a -tocopherol Furthermore, α 7 Rubus 2011 Society of Chemical Industry c -carotene had notable variations β and Vesa Virtanen b 11,12 MTT Agrifood Research Finland, Sotkamo Research Station, FI-88600Finland Sotkamo, Correspondence to: MariConsortium, Jaakkola, CEMIS-Oulu, Salmelantie University 43, FI-88600 Sotkamo, of Finland. E-mail: mari.jaakkola@oulu.fi Oulu, Kajaani University University of Oulu,Sotkamo, Finland Kajaani University Consortium, CEMIS-Oulu, FI-88600 Despite the fact that cloudberries have been found to ex- ∗ a b been developed inthere Finland, are Norwaycosts. still and North problems America, with but profitability due to the high press phenotypic variationis depending a on shortageposition their of habitat, of knowledge there cloudberries pertainingbest of to grown our the incloudberries knowledge, chemical has different only com- been habitats. the reported variation in To the in the literature. the seed oils of no studies arecomponents available or nutritional of value of cultivated the cloudberries.fore, There- annual the variation objectives of ofcomposition cloudberry this of study were cloudberriesthe to grown effect analyse in the ofof chemical different annual cloudberries habitats grown variationThe and in analyses on included the both theFurthermore, primary same and chemical the area secondary primary composition metabolites. in metabolitesvated and cloudberries northern antioxidants were analysed Finland. of and culti- comparedcloudberries. to those of wild Kalle Hoppula -tocopherol, anthocyanins and b α L.) is one of the most valuable berry-producing plants because of its nutritional L. grown in different habitats and have a beneficial fatty 6 L.; cloudberry; chemical composition; crop yield; habitats Rubus chamaemorus Ville Korpelainen, ∗ For example, cloudberries have been a 10 (Rosaceae) is one of the most valuable most probably brought about by phenolic : 1324–1330 www.soci.org 92 4,5 and possess antimicrobial activities against 3 These secondary metabolites, which are mainly 2012; 1,2 Rubus chamaemorus 7,8

Plants growing in different habitats express pheno- 9

2011 Society of Chemical Industry -linoleic, oleic and palmitic acids amounting to over 95% of all c between habitats. In particularly, cloudberriescompared grown to on those open grown on habitats shaded had sites.of higher In fatty content a colder of acids and citric rainy were acid summerexcept significantly the and in content less higher open of anthocyanins sites than and where in the yields unsaturation a were level quite warmer equal. and drierCONCLUSION: summer. Cloudberries Crop grown yields wereto in higher those open in habitats grown thefactors warmer showed on affecting summer, notable crop shaded yield differences sites. andanthocyanin chemical in Results concentrations. composition chemical of suggest cloudberry. composition that In when addition, the the compared amount amount of of rainfall may sunlight have an and effect temperature on Keywords: levels could be the main found to havebut long short shoots shootsences and and have small large been leavesreserves leaves attributed in in in to competitive open a shadytivation and areas. greater sites, methods resource-poor The need for environments. for differ- commercial Cul- large cloudberry food production have J Sci Food Agric Cloudberries typically inhabitbogs, forested peaty mires, moors,zone. and open wet spruce sphagnum forest within the boreal INTRODUCTION Rubus chamaemorus berry-producing plantsand because distinctive of flavour.source its Cloudberries of are nutritional an nutrientsE), properties important fibre, such dietary fatty ascompounds. acids antioxidant and vitamins also (C, non-nutrient bioactive A phenolic and Abstract BACKGROUND: Cloudberry ( Mari Jaakkola, Chemical composition of ripe fruits of (wileyonlinelibrary.com) DOI 10.1002/jsfa.4705 Research Article Received: 11 March 2011 Revised: 21 July 2011 Accepted: 14 September 2011chamaemorus Published online in Wiley Online Library: 14 November 2011 properties. The chemical composition andwas crop analysed yield over of two ripe consecutive fruits summers. of For comparison, cloudberry two grown clones wild ofRESULTS: in cultivated 10 cloudberries The habitats were in studied concentrations as northern well. of Finland citric and malic acids, acid composition: seeds isolated fromα cloudberries have linoleic, fatty acids. typic variation, suchleaf, as and differences rhizome in sizes. mean amount of seeds, ellagitannins with asuggested to exert high many beneficial health properties. content Cloudberries have of more ellagic antioxidants than acid,and many have commonly vegetables consumed been fruits human pathogens, compounds (mainly ellagitannins)such together as malic, with citric and organic benzoicfibres acids. acids Cloudberries than contain other more berries in general

