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Shahidul Islam, M. Jalaluddin and Navam Hettiarachchy

Bitter melon (Momordica charantia L.), an annual vegetable of Cucurbitaceae family, is a vegetable with important nutritional and medicinal qualities. Four adaptable lines/varieties were tested in replicated field trials for productivity, and biochemical and medicinal characteristics at the University of Arkansas at Pine Bluff. The total phenolic contents of the oven-dried and freeze-dried tissues, and seeds, ranged from 5.39–7.75, 6.72–8.02, 6.40–8.90, and 4.67–6.69 mg·g-1 on a dry weight basis, respectively. The total phenolic content of bitter melon from India green (IG), India white (IW), China green (CG) and China white (CW) varieties were 4.67–6.72, 6.03–8.02, 5.39–7.81, and 6.69–8.90 mg·g-1 dry material, respectively. The main phenolic acids in bitter melon flesh were gallic acid, gentisic acid, catechin, chlorogenic acid, and epicatechin. Bitter melon seeds had the phenolic acids, gallic acid, catechin, and epicatechin. The antioxidant activities of methanolic extracts from the bitter melons varieties IG, IW, CG, and CW ranged from 79% to 85%, 79% to 83, 80% to 85, and 79% to 86% inhibition, respectively. The antioxidant activities of the seed ranged from 79% to 84% inhibition. Methanolic extracts of freeze-dried flesh and seed from var. IW and CG showed very high antimutagenic effects against benzo(a)pyrene with Salmonella TA98 (92% to 100% inhibition) TA100 (79% to 86% inhibition), but lower antimutagenicity activities against sodium azide that ranged from 46% to 54% and 17% to 32% inhibition, respectively. The popular belief that bitter melon improves glucose tolerance in Type II diabetes and lowers blood cholesterol is being investigated. It has not been determined which alkaloids, polypeptides, or combination of chemicals found in bitter melon are responsible for the beneficial medicinal effect.

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Lavanya Reddivari and J. Creighton Miller Jr.

Antioxidants have been widely reported to play an important role in disease prevention. In addition to preventing cancer, stroke, heart diseases, and inflammation, they are also involved in immune surveillance. Since the per capita consumption of potatoes in the U.S. is about 137 lb, even moderate levels of antioxidants in this most important vegetable crop probably have an important human health benefit. About 75% to 80% of antioxidant activity in specialty potatoes is due to phenolics and carotenoids. The objectives of this investigation were to evaluate antioxidant activity and total phenolic and carotenoid content of specialty potato selections from the Texas Potato Variety Development Program, and to identify candidate compounds for cancer cell culture investigations. Potato tubers were also used to identify and quantify individual phenolics and carotenoids. Some 320 specialty selections were screened for antioxidant activity (AA), total phenolic content (TP) and carotenoid content (CC) using DPPH (2,2-Diphenyl-1-picrylhydrazyl), FCR (Folin-Ciocalteu Reagent) and colorimetric assays, respectively. After the initial screening, the top 10% were used for analysis of individual phenolics and carotenoids using HPLC. Wide variability for antioxidant activity, phenolic content, and carotenoid content was found among specialty potato selections, providing evidence for genetic control of theses traits. The specialty selection CO112F2-2P/P (purple flesh, purple skin) had the highest AA (832 μg trolox equivalents/g fw), TP (1553 μg chlorogenic acid equivalents/g fw) and CC (590 μg lutein equivalents/100 g fw). Chlorogenic acid (55% to 60%), caffeic acid (≈5%), gallic acid (18% to 20%), and catechin (18% to 20%) were found to be the most prevalent phenolic acids, and lutein and zeaxanthin were the most prominent carotenoids contributing to antioxidant activity. Gallic acid was identified as the candidate compound for use in cancer cell culture investigations.

