Û0.84 98.2Û0.63 97.4Û0.55 86.4Û0.17. 91.4Û1.81 97.9Û2.38 83.5Û3.63 99.5Û1.57 96.3Û5.33 99.4Û1.12 98.0Û1.99 99.3Û0.80 99.6Û1.57 103.4Û1.28 99.5Û0.78 100.4Û0.90 98.9Û1.66 94.0Û2.08 100.4Û0.88 98.6Û1.25 97.1Û1.36 95.6Û2.07 85.0Û2.55. 79.2Û0.21 93.3Û0.29 80.4Û1.60 93.8Û0.42 83.0Û0.32 95.6Û0.78 86.3Û0.52 94.8Û0.86 94.1Û0.17 95.6Û0.45 88.9Û0.47 95.6Û0.48 93.9Û1.90 78.5Û0.69 92.8Û0.68 94.6Û1.23 90.5Û0.51 91.4Û0.65 87.7Û0.77. 1). MeanÛstandard deviation.. Table 4. Concentration of the yellow colors quantified in various foods by HPLC Samples. No. of sample. Food color. Concentrations (mg/kg). Mean (mg/kg). Soft drink. 20. Y5 N.D.1)Ü17, 2.04, 5.22, 37.67 Y4 N.D.Ü11, 1.41, 1.68, 2.82, 4.14, 4.78, 5.84, 8.23, 23.61, 32.64 QY and ORII were not detected... Candy. 20. Y5 N.D.Ü8, 2.08, 4.17, 6.34, 6.89, 9.05, 10.98, 17.48, 19.71, 31.10, 34.91, 69.65, 90.41 Y4 N.D.Ü8, 4.56, 8.68, 9.40, 15.19, 17.41, 19.75, 21.66, 27.14, 44.18, 68.69, 70.76, 84.59 QY and ORII were not detected... 25.23 32.67. Y5. 12.74. 14.98 9.46. Jelly. 20. N.D.Ü10, 0.61, 0.90, 1.93, 1.93, 2.44, 13.53, 150.5, 22.33, 26.83, 41.89 N.D.Ü3, 0.93, 1.27, 3.40, 5.17, 7.72, 10.25, 11.30, 14.35, 18.38, 22.43, 22.94, 23.33, 28.15, Y4 38.45, 89.04, 154.19, 270.21 QY and ORII were not detected... Chewing gum. 20. Y5 N.D.Ü19, 21.77 Y4 N.D.Ü12, 1.39, 5.68, 6.45, 6.74, 8.38, 15.91, 16.32, 40.49 QY and ORII were not detected... 21.77 12.67. Ice cream. 20. Y5 N.D.Ü16, 0.57, 0.89, 7.95, 15.72 Y4 N.D.Ü10, 0.96, 1.16, 2.83, 3.36, 4.76, 5.44, 8.30, 13.78, 14.49 QY and ORII were not detected... 6.28 5.91. Cereal. 20. Y5 N.D.Ü12, 1.12, 2.26, 2.43, 3.27, 4.16, 33.96, 80.70, 144.61 Y4 N.D.Ü11, 2.05, 2.72, 6.67, 14.31, 17.18, 20.86, 125.80, 148.18, 157.25 QY and ORII were not detected... 34.07 55.00. 42.44. 1). N.D.: not detected.. Êâ ò 7 B& Codex~ &îÏïö "7 ÒÏï j & . R3º V&j ." òº ~¾ê ìî ¦ ÂB ^&æ 'ïêÛï² 7 R40 6&æ ® «~ *ÚöB & Ë ÒÏnêf ÒÏï ô~ . Óï 5 ï êÛ~ ï². 7öB B1f Îv ¦Â>î B2º "~¢ B Î ®öB, G3º Óïròf Òû, jÊâöBò ¦Â>î. (Table 6). B1~ ãÖ ß® "¶f Òûö ô ÒÏ>î . CodexöBº "~ 5 jÊÊö B1, B2 5 G3¢ 100 mg/kgræ & îÏ~ ® EUöBº B1" B2¢ jr zRW OËròö 100, "¶~ 300, ÊÛö 200 mg/kgræ & îÏ~ ®º(28) þöB ªC ò 7öB ÒÏV&j ." òº ìî ..

