Research Article A Study on the Quality Criteria of Some Mandarin …downloads.hindawi.com/archive/2014/982721.pdf · 2019. 7. 31. · Research Article A Study on the Quality Criteria

Research ArticleA Study on the Quality Criteria of Some Mandarin Varieties andTheir Suitability for Juice Processing

Erdal ALccedilam and Asiye AkyJldJz

Department of Food Engineering Cukurova University 01330 Adana Turkey

Correspondence should be addressed to Asiye Akyıldız asiye1cuedutr

Received 15 May 2014 Revised 10 August 2014 Accepted 11 August 2014 Published 26 August 2014

Academic Editor Soichiro Nakamura

Copyright copy 2014 E Agcam and A Akyıldız This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

In this study some composition properties of juices of different mandarin varieties (Robinson (R) Fremont (F) and Satsuma (S))were determined before and after pasteurisation 119871lowast 119886lowast 119887lowast and 119862lowast values of all varieties were increased after the pasteurisationprocess Degradation of ascorbic acid was calculated as 220 1686 and 2431 for R F and S samples respectively afterpasteurisationThe highest total carotenoid and phenolic contents were determined in S samples In general after the pasteurisationtreatment the total carotenoid content of juices was increased slightly but total phenolic contents were dramatically decreasedThe antioxidant activity of pasteurised samples was increased by approximately 6 The most abundant carotenoid and flavanoneglycoside compound was shown to be 120573-cryptoxanthin and hesperidin respectively in all samples The most popular fresh andpasteurised juice samples were made from the Robinson variety of mandarin with regard to taste smell and general impression

1 Introduction

Citrus has long been one of the largest world crops and itsjuice is one of the most appreciated by consumers One ofthe reasons for this is the high quality of their nutritionalcomposition Turkey is one of the major citrus-producingcountries of the northern Mediterranean region with asuitable climate and other ecological conditions and was thefifth largest world producer ofmandarin (872251metric tons)in 2011 [1]

Citrus fruits and their derived products have a beneficialeffect on human health Citrus fruits are attractive becauseof their nutritional and antioxidant properties [2ndash5] theprevention of diseases by nutrition is strongly supported [6]mainly due to the contribution of antioxidant compoundsincluding vitamin C phenolic compounds and carotenoids[5] Dhuique-Mayer et al [3] compared the influence ofvariety and species on the content of the main antioxidantmicroconstituents (flavonoids carotenoids and vitaminC) ofdifferent citrus fruits cultivated in the Mediterranean region

The production of single-strength citrus juice is increas-ingThe composition of fresh citrus juice is adversely affectedby industrial processing andor storage conditions Industrial

processing involves a number of different stages that result insome alterations from the original composition of fresh citrusjuice [7] Pasteurisation is essential to improve the shelf-life and safety of fruit juice Pectin methyl esterase (PME)is an enzyme of major impact in citrus juice If PME isnot inactivated enough this enzyme is caused losses in thenatural cloud quality of citrus juices PME is more thermallyresistant than vegetative spoilage microorganisms thereforethe target of pasteurisation of citrus juice is PME [8] Thedelicate fresh composition of citrus juices is easily changed bypasteurisation as the juice undergoes various compositionalchanges Pasteurisation has negative impacts on the quality ofmandarin juice such as the loss of colour flavour nutritionalvalue and as a result taste [9]

Some investigators [10ndash13] have monitored a gradualdecrease in certain aromas and an increase in undesirablecomponents during processing and storage However limitedreferences have been found for the effects of pasteurisationon the chemical composition of different mandarin cultivarjuices In order for processors to better understand thechanges that take place during the thermal processing ofmandarin juices quantitative information on the compo-nents present in both fresh and pasteurised mandarin juice is

Hindawi Publishing CorporationJournal of Food ProcessingVolume 2014 Article ID 982721 8 pageshttpdxdoiorg1011552014982721

