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Home , Cranberry, Hawaiian Vaccinium, Vaccinium reticulatum

Phytochemicals in of Hawaiian wild relatives

Hummer, K., Durst, R., Zee, F., Atnip, A. and Giusti, M. M. (2014), in fruits of Hawaiian wild cranberry relatives. Journal of the Science of Food and Agriculture, 94: 1530–1536. doi:10.1002/jsfa.6453

10.1002/jsfa.6453 John Wiley & Sons Ltd.

Version of Record http://cdss.library.oregonstate.edu/sa-termsofuse was Vaccinium ; A-type PACs 3 species were good V. calycinum Sm. Normal-phase HPLC and . The total PACs (as measured Vaccinium d to uroepithelial cells. and is in the public domain in the USA. V. calycinum ¯ ohelo; V. calycinum Sm. and Escherichia coli Allison Atnip c ) were about twice that in cranberry. Linus Pauling Institute, State University, Corvallis, Oregon 97331, USA US Department of Agriculture, ARS, Pacific(PBARC), Basin 64 Agricultural Nowelo Street, Research Hilo, Center 96720, USA Food Science and TechnologyParker Food Department, Science Building, The 2015 Fyffe Road, Ohio Columbus, OH State 43210, USA University, 110 Correspondence to: Kim Hummer, US Department of Agriculture,Clonal ARS, National Germplasm Repository, 33447USA. Peoria e-mail: [email protected] Road, Corvallis, Oregon 97333, US Department of Agriculture,33447 ARS, Peoria Road, National Corvallis, Clonal Oregon 97333, Germplasm USA Repository, 1 V. reticulatum Fruits of native North American contain high levels − c ∗ a d b could be a functional food. Additional examination of these health effects, itPACs is is of clear interest that for improved the health. regular dietary intakeof of PACs. Cranberry PACslinkages include as a higher compared percentageand to of have other A-type been PACs allieduropathogenic with containing preventing commodities, adhesion of P-fimbriated from cranberry have been implicated intract the prevention infections. of urinary Additional natural sources of PACs for human 1 could serve as a rich dietary source of PACs, comparable to or greater Vaccinium Ait.) contain high levels of phytochemicals such as Francis Zee, for pressed . Hawaiian berries had lower peonidin, quinic and citric b 1 − species is warranted. V. calycinum A- and B-type PACs 2 Though more research and 3 . A and B-type dimers were not differentiated in 0.10 mg catechin equivalents kg Vaccinium ± Vaccinium macrocarpon showed comparable or higher PAC levels than in cranberry. Cranberries had higher percentage of 1) glucose/fructose ratio compared with cranberry. Both Hawaiian ∼ d Robert Durst, (24.3 reticulatum V. reticulatum ∗ carbon skeleton centered around the Trolox equivalents kg a 6 1 : 1530–1536 www.soci.org Published 2013. This article is a U.S. Government work − –C calycinum 3 94 –C berries are rich in phytochemicals. One important 6 mol L V. calycinum 2014; µ and V. Vaccinium calycinum ; A-type linkage; ; phenolics; ORAC; FRAP; PAC; ‘ ; 90.2

± Vaccinium

Aside from their potential anti-adhesion properties, PACs have also shown other potential health benefits, including preventative roles in cancer and cardiovascularinflammatory disease, and antioxidant and properties. also display anti- are differentiated from one anotherPACs by are their linked linkages; by the C4/C8 B-type the or A-type C4/C6 linkage, which and is arebond characterized at more C2/C7. by common an than additional ester J Sci Food Agric A-type dimeric and trimerichave been PACs studied in because foods ofadhesion their such in potential as the to bladder cranberries inhibit and bacterial urinary tract. class is the proanthocyanidins (PACs)., Also PACs known as are condensed polymericpossessing members a of C the flavonoid family, INTRODUCTION Wild BACKGROUND: Cranberries ( Abstract (PACs). These polymeric condensationsevaluate phytochemicals of in fruit flavan-3-ol from Hawaiian cranberry monomers relatives, are associated with health benefits. Our objective was to cranberry