(7) ®"²æ B 36 ² B 6 ^ (2004). 898. Table 5. Concentrations of red colors quantified in various foods by HPLC Samples. No. of sample. Food color. Concentrations (mg/mg). Mean (mg/mg). 20. R40 N.D.1) 17, 0.81, 36.74, 46.26 R2 N.D. 16, 1.35, 3.91, 18.72, 24.98 R2G, R102, R106, Azo, R3, R104 and R105 were not detected.. 20. N.D.Ü5, 0.10, 0.82, 3.17, 8.80, 16.78, 24.77, 27.80, 27.92, 30.60, 32.61, 34.39, 63.80, 92.39, 143.92, 301.16 R2 N.D.Ü15, 4.35, 5.36, 6.08, 10.88, 12.98 R3 N.D.Ü17, 0.23, 0.47, 5.60 R2G, R102, R106, Azo, R3, R104 and R105 were not detected.. 20. R40 N.D.Ü10, 0.36, 3.16, 7.06, 19.21, 20.92, 27.30, 32.45, 61.23, 66.38 R2 N.D.Ü13, 1.54, 3.21, 4.27, 5.82, 13.67, 19.49, 196.85 R3 N.D.Ü19, 8.55 R2G, R102, R106, Azo, R3, R104 and R105 were not detected.. 22.84 34.98 8.55. Chewing gum. 20. R40 N.D.Ü16, 0.90, 2.72, 20.05, 64.87 R2 N.D.Ü16, 4.42, 7.51, 15.02, 21.28 R3 N.D.Ü14, 0.20, 0.24, 0.68, 1.81, 2.77, 4.43 R2G, R102, R106, Azo, R3, R104 and R105 were not detected.. 22.14 12.06 1.69. Ice cream. 20. R40 N.D.Ü15, 1.58, 3.64, 7.54, 14.24, 51.27 R2 N.D.Ü13, 4.47, 6.17, 6.47, 17.21, 23.97, 26.79, 34.98 R2G, R102, R106, Azo, R3, R104 and R105 were not detected.. 15.66 17.14. 20. R40 N.D.Ü17, 1.47, 45.78, 357.85 R2 N.D.Ü18, 3.70, 10.75 R3 N.D.Ü17, 0.42, 0.53, 54.57 R2G, R102, R106, Azo, R3, R104 and R105 were not detected.. 135.04 7.22 18.51. Soft drink. R40. Candy. Jelly. Cereal. 27.94 12.24. 53.94 7.93 2.10. 1). N.D.: not detected.. Table 6. Concentrations of blue and green colors quantified in various foods by HPLC Samples. Food color. Concentrations (mg/mg). Mean (mg/mg). 20. B2 N.D.1)Ü19, 0.78 G3 N.D.Ü19, 1.09 B1 N.D.Ü11, 0.18, 0.31, 0.50, 0.90, 1.11, 2.07, 2.21, 5.74, 7.61 GS, BBN, and PB were not detected.. 0.78 1.09 2.29. Candy. 20. B2 N.D.Ü15, 0.07, 0.44, 0.83, 1.65, 20.95 G3 N.D.Ü19, 0.09 B1 N.D.Ü5, 0.18, 0.87, 0.92, 1.23, 1.30, 1.39, 1.54, 1.95, 2.57, 2.76, 4.28, 8.16, 11.36, 14.16, 36.66 GS, BBN, and PB were not detected.. 4.79 0.09 5.96. Jelly. 20. B2 N.D.Ü19, 0.86 B1 N.D.Ü11, 0.05, 0.18, 1.12, 1.32, 2.31, 2.84, 3.88, 3.95, 7.43 GS, BBN, and PB were not detected.. 0.86 2.56. Chewing gum. 20. B2 N.D.Ü19, 0.15 B1 N.D.Ü11, 0.08, 0.91, 1.00, 1.01, 1.43, 5.15, 8.04, 8.37, 8.74 GS, BBN, and PB were not detected.. 0.15 3.86. Ice cream. 20. B2 N.D.Ü19, 0.63 G3 N.D.Ü18, 0.04, 0.21 B1 N.D.Ü14, 1.21, 1.37, 1.75, 2.36, 6.34, 8.43 GS, BBN, and PB were not detected.. 0.63 0.12 3.58. Cereal. 20. B1 N.D.Ü15, 0.91, 1.15, 2.37, 19.90, 21.43 GS, BBN, and PB were not detected.. 9.15. Soft drink. 1). No. of sample. N.D.: not detected..

