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'Hamlin' Orange Juice—Nutrients and Phytonutrients

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'Hamlin' Orange Juice—Nutrients and Phytonutrients Proc. Fla. State Hort. Soc. 123:207–212. 2010. A Comparison of Processed and Fresh Squeezed ‘Hamlin’ Orange Juice—Nutrients and Phytonutrients Jinhe Bai, Bryan L. Ford, John a. Manthey, and eLizaBeth a. BaLdwin* USDA-ARS Citrus and Subtropical Products Laboratory, Winter Haven, FL33881 AdditionAl index words. flavonoid, limonoid, alkaloid, carotenoid, ascorbic acid, total phenolic content ‘Hamlin’ orange juices were prepared using one of following methods: 1) freshly squeezed with a commercial food service squeezer (fresh), 2) freshly squeezed and pasteurized (fresh/pasteurized), and 3) processed with industrial extractor and pasteurized (processed). Samples were taken from the juices directly after extraction and again after 4 d of juice storage at 5 °C for analysis of flavonoids, limonoids, alkaloids, carotenoids, ascorbic acid and total phenolic content. Processed juice had higher levels of insoluble solids, but lower levels of peel oil in comparison to fresh juice regardless of pasteurization. The majority of flavonoid glycosides including hesperidin, narirutin, narirutin 4´-glucoside, 6,8-di-C- glucosyl apigenin and isosakuranetin rutinoside, which are rich in the albedo and segment membranes, occurred at the highest levels in processed juice. In contrast, the polymethoxylatedflavones associated with peel oil, such as quercetagetin hexamethyl ether, sinensetin, nobiletin, tetramethylscutellarein, heptamethoxyflavone and tangeretin occurred at the highest levels in the fresh juices. Limonoids (limonin glucoside, limonin aglycone, nomilin glucoside, nomilinic acid glucoside, and nomilin aglycone), alkaloids (feruloyl putrescine and an unknown alkaloid) and carotenoids (zeaxanthin, lutein, ß-cryptoxanthin, α-carotene and ß-carotene) occurred at higher levels in the processed juice than in the fresh juice regardless of pasteurization. The processed juice had higher total phenolic content but lower ascorbic acid content than the fresh juices. Thermal pasteurization increased the contents of the polymethoxylated flavones, but decreased the contents of carotenoids. During 4-d storage at 5 °C, 20% to 80% of hesperidin, narirutin, narirutin 4´-glucoside and isosakuranetin rutinoside precipitated in the processed juice but not the fresh juice. Orange juice is popular worldwide for flavor and nutrition Florida, Jan. 2010, and juiced by three different methods: fresh (Baldwin, 1993). The nutritional value of orange fruit is most squeezed (fresh) using a commercial food service juicer, fresh importantly linked not only to the high levels of vitamins and squeezed then pasteurized (fresh/pasteurized), and processed minerals, but also to the many phytonutrients, including the flavo- using an industrial extractor (set for single-strength juice) and noids (Gattuso et al., 2007; Nogata et al., 2006), limonoids (Maier finisher, and then pasteurized (processed). Pasteurization condi- et al., 1980; Manners et al., 2003), carotenoids (Dhuique-Mayer tions were 90 °C for 10 s with a flow rate of 1.2 L·min–1. Details et al., 2005; Dhuique-Mayer et al., 2007), ascobic acid (O’Neil for the materials and processing conditions can be found in Bai and Nicklas, 2008) and other phenolic compounds (Rapisarda et al. (2010). et al., 1999). After processing, all samples were cooled quickly to 5 °C with There is a perception by some that the nutritional value of orange an ice bath, and stored at 5 °C for 4 d. Three replicate samples per juice decreases during processing and storage, and that freshly treatment were taken at day 0 and day 4 after juicing. Analyses squeezed juice is more nutritious. The latter is also perceived as were carried out immediately or after frozen storage at –20 °C being less adulterated. In this study, the nutritional value of fresh or –80 °C. juice prepared with a commercial juicer – a type typically used in AnAlysis of insoluble solids, flAvonoids, limonoids, And restaurants and institutional food services- was compared to that AlkAloids. Juice samples were separated into pellets and super- of a processed juice extracted by an industrial extractor, passed natants by centrifugation at 27,000 gn for 30 min. Pellets were through a finisher, and thermally pasteurized. A portion of the resuspended in deionized water (equivalent volume as original fresh juice was also thermally pasteurized to differentiate the juice) and then centrifuged again at 27,000 gn for 30 min. The effects of extraction and finishing from pasteurization. final pellets were vacuum dried at 55 °C prior to being analyzed. At each step, the amount of juice, supernatant, and pellet were Material and Methods measured by weight to calculate the insoluble solids content. Supernatant samples were passed through a 0.45 μm PTFE source of fruit And processing methods. ‘Hamlin’ oranges filter and 50 μL of 0.13 mg·mL–1 mangiferin was added to 950 were harvested from a commercial orchard located in central μL filtered supernatant