relatives Kim Hummer, Phytochemicals in fruits of Hawaiian wild (wileyonlinelibrary.com) DOI 10.1002/jsfa.6453 Research Article Received: 6 May 2013 Revised: 27 August 2013 Accepted article published: 23 October 2013 Published online in Wiley Online Library: 20 November 2013 is needed to elucidate the mechanisms by which PACs display characteristic heterocyclic oxygen ring. PACsin are found many naturally foodsand such cocoa. as: Under cranberries,anthocyanidin acid compounds, , hydrolysis, hence grape these the term compounds proanthocyanidin. produce acids amounts and invert ( Keywords: Published 2013. This article is a U.S. Government work and is in the public domain in the USA. CONCLUSION: Berries of the phytochemicals in other wild coupled with fluorescence andused ESI-MS to detected determine total PACs; PACs.index the Spectrophotometric detectors colorimetric tests evaluated 4-dimethylaminocinnamaldehyde and phenolics, reverse-phase (DMAC) ,assays. HPLC and assay coupled organic was to acids. photodiode Antioxidant array capacity and was refractive determined byRESULTS: the ORAC and Antioxidant FRAP capacities of Hawaiian berries were high. The FRAP measurement for 454.7 M Monica Giusti sources of PACs; they contained phenolicsreticulatum and PAC monomers, A and B-type trimers, tetramers and larger polymers. than cranberries. These finding suggest that Hawaiian A-type dimers than did V. by DMAC) for reticulatum

1530 1531 . 4 1 15 , − PO fresh 2 Total 1 et al. 25 min, 12 − Column: = C. Mobile ◦ 12 – 10 fruit. and reported as 30 min, 35% A; 1 − C. Mobile phases: = 14 ◦ m) at 40 µ , pH 2.4 at 1 mL min 4 :acetic acid (v:v:v) 89:1:10 wileyonlinelibrary.com/jsfa 4 m) at 30 PO zero min, 2% A; time 2 PO µ fresh fruit. 3 1 = 4.6 mm 4 KH − × 1 − dithiothreitol (DTT). Quantification was water:H Trolox equivalents kg 1 1 4.6 mm 4 = − − 0–3 min, 5% A; time × and reported as mg cyanidin-3-glucoside = ; detector, 243 nm; mobile phase, 0.5% KH 1 fresh fruit. 0.07 mol L 13 mol L CN; B − 1 3 µ 30 min, 40% A, holding for 2 min then return to = − . Gradient: at time ORAC was measured using a Gemini SpectraMax XS = 1 CH − 16 = CN, B 35 min, 55% A, then return to starting conditions in 5 min. 3 fresh fruit. = CH 1 − Polymeric color was by the bisulfite bleaching method. Phenolic fingerprints were measured by HPLC-DAD. Column: = 20% A; time starting conditions in 5 min and measured at 520 nm. Antioxidant capacity Antioxidants were measuredoxygen-radical absorbance by capacity (ORAC) two assay of different Cao methods: the measured at 214 nm. Quantificationmethod was and by reported an as mg external kg standard by an external standard method and reported as mg kg equivalents kg andbytheferricreducingantioxidantpower(FRAP)assayofBenzieand Strain. w:v, pH 2.5 with 0.5 g L Anthocyanins and total anthocyanins Anthocyanins were measured by HPLC-DAD. anthocyanins were measuredmethod spectrophotometrically by 2005.02 AOAC fruit. Proanthocyanidins Extraction In Ohio, shipped frozen berries wereto blended with liquid obtain nitrogen acarefully fine weighed into powder. aadded Approximately flask, as 5 and g the 40at per mL extraction 300 of rpm sample solution. 66% for was Thesethe acetone 24 samples h, aqueous was were filtered extractto through stirred was further Whatman collected. #1 purifyusing A paper, 20% liquid–liquid the and (v/v) aqueous extraction evaporation. ethyl The fraction acetate, extract wasacidified which was water brought was then and to removed performed further30 100 mL mL by purified of in rotary using sample 0.01% a0.01% HCl was HCl acidified C18 water, loaded and SPE eluted onto with cartridge. were methanol. The the samples then cartridge, rotary washed10 mL evaporated with of to 30% dryness, methanol. All and 10 re-dissolved mL of in each sample were then mg gallic acid equivalents (GAE) per kg fresh fruit. Prodigy ODS-3 (250 Phenolics and total phenolics Total phenolicsFolin–Ciocalteu were spectrophotometric method measured by a modification of the plate reader and FRAP used a Gemini(Molecular SpectraMax 190 Devices, plate Sunnyvale, reader CA,are USA). reported Results as for both assays phases: A Gradient: attime time Data were collected at 260, 280 andquantified 320 nm. by Chlorogenic an acid was externalkg standard method and reported as mg at 1 mL min A Analysis of C VitaminCwasmeasuredbyHPLC-DAD.Column:SpherisorbODS-1; flow, 0.7 mL min Synergi Hydro-RP 80A (250 9 V. C. 4,5 ◦ )to and 4 g 1 × and fruit in − fresh fruit. 