1324 1325 1 C, − ◦ ). Cmin 24:1 ◦ to 210 ) for 5 min. –C g 1 − 4:0 × -dehydroascorbic C, 60 min). Water L ◦ Cmin ◦ Metaphosphoric acid -dehydroascorbic acid L 19,20 wileyonlinelibrary.com/jsfa 250 mm, Agilent Technologies). m; Phenomenex, Torrance, CA, 17 µ × m, polyethylene glycol TPA; Agilent µ -ascorbic acid and L) were injected in a split mode (ratio L µ 30 . ) buffer (pH 2.4) using a Zorbax Eclipse 1 − 200 mm, 5 m, 4.6 mm C, where it was held for 5 min. Analysis was -dehydroascorbic acid. Therefore vitamin C ◦ µ L and multiplying the obtained nitrogen content × ) was added to the filtered extract (1 : 1, v/v), C for 2 min, then increased by 15 was determined measuring total nitrogen by the 1 ◦ content was determined by the Folin–Ciocalteu 16 were extracted from the berries (5 g, fresh weight − compounds were extracted from freeze-dried is very sensitive to light and temperature and C and helium (purity 6.0; AGA, Espoo, Finland) was ◦ to 220 in which gallic acid was used as a calibration 1 − (117.5 mmol L 18 C; this was further increased by 2 ◦ 4 msyringefilter.Anthocyaninswereidentifiedandquantified µ PO Cmin -ascorbic acid, dithiothreitol (DTT) treatment was performed L 2 ◦ Total phenolic Fat-soluble Anthocyanins Vitamin C Proteincontent 45 . DTT (40 mmol L method, 30 : 1) at 250 to 190 performed using electron impact (EI)were and identified scan by authentic mode. methylated Fatty fatty acids 461, acid standard supplied (GLC- from Nu-chekcontained a Prep mixture Inc., of 32 Elysian, methylated fatty MN, acids USA), (C which (10%, 5 mL) wasflask added (25 mL) to and the the samplewith sample was water shaken and (1g, well. the The fw) sample flask was in was centrifuged filled a (1808 volumetric for the samples. Each sampleincubation, was but analysed before only and thethat after DTT-treated gives DTT the value total wasthan amount reported, of the since vitamin value C determinedmodified and before was from always treatment. previous higher The publications. method was by a general nitrogen-to-protein conversion factor of 6.25. (fw)) by 0.01% (v/v) HCl in methanol(20 (10 min).Extractionwasrepeatedtwiceandthethreeextractswere mL) in an ultrasound bath combined. One millilitre of the extractsand then was dissolved in dried 0.01% HCl under water (1 nitrogen mL)0 and filtered through a USA) as described in detail earlier. by high-performance liquid chromatography (HPLC) (Agilent 1100 Series HPLC, Agilentcolumn (2.00 Technologies) mm using a Hyperclone ODS after which thethe reduction dark reaction at was roomthen allowed temperature analysed by to overnight. HPLC (Agilent A proceed 1100 Series)array sample in connected to aliquot detector a was diode (245 nm).KH An isocratic run was performed with standard inJapan) spectrophotometric measurement. (UC-1601; Shimadzu, Kyoto, as where it was held for10 20 min, and then finally increased again by cloudberry samples (1 g) by Soxhletyield extraction of in total hexane fat and was the acid profile determined was analysed by from samples a after conversion gravimetric ofmaterial the method. to lipid the Fatty corresponding methyl esters (FAME). Driedextract Soxhlet was dissolved in toluene (110% mL) and (v/v) transmethylated with borontrifluoride in methanol (100 was measured as a sum of used as a carrier gas.was The gas held chromatograph at oven temperature 60 Kjeldahl method XDB-C18 column (5 (1.6 mL) and hexane (1.6and mL) mixed were added well. todissolved The the in cooled hexane organic sample (0.7chromatograph mL). layer (HP6890; Agilent FAMEs Technologies, was were Inc., Palo analysed Alto, separated,USA) CA, using connected dried a to gas and Technologies). a The mass compounds selective werecolumn detector (25 separated m, (HP5973; 0.20 on mm, Agilent 0 anTechnologies). HP-FFAP Samples (1 acid. To prevent oxidation and to recover oxidizes easily to L. www.soci.org 2011 Society of Chemical Industry c ,moderateamountof E). Sampling site areas  ++ 300300 12 35 16 28 28  Rubus chamaemorus > > depth cm 2006 2007 Peat layer 22 ◦ , few small trees; Low 200 46 10 High 60 17 10 High 200 34 12 HighHigh 150 80 80 74 22 12 HighHighHigh 150 100 332 100 n.a. n.a. 0 360 220 + Description Crop yield (kg/ha) : 1324–1330 N, longitude 30 of the sample was measured by a gravimetric 50 m, with specified characteristics (Table 1). 