Physiological Effects of Curcumin Extracted by Supercritical Fluid from Turmeric (Curcuma longa L.)

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(1)KOREAN J. FOOD SCI. TECHNOL. Vol. 36, No. 2, pp. 317~320 (2004). ©The Korean Society of Food Science and Technology. ;(Curcuma longa L.)b¦8 .ªê FÚ ºÂ curcumin~ Ò9 ;ß*ÁË51,*Áã\ ()JÂ8, 1ÏÎv ®". Physiological Effects of Curcumin Extracted by Supercritical Fluid from Turmeric (Curcuma longa L.) Seung-Hyeon Jung, Kyu-Seob Chang1,*, and Kyung-Hee Ko Ottogi Co. Ltd. Department of Food Science and Technology, Chungnam National University. 1. Physiological effects of curcumin, major yellow-colored pigment in tumeric (Curcuma longa L.), extracted by traditional extracting methods, ethanol and hot-water extractions, and supercritical fluid extraction (SFE) using supercritical carbon dioxide as new extracting method. Antioxidative activity of ethanol extract was higher than those of SFE and hot-water extracts. Results of Ames mutagenicity test on SFE, ethanol, and hot-water extracts revealed no mutagen in the extracts. Antimutagenicity rates of SFE, ethanol, and hot-water extracts against direct mutagen, 2-nitrofluorene (2-NF), were 20.1, 9.3, and 15.2%, respectively. Antimutagenicity rate of SFE extract against TA98 derived from indirect mutagen, 2-acetamidofluorene (2-AF), was 12.2%, whereas none was observed in ethanol and hot-water extracts. Nitrite-scavenging ability of SFE extract was higher than those of ethanol and hot-water extracts. Key words: physiology effects, curcumin, turmeric, supercritical fluid extraction. *. . .ªê FÚ(3,4)¢ Ï ºÂ»f bî~ Vç" ç~ çãêæ6 critical point ç~ {K" Nê¢ J; 2b ç~ ÏK" Vç~ {Öê>f 6ê~ ßWj æî ² ~ ³ ºÂ" F' ºÂ &Ë~² ~º O». . $ .ªê FÚº " Özê² _f Özê²f ï~ Ï ;W>V r^ö Ï ºÂ»ö j F W Ï ~ º *þ ìj öò jî¢ çN¦"öB ºÂ · Úî > ®V r^ö Âb $º ®" ? ö "6 bî~ ºÂö FÏ O» . ¾ jçræ ;~ ÒW bî curcuminö & Öë' wÏj * .ªê ºÂ~ ' {ã>æ p~b, ;~ .ªê ºÂb" ºÂ O»b ºÂB ; ºÂb~ Ò' ß Wj jv º ; . V¢B º .j. þj Û ' .ªê ºÂb ºÂ curcumin" > 5 öêR ºÂ curcumin~ ÒW Î"¢ ÒÁj v~ .ªê FÚ ºÂ curcumin~ Ï&ËWj rj ¶ ~& .. ;f ;"(Zingiberaceae)~ ;³(Curcuma)b ª~> º j bB ¶ ö V¢ 30~70«~ ëã« Ò C® ®æò ;{ «~ >º rJææ p ® b, &¦ª &ç ;º jÝ~ (1). ;f ËVWª . º curcuminoidsö ~ Oï Î"(2)& 7º ©b ï& >ÚN Ëò ºÂ ~ &ì ;ªòbê ®~ ïçj . > ®Ú ºÂö & & Ëòö j 'î . ¾ " ;~ ÒWbî curcumin ÖzW, òæW, zÎ", "Ã &æ VËW C&æB ~ª¢¢ 7b B & ê¯> ® . ö V¢ ;~ ÒW bî ºÂª¢ê .ªê F Ú¢ Ï ;FWª 5 curcuminoids ºÂ Öë' wÏ G öB Ö 7º~Ò¢ ÒòB .. *Corresponding author : Kyu-Seob Chang, Department of Food Science and Technology, College of Agriculture and Life sciences, Chungnam National University, 220 Gung-dong, Yusung-gu, Daejon 305-764, Korea Tel: 82-42-821-7876 Fax: 82-42-822-2153 E-mail: [email protected]. Òò 5 O» ò*` þöB ÒÏ ;f 2001j êöB ÒVB Madras Ö finger;j ªê~ rª 2Ö~ö &/ ç 4oC ï 317.