as an internal standard. For pellet sample preparation, 100 mg well-ground dry pellet with 3.0 mL of di- Mention of a trademark or proprietary product is for identification only and does methyl sulfoxide (DMSO) was placed in a Teflon gasket screw-top not imply a guarantee or warranty of the product by the U.S. Department of Ag- test tube and shaken for 18 h with an orbital shaker (VSOS-4P; riculture. The U.S. Department of Agriculture prohibits discrimination in all its Pro Scientific, Oxford, CT) at 120 rpm at room temperature. programs and activities on the basis of race, color, national origin, gender, religion, The extraction was then passed through a 0.45 μm PTFE filter age, disability, political beliefs, sexual orientation, and marital or family status *Corresponding author; phone: (863) 293-4133, ext. 120; email: Liz.Baldwin@ and mangiferin was added as internal standard prior to analysis ars.usda.gov by HPLC-MS. Proc. Fla. State Hort. Soc. 123: 2010. 207 A Waters 2695 Alliance HPLC (Waters, Medford, MA) con- peel oil. Peel oil content was analyzed according to Scott- nected in parallel with a Waters 996 PDA detector and a Waters/ Veldhuis bromate titration method (Scott and Veldhuis, 1966). Micromass ZQ single quadrupole mass spectrometer equipped stAtisticAl AnAlysis. SaS Version 9.1 (SAS Institute, Cary, with an electrospray ionization source was used for the analysis. NC) was used for analysis of instrumental analytical data. Each Compound separations were achieved with a Waters XBridge quality attribute with three replicateswas analyzed using analysis C8 column (4.6 × 150 mm), using solvent gradient conditions of variance (ANOVA). The treatment means were separated at as reported previously (Manthey, 2008).Data handling was done the 0.05 significance level by least squares means test (LSD). with MassLynx software version 3.5 (Micromass, Division of Waters Corp., Beverly, MA). Post column split to the PDA and Results and Discussion mass ZQ detector was 10:1. MS parameters were as follows: ionization mode, ES+; capillary voltage 3.0 kV; extractor voltage insoluble solids And peel oil. Processed juice had 0.52% 5 V; source temperature 100 °C; desolvation temperature 225 °C; (w/w) insoluble solids (precipitated pellet material), 37% higher –1 –1 desolvation N2 flow 465 L·h , cone N2 flow 70 L·h ; scan range than in fresh juice (Table 1). The industrial extractor introduced m/z 150–1600; rate 1 scan/s; cone voltages 20 and 40 V. more albedo and segment membrane tissues into the juice, although Quantification of flavonoids and other secondary metabolites the finisher removed some of the juice vesicle walls. Fresh juice were made using either ESI-MS mass extracted Total Ion Chro- had 0.091% juice oil, over 4-fold higher than the processed juice matograms (TIC) obtained in scanning mode by the ZQ mass (Table 1). The thermal pasteurization did not change insoluble spectrometer, or by single ion response (SIR) mode. To normalize solids and peel oil content in the fresh juice (Table 1). These the mass spectrometer response during sequential runs, an inter- characteristics markedly influenced certain nutritional qualities nal standard, mangiferin, was additionally measured at 423 m/z. of the juices. AnAlysis of cArotenoids. Carotenoids in pellet were analyzed flAvonoids. Eleven flavonoids, including five flavonoid -gly using an HPLC. Juice samples (30 mL) were centrifuged at 10,000 cosides and six polymethoxylated flavones were measured. The gn for 15 min. The pellet extracts were collected by dissolving flavanones, including hesperidin, narirutin, narirutin 4´-glucoside, pellets in acetone. The solution was injected into an HPLC (20 and isosakuranetinrutinoside exhibited more than two times higher μL loop) equipped with an YMC carotenoid column (Waters). concentrations in processed juice compared to fresh-squeezed Elution conditions included a three solvent gradient composed juice in both serum (Fig. 1) and pellets (Fig. 2). The contents initially of water/methanol/methyl tert-butyl ether (4/81/15, v/v/v), of these components in the pellets markedly increased during and changed with linear gradients to 4/6/90 (v/v/v) by 60 min at 4-d storage in the processed juice (Fig. 2), while decreased in a flow rate of 1 mL·min–1, at 30 °C. Compounds were detected the serum (Fig.1). The decreases in the levels of hesperidin and using a photo diode array (PDA) detector scanning 200–700 nm. isosakuranetin rutinoside in the processed juice serum were so AnAlysis of Ascorbic Acid (AsA) And dehydroAscorbAte great that after 4-d storage, the contents of these two compounds (dhA). Juice samples were blended with cold (4 °C) 6% tri- were lower than in the fresh juices (Fig. 1), indicating that the chloroacetic acid (TCA) (v/v, 1:1) at a setting of 4 for 30 s us- composition of the cloud of fresh juice may be different from ing a homogenizer (model PT 10/35, Brinkmann Instruments, that of processed juice, or the high content of oil in the fresh Switzerland). The homogenate was centrifuged for 20 min at juice stabilized turbid juice systems.
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