92  × Cloudberry fruits were mixed well and stored in a 21 +  ++ ++ ++ ++ ++ C) until analysed. From the frozen fruits (the whole 2012; Tree stand Undergrowth +++ +++ ◦ 58 , dense forest. n.a., not analysed. Sampling site description and crop yields of the studied ◦ 13,14 20 15 − +++ Moisture content trees; Tree stand: 0, open area; 1 Sampling site Table 1. cloudberry populations in 2006 and 2007 56 7 0 Low 3 4 0 Low 2 8 9 10 Chemical composition of ripe fruits of Laboratory analyses Frozen samples (200analysis. g) The were wholefrom thawed set the same of and sample chemical homogenate.at homogenized Each least analyses twice, analysis except for was were in repeated thelaborious, performed and analysis in of the analysis dietary of fibre, fatty acids, whichused where as is rapeseed a very oil control was sample to validate the assay in each samplemethod. set. crop), three separate random samplestaken (200 in g each) 2006 per and sitefor one were analysis (200 g) of randomsamples chemical sample composition. (three per The replicates) sitebetween results in were habitats, 2007 of whereas used the year annual tocalculating 2006 variation arithmetic study was means analysed of the by all samplesyear. differences collected Therefore in the a number specific of replicatesvariation was in nine the in analysis 2007 (no of crop annual 30 was in obtained 2006. in site eight) and J Sci Food Agric Plant material Wild cloudberry sampleslocated in were Ulvinsalo Strict collected Nature Reserve, Kuhmo,(latitude at eastern 63 Finland 10 sampling sites MATERIAL AND METHODS freezer ( measured 50 m In each sampling site thelasted entire from yield mid was July harvested. todepending The mid on season August light and and ripeningopen temperature. of sites fruits Harvesting approximately varied took 5–14 placedepending days on on earlier the than year. forestedin Crop summer areas, yields 2006 were and observed 2007.Fjordgull) for Cultivated were each cloudberries site obtained (Fjellgull from and Eastern a Finland, greenhouse approximately 40 located km in fromThe the Kuhmo, greenhouse wild habitat was sites. unheatedpeat and at the humification stage growingwas H2–H4. substrate harvested was In the inthe greenhouse, June, the wild approximately yield habitat 4Fjellgull sites. weeks and In earlier the Fjordgull thanwere greenhouse, together used. in the with female the cultivars male cultivar Apollo . et al : 1324–1330 ), where also , respectively. 1 92 (Table 2). The 1 − (marsh tea) and − 1 − (relative standard 2012; 1 − 300 cm) and lowest at 7gkg > ± , respectively (Table 2). This 1 Ledumpalustre − J Sci Food Agric 2and73g spp. (sphagnum) and other mosses ± and analysis of variance (ANOVA) procedures Sphagnum 26%) between sampling sites (Table 2). Variation U = spp.), wild grass species, values obtained in 3temperatures h that intervals, were dailytemperaturesthatwerecalculatedasameanofdailytemperatures, minimum recorded and separately, maximum averagemonthly monthly sunshine duration and monthly rainfall. Statistical analyses Statistical analyses were performedvariationsinchemicalcompositionbetweenconsecutiveyearsand with SPSS Statisticsbetween 18. open and The variously shaded habitats were evaluated using Mann–Whitney and the correlations between concentrations of themeasured analytes using were non-parametric Spearman’s rho correlation test. RESULTS AND DISCUSSION Sampling sites Tree stand,availability undergrowth of and lightconditions as surface of well the geometry5 as plants. represented affect the In open humidity the stand, this sites whereas and the study, with other temperature sites sampling lowfrequency were of shaded undergrowth sites trees by and moderate and 4 longer orand undergrowth no high and (Table 10 1). tree had Sites 2, moderatewhereas 3, amounts 9 sites of trees 1 andhad and high moderate 7 undergrowth, amounts were ofsite trees covered 8 and with had low only undergrowth, dense aand5hadmainly whereas forest. few small Site trees 6 within high the undergrowth. undergrowth, Sites in 4 other sites addition had taller to undergrowth consisting which ofSphagnum mosses site (including 5 hadother grass. suffrutices The (subshrubs) asgeometry dominant in plant all species. sites Surface waswith mainly uneven the with natural exception hummocks, hummocks, of and site sites 10, six which waslayers and were located deepest on eight, at a open steep which sites slope. 