(2) 318. ®"²æ B 36 ² B 2 ^ (2004). Ë &~B Òò ÒÏ~& . .ªê ºÂbf .j. þj Û curcumin ºÂ >N ¸~~ {K 250 bar, Nê 40oC, Ï (öêR) F³ 3 mL/min öB 3* ºÂ~ Ï öêRj &~ 200 mL ;Ï~&, öêR º Âbf öêR 100 mLö ;ª 5 gj Î&~ 70oC Nêö B 2* ÿn ºÂ~&, > ºÂbf Ã~> 200 mLö ; ª 10 gj Î&~ 100oCöB 2* ÿn ºÂ~& . Î ºÂbf 12,000 rpmb 10ª* öªÒ ê membrane filter (0.45 µm) "~ òÏb ÒÏ~& . HPLCö ~ curcumin ï ª+ *¾Ò¢ ò¢ HPLC(Agilent, 1100 Series, Hewlett Packard, USA) curcumin~ ïj ªC~& . Columnf Zorbax Eclipse C18(4.6 mmÜ150 mmÜ5 µm)j ÒÏ~&, Mobile phaseº Methanol 100%, F³f 0.8 mL/min b ~&. . DAD(UV 424 nm) ¦Â~&, Injection volumef 10 µL & . Standard curcuminf SigmaÒ~ curcuminj ÒÏ~& . Öz Î Rancimat(Model743, metrohm, Switzerland)¢ Ï conductometric determination method(CDM)(5)b ï&~& . >w ö ò Öæ¢ 3 g ê 120oC .B aluminium heating blockçöB * 20 L~ "B V¢ "«~ Öz V . ÖzKf ºÂbj Î&~æ pf Fæò¢ & ~ FêV*(hour)j êÖ~& . òæ9 5 òæ9(Mutagenicity and antimutagenicity test) þ": Maron" Ames(6)~ O»ö V¢ þ~& . Salmonella typhimurium TA98(KCTC2053), Salmonella typhimurium TA 100(KCTC2054)º F*;î j { ê þö ÒÏ~& . S9 mixture *`: *7 òæöj WzÊV *B Maron" Ames(6)~ O»ö V¢ B~& . S9 mixtureº S9 fraction(10%)j MgCl-KCl"(2%), 1 M glucose-6-phosphate (0.5%), 1 M NADP(4%), 0.2 M phosphate buffer(pH 7.4)f >f b~ þö ÒÏ~& . òæ9 þ Maron" Ames(6)~ O»ö V¢ þ~&º, Ò cap tubeö ò³ê~ Ã&ö V òæWj {~V * ªCòB O»ö V¢ B sample Ïj ³êê preincubationÎ ê ©j top agar 2 mLf b ê ² ï6Væ(minimal glucose agar plate)ö êö~ 37oCöB 48 * V· ê Væç~ revertant colony Q¶¢ ê>~& . r spontaneous colony~ B> 2V ç òæW ®º ©b 6 . òæ9 þ Maron" Ames(6)~ O»j Ï~ preincubation mutagenicity test ¦Æ~& . ÒÏ æöf ç7æöb 2-nitrofluorene(3 µg/plate; Aldrich Co.; 2-NF)j ÒÏ~& *7æöbº 2-acetamido fluorene(10 µg/plate; Sigma Co.; 2-AF)j ÒÏ~& . òæ. ÛB Î"(inhibition rate)º r~ j Ï~ êÖ~& . Inhibition rate(%) = [(a − b)/(a − c)]Ü100 a: òæöö ~ FêB æ òæ> b: ò¢ ¾Ò~&j r~ æ òæ> c: òæö" ò& ìj ãÖ~ ¶æ òæ> Nitrite scavenging activity: nitrite ªËf Kato(7)~ O»b þ~& . 1 mM NaNO2 Ï 1 mLö òÏ 0.6 mLj &~ Vö 0.1 N HCl 5 0.2 M Ö jÏÏj ÒÏ~ >wÏ~ pH¢ '' 1.2, 4.2 5 6.0b ; ê >w Ïj 10 mL B~& . Ïj 37oCöB 1*ÿn > wÎ ê 1 mL¢ ~ Vö 2% acetic acid 5 mL" Griess £ 0.4 mLj &~ 15ª* NöB O~Î r 520 nmöB 7ê¢ G;~ º~º nitriteïj ~, nitrite ªËj r" ? òÏj ^& ãÖf Î&~ æ pf ãÖ~ nitrite WªN(%) ¾æÞ . A–C Nitrite ªN(%) = 1 – -------------- × 100 B. A: 1 mM NaNO2 Ïö òÏj Î&~ 1* O~ Î ê~ 7ê B: 1 mM NaNO2 Ï~ 7ê C: ò¶Ú~ 7ê. Ö 5 V HPLCö ~ curcumin ïª+ .ªê ºÂ, öêR ºÂ, > ºÂö ~ ' ºÂb~ curcumin ïj HPLC ªC Ö", .