2 Journal of Food Processing

needed Such information will provide a list of varieties thatare more appropriate for the production of juice

The main objective of this study was to compare thechemical composition of three mandarin cultivars (Robin-son Fremont and Satsuma) that are currently grown inTurkey and to determine the chemical changes that occur asa result of pasteurisation

2 Materials and Methods

21 Juice Processing Robinson (R) (Citrus reticulata hybridof Clementine times Orlando) Fremont (F) (Citrus reticulatahybrid of Clementine times Ponkan) and Satsuma (S) (Citrusunshiu) mandarin varieties grown in the Cukurova Regionof Turkey were used in this study After being washed man-darins were cut into two halves and pressed using a bench-scale automatic squeezing machine (CANCAN Turkey)Mandarin juices were passed through 1mm stainless steelsieves to remove seeds and coarse pulp Juicewas immediatelyprocessed by heat pasteurisation The study was carried outwith three replications

22 Heat Pasteurisation The bench-scale system designed inthe Department of Food Engineering of Cukurova University(Adana Turkey) was used for heat pasteurisation applications[14] As a result of the preliminary experiments heat wasapplied at 95∘C for 15 s for pasteurisation Heat-pasteurisedsamples were placed in amber coloured bottles for furtheranalyses Pasteurised samples were called Robinson (RP)Fremont (FP) and Satsuma (SP) in future analyses

23 General Properties of Mandarin Juices Total acidity anddry matter of juice samples were carried out according toAOAC [15] Brix and pH of mandarin juices were analysedwith an Abbe refractometer (Carl Zeiss Jena Germany) andWTW pH-meter (Weilheim Germany) respectively

24 Colour Measurement Colour (CIE 119871lowast 119886lowast 119887lowast) analysiswas conducted by the Colour Flex HunterLab instrument50mL of juice was transferred to the 20mm Glass OpticalCell Light Path and analysed The results were given accord-ing to the CIELAB colour system In this system 119871lowast defineslightness (0 black 100 white) 119886lowast denotes the redgreen value((+) red (minus) green) and 119887lowast denotes the yellowblue value((+) yellow (minus) blue) In addition the following formulaswere used for the calculations of Huelowast 119862lowast and Δ119864lowast colourvalues

Huelowast = arctan(119887lowast

119886lowast)

119862lowast

= radic(119886lowast

)2

+ (119887lowast

)2

Δ119864lowast

= radic(Δ119871lowast)2

+ (Δ119886lowast)2

+ (Δ119887lowast)2

(1)

25 Determination of Browning Index 5mL of each varietyof mandarin juice was mixed with 5mL ethyl alcohol (95)in teflon tubes and then centrifuged (4000 rpm 10min at

4∘C) The supernatant was passed through a 045 120583m teflonmembrane filter and the absorbance of the supernatant wasobtained at 420 nm in a spectrophotometer (Perkin ElmerLambda 25-UVVIS USA) [16]

26 Determination of Hydroxymethylfurfural (HMF) HMFextractions of samples were carried out according to themethod reported by Gokmen and Acar [17] The bestchromatographic conditions were determined as a result ofpreliminary experiments as follows 20120583L of supernatantwas injected into the C18 ACE (46 times 250mm) columnthe column was maintained at 30∘C with a flow rate of05mLmin and the photodiode array detector was set at285 nmMethanolwateracetic acid (20791 vvv) was usedas mobile phases

27 Antioxidant Activity The antioxidant activity of themandarin juices was evaluated using the DPPHlowast free radical-scavenging method The DPPHlowast free radical-scavengingactivity measurements were carried out according to the pro-cedure of Klimczak et al [18] with some modifications 5mLof mandarin juices was mixed with 5mL of methyl alcohol(80) in teflon tubes and then centrifuged (4000 rpm 10minat 4∘C) Briefly 01mL of supernatant was added to 246mLof11-diphenyl-2-picrylhydrazyl radical (DPPHlowast 0025 gLminus1 in80 methyl alcohol) and mixed by vortex After incubationfor 10min in the dark the absorbance of the sampleswas measured at 515 nm using the spectrophotometer Theantioxidant activity was expressed as the percentage declineof the absorbance as follows