(8) ® 7 æï²~ ò*¾ÒO» {ã 5 Fï ªC. *Ò ÖÒ¾¢ ®Î&b* 7 Ïï²~ ÒÏV&f & ç®ò ;^ ® îÏïf B~ ®æº pb¾ ¢¢ îÏSï(ADI: acceptable daily intake) J;>Ú ®, Codex, EU öBº &ç®" þ &îÏïj B~ ®bæ Nê ®Î&b*ê Bz º^¢ >' ©b '> ö Ö"& '.® ÏF > ®j © b V& .. º. £. º ®*ö >B ® 7 æï²~ ªCO» j BF, j~ ® ¦ÒV&öB ;{~ ö ¦Ò¢ >¯ > ®ê V~ ÿö ¢ Û Ïæï² ~ Sï Ò, *ê ï& 5 ÒÏï B ;þ»~ Ï, Ûç^B¢ &j "' "¢ {~º Ï~ ¶ >¯>î . ÖÒ¾¢öB ®ö ÒÏ îÏB æï² 9«" ®îÏ æï² 10« C 19«ö & N3 ³ ÚîÆ¢b¢ 'Ï~ ªC~& . 7öB FÛ 7 Óïrò, Òû, JÒ, *¯ò, jÊâ 5 "~ 6 « 120®Ïj &çb ' ®ê Ïæï²~ ºÂ" ²>N~ 'j {ã~&b, ¢ Æ& ® 7 æ ï²~ ïj Ò~& . Óïròf Òû, JÒ~º bö C~¾ ÏÎ ê pH¢ 5-6b ;~ ÒÏ~& * ¯ò" jÊâ~º b" b ê ;bj B~ pH ¢ 5-6b ;~&b¾ jÊâ" "~~ ãÖ ÊÖ ê ï²& FB òº pH¢ 11-12 ;~&j r ÎN' b ºÂ > ®î . "~º Ö $º CFör æ Oî Wªj Ï~ B r ºÂ~& . ® «~ê Î ®ö Î& 19« æï²~ ²>Nf Óïrò 90.397.9% &Ë ¸~, ®öBº Îv 80%çî . 7öB FÛ> ®º Ú 5 >«® 7 ï²~ ïj ;ï Ö", ®ê ï²~ ï ³êº*º '' Óïrò 0.78-27.94 mg/kg, Òû 0.94-53.94 mg/kg, JÒ 0.86-42.44 mg/ kg, *¯ò 0.15-22.15 mg/kg, jÊâ 0.12-39.17 mg/kg, " ~ 7.22-135.04 mg/kg~ º*î ¦ÂB ï²º Ò" ¢~~& .. ^. ò. 1. Song CC, Tak HJ. Food Additives. Jisungsa, Seoul, Korea. pp. 198-208 (1998) 2. Mun BS. Food Additives. Suhaksa, Seoul, Korea. pp. 134-149 (1998) 3. Korea Food and Drug Administration. Food Additives Code. Seoul, Korea. pp. 179-209 (1998) 4. Kwon HS, Kim IB, Lee JO. Study on the determination of mixed color in foods (II). Report NIH 15: 445-448 (1981) 5. Kim KS, Kim BY, Shin KB, Kim IB, Yoo SY. Study on the determination of mixed coloured in foods (V). Report NIH 18: 393-398 (1981) 6. AOAC. Official Methods of Analysis of AOAC Intl. 16th ed. Association of Official Analytical Communities. Gaithersbrug, MD, USA (1995) 7. Lee JO, Yoon HJ, Lee CH, Park SK, Lee CW. A study for analytical method of synthetic color additives in foods (I). Annual Report KFDA 2: 107-115 (1998) 8. Huest HJ, Mckim JMA, Martin RA. Determination of tartrazine. 899. in food products by HPLC. J. Food Sci. 46: 419-420 (1981) 9. Lawrence JF, Lancaster FE, Conacher HBS. Separation and detection of synthetic food colors by ion-pair high performance liquid chromatography. J. Chromatogr. A 210: 168-173 (1981) 10. Puttemans ML, Dryon L, Massart DL. Isolation, identification, and determination of food dyes following ion-pair extraction. J. Assoc. Off. Anal. Chem. 65: 735-745 (1982) 11. Chudy J, Crosby NT, Patel I. Separation of synthetic food dyes using high-performance liquid chromatography. J. Chromatogr. A 154: 306-312 (1978) 12. Pharmaceutical Society of Japan. Standard Methods of Analysis for Hygienic Chemists. Ginin, Tokyo, Japan. pp. 512-547 (1995) 13. Park SK, Lee CH, Park JS, Yoon HJ, Kim SH, Hong Y, Lee JO, Lee CW. Simultaneous analytical techniques for determination of 8 synthetic food colors in foods by HPLC. Anal. Sci. Technol. 