1 , isocratic at ripen bright m) at 30 − 2 ) µ 3 and is in the public domain in the USA. was collected from V. calycinum Ca(NO C, and shipped on dry V. reticulatum ◦ 1 − 4.6 mm 4 V. calycinum C. Quantification was by and 20 has a range of skin colors ◦ × − , pH 2.4 at 0.7 mL min were harvested from the Island 4 V. reticulatum PO 2 ¯ have not been well described. The ohelo, are wild cranberry relatives. KH V. reticulatum V. reticulatum 1 − ¯ ohelo species matches that of the skin. : 1530–1536 Published 2013. This article is a U.S. Government work V. calycinum 94 2014; fruits differ in content and in aglycone , column temperature 85 1 The internal color of the Hawaiian species is cause Brix was measured on a Leica Auto Abbe refractometer − ¯ , commonly called ‘ ◦ ohelo species taste bland and have sour, bitter or low 7,8 Vaccinium calycinum Unlike cranberry fruit, which have white flesh under skin, 6 Vaccinium Several subtropical Hawaiian species, Titratable acidity and pH were measured with a Brinkman For analysis, samples were pressed to extract Spectrophotometric assays were performed on a Beckman DU- comparison with that of cranberry. profiles. Non-volatile organic acids Acids were analyzed by HPLC using AOAC method 986.13. model 10500 (Buffalo, NY, USA). for interest about theirdiffer phytochemical from content the and cranberry.native how it Phytochemicals may for fruitobjective of of Hawaiian thiscomposition, study including was PAC,antioxidant to total capacity phenolic, determine for anthocyanins the and phytochemical the fruit flesh color of ‘ J Sci Food Agric Column: Synergi Hydro-RP 80A (250 calycinum MATERIALS AND METHODS Sampling procedure In 2009, fruit from wild of Fruits of ‘ Phytochemicals in ohelo berriesconsumption could provide similartemperate benefit zone fruit, where are cranberries, unavailable. a www.soci.org Mobile phase: 27.2 g L Metrohm autotitrator (Herisau, Switzerland)pH consisting meter, of a 614assembly. 605 impulsomat, 655 dosimat and TT48 titration (Table 1). Juice was obtained byin squeezing fully a thawed hand fruits garlic press, and then centrifuging (5 min at 4000 ice to the Linus Pauling Institute, Oregon State University,Oregon, Corvallis, for phytochemical analysis, and toTechnology the Department Food of Science The and OhioOhio State for University, PAC Columbus, analysis. Samples ofa cranberry reference fruit comparison. were analyzed as Acidity, titratable acidity, and sugars Sugars werecation analyzed exchange by column (Hercules,Mobile HPLC phase: CA, water containing USA) using 200 mg and L a RI detection. BioRad HPX 87-C 640 spectrophotometer (Beckman-Coulter, Brea, CA, USA). Phytochemical analyses In Oregon,analyses high-performance were performed liquid on ana HP chromatography (Agilent) built-in 1090 diode-array (HPLC) equipped with detector(RI) (DAD) detector or (1047A) addedsoftware and (Agilent refractive Technologies, processed index Santa Clara, with CA, USA). Chemstation A.08.03 separate solids. the external standard method and reported as mg kg 0.7 mL min three locations on the Islands offruit Hawaii from and two three on wild . Inof 2010, Hawaii. Fruit samples were frozen at from yellow to dark blue–purple;red. fruit of acid qualities. The fruit of A 17 3), et al. = , 2010 0.7 1.1 n 29.1 1.63 5.6 90.2 0.05 1.1 12.8 0.09 0.40 0.004 0.036 0.063 0.40 0.10 0.008 0.024 0.007 0.003 0.000 0.009 0.015 