Antioxidant activity () = (119860control minus 119860 sample

119860control) times 100

(2)

where 119860control is the absorbance of the control and 119860 sample isthe absorbance of the sample

28 Determination of PME Activity For the measurementof PME activity 10mL of mandarin juice was mixed with20mL of 1 pectin-salt substrate (01M NaCl) and incu-bated at 30∘C The solution was adjusted to pH 70 with20NNaOH and the pH of the solution was readjusted to77 with 005NNaOH After the pH reached 77 010mL of005NNaOH was added Time was measured (1199051015840) until thepH returned to 77 PME activity () was calculated as follows[19]

PME Activity (119860) =(005NNaOH ) (010mLNaOH)(1199051015840) (10mLsample)

Residual PME Activity () = (119860119905

1198600

) 100

(3)

where 1199051015840 is time in min 1198600is initial PME activity and 119860

119905is

PME activity after pasteurisation

29 Determination of Total Carotenoid Content Totalcarotenoid determination was carried out according to the

Journal of Food Processing 3

previously described method of Lee and Castle [20] withsome modifications 5mL of juice and 10mL of hexanesolution (hexanemethanolacetone 502525 vv with01 BHT) were mixed and then centrifuged for 10min4000 rpm at 4∘CThe supernatant phase was used to measurethe absorbance (450 nm) using a spectrophotometer Totalcarotenoids were calculated using the extinction coefficientof 120573-carotene (11986412 = 2505)

210 Determination of Carotenoid Compounds Pigmentextraction from juices and saponification procedures werecarried out according to the previously reported method ofMelendez-Martınez et al [21]

HPLC analyses were carried out by means of a Shi-madzu LC-20AT (Japan) system consisting of a quaternarypump a column temperature control oven (CTO-10AS)an autosampler unit (SIL-20A) a degasser module (DGU-20A5) and a photodiode array detector (SPD-M20A) 50 120583L

of supernatant was injected into the C30 ProntoSIL (50 120583m46 times 250mm) columnThe column was kept at 20∘C and theflow rate was 1mLmin The photodiode array detector wasset to 450 nm Methanol (A) methyl-tert-butyl ether (B) andultrapure water (C) were used as mobile phases Accordingto the preliminary experiments the best gradient elution wasas follows 0min 90 A + 5 B + 5 C 5min 95 A + 5B 40min 75A + 25B 55min 55A + 45B 60min90A + 5B + 5C 65min 90A + 5B + 5C

The carotenoid compounds were identified by comparingtheir UV-visible spectra and retention times with that ofcorresponding standards Quantification of carotenoid com-pounds was carried out at 450 nm using external standardmethod

211 Determination of Ascorbic Acid Content Ascorbic aciddetermination was performed using the HPLC method andextraction procedure according to Lee and Coates [22] TheHPLC column was maintained at 25∘C and the flow ratewas 05mLmin 10 120583L of supernatant was injected into theC18 XTerra (Waters 46 times 250mm) columnThe photodiodearray detector was set at 244 nm and 2 KH

2PO4(pH 24)

was used as the mobile phase

212 Determination of Total Phenolic Content The totalphenolic compounds in the study samples were measuredusing the Folin-Ciocalteu method with some modifications[23] For the measurement of phenolic compounds 5mLof mandarin juice was mixed with 5mL of 80 methanolin teflon tubes and the tubes were then centrifuged at4000 rpm for 20 minutes at 4∘C (Heraeus Bofuge Primo RGermany) For the analysis 100 120583L of appropriately dilutedsample (various concentrations) or standard solution wasmixed with 100 120583L Folin-Ciocalteu reagent and 3000 120583Ldeionised water and mixed thoroughly After incubation for10min at room temperature 100 120583L of 20 Na