ªê ºÂbf 1.89%, öêR ºÂbf 2.55%, > ºÂbf 0.004% > ºÂb ~ ï &Ë Ô² ¾æÒ . º curcumin ²>W~ b ö æ pº bîæ ¢>' > ºÂ öBº º Â>æ pº ©b 'B . ÖzÎ .ªê, öêR, > ºÂ ºÂbj Öæö 0.1%, 0.5% ³ê Î&~ Rancimatb Fê V*j G; Ö"º Table 1" ? . ºÂb 7 öêR ºÂb Öz Î"& ² ¸ ² ¾æÒb¾, Î& ³ê~ Ã&ö V FêV*~ Nº ~& . ºÂO»ö V ò*~ Öz Î"º jÝ FêV* Ã& ·çj ¾æÚî . Table 1. Antioxidative effects of the turmeric extracts at concentration (0.1, 0.5%) on beef tallow Concentration. Extracts. Induction periods (hr). 0.1%. Control SFE extracts Ethanol extracts Water extracts. 1.17 2.04 2.19 1.94. 0.5%. Control SFE extracts Ethanol extracts Water extracts. 1.17 2.10 2.22 2.10.

(3) ;(Curcuma longa L.)b¦V .ªê FÚ ºÂ curcumin~ ÒW Table 2. Reverse mutation test of turmeric extracts in Salmonella typhimurium TA98 Dose of extracts (mL/plate) 1). His+Revertants/plate. Table 4. Antimutagenic effects of turmeric extracts against the mutagenicity of 2-nitrofluorene (2-NF) in Salmonella typhimurium TA98. TA98 59Û17. Spontaneous SFE extracts. 00.05 0.1 0.2. 46Û10 43Û06 30Û07. Ethanol extracts. 00.05 0.1 0.2. 59Û14 68Û10 38Û07. Water extracts. 00.05 0.1 0.2. 47Û06 60Û01 78Û12. His+Revertants/plate TA98 57Û13. SFE extracts. 00.05 0.1 0.2. 35Û08 45Û06 46Û08. Ethanol extracts. 00.05 0.1 0.2. 67Û07 64Û04 52Û14. Water extracts. 00.05 0.1 0.2. 50Û10 76Û16 62Û15. 1). The values are meanÛS.D. of 4 replications.. n(8)f zêR, öj^rÞ, Öj Ï ºÂ ; ºÂbj 1,500 ppm ³ê ;Fö Î&~ þ Ö" & ~ FêV* 22.6* îb, zêR ºÂbj Î& öº FêV* 26.9*, ö j^rÞ ºÂbf 25.6 *b Ã&~&b¾ Ö ºÂbf FêV* 22.7*b &f FÒ Ö"¢ Öz Î"& ìî ~& . ; (9)f ;~ öêR ºÂbj BFö &³ê 0.02-0.10% º*f ³ê 0.1-0.5% º* ³ê¢ Ò ~ ÖzWj Ò Ö" ³ê& Ã&ö V¢ ÖzW Ã&j : ®b¾ öBº ³ê Ã&ö V Öz Î"º £~& . ;~ Öz Î"º phenolV ¢ &æ ®º curcumin 5 ËVWª ÖzWö &7 & N ®º ©b 6B . òæ9 5 òæ9 ; ºÂ>~ òæ9: öêR ºÂb, > ºÂb, . ªê ºÂb~ òæW FZ¢ { Ö" Table 2, 3f ? ; ºÂb f his+ revertant colony>~ Ã6 &, TA98, TA100~ ¶òæ > 59, 57 2V ç Ã& Î ºÂbf ìº ©b { >î . V¢B ;ºÂb ö >Ú òæ¢ FB~æ pº ©b 6>î .. Inhibition rate (%). Spontaneous1) 2-nitrofluorene SFEextracts Ethanol extracts Water extracts. 45Û2 810Û28 656Û83 738Û34 693Û92. 20.1 09.3 15.2. Table 5. Antimutagenic effects of turmeric extracts against the mutagenicity of 2-acetamidofluorene (2-AF) in Salmonella typhimurium TA98. Table 3. Reverse mutation test of turmeric extracts in Salmonella typhimurium TA100. Spontaneous. Revertants/plate TA98. The values are meanÛS.D. of 4 replications.. The values are meanÛS.D. of 4 replications.. 1). Dose of extracts (0.1 mL/plate). 1). 