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J. Emilio Villarreal, L. Lombardini and L. Cisneros-Zevallos

The objective of this study was to evaluate kernels of different pecan (Carya illinoinensis) cultivars for their antioxidant capacity and characterize the nature of the antioxidant compounds. Nuts collected from four pecan cultivars `Cheyenne', `Cape Fear', `Desirable', and `Pawnee' were shelled, chopped and analyzed for their antioxidant capacity (AC), and for their phenolic, tannin, and vitamin C content. AC was measured using one spectrophotometrical [DPPH (2,2-diphenyl-1-picrylhydrazyl)] and one fluorescence method [ORAC (Oxygen Radical Absorbance Capacity)]. Total phenolic and tannin content were determined using spectrophotometrical assays. Finally, ascorbic and dehydroascorbic acid were determined using a high performance liquid chromatograph. Both AC methodologies, DPPH and ORAC, gave similar results with marked differences between cultivars. `Desirable' had the highest antioxidant capacity (47,747 μg TEq/g DW with DPPH method) followed closely by `Cheyenne' (36,192 μg TEq/g DW) and, with smaller amounts, by `Cape Fear' and `Pawnee' (16,540 and 13,705 μg TEq/g DW, respectively). Total phenolic content showed a similar trend but `Pawnee' showed a higher phenolic content than `Cape Fear'. `Cheyenne' had the highest amount of tannins, 9,114 μg/g DW, followed by `Cape Fear', `Pawnee' and `Desirable' with 7,764, 6,043 and 5,508 μg/g DW respectively),. `Cheyenne' had also the highest vitamin C content, up to ≈10-fold greater than `Cape Fear' and `Pawnee', the highest difference within the antioxidants analyzed. There is the need to determine the phenolic profile and degree of polymerization of tannins, their contribution to the AC and how they are affected by horticultural practices in order to better understand the nutraceutical potential of each cultivar.

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R.E. McDonald, W.R. Miller and T.G. McCollum

Irradiation is being evaluated as a quarantine treatment of grapefruit (Citrus paradisi, Macf.), but it can cause damage to the fruit. We wanted to determine if pre-irradiation heat treatments would improve fruit tolerance to irradiation as they improve tolerance to low-temperature injury. `Marsh' grapefruit were harvested from interior and exterior canopy positions and irradiated at 0 or 1.0 kGy at a dose rate of 0.148 kGy·min-1 before storage for 4 weeks at 10 °C. Following storage, pitting of flavedo tissue was the most evident condition defect noted as a result of irradiation. Pitting was noted on 15% and 27% of irradiated interior and exterior canopy fruit, respectively, whereas there was no pitting on non-irradiated fruit. Temperature conditioning before irradiation decreased the susceptibility of fruit to damage. Pitting was 26%, 19%, and 17% when fruit were held 2 h at 20 (ambient), 38 or 42 °C, respectively. There was a marked increase in phenylalanine ammonia-lyase (PAL) activity following irradiation. Maximum activity (≈18-fold increase) was attained 24 h after irradiation. Irradiation-induced PAL activity was reduced significantly by temperature conditioning at 38 or 42 °C. Exterior canopy fruit flavedo contained higher levels of total phenols, including flavanols and coumarins, compared with interior canopy fruit. The deposition of lignin was not related to canopy position. Neither irradiation nor heat treatment had an effect on total phenols or lignin deposition. It seems that irradiation causes a stress condition in the fruit, which leads to pitting of peel tissue. Heat treatment before irradiation reduced the damaging effects of irradiation.

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R.E. McDonald, W.R. Miller and T.G. McCollum