2CO3solution

was added followed by immediate mixing the solutionwas further incubated at room temperature for 2 h in thedark The absorbance of the mixture was then measured at765 nm using a spectrophotometer (Perkin Elmer Lambda

25-UVVIS USA) Gallic acid was used as the standard andthe total phenolic compounds of the samples were expressedin milligrams per L of gallic acid equivalents (mgGAEL)

213 Determination of Phenolic Compounds For the mea-surement of phenolic compounds 5mL of mandarin juicewas mixed with 10mL of 80 methanol in the teflon tubesand sonicated (Bandelin Sonerex Germany) at room temper-ature (sim25∘C) for 15 minutes Then tubes were centrifugedat 4000 rpm for 10 minutes at 4∘C (Heraeus Bofuge PrimoR Germany) Finally the supernatant was passed through a045 120583m teflon membrane filter and injected into the HPLCinstrument [14]

HPLC analyses were carried out by means of a Shi-madzu LC-20AT (Japan) system consisting of a quaternarypump a column temperature control oven (CTO-10AS)an autosampler unit (SIL-20A) a degasser module (DGU-20A5) and a photodiode array detector (SPD-M20A) 20120583L

of supernatantwas injected into theC18XTerra (Waters 46 times250mm) column The column was kept at 30∘C and the flowrate was 08mLmin The photodiode array detector was setto 280 and 320 nm 2 formic acid (A) and 100 ACN (B)were used as mobile phases According to the preliminaryexperiments the best gradient elution was as follows 0min100A 10min 95A + 5B 25min 90A + 10 55min80A + 20B 70min 55A + 45B 90min 100B95min 100A

The phenolic compounds were identified by comparingtheir UVndashvisible spectra and retention times with that ofcorresponding standards Quantification of phenolic com-pounds was carried out at 280 and 320 nm using externalstandardmethod Calibration curves were obtained using thecommercial standards of the concentrations normally presentin mandarin obtaining regression coefficients (1198772) above0997 in all cases

214 Sensory Analysis Juices were evaluated using a graphicscale testThe panel was composed of eighteen assessors fromour department Five sensory parameters (colour turbiditytaste smell and general impression) were established formandarin juices each of which was given a score out of ten

215 Statistical Analysis The software SPSS 20 for Windows(SPSS Inc Chicago IL USA) was used for analysis ofvariance (ANOVA) and Duncanrsquos multiple comparison testin order to determine significant differences between thetreatments Each experiment was repeated at least threetimes

3 Results and Discussion

31 General Properties of Mandarin Juices The general prop-erties of the mandarin juices and the effects of pasteurisationon these properties are shown in Table 1 Brix of mandarinjuices changed between 1247 plusmn 006ndash1360 plusmn 026 with thebiggest value found for the F sample Total drymatter of freshand pasteurised mandarin juices ranged between 1295 plusmn037ndash1416 plusmn 001 and there was no difference between


Table 1 General properties and PME activities of mandarin juices

R RP F FP S SPBrix∘ 1265 plusmn 013c 1247 plusmn 006c 1360 plusmn 026a 1350 plusmn 025a 1303 plusmn 012b 1252 plusmn 003c

Total dry matter () 1295 plusmn 037b 1325 plusmn 006b 1386 plusmn 028a 1416 plusmn 001a 1308 plusmn 015b 1297 plusmn 001b

pH 319 plusmn 001bc 322 plusmn 005b 317 plusmn 001c 327 plusmn 001a 312 plusmn 001d 318 plusmn 001c

Total acidity (g100mL) 104 plusmn 001de 102 plusmn 001e 111 plusmn 002c 107 plusmn 001cd 153 plusmn 004a 146 plusmn 003b

