Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /., No.1, -/+ -// (,**1) 351. a, b,

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1 39 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /., No.1, -/+-// (,**1) 351 ao bo * * A Simple Spectrophotometric Method for the Estimation of a-carotene, b-carotene and Lycopene Concentrations in Carrot Acetone Extracts Masayasu Nagata, Yuji Noguchi, Hidekazu Ito, Shunsuke Imanishi and Keita Sugiyama* National Research Institute of Vegetables and Tea Science, National Agriculture and Food Research Organization, -0* Kusawa, Ano, Tsu, Mie /+.,-3, * National Agricultural Research Center for Hokkaido Region, National Agriculture and Food Research Organization, + Hitsujigaoka, Sapporo, Hokkaido *0,2/// We developed a simple and e#ective method for measuring concentrations of a-carotene, b-carotene and lycopene in acetone extracts of common carrots and Kintoki red carrots. This method utilizes the di#erences in visible absorption spectra for each of these pigments. Concentrations of each pigment can be determined by measuring the absorbance at..-,.1/,.3, and /*/ nm, i.e., A..-, A.1/, A.3, and A /*/, respectively, and performing calculations. The equations, with correlation coe$cients between prediction and measurement (r), and the standard errors of prediction (SEP), are as follows : a-carotene (mg/l)*.2.1 A..--.,+2A.1/+..33A.3,-./+3A /*/*.++3 (r*.30/, SEP *.,-+) ;b-carotene (mg/l)+..22a a.3,,.-/, A /*/*.*32 (r*.3.0, SEP*.,,2) ; lycopene (mg/l)*.,/0 A A.3,/.*22A /*/*.,-1 (r*.330, SEP*.+-3). A standard spectrophotometer is su$cient for this method, and concentration calculations can be made within minutes of taking the absorbance measurements. (Received Dec. +-,,**0 ; Accepted May,,,**1) Keywords : carrot, carotenoids, spectrophotometric quantification, equation, quality : /+.,-3, -0* * *0,2/// + Corresponding author masayasu.nagata@a#rc.go.jp A +0 A +,1., + A,-./ a, b, bo 0 ao bo 1 ao bo + ao bo 2 bo ao bo - HPLC n,. n-, n/0 - g -/ 3 +** ml UV-Spec -/*1/* nm + nm HPLC

2 352 /. 1,** Table + Pigments Absorption coe$cient of three pigments Absorption coe$cient (E + + cm)..- nm a.1/ nm b.3, nm c /*/ nm d a-carotene b-carotene lycopene,1+,,,/1,,12,,//,,// -.+, ,,,,2,-0 /./ -+/* a Di#erential absorption spectrum between a-carotene and b-carotene has a maximum at..- nm4 b a-carotene and b-carotene have the same absorption coe$cients at.1/ nm4 c Di#erential absorption spectrum between a-carotene Fig. + Calculated absorption spectra (+, + cm) of a- carotene, b-carotene and lycopene in acetone and b-carotene has a minimum at.3, nm4 d Lycopene has an absorption maximum at /*/ nm4 A..-,1+,C a,,/1c b,,12c L Eq. LC-+*A L- 1.// -/*1/* mn TSKgel ODS-+,*T,/* mm..0 mm ID + mlmin HPLC, ao bo +* + + cm Fig. + ao bo..- nm ao.3, nm bo 1 /*/ nm ao bo /*/ nm ao bo.1/ nm. Table +, ao bo HPLC Table cm Eq. A.3,3.1C a+02,c b,,,2c L Eq. A /*/,-0C a/./c b-+/*c L Eq. A..- A.3, A /*/..-.3, /*/ nm C a, C b, C L ao bo mgl Eq. a-carotenemgl1.*.a..-+*.++a.3,,.*0a /*/ Eq. b-carotenemgl..,.a *a.3,0.1*a /*/ Eq. LycopenemgL*.,+A A.3,..+2A /*/ Eq. bo HPLC bo,, n,2 n,2 Eq. HPLC Fig., *.3/* Eq. HPLC *.-/+ *.*32 Eq. Eq. Standard error of calibration, SEC *.,,+ b-carotenemgl+..22a a.3,,.-/,a /*/*.*32 Eq.