1). Dose of extracts (mL/plate). 319. Dose of extracts (0.1 mL/plate). Revertants/plate TA98. Inhibition rate (%). Spontaneous1) 2-acetamidofluorene SFEextracts Ethanol extracts Water extracts. 035Û03 136Û12 124Û24 157Û17 149Û26. 12.2 -. 1). The values are meanÛS.D. of 4 replications.. ç%æöö & òæ Î: ^Ú~ DNAö ç7 'b ¶çj "Ú òæ¢ FB~º bî rJê æ ö 2-nitrofluorene(3 µL/plate; 2-NF)b FêB Salmonella typhimurium TA98ö & ; ºÂb~ òæNf Table 4f ? . 3«~ ºÂb 7 .ªêºÂb~ &N 20.1% &Ë ¸f òæNj ¾æîb, öêR" >ºÂbf '' 9.3%, 15.2%~ òæNj ¾æÚî . (10)º ç 7æö 2-NF FêB Salmonella typhimurium TA98öB ; ~ > ºÂbj 0.1 mL ¾Ò òæ>& 924öB 719 6²~ 23%~ òæ ÛBÎ"¢ : ®Ú þÖ". ² ¸f Ö"¢ ¾æÚî . *%æöö & òæ Î: Table 4ö *7æö 2-AFö FêB TA98~ òæNj ¾æÚî . 2-AF F êB TA98f SFE extract¢ 0.1 mL/plate ¾Ò 12.2%~ òæNj &b, $ rzR ºÂb" >ÏW ºÂb f *7æö 2-AFö FêB TA98ö 0.1 mL/plate ¾Ò òæ Î"& ¾æ¾æ p~ . Soni (11)f tumeric~ rzR 5 >ÏW ºÂböB S9&Ò aflatoxinö ~ FêB Salmonella typhimurium TA98, TA100öB 80%~ òæ Û BÎ"& ®rj ~&, Nagabhushan (12)f tumericj rzR ºÂ ºÂb~ ³ê& 125 µg/plateöBº benzo(α) pyrene, dimetyl benzanthracene æööBº 50%~ òæ ÛBÎ"& ®îb, 500 µg/plate ³êº benzo(α)pyrene~ ã Ö 75%, dimethylbenzanthraceneº 80%~ òæ ÛBÎ"& ®îrj ~& . Nitrite scavenging activity ;ºÂb 3«j ÒÏ~ nitrosamineW öbî nitrite ~ ªËj { Ö"º r" ? . pH 1.2 ~öB >wÎ ãÖ &b ÒÏ vitamin C 10 mg~ nitrite ª.

(4) ®"²æ B 36 ² B 2 ^ (2004). 320. Nf 99.60Û0.21%&, .ªê ºÂbf öêR, >º Âbö j~ ¸² ¾æÒb, r ªNf 95Û0.25%&. . öêR ºÂbf nitrite ªN 74.3Û0.37%¢ ¾æîb, >ºÂbf 42.8Û0.63% ºÂb 7 &Ë Ôf ªNj ¾ æÚî . f (13)f ;j 25oC b" >¢ Ï~ º Â ºÂb~ nitrite ªNj G; Ö" 25oC bj Ï ºÂbf 35% ç, > ºÂbf 40% ç~ ªNj ¾ æÚ þ~ > ºÂb" FÒ Ö"¢ ¾æî . pH 4.2öB >wÎ ãÖ & ÒÏ vitamin C 10 mg~ nitrite ªN 88.1Û0.13%&b, .ªê ºÂbf 61.4Û 0.41%, öêR ºÂbf 45.4Û0.52%, > ºÂbf 35.7Û 0.35%~ ªNj ¾æî . pH 6.0öB >wÎ ãÖ & ÒÏ vitamin C 10 mg~ nitrite ªN 78.8Û0.32% &, .ªê ºÂbf 48.6Û0.55%, öêR ºÂbf 36.9Û 0.26%, > ºÂbf 18.3Û0.71%~ ªNj ¾æî . Nitrite ªNf pH& 1.2¢ r &Ë Ö>~&, pH& ¸jî> ª N Ô~ . Ö"º f (13) ; ºÂb" . ö ò Ëò ºÂb~ nitrite ªN pH 1.2öB Î" & ¸~ º Ö"f Kyrtopoulos (14) N-nitroso compounds ;Wö & öB nitrosamin f pH 2.5-3.0 ' b pH~' pH& Ôj> nitrite ªN Ã& º f ÿ¢ ãËj ¾æî . þ~ pH 1.2, 4.2, 6.0 ÎvöB .ªê ºÂb öêR ºÂb, > ºÂb nitrite ªN Ö>~& .. º. £. Curcumin~ ºÂÎNf öêR ºÂO» &Ë ¸~, ÖzÎ"º öêR, .ªê, > ºÂb Bb ¾æÒ . Ames ~ mutagenicity test¢ Ö" ''~ ºÂbf òæ ö ìº ©b {>î, TA98ö ç7æö 2-NF ¾Ò .ªê ºÂb~ &N 20.1% öêR" > ºÂb ö j~ &Ë ¸f òæ Î"¢ ¾æî . $ *7 æö 2-acetamidofluorene(10 µL/plate; Sigma Co.; 2-AF)ö & òæ Î"¢ rj Ö" 2-AF FêB TA98f . ªê ºÂbj 0.1 mL/plate ¾Ò òæ~ Î"º 12.2%&b, rzR ºÂb" > ºÂb~ ãÖº òæ Î"& ¾æ¾æ p~ . Nitrite~ ªË~ ãÖö ®ÚB ê öêR 5 > ºÂbö j~ .