PME activity () 100a 559 plusmn 028c 100a 1387 plusmn 069b 100a 1426 plusmn 060b

Yield () 3667 plusmn 025b mdash 4348 plusmn 029a mdash 3575 plusmn 014b mdashSuperscript lowercase letter show the significant differences with each row (119875 lt 005) plusmn standard deviationR and RP fresh and pasteurized juice of Robinson respectively F and FP fresh and pasteurized juice of Fremont respectively S and SP fresh and pasteurizedjuice of Satsuma respectively

Table 2 Color properties and HMF concentration of mandarin juices

R RP F FP S SP119871lowast 3043 plusmn 148b 3308 plusmn 025a 2843 plusmn 047c 3159 plusmn 022b 3148 plusmn 034b 3346 plusmn 017a

119886lowast

minus173 plusmn 033e minus111 plusmn 009d minus054 plusmn 009c minus102 plusmn 004d 359 plusmn 028b 483 plusmn 011a

119887lowast 1665 plusmn 114bc 1680 plusmn 022bc 1617 plusmn 077c 1745 plusmn 021b 1892 plusmn 024a 1981 plusmn 021a

Huelowast 9600 plusmn 157a 9379 plusmn 027b 9194 plusmn 039c 9334 plusmn 010b 7926 plusmn 069d 7631 plusmn 039e

119862lowast 1675 plusmn 110cd 1684 plusmn 022cd 1618 plusmn 077d 1748 plusmn 021c 1926 plusmn 028b 2039 plusmn 019a

Δ119864lowast 284 plusmn 131a 353 plusmn 020a 250 plusmn 029a

Browning index (abs)lowastlowast 016 plusmn 000bc 014 plusmn 001c 016 plusmn 001bc 014 plusmn 003c 022 plusmn 000a 018 plusmn 001b

HMF (ppb) nd 713 plusmn 027b nd 462 plusmn 084c nd 1069 plusmn 046alowast

Superscript lowercase letter show the significant differences with each row (119875 lt 001) plusmn standard deviation lowastlowastAbsorbanceR and RP fresh and pasteurized juice of Robinson respectively F and FP fresh and pasteurized juice of Fremont respectively S and SP fresh and pasteurizedjuice of Satsuma respectively

fresh and pasteurised samples (119875 gt 005) The pH and totalacidity of juice samples were between 312 plusmn 001ndash327 plusmn 001and 102 plusmn 001ndash153 plusmn 004 g100mL respectively After thepasteurisation process the acidity of juices was found to besignificantly decreased (119875 lt 005) Finally the highest yieldwhich is a very important factor for citrus juicing industrieswas calculated for the Fremont (4348 plusmn 029) variety

32 Colour Properties and HMF Content of Mandarin JuicesColour values of fresh and pasteurised juices are given inTable 2 119871lowast values were determined to be 3043 2843 and3148 for R F and S respectively 119871lowast values of pasteurisedjuices were increased after the pasteurisation process (119875 lt005) and found to be 3308 3159 and 3346 for RP FP andSP respectively The 119886lowast and 119887lowast values of mandarin juicesranged betweenminus173ndash483 and 1617ndash1981 respectively Afterthe pasteurisation process 119886lowast and 119887lowast values of all varietieswere increased as was the colour value of 119871lowast (119875 lt 005)

Huelowast 119862lowast and Δ119864lowast colour values were derived using thevalues of basic CIELAB colour parameter (119871lowast 119886lowast 119887lowast) Thefirst of these Huelowast values is described with basic colours(red yellow green and blue) being closer to the colourof the object The 119862lowast value defines the saturation of thecolour object The point of intersection with Huelowast and 119862lowastvalues is known as the colour of the object (Figure 1) FinallyΔ119864lowast value is given for the total colour difference after any

operationHuelowast values were calculated as 9600 plusmn 157 9379 plusmn 0279194 plusmn 039 9334 plusmn 010 7926 plusmn 069 and 7631 plusmn 039