4 and did Peat 5 ( not have deviation, RSD in the contents ofsites other was primary not metabolites high: between RSDswere sampling of 16%, proteins, 10% dietary and fibre 4%, andweight respectively, dry basis. when weight Calculated calculated on a on dry afibre weight fresh was basis the significantly RSD for lower(14%). dietary (6%) and a little lower for proteins Malic acid and citric acid There were significant differences in the amountcitric of acid malic analysed acid from and different habitatsthere (Table was 2). a Furthermore, significant negative correlation in the concentrations is reasonable since dietarycontributing fibre to is dry one weight. of Thedietary the average fibre main (fw) content were components of 16 proteins g and Total fat content varied from 4 to 10 g kg protein and dietary fibrebasis content was calculated lowest: on 10 a and fresh 63 g weight kg lowest dry weight was in sampling site 8 (150 g kg sites 1 (60 cm) and 3layers were (80 100–200 cm) (Table cm 1). deep. At the other sites the peat Composition of cloudberries grown in different habitats Primary metabolites The dry weights of analysed cloudberriessampling collected from sites different were commonly 170–180 g kg - α C, in ◦ www.soci.org M Jaakkola m, 4.6 mm 99%, Fluka, µ 2011 Society of Chemical Industry hydrochloric > c 1 − ) for 10 min and g ) using an ORH-801 × 1 − -tocopherol (purity 97%; α -tocopherol, except for the The extract was evaporated α 300 mm, Transgenomic, Norlab 21 were extracted from cloudberry using an enzyme kit (total dietary × -Carotene was analysed from the 22 were extracted from cloudberries β (0.00125 mol L 4 SO 2 tocopherol - α was analysed by the enzymatic–gravimetric -Tocopherol (in 0.5 mL acetone) was determined α ). The mobile phase consisted of methanol and water -Carotene was analysed only in 2006. Samples were 1 -ascorbic acid standard (p.a., Merck KGaA, Darmstadt, β L − Carotene and - 150 mm, Agilent Technologies) column with isocratic run Dietary fibre Citric acid and malic acid β -carotene (purity 97%; BioChemica, Fluka). method of AOAC International (10 g fw, collected inhigh-performance 2006 disperser (Ultra and Turrax, 2007) IKA, infor Staufen, 80% 2 Germany) min. ethanol The (20 mL) extract by was centrifuged (1808 filtered. Extraction was repeated twice andcombined all in three a extracts 50 were mLethanol. volumetric Samples flask, of which 1 was mL filledwith were with combined purified 80% CH(EC) by (Isolute, solid-phase 500 extraction mg,Oy, 3 Vantaa, mL; Finland) Biotage and AB, Norlab SAXNorlab Oy) (Isolute, columns 500 activated mg, with 3 methanolwere mL, and water. eluted Biotage Columns with AB, water toand remove malic sugars, acid after were which eluted citric with acid 3 mL of 1 mol L Germany). (95 : 5, v/v). by FD using 292 nm excitationConcentrations in and samples 324 were calculated nm by an emission external wavelength. standard method using the calibration curve for Sigma-Aldrich, Switzerland). acetone extract after concentration and hydrolysis, modifiedthe from previous published method. wileyonlinelibrary.com/jsfa Quantitative results were calculated usingauthentic a calibration curve for and the sample was dissolvedevaporated in to hexane, 1 washed mL. with An aliquot waterin of and the a sample methyl (0.5 mL) tributyl was ethertemperature diluted (MTBE, overnight 25 (in mL) darkness) andhydroxide with saponified (30% methanolic at w/v, potassium room 1.25 mL).washed The three organic times phase with 50 was mL separated, sulfate, water, dried in filtered, anhydrous sodium andsample evaporated was to dissolved dryness.detection in (DAD) Finally, (Agilent 1 the mL 1100)identical dried analysis. acetone to The for the HPLC HPLC–diodedetection method method was (DAD used array at for 450by nm). an Quantitative external results standardβ were method obtained using the calibration curve of acid in aSamples volumetric were filtered flask and analysed (5 by mL),connected HPLC (Agilent to which 1100 a Series) was diodewas filled array performed detector with with (210 H water. nm). The isocratic run Oy). Calibration curves werestandards prepared (p.a., Merck with KGaA) authentic and citric malic acid acid (purity ion-exclusion column (6.5 mm Sigma-Aldrich, Buchs, Switzerland). (5 g, fw) by acetone (15 mL) in an ultrasound bath (4 Weather and climate Weather and climate conditions wereMeteorological obtained from Institute. the Finnish weather Statistics stations located were closestand collected to Sotkamo, the Oulu from samplingtemperatures province, sites two Finland). derived (Kuhmo Statistics as included arithmetic daily means of eight measured fibre, assay kit; Megazyme International Ireland Ltd, Bray, Ireland). darkness). Tocopherol was analysedcrude from acetone extract thefluorescence by detector subsample (FD), HPLC using an Eclipse (1 (Agilent XDB-C8 mL) (5 × 1100) of connected the to(1 a mL min extracted twice and three supernatants were combined.

