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Food Sci. Technol. Res., ++ (+), +-ῌ+2, ,**/ Technical paper Quantitation of Carotenoids in Raw and Processed Fruits in Japan +ῌ + + + + Masamichi YANO , Masaya KATO , Yoshinori IKOMA , Akemi KAWASAKI , Yoshino FUKAZAWA , + + , - . Minoru SUGIURA , Hikaru MATSUMOTO , Yumiko OOHARA , Akihiko NAGAO and Kazunori OGAWA + Department of Citrus Research Okitsu, National Institute of Fruit Tree Science, .2/ῌ0 Okitsunakacho, Shimizu-ku, Shizuoka-shi, Shizuoka .,.ῌ*,3,, Japan , Saga Prefectural Fruit Tree Research Station, 3+ Teraura, Ogi-cho, Ogi-gun, Saga 2./ῌ**.+, Japan - National Food Research Institute, ,ῌ+ῌ+, Kannondai, Tsukuba-shi, Ibaraki -*/ῌ20.,, Japan . Okinawa Subtropical Station, Japan International Research Center for Agricultural Sciences, +*3+ῌ+ Maezato, Kawarabaru, Ishigaki-shi, Okinawa 3*1ῌ***,, Japan. (Present address, Department of Citrus Research Okitsu, National Institute of Fruit Tree Science, .2/ῌ0 Okitsunakacho, Shimizu-ku, Shizuoka-shi, Shizuoka .,.ῌ*,3,, Japan) Received June ,+, ,**.; Accepted October ,1, ,**. To obtain the quantitative and qualitative data available for estimating the intake of carotenoids from fruits in Japan, carotenoids were analyzed with reversed phase high-performance liquid chromatography (HPLC). Ten carotenoids were examined in 1/ raw fruits and +/ processed fruits, all of which were harvested or purchased in Japan. Phytoene was detected in /2 of 3* fruit samples; z-carotene, in /* of 3* ; lycopene, in +- of 3*; a-carotene, in +2 of 3*;lutein,in/0 of 3*; b-carotene, in 2* of 3*; b-cryptoxanthin, in 02 of 3*; zeaxanthin, in /2 of 3*; all-trans-violaxanthin, in // of 3*;and 3-cis-violaxanthin, in .1 of 3* samples. Citrus fruits of the mandarin type (Satsuma mandarin and its hybrids, such as tangor) were rich in b-cryptoxanthin, b-carotene, all-trans-violaxanthin, and 3-cis-violaxanthin; ‘Star ruby’ grapefruit in lycopene; loquat, Japanese persimmon, and peach in b-cryptoxanthin, b-carotene, all-trans-violaxanthin, and 3-cis-violaxanthin; mango in b-carotene, all-trans-violaxanthin, and 3-cis-violaxanthin; acerolas in phytoene; passion fruits in z-carotene. Carotenoid levels in common fruits, such as apple, grape, lemon, pear, strawberry, kiwifruit, cherry, pineapple, and banana, were low. Keywords: carotenoid, fruit, phytoene, lycopene, b-carotene, b-cryptoxanthin, violaxanthin Introduction ,**.a; Sugiura et al., ,**.b). The USDA-NCC Carotenoid Carotenoids are derived from phytoene after steps of Database is a valuable resource for this research. How- dehydrogenation, cyclization, hydroxylation, oxidation, ever, there may be some di#erences in the carotenoid and epoxidation (Fig. +). They have several health bene- composition of fruits and vegetables grown in Japan and fits, such as provitamin A activity, scavenging of free those used for the establishment of the USDA-NCC radicals, enhancement of gap junctions, immunomodula- Carotenoid Database. For example, the characteristics of tion, and modification of enzyme activity involved in cultivars, climate, and conditions for growing fruit in health and disease (Faure et al., +333). Japan may produce carotenoid profiles that are di#erent Therefore, a large number of epidemiological studies from those in the USDA-NCC Carotenoid Database. Ac- have been conducted on the relationship between caro- curate carotenoid data is important when analyzing the tenoid intake or serum-carotenoid levels and the risk of relationship between carotenoid intake from fruit grown various chronic diseases (Ford et al., +333; Yuan et al., ,**+ in Japan and the risk of various chronic diseases. Thus, ; Zeagers et al., ,**+). These studies require that the com- the objective of this study was to characterize the position of the foods being tested for individual carotenoid content in various raw and processed fruits carotenoids are known. The USDA-NCC Carotenoid Data- consumed in Japan. A total of 1/ fresh fruit and +/ base (Holden et al. +333) provides the most valuable re- processed fruit samples were collected in Japan. A total source for this purpose worldwide. We have been inter- of +* carotenoids, such as phytoene, z-carotene, lycopene, ested in the e#ects of carotenoids, particularly in domes- a-carotene, lutein, b-carotene, b-cryptoxanthin, zeaxanthin, tic fruits, on the risk of various chronic diseases in Japan all-trans-violaxanthin, and 3-cis-violaxanthin were deter- (Sugiura et al., ,**,a; Sugiura et al., ,**,b; Sugiura et al., mined by means of reversed phase HPLC analysis. * To whom correspondence should be addressed. Materials and Methods E-mail: ym0*2,@a#rc.go.jp Carotenoids Carotenoids used in this study are as 14 M. YANO et al. tracts by adding NaCl-saturated water. The pigments repartitioned into the diethyl-ether phase were recovered and dried. Subsequently, the residue was re-dissolved in / mL of a methyl tert-butyl ether (MTBE): methanol (+: + v/v ) solution. Percent recoveries of carotenoids under the condition of this sample preparation procedure were 32.,ῌ for b-car, +*/./ῌ for b-cry and 23.2ῌ for z-car. An aliquot (,* mL) was separated by a reverse-phase HPLC system (Jasco) fitted with a YMC Carotenoid S-/ column of ,/*῍..0 mm (i.d) (Waters, Milford, MA) at a flow rate of + mL minῌ+. The eluent was monitored by a photodiode array detector (MD-3+*, Jasco). The sample was analyzed with three di#erent gradient elution sched- ules. To assay t-vio, c-vio, lut, b-cry, a-car, and lyc the gradient elution schedule consisted of an initial -* min of 3/ῌ (v/v) methanol, +ῌ (v/v) MTBE, and .