0.06 0.08 0.92 0.05 1.35 0.02 0.1 0.01 0.4 0.02 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± m pore size : 1530–1536 Sm.; 3 mm Cyano µ . Absorbance 94 × × V. calycinum 2014; V. calycinum ¯ ohelo ( J Sci Food Agric m polypropylene filters (Thermo Fisher . , 2009 9.1 23.7 64.4 117.6 5.0 92.3 66.5 454.7 0.60 0.143 4.64.1 65.84 13.82 3.0 26.4 µ 1 0.80 2.05 2.67 2.88 0.16 1.31 0.22 2.24 0.30 24.3 0.2 0.81 0.0010.0900.157 0.012 0.197 0.444 2.10 9.49 0.131 0.447 0.02711.140.073 0.089 0.018 10.71 0.0290.0030.015 0.201 0.080 0.031 0.010 0.061 0.21 0.85 0.48 6.69 0.29 3.03 2.00.21.10.1 2.8 0.27 3.7 0.40 0.19 0.77 − 5), highbush ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± = 5.2 0.9 2.8 1.2 n 41.0 11.4 1.8 47.8 250 mm Develosil Diol column, 5 0.97 144.2 0.136 0.188 0.053 16.50 0.229 0.056 0.050 0.015 0.014 × Sm.; V. reticulatum Species and sample year analysis was adapted from Wallace and Giusti. Fluorescence of the compounds was monitored at excitation were filtered through 0.45 Scientific, Waltham, MA) prior towas run HPLC-MS in analysis. duplicate. Each sample of the proanthocyanidins was also monitored at 280 nm using and emission wavelengths of 230detector was and set 320 nm. to low The sensitivity fluorescence with a gain of 4 normal phase 4.6 (Phenomenex, Torrance, CA) was connected toSecurityGuard a 4 column. Solvent Aacid was and 95:3:2 solvent B MeOH:water:acetic wasof 2% proanthocyanidin acetic acid extracts in100–80% was acetonitrile. Separation achieved B in with 550–0% min, a B 80–60% in gradient 35 min, B of holding at inThe 0% column 15 B was min, in re-equilibriated 40 60–50% for min, 10 0–100% minflow B B between rate in samples. in was 45 The min. 0.8 25 mL min, min , 2009 ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ ∗ 0.30 11.3 ∗ . ¯ ± ohelo ( ——Trace www.soci.org K Hummer 0.0020.3520.462 0.003 0.225 0.456 0.250 0.108 0.131 0.090 0.003 0.126 0.350 mnylon µ V. macrocarpon and is in the public domain in the USA. V. reticulatum Published 2013. This article is a U.S. Government work SD) of low-growing 0.05) to ± < P ) 86.5 110.3 ) 107.6 1 1 1 − − Ait.) as standard − ) 12.8 11.8 ) 12.2 extractedpressed 2.55 — 3.85 1.46 1 1 − 1 1 − ) — 2.64 − − 1 − ) 0.930 1 1 − 1 − Trolox equiv kg Trolox equiv kg − 1 V. macrocarpon 1 1 1 1 1 1 − − − − − − 1 − 1 1 − Chemical profile of fruit (mean − − mol L mol L µ µ Cranberry value significantly different ( Brix/acid ratio 0.380 Brix 9.58 6.74 and cranberry ( Quinic, g kg Pnd-3-gluPnd-3-arabCyd/pndPnd-gal/pnd-arab 4.4 13.0 2.96 0.78 Total sugars, g kg Malic, g kg Shikimic, g kg Citric, g kg Fumaric, g kg FRAP ( Sucrose, g kg Anthocyanin conc., mg kg Total acids, g kg Glucose, g kg Fructose, g kg Glucose/fructose 3.26 Chlorogenic acid (mg kg Anthocyanin conc., mg kg (mg kg ∗ ◦ Parameter ◦ Table 1. PAC (mg catechin equiv kg ORAC ( Quinic/malic 1.46 1.38 Citric/malic 1.40 0.23 Pnd-3-galCyd-gal/cyd-arab 38.5 1.82 Cyd-3-glu 1.3 2.3 pH 2.70 3.27 % Polymeric color pressed — 17.4 Total phenolics (mg GAE kg Cyd-3-gal 27.1 Cyd-3-arab 14.9 Titratable acidity (as citric) g kg wileyonlinelibrary.com/jsfa loaded onto 3 g of Sephadexin LH-20 which 30% had beenMeOH equilibrated MeOH and overnight. elutedacetone:water:acetic Samples acid. with were The an samplesto washed were extraction dryness rotary with solution evaporated andSamples for of 30% brought LC-MS analysis 70:29.5:0.5 to were filtered through 5 mL a 0.2 in the extractionLC-PDA-fluorescence-MS solution. analysis Samples were analyzed usingUSA) HPLC equipped (Shimadzu, with Columbia,CTA-20A LC-20 AD MD, column pumps, oven, SIL-20 ACphotodiode array coupled (Shimadzu), auto RF-10AXL to fluorescence sampler, (Shimadzu), and an mass spectrometer LCMS-2010, (Shimadzu) detectors, SPD-M20A andSoftware LCMS Solution (Versionconducted 3.0; on Shimadzu).equipped a with electrospray Mass ionization quadrapole (ESI) interface. spectrometry ion-tunnel All samples mass was spectrometer filter.