1326 1327 30 13 08 75 33 04 5 ver- ...... ,fw). 1 − In addition to 41 0 11 0 11 0 20 0 30 0 30 ...... 27 48 0 06 0 61 0 27 0 26 0 62 ...... wileyonlinelibrary.com/jsfa (Table 2), which is comparable 1 − fruit size affects the anthocyanin Fruit size was not measured in this 07 0 04 0 98 0 25 0 68 0 30 28 ...... 29 Anthocyanin results are presented as 1 , respectively. The anthocyanin content varied 16 0 06 0 57 0 15 0 26 0 02 1 ...... − 26 0 08 0 51 0 43 0 26 0 20 ...... Sampling site 18–20 mg kg vitamin C and proteinsphenolics may by interfere the with Folin–Ciocalteuof the phenolic method. assay analysis However, were of not thecloudberry altered total results sample when (data sugar not was shown) added andC to the contents the in protein samples and were vitamin similar.environmental Further studies effects of climate on and however, cloudberry to confirm ellagitannins these conclusions. are needed, Anthocyanins Cloudberry anthocyaninscyanidine-3-rutinoside, cyanidin-3-sophoroside consist and3-(2-glucosylrutinoside). of cyanidine- cyanidine-3-glucoside, study. Vitamin C Ascorbic acid concentrations between the habitatsberry varied of from wild 0.64 cloud- to 1.0 g kg However, taking into accountdifference the between deviation habitats was within insignificant. habitats, the a sum ofglucoside all equivalents. detected Our anthocyaninsand 3) results calculated were closely as onsus related cyanidin-3- anthocyanins to earlier (Tables 2 published data: 5–27 between sampling sites inthe 2006 anthocyanin (Table content 2). has Extensivetween been variation reported different in also bilberry within populations and in be- Finland. environment and genotype, with the results of an earlier publication (0.50–1.50 g kg concentration in the plant. 02 0 06 0 12 0 49 0 23 0 41 ...... , 1 − L. www.soci.org 28 0 02 0 79 0 19 0 17 1 80 ...... 2011 Society of Chemical Industry c 3) grown in 10 different habitats in 2006 = (fw) and therefore n 1 24 20 0 18 0 41 0 44 0 01 0 11 ...... − Rubus chamaemorus 01). When citric acid content . 0 07 0 04 0 47 0 37 0 03 0 00 ...... The levels of analysed citric acid , fw). Malic acid dominated over < 1 respectively. In our study the total , respectively. 23 − 1 2 P , − 1 According to the literature the total − 25 )64566038414960575948 ) 2.26 2.51 2.83 2.70 3.00 2.50 2.70 2.38 2.48 2.95 111184 6 5 8 )161216118146 1 1 1 ) 0.79 0.68 0.90 1.48 0.90 0.90 1.00 2.19 2.43 0.94 : 1324–1330 − − − ) 0.69 0.82 0.68 0.94 0.64 0.64 0.87 0.74 0.98 1.01 1 ) 72697464718284637576 1 − 1 92 ) 173 168 164 166 168 181 178 154 175 169 − 1 − ) 6.01 7.81 6.51 5.50 6.56 9.31 7.34 7.27 9.48 7.46 ) 4.62 2.64 5.48 7.52 6.48 2.76 3.93 2.12 3.13 3.85 1 1 − ) 16171712171616101916 ) 6.9 9.9 6.9 6.3 4.8 4.4 5.3 5.4 8.1 8.4 − − 1 1 − − and 0.6 g kg 2012; Chemical composition of cloudberries ( (fw) in fruits. 126 − 1 300 cm) compared to the other sites. The availability of − > Ellagitannin content in cloudberry has been reported to be SD 5 2 3 10 4 6 11 4 3 5 SD 0 SD 2442212231 SD 0 SD 1323011110 SD 0 SD 16 23 3 13 7 8 9 24 15 20 SD 0 SD 0 SD 1 SD 25110235323 1 Crtn(gkg -Carotene(mg - Tocopherol (mg kg 3gkg Anthocyanins (mg kg Ascorbic acid (g kg α Citric acid (g kg Malic acid (g kg Dry weight (g kg Dietary fibre (g kg β nlts12345678910 Analytes Proteins (g kg Table 2. Phenolics (total) (g kg Total fat (g kg Chemical composition of ripe fruits of ∼ phenolic content in fruits was 2.2–3 g kg J Sci Food Agric Total phenolics The main phenolic compound group in cloudberryand is the ellagitannins content of anthocyanins is comparatively low (0.02 g kg of citric acid and malic acid ( was low, it was compensated bycontributing higher malic to acid concentration, similareach total growing site malic (8–12 and g kg citric acid contents in citric acid in allFurthermore, sampling the sites total except malic in andin citric sampling these acid sites sites contents 4 (4 were and and higher were 5) 5. the than only in sites other with open sites.layer populations Sampling and ( sites with 4 a and thicker peat 5 and malic acid contents in cloudberrypublished were data: comparable 4 and to 8 earlier g kg can be assumedPhenolicscontentdidnothavesignificanthabitat-relatedvariation to represent mainly(Table the 2): the ellagitannin variance within the content. habitatthe was higher habitats. than between Ellagitannins are notas as most widely studied other compounds accumulation plant-derived and variation polyphenolssuggest are and that biosynthesis not of therefore ellagitannins well couldcloudberry. their be known. On quite the stable These in other hand, results some compounds such as sugars, ellagic acid content in driedis leaves 70 and g kg fresh fruits of cloudberry fw). light was higher in the opencitric sites, acid which concentration could in explain the thoseaccumulatedrapidlyalongwithfruitgrowthandripening,whereas higher sites. In blackcurrant, citric acid malic acid content decreased. . 