3 41 : 353 Fig., Scatter plots and correlation of calculated b-carotene concentration with the Eq. (/) versus measured concentration Eq. HPLC Y, X Y+.*-,X*.**. *.3.0 Standard error of prediction, SEP *.,,2 Fig. - Eq. Eq. *.331 SEC *.++2 Fig. - Scatter plots of calculated b-carotene concentration with the Eq. (1) versus measured concentration SEP, Standard error of prediction. LycopenemgL*.,/0A A.3, /.*22A /*/*.,-1 Eq. Eq. HPLC Y X Y+.***X*.*,. *.330 SEP *.+-3 Fig.. mgl *.-* *.+. ao Eq. HPLC r*.-32 Eq. ao Eq. 1 a-carotenemgl*.32.a..--.*3+a.1/,.1/2a.3,*.,33 Eq. Table + + mgl..-.1/.3, nm *.,,12 * -.+, *.,,,2 A..- A..- Fig.. Eq...- nm *.,,12 A..- *.,,12*.,/0A A.3,/.*22A /*/*.,-1 Eq. a-carotenemgl*.2+.a..--.*3+a.1/ +...*A.3,-.-2*A /*/*.+/* Eq. HPLC *.302 SEC *.,,1 +.*.+ *.,1/ Eq. Scatter plots of calculated lycopene concentration with the Eq. (2) versus measured concentration a-carotenemgl*.2.1a..--.,+2a.1/ +..33A.3,-./+3A /*/*.++3 Eq.

4 354 /. 1,** Fig. / Scatter plots of calculated a-carotene concentration with the Eq. (++) versus measured concentration Eq. X HPLC Y Y*.310X*.**+ *.30/ SEP *.,-+ Fig. / ao *.02 *..3 - ao bo /*/ nm Fig. + ao Eq. ao ao Eq. 2 Eq. 1 Eq. Eq. mgl *.-* *.+. *.-* ao *.02 *..3, 3/ ++ ao bo SEP *.,-+ *.,,2 *.+-3 ao bo *.. mgl *.- mgl 3/ ao * HPLC +,*.* HPLC - ao bo a b +,, +, A ao bo A A bo +0. ao bo..-.1/.3, /*/ nm A..- A.1/ A.3, A /*/ r SEP a-carotenemgl*.2.1a..- -.,+2A.1/+..33A.3,-./+3A /*/*.++3r*.30/SEP *.,-+ b-carotenemgl+..22a a.3,,.-/,a /*/*.*32r*.3.0SEP*.,,2 Lycopenemg L*.,/0A A.3,/.*22A /*/*.,-1r*.330SEP *.+-3

5 43 : /+1 +, pp.1,1/,**0 Van den Berg, H., Faulks, R., Granado, H.F., Hirschberg, J., Olmedilla, B., Sandmann, G., Southern, S. and Stahl, W., The potential for the improvement of carotenoid levels on foods and the likely systemic e#ects. J. Sci. Food Agric., 2*, 22*3+, (,***). Delgado-Vargas, F., Jimenez, A.R. and Paredes-Lopez, O., Natural pigments : carotenoids, anthocyanins, and betalains-characteristics, biosynthesis, processing, and stability. Critical Rev. Food Sci. Nutr.,.*, +1-,23 (,***). Roussell, R., Raley, Lee. and Hofsommer, H.J., Application of diode array detection with C--* reversed phase column for the separation and identification of saponified orange juice carotenoids. J. Agric. Food Chem.,..,,+10,+2+ (+330). / Kato, M., Ikoma, Y., Matsumoto, H., Sugiura, M., Hyodo, H. and Yano, M., Accumulation of carotenoids and expression of carotenoid biosynthetic genes during maturation in citrus fruit. Plant Physiol., +-., 2,.2-1 (,**.) ,/ 3,2 +33, 1 Nagata, M., Noguchi, Y., Sugiyama, K., and Imanishi S., A Simple Method for the Estimation of alpha- and beta- Carotene in Carrots. Acta Horticulturae, (submitted). 2 -/ -+/-,* /**/*0,**, +* pp.,*/,,3 +32* ++ SAS pp. +, , Bradely, G.A. and Rhodes, B.B., Carotenes, xanthophylls, and color in carrot varieties and lines as a#ected by growing temperature. J. Amer. Soc. Hort. Sci., 3., 0-0/ (+303). +2 +, /,