1532 1533 ∗ ∗ V. calycinum 6.52 23.10 10.22 6.85 and ∗ ∗ The DMAC reagent was wileyonlinelibrary.com/jsfa V. reticulatum 18 , Vaccinium macrocarpon prepared immediately before use. A standard curveusing was prepared a catechinUSA). standard The standard from curve was prepared Sigma–Aldrich and measured prior (St to the Louis, MO, Preparation of 4-dimethylaminocinnamaldehyde Quantitative analysisproanthocyanidins was using achieved the(DMAC) by 4-dimethylaminocinnamaldehyde spectrophotometric measuringdescribed for method by total under Wallace and the Giusti. conditions V. reticulatum V. calycinum . z / m 1. Full- − C; focus lens ABCDEFGH I ◦ and is in the public domain in the USA. 289, 577, 579, 863, z 3.6 V; main rod bias / − m 13.37 16.97 24.99 19.61 16.73 15.36 25.64 19.83 27.70 19.03 flow was diverted to the MS 1 ; block temp 200 − 1 − ∗ z V. macrocarpon 50 V; pre-rod bias / − m : 1530–1536 Published 2013. This article is a U.S. Government work 94 2014; Approximate HPLC % peak areas of identified proanthocyanidin constituents in HPLC-fluorescence chromatogram and MS peaks of the cranberry sample. Approximated % areas due to poor resolution of fluorescence data. 2.5 V; entrance lens 3.5 V; detector voltage 1.5 kV; scan speed 2000 amu s based on retention time and mass spectrometry data ∗ The letters A through I designate sample numbers for fruit from separate plants collected from the Big Island of Hawaii. B-type trimersTetramersPentamers 865Oligomers 1151/1153 1493/1441 ND 1.99 3.37 8.06 22.75 10.71 18.01 10.66 9.50 6.49 15.37 12.14 11.34 16.95 10.73 11.47 17.47 13.63 11.46 15.26 10.07 12.23 14.58 5.59 10.89 15.19 12.67 19.42 11.17 Sample MonomersA-type dimersB-type dimersA-type trimers 289 575 577 863 15.13 15.66 11.09 15.48 2.97 1.17 16.77 11.93 9.11 7.34 16.77 13.58 9.44 4.27 17.47 10.67 17.62 4.66 6.48 13.96 22.16 7.45 1.68 16.19 13.10 25.52 3.44 ND 10.84 14.89 7.51 12.82 ND 9.69 12.72 10.76 ND 0.20 Table 2. ND, not detected. J Sci Food Agric Figure 1. the PDA detector. A 0.25 mL min Phytochemicals in ohelo berries www.soci.org detector. Analysis was performed undernebulizing negative-ion gas mode flow with at 1.5 L min − scan total ion monitoring was performed from 200 to 2000 − Selective ion monitoring was performed at 1151, 1153, 1439 and 1441). Samples were run in duplicate. had et al. ¯ ohelo : 1530–1536 V. calycinum 94 V.calycinum and 2014; and V. reticulatum J Sci Food Agric V.reticulatum ranged from simple, for the yellow fruit, was almost completely composed of chlorogenic acid V. reticulatum ¯ ohelospecieswererichsourcesofphenolics.Chromatograms Chemically extracted samples of in extracted samples of thelower Hawaiian than that species of were cranberry. 32–79% Anthocyanins and total anthocyanins Chromatograms of the anthocyanin profiles of extracted ‘ fruits wereexceptions. similar The cranberry profile to included peaks of cyanidinsand those (cyd) peonidins found (pnd) glycosylated(glu) in and arabinose with (arab). Both cranberry galactose (gal),significant with quantities glucose of some cyd-3-gal and3-glu. cyd-3-arab These but species had lower low cyd- quantities peonidin of content, with pnd-3-gal very and limited pnd-3-glu pnd-3-arab (Table 1). and smaller amounts of had two topressed three samples times (data as notskin, much shown). total particularly This anthocyanin in indicatesanthocyanins as than that dark found did the in samples, the the fruit fruit contains flesh. larger amounts of Total phenolics and phenolic profiles Both‘ of phenolic fingerprints of pressedThose cranberry for fruit were complex. to moderate, for thepurple–blue red fruit. Extracted fruit fruit samples were samples, more complexthat to