7 7 ), 1 − et al Rainfall 05) and . 0 42 : 1324–1330 < 92 P , fw) was very 1 − 2012; On the other hand, in 2007, and there were 2gkg 38 geographical variation, 1 ± 05) than berries collected − . 0 40,41 < -linolenic acids ( α P J Sci Food Agric 10)gkg ± ), whereas in summer 2007 a crop was C higher in July 2006 than in July 2007. 1 ◦ − Average water content of cloudberries was 830 37 in 2006 and 850 ( Rainfall in July 2007 was two times higher than in July 1 39 − 05)(Table 3).Itiswellknownthatlowtemperatureincreases . 9 0 ). 1 < 10)gkg − The mean amount of total lipids (7 Significant difference in the annual mean values of cloudberries P ± in the summer of 2007. Even moredaily striking, temperature the was average 3.5 maximum 2006; therefore water excess could possiblythe fruit increase and the induce stress anthocyanin of biosynthesis. Rainfall in July 2007conclusion, the was mean over temperature twofoldrainfall of higher was June than and higher insummer July 2007 and 2006. than was in the In lower, 2006. However, sunshineweather climatic stations data are duration collected only from indicative was of thearea, shorter climate owing in to the in probable sampling local differences in weather conditions. Crop yield Compared to summer 2006 the crop yields decreasedanalysed in sampling 2007 sites in except all in site 4,slightly where the (Table yield 1). increased However,low in also in this summer 2006. samplingyields Sampling in site sites 2007, but 9 the unfortunately and the yield 10 totalfrom had crop those was was high sites not crop in calculated 2006.summer In 2006 site (332 8, kg the ha crop yield was very high in and in July 2006temperatures the were number markedly of higher. warmother days However, parameters affecting and there the seed daily are oil composition maximum several in plants,as such maturation level, water deficiency, and interaction of latitude, elevation and temperature. Chemical composition Annual variation in the chemicalmeasured composition as of arithmetic cloudberry mean values was in of two all consecutive sampled years cloudberries (Tableand 3). secondary The metabolites contents were of surprisinglythe both equal. For primary mean example, results ofvery vitamin consistent C (Table analysed 3),of in especially other 2006 plants. when and compared 2007 to were studies no significant differences between habitats. Annual deviationinsignificant was but might be related to the higher rainfall in 2007. was found only in the content ofof anthocyanins. anthocyanins The was concentration higher in summer 2007( than in summer 2006 the anthocyanin content in plant cells. excess water has been reportedflavonoids. to cause several-fold increase in consistent in year-to-year comparisonwas a (Table difference in 3). the fatty However, acid composition between there thecollected samples in years 2006 andhigher 2007. proportion Berries of collected linoleic in and 2007 had a ( which might be explainedflowering by time. a A favourable comparison microclimatecrop during yields of indicates sampling that site yieldsmore equal description in in the and both open years, whereas sitesclearly in (4 lower shaded sites and in the 5) 2007 yields were than were to in the 2006. average crop Crop yield yieldsha reported were for comparable wild cloudberry (20–50 kg not obtained at all. Sampling10 in site 2007 8 had in exceptionally year high 2006 crop and yields (over sites 200 9 kg and ha in July 2007 was twice as high as in July 2006; therefore water a lower proportion of oleic acid ( in 2006 (Fig. 1).decrease Previous will decrease the studies degree of have saturation of shown oils in that the seed, temperature 31 33 05) . 0 2–3) C) than < www.soci.org M Jaakkola ◦ = although -carotene P n β Cultivated cloudberry 33 -carotene. 2011 Society of Chemical Industry Furthermore, Cucumis melo β c -carotene and 9) compared to ,fw, 31 -carotenefound α 1 -carotene varied = β − β n -tocopherol in plants Hippophae rhamnoides -tocopherol content in α α 32 and Wild C higher in 2006 (14.4 cloudberry , fw) between sampling sites 30) and 2007 ( ,fw). ◦ 1 1 -tocopherol concentrations with − = − α n ) 53 9 47 10 39 1 ) 10 4 27 13 14 9 -tocopherol content was lower ( ) 2.6 0.4 2.4 0.5 1.9 0.3 1 1 1 C was 12 in the summer of 2006 and only six − 30 mg kg α − ) 0.80 0.14 0.79 0.15 0.56 0.1 Theconcentrationlevelsof − ◦ 1 ∼ Brassica oleracea ) 170 4 150 8 150 18 − -carotene levels were found in berries collected 1 ) 7.3 1.3 7.0 1.1 7.2 0.3 ) 4.3 1.8 4.5 1.4 4.6 1.4 β 1 -carotene could occur due to hydrolysis of the 1 − )162142154 ) 7.0 1.0 6.0 1.7 8.0 1.7 − − 34–36 1 1 β − − C), and the number of warm days with a maximum ◦ Annual variation in the chemical composition of wild - Tocopherol (mg kg Carotene Total fat (g kg