ῌ (v/v) water followed by a linear gradient of 0ῌ (v/v) methanol, 3*ῌ (v/v) MTBE, and .ῌ (v/v) water for 0* min (method A). To assay z-car, and b-car, the initial solvent composition consisted of /*ῌ (v/v) methanol, .0ῌ (v/v) MTBE, and .ῌ 0ῌ Fig. +. Carotenoid biosynthetic pathway. (v/v) water followed by a linear gradient of (v/v) methanol, 3*ῌ (v/v) MTBE, and .ῌ (v/v) water for 0* min (method B). Zea was assayed by the gradient elution follows: a-carotene (a-car), lutein (lut), and all-trans- schedule of Rouse# and Raley (+330). The initial compo- violaxanthin (t-vio) were obtained from DHI Water and sition consisted of 3*ῌ (v/v) methanol, /ῌ (v/v) MTBE, Environment (Horshholm, Denmark), lycopene (lyc), b- and /ῌ (v/v) water followed by a linear gradient of 3/ῌ carotene (b-car), and zeaxanthin (zea) were obtained from (v/v) methanol, /ῌ (v/v) MTBE for +, min, 20ῌ (v/v) Extrasynthese (Genay, France), b-cryptoxanthin (b-cry) methanol, and +.ῌ MTBE for 2 min, 1/ῌ (v/v) methanol was obtained from Sokenkagaku (Tokyo). Phytoene and ,/ῌ (v/v) MTBE for +* min, and /*ῌ (v/v) methanol (phy) and z-carotene (z-car) were isolated from phy- and /*ῌ (v/v) MTBE for ,* min (method C). producing Escherichia coli or z-car-producing E. coli (Kato The peaks were identified by comparing their specific et al, ,**.). Nine-cis-violaxanthin (c-vio) was prepared retention times and absorption spectra with the authentic from the flavedo of Satsuma mandarin (Kato et al., ,**.). standards. Standard curves for the carotenoid quan- Sampling and Sample Preparation Ripe samples of tification were prepared with those of the authentic raw fruits were harvested from trees at the National standards at ,20 nm for phy, .** nm for z-car, ./, nm for Institute of Fruit Tree Science (Okitsu), Shimizu-Okitsu t-vio, c-vio, lut, b-cry, a-car, lyc, and zea, and ./- nm for Shizuoka-shi Shizuoka, the National Institute of Fruit b-car. The carotenoid concentration was estimated by Tree Science, Tsukuba-shi Ibaraki, Saga Prefectural Fruit the standard curves and expressed as milligrams per +** Tree Research Station, Ogi-cho Saga, and the Okinawa g fresh weight. Subtropical Station, Japan International Research Center for Agricultural Sciences, Ishigaki-shi, Okinawa, all of Results which are in Japan. The rest of the fruit samples were The carotenoid content of ninety fruit samples was purchased at local Japanese markets. For each sample, - determined. Qualitative and quantitative data of these to / fruits of average size were selected. Each fruit was fruit samples are presented in Table +. Phytoene was homogenized and / g samples were removed for analysis. detected in /3 of 3* fruit samples. The high phy levels Carotenoid analysis was performed in triplicate. were found in acerolas (++..12 mg/+** g fresh wt.), dried Carotenoid quantification For carotenoid analysis, apricots (,.2+. mg/+** g fresh wt.) and tangor cv. ‘Youko’ samples were homogenized in .*ῌ (v/v) methanol con- (,./-1 mg/+** g fresh wt.). Zeta-carotene was detected in taining +*ῌ (w/v) magnesium carbonate basic. Pigments /* of 3* fruit samples. High z-car levels were found in were extracted from the residues using an acetone: meth- raw passion fruit (,..13 mg/+** g fresh wt.) and passion anol (1: - v/v ) solution containing *.+ῌ (w/v) ,,0-di-tert- fruit juice (+.+*- mg/+** g fresh wt.). Lycopene was butyl-.-methylphenol and partitioned into diethyl ether detected in only +- samples. High lyc levels were found by using NaCl-saturated water. The diethyl ether layer in watermelon (0.+2. mg/+** g fresh wt.), guava (..-2- mg/ was mixed with ,*ῌ (w/v) methanolic KOH for saponific- +** g fresh wt.), papaya cv. ‘Fruit tower’ (,..2+ mg/+** g ation. The sample was shaken and wrapped in alumi- fresh wt.), and grapefruit cv. ‘Star ruby’ (+.203 mg/+** g num foil to protect it from light. Prior to capping, the fresh wt.). Alfa-carotene was detected in only +2 sample was gently blanketed with nitrogen, before being samples, but no samples showed high levels (῎+ mg/+** g closed and placed in the dark overnight at room tempera- fresh wt.). Similarly, although lut was detected in /2 of ture. Water-soluble extracts were removed from the ex- 3* samples, no samples showed high levels of lutein (῎+ Quantitation of Carotenoids in Raw and Processed Fruits in Japan 15 Carotenoid contents of raw and processed fruits.