13: 378-384 (2000) 14. Park SK, Hong Y, Jung YH, Lee CH, Yoon HJ, Kim SH, Lee JO. Optimization of HPLC method and clean-up process for simultaneous and systematic analysis of synthetic color additives in foods. Korean J. Food Sci. Technol. 33: 33-39 (2001) 15. Ministry of Health, Labor and Welfare. Analytical Methods of Food Additives in Foods. Kangdamsa, Tokyo, Japan. pp 260-307 (1993) 16. Kuwano K. Mitamura T., Ion-pair high performance liquid chromatographic separation and determination of food coal-tar dyes. J. Food Hyg. Soc. Japan 27: 278-282 (1986) 17. Boley NP, Crosby NT, Johnson AE, Roper P, Somers L. Determination of synthetic colours in foods using high-performance liquid chromatography. Analyst 105: 589-599 (1980) 18. Japan Society of Food Sanitation. Determination of Food Additives in Foods. A Guide to Examination of Food Sanitation. Japan. pp. 135-177 (1989) 19. Kim JH, Kim BH, Kim GS, Oh SK, Han SU, Lee DH, Determination and separation of food tar dyes by HPLC. Report SIHE 29: 122-125 (1993) 20. Tonogai Y, Kingkate A, Halilamian C. Quantitative determination of colorants in dried shrimp and shrimp paste using ion-exchange extraction and high performance liquid chromatography. J. Food Prot. 46: 592-595 (1983) 21. Hann JT, Gilkison IS. Gradient liquid chromatographic method for the simultaneous determination of sweeteners, preservatives and colours in soft drinks. J. Chromatogr. A 395: 317-322 (1987) 22. Lau OW, Poon MMK, Mok SC, Wong FMY, Luk SF. Spectrophotometric determination single synthetic food colour in soft drinks using ion-pair formation and extraction. Int. J. Food Sci. Technol. 30: 793-798 (1995) 23. Oh SK, Park HH, Chung SY, Choi HY, Park SB. A study on food additives : For contents of tar dyes in ice creams. Report SIHE 13: 49-55 (1977) 24. Tsuji S, Shibata T, Okamoto, K, Takeda K, Fujiwara Y, Ito Y. Preparation of sample solution using pronase treatment for determination of food coal-tar dyes in foods. J. Food Hyg. Soc. Japane 36: 68-76 (1995) 25. Puttemans ML, Dryon L, Massart DL. Evaluation of thin layer, paper and high performance liquid chromatography for identification of dyes extracted as ion-pair with tri-n-octylamine. J. Assoc. Off. Anal. Chem. 65: 730-736 (1982) 26. Kim JH, Kim BH, Oh YH, Kim GS, Cho TH, Oh SK, Han SU, Lee DH. Studies on the stability of food tar dyes. Report SIHE 30: 91-94 (1993) 27. Tsunoda K, Inoue N, Aoyama M, Hasebe A. Rapid analysis of the food colors in food stuffs using polyamide. J. Food Hyg. Soc. Jpn. 28: 473-479 (1987) 28. Korea Institution of Food Sanitation. Study on the application of chemically synthetic food additives in foods. Ministry of Health and Welfare. Korea. pp. 194-236 (1997) (2004j 10ú 14¢ 7>; 2004j 11ú 11¢ j).

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