Dry weight (g kg Anthocyanins (mg kg α Ascorbic acid (g kg AnalyteTotal phenolics (g kg 2006 SD 2007 SD Mean SD Table 3. cloudberries collected in 2006 ( Proteins (g kg Citric acid (g kg Malic acid (g kg mean concentrations of analytes in cultivated cloudberries ( - Carotene is the predominant carotenoid in cloudberries, - - Tocopherol content in different sites was comparatively -tocopherol content determined in this study was slightly higher β Annual variation Climate and weather Climatic and weathercloudberries conditions as affect well the ascompounds. maturation the Sunshine biosynthesis rate duration of of in thein July plant’s 2007 was (189 bioactive h) significantly2007 than shorter the in sunshine 2006 durations were (280 comparableaverage h), (about June whereas 300 temperature h). in was The June 2 2006 and sample. Hydrolysis was, however, performed to obtain better HPLC chromatograms with less background noise. (seabuckthorn). wileyonlinelibrary.com/jsfa lutein) were notconsiderably analysed. Concentration (0.68–2.40 mg of kg and therefore other cloudberry carotenoids ( β In this study,those correlation of other antioxidants of was notanalysis detected. showed However, statistical that Vitamin E α constant (Table 2). The biosynthesis of is influenced by several different factors,harvest, and such geographical as and seed climatic conditions. maturity at in cloudberries grown in open habitats thanα in other habitats. The compared to earlier publishedFinnish cloudberry data ( of in this study are comparable to the earlier reported concentration in Finnish cloudberry: 1.40 mg kg there has been found toand be other an antioxidants interaction such between tocopherols as ascorbic acid and temperature over 25 some losses of (Table 2). Highest from sites 8 and 9.crop Interestingly, these yields. sites also High gavereported the to variation highest be found in also in the(muskmelon) other plants, fruit, carotene such as in content has been in 2007 (12.3

1328 1329 and :6178–6187 Fragaria 52 -carotene has β Significant correlations ¨ onen AR,Identificationand ¨ orr ¨ ainen for help in manuscript wileyonlinelibrary.com/jsfa but it has not been possible 7,38,39 J Agric Food Chem -tocopherols detected could be 34–36 α species (family Rosaceae). ¨ a-Riihinen KR,Kamal-Eldin AandT ¨ att -carotene varied significantly between habitats, whereas quantification of phenolic compounds in berries of Rubus (2004). ¨ a β The crop yields in the open sites were more equal in 2006 Taken together, this study suggests that the amount of sunlight Results for cultivated cloudberries suggest that the composition 1M and the concentrations of vitamin C werebetween surprisingly consistent different even years.been Extensive reported variation in other of studies, REFERENCES ACKNOWLEDGEMENTS The authors would likePatrikainen, to Ms thank Riitta Mr Lotvonen,Karppinen for excellent Mauri Ms technical assistance. We Heikkinen, Kirsi are also grateful Ms KuvajaDr to Adama Tiina M and Sesay and Ms Dr Pekka Kilpel Sanna preparation. This work was funded byfinanced by the the Interreg EU. III programme to explain these observations by any specific characteristics related to habitat. However, this wouldstudies. be an intriguing topic for further and 2007 thanhabitats in were higher other in 2006total sites. than in yields The 2007 of and crop the thereforecarotene, also tocopherol, studied yields total the bioactive phenolics) in were compounds higher2007. the in (anthocyanins, 2006 The than concentrations shaded in of primary metabolitesand were between consistent habitat andexcept annual in differences the were saturationmetabolites insignificant, level of had fatty more acids,differences were whereas variation. found secondary The inin the most the saturation concentrations level significant of of anthocyanins.were annual fatty The acids mainly observed and differences explainedthe by summers variation ofpreviously in 2006 weather reported and conditions effectsthese parameters/metabolite in 2007 levels. of and sunlight they and agreed temperature with on the related to the lowerenvironment, stress resulting of plants in inmetabolites decreased the protecting biosynthesis isolated plant of greenhouse againstclones secondary stress. of However, cultivated onlyvariation two varieties could likely were result inwild analysed, larger populations. variations and between different the genetic and temperature levels couldcrop be yield the and main chemicalthe factors composition affecting of amount the cloudberry. In ofconcentrations. addition, rainfall To maythe evaluate have environment more an andcloudberries, accurately climate effect a the effects more onas impact amount detailed on anthocyanin of undergrowth, analysis of the light level ofneeded. and composition temperature Nevertheless, sampling would of be this siteson work such provides the valuablecloudberries information mean and their annual concentrations and habitat-related variation. of nutritional components in between primary and secondarythis metabolites study, although were the secondary not metabolites are found biosynthesized from in primary metabolites. and nutritional value of cultivated cloudberriesto wild were cloudberries. comparable The slightly lower amounts(ellagitannins), of total vitamin phenolics C and and L. www.soci.org 43 2011 Society of Chemical Industry -tocopherol, α c Rubus chamaemorus -tocopherol was lower in the open α : 1324–1330 9). 