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    G C A T T A C G G C A T genes Article Characterization of the Role of the Neoxanthin Synthase Gene BoaNXS in Carotenoid Biosynthesis in Chinese Kale Yue Jian 1,2,†, Chenlu Zhang 1,†, Yating Wang 1,†, Zhiqing Li 1, Jing Chen 1, Wenting Zhou 1, Wenli Huang 1, Min Jiang 1, Hao Zheng 1, Mengyao Li 1 , Huiying Miao 2, Fen Zhang 1, Huanxiu Li 1, Qiaomei Wang 2,* and Bo Sun 1,* 1 College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China; [email protected] (Y.J.); [email protected] (C.Z.); [email protected] (Y.W.); [email protected] (Z.L.); [email protected] (J.C.); [email protected] (W.Z.); [email protected] (W.H.); [email protected] (M.J.); [email protected] (H.Z.); [email protected] (M.L.); [email protected] (F.Z.); [email protected] (H.L.) 2 Department of Horticulture, Zhejiang University, Hangzhou 310058, China; [email protected] * Correspondence: [email protected] (Q.W.); [email protected] (B.S.); Tel.: +86-571-85909333 (Q.W.); +86-28-86291840 (B.S.) † These authors contributed equally to this work. Abstract: Chinese kale (Brassica oleracea var. alboglabra) is rich in carotenoids, and neoxanthin is one of the most important carotenoids in Chinese kale. In this study, the function of the neoxanthin synthase gene (BoaNXS) in Chinese kale was investigated. BoaNXS, which had a 699-bp coding sequence, was cloned from the white flower cultivar of Chinese kale and was expressed in all developmental stages and organs of Chinese kale; its expression was highest in young seeds.
  • Petition to Include Synthetic Crystalline LYCOPENE at 7 CFR 205.605

    Petition to Include Synthetic Crystalline LYCOPENE at 7 CFR 205.605

    Petition to Include Synthetic Crystalline LYCOPENE at 7 CFR 205.605 Item A This petition seeks inclusion of Synthetic Crystalline LYCOPENE on the National List as a non-agricultural (non-organic) substance allowed in or on processed products labeled as “organic” or “made with organic (specified ingredients),” at §205.605(b). Item B 1. The substance‟s chemical or material common name. Lycopene is a naturally occurring aliphatic hydrocarbon of the carotenoid class. Lycopene is the most abundant carotenoid in ripe tomatoes and comprises 80-90% of the total pigment. Lycopene contains thirteen double bonds. The all-trans isomer is predominant in tomatoes and other natural sources. Storage, cooking, food processing, and exposure to light may result in some isomerization of the all-trans form to various cis forms including the 5-cis, 9-cis, 13-cis, and 15- cis forms. Synthetic crystalline lycopene is predominantly the all trans-lycopene (>70%) with some 5-cis- lycopene and other cis isomers. Other chemical names for lycopene are ψ,ψ-carotene and (all-E)-all-trans-lycopene. It is commonly known as all-trans-lycopene. The systematic name for the all-trans-lycopene is (all- E)-2, 6, 10, 14, 19, 23, 27, 31-octamethyl-2,6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 30- dotricaconta-tridecaene. 2a. The name and address of the manufacturer/producer of the substance. Synthetic crystalline lycopene is produced by: BASF SE Carl-Bosch-Straße 38 67056 Ludwigshafen/Germany 2b. The name, address and telephone number and other contact information of the petitioner. International Formula Council 1100 Johnson Ferry Road NE, Suite 300 Atlanta, GA 30342 Contact: Mardi Mountford, Executive Vice President Phone: (678) 303-3027 Email: [email protected] Petition to Include Synthetic Crystalline LYCOPENE at 7 CFR 205.605 3.