ªê ºÂb ¸f ª Nj ¾æîb pH 1.2öB &Ë ¸f ªNj & . . f ? .ªê¢ Ï ºÂ curcuminf ÖzÎ", nitrite ªË, òæÎ" ö & ÒW VËj &b, V~ öêR¾ > ºÂ»ö j~ FÒ~¾ z ¸f ÒW Î"& ¾æ¾Vê ® .. ^. ò. 1. Purseglove JW, Brown EG, Green CL, Robbins SRJ. Spices. Longman Inc., New York, NY, USA. pp. 100-286 (1981) 2. Taka H. The characterization and application of Curcuma longa. L extracts. New Food Ind. 40: 7-15 (1998) 3. Rizvi SSH, Benado AL, Zollweg JA, Daniels JA. Supercritical fluid extraction fundamental principles and modeling methods. Food Technol. 40: 55-65 (1986) 4. Rizvi SSH, Daniels JA, Benado AL, Zollweg JA. Supercritical fluid extraction operating principles and food applications. Food Technol. 40: 57-64 (1986) 5. JOCS. Standard methods for the analysis of fats, oil and related materials. Japan Oil Chemists Society, Nihonbashi, TK, Japan (1996) 6. Ames BN. Dietary carcinogen and anticarcinogens. Science 221: 1256-1264 (1983) 7. Kato H, Lee IE, Chuyen NV, Kim SB, Hayase F. Inhibition of nitrosamine formation by nondialyzable melanoidins. Agric. Bio. Chem. 51: 1333-1338 (1987) 8. Ahn CK. Antioxidative effects of spices and their synergism with catechin and ascorbic acid. PhD thesis, Sookmyung Women's University, Seoul, Korea (1998) 9. Kang WS, Kim JH, Park EJ, Yoon KR. Antioxidative property of turmeric (Curcumae Rhizoma) ethanol extract. Korean J. Food Sci. Technol. 30: 266-271 (1998) 10. Lee JS. Physiological effects of Instant curry and curry raw materials. PhD thesis, Sungkyunkwan University, Seoul, Korea (2002) 11. Soni KB, Lahiri M, Chackradeo P, Bhide SV, Kuttan R. Protective effect of food additives on aflatoxin-induced mutagenicity and hepato carcinogenicity. Cancer Lett. 115: 129-133 (1997) 12. Huang MT, Smart RC, Wong CO, Conney AH. Inhibitory effect of curcumin, chlorgenic acid, caffeic acid and ferulic acid on tumor promotion in mouse skin by 12-O-tetraphorbl-13-acetate. Cancer Res. 48: 5941-5946 (1988) 13. Kim JH, Park KM. Nitrite scavenging and superoxide dismutaselike activities of herbs, spices and curry. Korean J. Food Sci. Technol. 32: 706-712 (2000) 14. Kyrtopoulos SA. Ascorbic acid and formation of N-nitroso compounds: possible role of ascorbic acid in cancer prevention. Am. J. Clin. Nutr. 45: 1344-1350 (1987) (2003j 10ú 21¢ %>; 2004j 2ú 25¢ j).

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