92 = n 2012; Composition (%) of fatty acids in cloudberries collected in 2006 30) and 2007 ( The same profile in malic acid and citric acid concentrations = -carotene, vitamin C and anthocyanins in cultivated fruits were n Chemical composition of ripe fruits of J Sci Food Agric Studied growing sites differed in the levelof of tree undergrowth, stand and which height affectsuch the as growing conditions availability ofgrown of plants, in sunlight habitats and havingdiffered temperature. no from Cloudberries tree otherdifferencesincloudberriesgrownintheareaswithlowtreestandor stand studied and low cloudberrydense undergrowth forest populations, were not whereas found. However,one all shaded (site habitats except 6)that had the undergrowth high could contribute undergrowth notablyhabitats to and shadiness. having Two the no results tree standmore may and easily imply thicker altered peat by layer climaticopen of and sites soil weather had were changes. different Thosethan concentrations two other of sites: malic the andacid total citric dominated content acids over of malic acids acidthe was in higher higher open and sites. temperature This citric andFurthermore, can the amount be amount due of of to lightsites in than in the shaded open sites.characteristics sites. Other correlations on of the the sampling levelsfound site in of this study. bioactive However, the compounds concentrations of were anthocyanins not CONCLUSIONS was observed in 2006(4 and and 2007 5) (results differed notgrowing from shown): site other open clearly sites sites affectedThis the (Table 2). is measured This interesting acid indicates concentrations. sinceaffect the that several amount of factors, acids in especiallytotal the organic berries. temperature, acids Generally, decreases the during amount the of ripening of berries, excess rather than deficiency andresulted lower in the temperature decrease could in have saturationon of the fatty acids. effect Earlier of studies excessfrom water the literature. on oil composition were not found Figure 1. ( Cultivated cloudberry Protein, water, total fat, malic and citriccloudberry acid contents were in cultivated comparableConversely, with the concentrations wild of cloudberries total phenolics, (Table 3). warm climate has been found to result in lower acidin concentrations apple and grapes. 75–80% of the concentrationsThis measured might from be wild due to cloudberries. the isolatedfruits growing conditions of and/or cultivated geneticcloudberries. differences between wild and cultivated β . L.) L.) Acta et al Rubus J Berry J Agric L. Cuphea :117–122 :160–167 17 13 : 1324–1330 Phytochemistry 92 :100–107 (2004). Acta Agric Scand B Can J Biochem Cell ¨ onen AR, Contents 10 Prunus avium ¨ 2012; orr :541–549 (1994). Vaccinium myrtillus cultigens and seasons. FoodResInt Rubus chamaemorus :31–57 (2002). 194 :891–899 (2003). :279–287 (2008). 21 :67–80 (2003). 48 -carotene, and chlorophyll 27 16 β :108–111 (1992). 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(1997). active metabolites: a review. from 44 wild, edible Finnish berries. and Katajisto JK, ¨ orr 2T 3 Halvorsen BL, Holte K, Myhrstad MC, Barikmo I, Hvattum E, 4 Puupponen-Pimi 6 Saarivirta M and Kreula M, The contents of water-insoluble dietary 5 Puupponen-Pimi 9ThiemB, 8 Johansson A, Laakso P and Kallio H, Characterization of seed oils of 7 Johansson AK, Kuusisto PH, Laakso PH, Derome KK, Sepponen PJ 22 AOAC, Total, soluble, and insoluble dietary fiber in foods, in 20 Maeda Y, Ochi S, Masui T and Matubara S,21 Liquid-chromatographic Zorn H, Breithaupt DE, Takenberg M, Schwack W and Berger RG, 19 Lee HS and Coates GA, Liquid-chromatographic determination of 18 Wrolstad RE, Acree TE, Decker EA, Penner MH, Reid DS, Schwartz SJ, 17 Pap N, Pongr 16 Crude protein–Kjeldahl method, boric acid modification, in 11 Nilsen G, Role of plant hormones in parthenocarpic development of 12 Hoppula K, Kajalo M and Pirinen H, The price13 of Rapp K the and Martinussen I, cultivated Breeding cloudberry ( 15 AOAC, Loss on drying (moisture) at 95 10 Ågren J, Seed size and number in 14 Rapp K, Selection and variety development for increased yield in wileyonlinelibrary.com/jsfa

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