Irradiation is being evaluated as a quarantine treatment of grapefruit (Citrus paradisi Macf. `Marsh'), but it can cause damage to the fruit. Research was conducted to determine if preirradiation heat treatments would improve fruit tolerance to irradiation as they improve tolerance to low temperature injury and to determine if canopy position influenced fruit tolerance to irradiation. Initially, grapefruit were irradiated at 0 or 2.0 kGy at a dose rate of 0.14 kGy·min-1 and selected biochemical changes were monitored over time. There was a marked increase in phenylalanine ammonia-lyase (PAL) activity following irradiation. Maximum activity (≈18-fold increase) was attained 24 hours after irradiation. Subsequently, grapefruit were harvested from interior and exterior canopy positions and irradiated at 0 or 1.0 kGy at a dose rate of 0.15 kGy·min-1 before storage for 4 weeks at 10 °C. Following storage, pitting of flavedo was the most evident condition defect noted as a result of irradiation. Pitting was observed on 27% and 15% of irradiated exterior and interior canopy fruit, respectively, whereas there was no pitting on nonirradiated fruit. Heat treatment before irradiation decreased susceptibility of fruit to damage. Pitting was 26%, 19%, and 17% when fruit were held 2 hours at 20 (ambient), 38 or 42 °C, respectively. Irradiation-induced PAL activity was reduced by temperature conditioning at 38 or 42 °C. Exterior canopy fruit flavedo contained higher levels of total phenols, including flavanols and coumarins compared with interior canopy fruit. Deposition of lignin was not related to canopy position. Neither irradiation nor heat treatment had an effect on total phenols or lignin deposition. Generally, cholesterol, campesterol, stigmasterol, β-sitosterol, and isofucosterol were found to be higher in four steryl lipid fractions in exterior canopy fruit compared with interior canopy fruit. Irradiation increased campesterol in the free sterol and steryl glycoside fractions and decreased isofucosterol in the free sterol fraction. Heat treatments had no effect on individual sterol levels. It seems that irradiation causes a stress condition in the fruit, which leads to pitting of peel tissue. Heat treatment before irradiation reduced damaging effects of irradiation.

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Bruno Defilippi*, Abhaya Dandekar and Adel Kader

To understand the role of ethylene in overall flavor of apple fruits, ethylene production, and action were reduced using apple trees lines transformed for suppressing activity of ACC-synthase or ACC-oxidase enzymes, and 1-methylcyclopropene (1-MCP), an ethylene action inhibitor. A major reduction in ethylene biosynthesis and respiration rates was measured in fruits from these treatments. As expected, we found differential levels of dependence of flavor components on ethylene biosynthesis and action. Regarding aroma production, an ethyleneassociated event, headspace analysis showed a reduction in ester production in the ethylene-suppressed lines and in the apples treated with 1.0 μL·L-1 1-MCP for 20 hours at 20 °C. However, no major differences were observed in concentrations of alcohol and aldehyde volatiles. Other flavor metabolites that showed an ethylene-dependent pattern were organic acids and sugars. Malic acid degradation was significantly reduced under ethylene suppressed conditions, showing a recovery after exposing the fruit to ethylene. Sucrose and fructose concentrations were influenced by suppression or enhancement of ethylene. Total phenolics and individual phenolics showed an ethylene-dependent behavior only when ethylene biosynthesis was reduced, but not when ethylene action was affected. These results suggest that the regulatory mechanisms of aroma biosynthesis in apple are under partial ethylene regulation. Therefore, we are using the ethylene suppressed apple fruits study the channeling and regulation of other metabolic pathways that lead to the manifestation of a complex trait like fruit quality.

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J. Matt Fulkerson and Douglas D. Archbold*

Modified atmosphere (MA) storage of blackberries may maintain quality and increase storage life, but there is limited information about how eastern thornless cultivars respond to MA's. Because there is also a growing interest in the health benefits of antioxidants in blackberries, it would be useful to know how those levels might change during MA storage. In 2002, the eastern thornless blackberry variety Chester was stored in MA; treatments included a control, or initial levels of 20% CO2 or 5% O2. Color, pH, firmness, fresh weight, soluble sugars, titratable acidity, total antioxidant capacity, and the levels of major classes of compounds with antioxidant activity of the fruit were measured at harvest, after 1 week of MA storage at 4 °C, and after 3 additional days at room temperature to simulate common industry practices and grocery display. Total antioxidant capacity was measured using the FRAP (ferric reducing/antioxidant power) assay, and total phenolics and anthocyanins were measured spectrophotometrically. In 2002, soluble sugar levels, fresh weight, titratable acidity, and all classes of antioxidants decreased from day 0 to day 7 to day 10 while the pH increased. Color values did not change. The only trait that differed among treatments was berry firmness; from day 0 to day 7 control fruit was the most firm, and those from the high CO2 treatment were the least firm. Studies were continued in 2003 with the addition of two more eastern thornless blackberry cultivars, Hull Thornless and Triple Crown, and these results will also be presented.