Yellow

Green

Blue

Red

R F S

minusalowast +blowast

minusalowast minusblowast

+alowast +blowast

+alowast minusblowast

Croma lowast

Huelowast

minusalowast +a

lowast

minusblowast

+blowast

Figure 1 Graph of color according to CIELAB scale (pasteurizedjuice of Robinson (R) Fremont (F) and Satsuma (S))

for R RP F FP S and SP samples respectively Accordingto the results after pasteurisation the Huelowast value of Fremontwas increased but the Huelowast values of Robinson and Satsumawere decreased (119875 lt 005) 119862lowast values ranged between1618 plusmn 077ndash2039 plusmn 019 for mandarin juices and werehigher for S than for R and F samples In addition this valuewas slightly increased after pasteurisation treatment for all


Table 3 Bioactive compound concentrations and antioxidant activities of mandarin juices

R RP F FP S SPAscorbic Acid (mgL) 54682 plusmn 530a 53481 plusmn 789a 36272 plusmn 1302b 30155 plusmn 4545c 28225 plusmn 2641c 21363 plusmn 892d

Total carotenoid (mgL) 1720 plusmn 041e 1736 plusmn 084e 2006 plusmn 090d 2243 plusmn 095c 3032 plusmn 176b 3311 plusmn 174a

Total phenolic (mgL) 31665 plusmn 1398ab 29110 plusmn 2036b 29843 plusmn 2602b 23760 plusmn 1451c 33623 plusmn 839a 26017 plusmn 1106c

Antioxidant activity () 7769 plusmn 073c 8424 plusmn 053a 7825 plusmn 040bc 8419 plusmn 082a 7911 plusmn 059b 8513 plusmn 038a

Carotenoid compounds (mgL)120573-cryptoxanthin 629 plusmn 121c 665 plusmn 088c 899 plusmn 078b 972 plusmn 173b 1938 plusmn 100a 2104 plusmn 140a

120572-caroten 005 plusmn 002b 005 plusmn 001b 016 plusmn 004a 016 plusmn 001a 013 plusmn 003a 014 plusmn 001a

120573-caroten 035 plusmn 005c 035 plusmn 006c 062 plusmn 011ab 072 plusmn 015a 048 plusmn 007bc 052 plusmn 007bc

lutein 019 plusmn 005c 022 plusmn 005bc 045 plusmn 003b 040 plusmn 002bc 037 plusmn 012bc 082 plusmn 028a

zeaxanthin 029 plusmn 012bc 033 plusmn 006bc 022 plusmn 001c 028 plusmn 003bc 047 plusmn 014b 087 plusmn 023a

Phenolic compounds (mgL)gallic acid 173 plusmn 014a 184 plusmn 014a 009 plusmn 001c 047 plusmn 060cb 075 plusmn 004b 092 plusmn 021b

vanillic acid 211 plusmn 016c 180 plusmn 003c 312 plusmn 025ab 322 plusmn 014ab 297 plusmn 064b 374 plusmn 053a

chlorojenic acid 326 plusmn 013c 331 plusmn 005c 532 plusmn 034b 479 plusmn 029b 789 plusmn 143a 772 plusmn 048a

caffeic acid 113 plusmn 016b 104 plusmn 001b 149 plusmn 011b 156 plusmn 002b 443 plusmn 076a 427 plusmn 026a

PAEElowastlowast 418 plusmn 039c 432 plusmn 007bc 472 plusmn 048b 658 plusmn 028a 026 plusmn 003d 042 plusmn 006d

p-cumaric acid 069 plusmn 053ab 021 plusmn 008b 070 plusmn 040ab 023 plusmn 024b 129 plusmn 039a 066 plusmn 004b

syringic acid 124 plusmn 085bc 014 plusmn 001c 269 plusmn 019ab 171 plusmn 121ab 288 plusmn 050a 178 plusmn 126ab

sinapic acid 012 plusmn 008c 000 plusmn 000d 029 plusmn 002b 037 plusmn 003a 014 plusmn 003c 010 plusmn 002c