than of pressed. complex The chromatogram for of the the extractedV. fruit calycinum dark sample of sample E, collected from the Big Island of Hawaii. V. www.soci.org K Hummer V. reticulatum ¯ ohelo species and is in the public domain in the USA. ¯ ohelo species had Published 2013. This article is a U.S. Government work fruit skin color range from yellow, V. reticulatum ripened only bright red. Fruits of both species tasted HPLC-fluorescence chromatogram and MS peaks of Brix, lower acidity and titratable acidity, and thus had Brix/acidratiosthandidcranberry(Table1).Fromextracted ◦ ◦ fresh fruit using the standard curve. 1 − Juice from pressed berry samples of the ‘ were similar to those of cranberries,(Table but the 1). quantities Cranberries were lower arerelative typified to the by level of high other berry quinic crops. The and amounts of citric quinic acid acids Non-volatile organic acids The non-volatile organic acid profiles of berries of ‘ berries, the sucrose levelsglucose/fructose were ratio low of in cranberries was all aboutthe species 3.5 Hawaiian while evaluated; species those were the of more typicalother of fruits an (Table 1). invert Sorbitol ratio was found not present in of in extracted berries samples of the Hawaiian species. higher wileyonlinelibrary.com/jsfa RESULTS AND DISCUSSION Acidity, titratable acidity, and sugars Ripe fruit of red-speckled, salmon, bright red, dark red, to dark purple. The samples because the color of thethen standard faded over solution time. increased The spectrophotometer and was set toa zero using methanol blank. Absorbance was measured atproanthocyanidins 640 nm. were The total calculated askg mg catechin equivalents Figure 2. bland and had some sour, bitterof or cranberry low were tart. acid qualities, while those calycinum higher

1534 1535 1 − had may species. (Table 2, Vaccinium 21 V. reticulatum ¯ V. calycinum ohelo V. calycinum Vaccinium and The PAC content of V. calycinum was not shown to be a 20 or wileyonlinelibrary.com/jsfa V. reticulatum V. calycinum V. reticulatum 21 Based on these results, both Interestingly, cranberries showed a higher percentage of A-type A-type dimers were found to be present in the most popular quantificationgreatly methods between analysts (including and DMAC) samples. vary source of A-type proanthocyanidinshigher (Table percentage 2). of largersamples. polymers In as compared fact, to the(other the cranberry than other samples dimers) were thanMore lower were research in the is other polymers needed two onsmaller the polymers effects of (dimers, larger trimers, polymerspotential tetramers) versus health with benefits of regard the various to berries. the serve as a rich dietary source of PACs, comparable to cranberries. Fig. 1, Fig. 2 and Fig. 3). Theare anti-adhesion properties attributed of cranberries to thetype relatively high dimers, percentage since ofPAC-containing the these commodities health such rare as A- benefit cocoa or is grapes. not observedsamples, in though at other lower levels compareda to cranberry. rich Although source of total PACs, dimers as compared to CONCLUSION In summary, whilespecies were many similar to phytochemicals those foundwas of in cranberry ‘ observed. some distinction Both Hawaiian species were rich sources of sample. cranberries is reported to vary between 4.0 and 40.0 mg kg sample F, collected from the Big Island of Hawaii. 