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Vicky W. Lee, H.P. Vasantha Rupasinghe* and Chung-Ja Jackson

Apples are excellent sources of dietary phenolics, in particular flavonoids and chlorogenic acid, which are potent antioxidants that may play important roles in the prevention of chronic diseases. This study investigated the major phenolics profiles of apple fruit in relation to (1) the distribution among 8 Ontario-grown cultivars, (2) the different fruit parts, and (3) the effect of processing of fresh-cuts. In addition, total antioxidant capacity (TAC) and total phenols content (TPC) were measured in apples by spectrophotometric assays. Flavonoids and chlorogenic acid were quantified using HPLC/PDA. Vitamin C was quantified using HPLC/Fluorescence. TAC, TPC and flavonoids levels were the highest in Honey Crisp and Delicious, moderate in Idared, Spartan, Granny Smith, and Cortland, and the lowest in Crispin and Empire. Apple peel contained 2 to 10-fold higher TAC, TPC and total of 10 major phenolics than that of core and flesh indicating peeling of apples during processing could reduced significantly the nutritional quality of fresh-cut apples. Dihydrochalcone (phloridzin) and chlorogenic acid levels were 2 to 21-fold higher in apple core than skin and flesh. TAC levels and vitamin C contents could be increased up to 3-fold and 14 to 20-fold, respectively by the post-cut dipping treatment with an ascorbic acid-based antioxidant formula. The phenolic profiles of sliced apples were stable up to 21 days at 4°C.

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M. Elena Garcia, C.R. Rom and J.B. Murphy

Two experiments were conducted to determine the effects of leaf age and shading on the phenolic content and composition of apple foliage. In the first study, it was determined that the phenolic content of `Liberty', at increasing leaf developmental stages, was leaf age—dependent. Early during leaf development, there was an increase in the phloridzin (the primary glycoside identified) and in total phenolics, reaching a maximum when the leaf is 6 days from 20-mm blade length. After this stage, the phenolic content decreased with increasing leaf age. In the second study, the leaves of two cultivars, `Liberty' and `Starkspur Law Rome', were tagged weekly when the leaf was two-thirds unfolded. Three weeks after budbreak, the trees were placed under three shade cloth treatments (0%, 60%, and 90% shade). After 4 weeks under the shade treatments, the tagged leaves were collected to determine their phenolic content. Shade significantly affected the foliar phenolic content. Leaves in 0% shade had the highest phenolic content, whereas the lowest content was found in leaves exposed to 90% shade. There was a significant leaf age × shade interaction. The phenolic content decreased with increasing leaf age except for those leaves whose development occurred before the experiment was started. The results indicate that light and leaf developmental stage are important factors in determining the phenolic content of apple leaves, but shading appears to have a stronger influence than leaf developmental stage. E-mail mgarcia@zoo.uvm.edu; phone (802) 656-2824.

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Artemio Z. Tulio Jr., Ann M. Chanon, Nithya Janakiraman, Mustafa Ozgen, Gary D. Stoner, R. Neil Reese, A. Raymond Miller and Joseph C. Scheerens

This study was conducted to determine the effects of postharvest storage temperatures on the antioxidant capacity, anthocyanin compounds, phenolic constituents, and physico-chemical properties of black raspberries. Fresh `MacBlack' berries were stored at 4, 12, 20, and 28 °C for up to 11, 6, 4, and 3 days, respectively. Results showed that higher storage temperatures promoted tissue deterioration (cellular leakage), fungal growth, and moisture loss. The levels of the two major anthocyanins, cyanidin 3-rutinoside and cyanidin 3-xylosylrutinoside, increased by up to 2.7- and 1.9-fold, respectively, with increasing storage temperatures. The antioxidant capacity of berries, as measured by FRAP and ABTS assays, increased by up to 1.5- and 1.4-fold, respectively, which was accompanied by increases in soluble solids, total sugars, total phenolics, and total anthocyanin contents. Our findings indicate that postharvest storage at higher temperatures increases the level of bioactive compounds and antioxidant capacity in black raspberries, but this increase may be due in part to moisture loss and sugar metabolism. Storage at 4 °C maintained the level of bioactive compounds and antioxidant capacity present at harvest and prolonged the effective shelf life of the product. Further studies of black raspberry bioactive components as influenced by postharvest conditions and processing procedures (e.g., IQF, freeze-drying, air-drying) are warranted.