1 V. − were Trolox 1 − V. calycinum mol L and is in the public domain in the USA. µ V. calycinum -caffeoylquinic acid) 90.2 O ± were and were sources of PACs type -caffeoylquinic acid) and 9–25% showed comparable PAC levels, O was 454.7 samples had equivalent or slightly lower V. calycinum V. reticulatum : 1530–1536 Published 2013. This article is a U.S. Government work and total PACs were approximately twice as high 94 V. calycinum samples were also dominated by chlorogenic fruit compared with 107.6 for the cranberry V. reticulatum 1 − 2014; V.reticulatum for the pressed cranberry sample was 12.8 mg kg were approximately nine times that amount (Table 1). HPLC-fluorescence chromatogram and MS peaks of 19 V. calycinum V. reticulatum V. reticulatum Quantitative measurements of PACs are difficult to standardize. The total phenolics, as measured by the Folin–Ciocalteu Though recent improvements have been made, even the as the total PACs in(Table cranberry 1). as determined by the DMAC assay A and B (Tablesource 2). of As compared PACs, towhile cranberry, a well-known rich in the upper range of that for cranberry. The FRAPpressed measurement for sample of Proanthocyanidins Both (Table 1). J Sci Food Agric berry. Other in approximately equal amountsThe (30–43% of chlorogenic acid). levels (Table 1). Thecalycinum total phenolics in the pressed samples of Antioxidant capacity The ORAC capacities for and (tentatively) cryptochlorogenic acid (4- Figure 3. Phytochemicals in ohelo berries www.soci.org equivalents kg method acid and contained small amounts11–16% of neochlorogenic acid other (5- chlorogenic isomers, cryptochlorogenic acid, with additional unidentified phenolics. , et al. Official :15–23 ,ed.by JAmSoc . Handbook Vaccinium J Food Sci 299 : 1530–1536 , 18th edition, Am J Enol Vitic 94 , Multi-laboratory Handbook of Food Vaccinium :303–311(1993). et al 2014; 14 :1473–1478 (2010). MethodsEnzymol :519–525 (2002). Handbook of Food Analytical 90 50 Official Methods of Analysis of the :C619–C625 (2010). , 15th edition. AOAC, , 75 J Sci Food Agric ed. by Wrolstad RE and Schwartz, SJ. :613–617 (2004). FreeRadicalBiolMed J Sci Food Agric Handbook of Food Analytical Chemistry , ed. by Wrolstad RE and SJ Schwartz. John 134 J Food Sci J Agric Food Chem . JNutr :746–749 (1988). Ribes , ed. by Wrolstad RE. John Wiley & Sons, New York, pp. 113 ,and :144–15821 (1965). :C690–C696 (2010). proanthocyanidins in commonconsumption foods and estimations of normal validation of a standard method forin quantifying cranberry powders. proanthocyanidins phosphomolybdic–phosphotungstic acid reagents. 16 Hort Sci of Tropical Agriculture and HumanManoa (2011). Resources, University Hawaii at aglycone, and aglycone–sugarNorthern content American in species the of fruits four sections of in temperate juice: Citric acid, malic acid, tartaric acid, Method 986:13, in Methods of Analysis of theDC (1990). AOAC and bioactive foodChemistry components, in 33–45 (2005). purification ofAnalytical anthocyanins Chemistry (unitWiley F1.1), & Sons, New in York, pp. 7–18 (2005). anthocyanins by UV–visible spectroscopyof (unit Food F1.2), Analytical in Chemistry John Wiley & Sons, New York. pp. 7–18 (2005). content of fruit ,the beverages, pH natural colorants, differential andAssociation method, of wines Official in Analytical by Chemists International ed. by Horowitz H. AOAC, Washington DC (2005). food components, in Wrolstad RE. John Wiley & Sons, New York. pp. 463–470assay (2005). for antioxidants. Direct measure of total antioxidantmodified version activity for simultaneous of measurement of biological total antioxidant fluidspowerandascorbicacidconcentration. and (1999). HPLC–fluorescence–electrosprayquantification of MS procyanidins75 in cranberry characterization extracts. the and 4-dimethylaminocinnamaldehydeproanthocyanidins. assay for flavanols and phenolics, and antioxidant capacity in diverse small fruits: Rubus 7 Ballington JR, Kirkman WB, Ballinger WE and Maness EP, Anthocyanin, 9 Association of Official Analytical Chemists, Acids in cranberry and apple 8 Moyer R, Hummer K, Finn C, Wrolstad R and Frei B, Anthocyanins, 21 Gu L, Kelm MA, Hammerstone JF and Beecher G, Concentrations of 19 Singleton VL and Rossi JA,20 Colorimetry Prior of L, Fan total E, Ji phenolics H, Howell with A, Nio C, Payne M, 10 Durst, RW and Wrolstad, RE, Pigments, colorants, flavors,11 texture, Rodriguez-Saona LE and Wrolstad RE, Extraction,12 isolation, Giusti and MM and Wrolstad RE, Characterization and measurement of 13 Lee J, Durst R and Wrolstad R, Total monomeric anthocyanin pigment 14 Waterhouse AL, Pigments, colorants, flavors, texture, and15 bioactive Cao G, Alessio HM and Culter RG, Oxygen-radical16 absorbance capacity Benzie FF and Strain JJ, Ferric reducing antioxidant power assay: 17 Wallace TC and Giusti18 Wallace M, TC Extraction and Giusti and M, Evaluation normal-phase of parameters that affect V. were and ‘Kilauea’ www.soci.org K Hummer , : 2281–2291 V.calycinum et al Cultivars from V. reticulatum 66 and is in the public domain in the USA. V. calycinum :1225–1228 (2000). V. reticulatum Published 2013. This article is a U.S. Government work 63 :1094–1117 (2000). 80 Bot Jahrb Syst Pflanzengesch Phytochemistry J Nat Prod . A and B-type dimers were not , Vaccinium reticulatum, . Braun-Brumfield, Inc. Ann Arbor, pp. . The total PACs for ¯ ohelo J Sci Food Agric berries as a functional food. More research :375–510 (1941). 71 V. reticulatum V. calycinum weresimilartothatofcranberrywhile Plants of Hawaii National Park. Illustrative of Plants and Vaccinium , Fruit, Nut and Beverage Crops – May 2011 – FN-18. College showed comparable or higher total PAC levels as well Hawaii and ‘Red Button,’ Two ‘ Pflanzengeogr 240–245 (1984). trimers from cranberryp-fimbriated that Escherichia coli. inhibit adherence of uropathogenic Leahy M, A-type cranberrybacterial proanthocyanidins and anti-adhesion uropathogenic (2005). activity. Customs of the South Seas compounds: Nature, occurrence,nutrition and health. dietary intake, and effects on V.calycinum When compared to cranberry, both 6 Zee F, Keith L, Follett P, Bassil N, Reed B, Strauss A, 5 Sleumer H, Vaccinioideen studien. 2 Foo LY, Lu Y, Howell AB3 and Howell AB, Reed Vorsa JD, Krueger CG, N, Winterbottom R, Cunningham DG A-type and proanthocyanidin 4 Degener O, 1 Santos-Buelga C and Scalbert A, Proanthocyanidins and -like wileyonlinelibrary.com/jsfa REFERENCES This research is supported by aAgricultural reimbursable Research agreement between Service and the CooperativeEducation State Research & ExtensionAgriculture Service (SCRI (CSREES),and Grant U.S. ARS Project CRIS Department#04515115, Project and Number of ARS CRIS Number 5358-21000-03800D. The 2008-51180-04873), 5320-21000-012-06R, authorslike would Accession to thank TheCenter Ohio for Agricultural partially Research fundingStuart and Nakamoto, this University Development of Hawaii, research. for providing We fruit samples. acknowledge Dr. ACKNOWLEDGEMENTS phenolics. Their non-volatileto organic that acid of profiles cranberry,were were but lower. the similar Both amounts Hawaiianratios of than quinic species did and had cranberry. citric lowerand The acids ORAC glucose/fructose capacities for FRAP was higher than that for cranberry. approximately twice as highby as the that DMAC assay in (Table cranberry 1).for Hawaiian as These finding determined suggest the potential will be needed to determine their health effects. differentiated in calycinum as for monomers, trimers andThe higher-order compounds cranberries (Table 2). showed aas higher compared percentage to of A-type dimers

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