o-cumaric acid 034 plusmn 006b 028 plusmn 005ab 018 plusmn 005b 024 plusmn 002ab 063 plusmn 014a 055 plusmn 007a

ferrulic acid nd nd ndhesperidin 20845 plusmn 435b 12648 plusmn 611c 22623 plusmn 2086b 14978 plusmn 329c 28295 plusmn 2893a 21773 plusmn 597b

neohesperidin 040 plusmn 014ab 000 plusmn 000b 036 plusmn 006ab 159 plusmn 173a 045 plusmn 009ab 176 plusmn 094a

eriocitrin nd nd ndneoeriocitrin nd nd ndnaringenin 058 plusmn 003a 043 plusmn 010b 061 plusmn 002a 060 plusmn 000a 064 plusmn 003a 064 plusmn 002a

naringin nd nd ndquercetin nd nd ndisoquercetin 073 plusmn 007b 069 plusmn 000b 087 plusmn 04a 091 plusmn 002a 067 plusmn 002b 067 plusmn 001b

rutin nd 357 plusmn 256b 564 plusmn 026a ndluteolin 010 plusmn 002b 013 plusmn 007b 018 plusmn 004b 012 plusmn 004b 038 plusmn 008a 038 plusmn 003a

kamferol 242 plusmn 046a 290 plusmn 117a 022 plusmn 000b 040 plusmn 001b 064 plusmn 042b 016 plusmn 003b

apijenin nd nd ndSuperscript lowercase letter show the significant differences with each row (119875 lt 005) plusmn Standard deviation lowastlowastProtocatechuic acid ethyl esterR and RP fresh and pasteurized juice of Robinson respectively F and FP fresh and pasteurized juice of Fremont respectively S and SP fresh and pasteurizedjuice of Satsuma respectively

varieties (119875 lt 005) Δ119864lowast values of samples ranged between250 plusmn 029ndash353 plusmn 020 after the pasteurisation treatmentand the highest value was calculated for the Fremont variety(119875 gt 005)

Browning indexes of samples were determined as anabsorbance between 014 plusmn 001ndash022 plusmn 000 and weredecreased in pasteurised juices of all varieties (119875 lt 005)HMF was not detected in fresh mandarin juices but HMFconcentrations of RP FP and SP were found to be 713 plusmn027 462 plusmn 084 and 1069 plusmn 046 ppb respectively afterpasteurisation treatment

33 Bioactive Compounds and Antioxidant Activities ofMandarin Juices Bioactive compound concentrations and

antioxidant activities of mandarin juices are given in Table 3Ascorbic acid concentrations of mandarin juices rangedbetween 21363 plusmn 892ndash54682 plusmn 530mgL with the highestconcentration observed in R samples (119875 lt 005) Afterpasteurisation ascorbic acid degradation was calculated as220 1686 and 2431 for RP FP and SP samples respec-tively Total carotenoid and phenolic contents of juices rangedbetween 1720plusmn041ndash3311plusmn174 and 23760plusmn1451ndash33623plusmn839mgL respectively The highest total carotenoid andphenolic contents were determined in S samples (119875 lt005) In general after the pasteurisation treatment the totalcarotenoid content of juices was increased slightly but thetotal phenolic contents decreased dramatically Hayat et al[24] stated that the total phenolic acid content in mandarinpomace was decreased with increasing microwave power


Table 4 Sensory analyses results of mandarin juices

R RP F FP S SPColor 867 plusmn 060a 874 plusmn 073a 892 plusmn 056a 890 plusmn 044a 875 plusmn 054a 896 plusmn 033a

Turbidity 839 plusmn 064a 891 plusmn 033a 895 plusmn 071a 880 plusmn 036a 862 plusmn 045a 847 plusmn 057a

Taste 831 plusmn 060ab 737 plusmn 027b 853 plusmn 031a 553 plusmn 073c 809 plusmn 026ab 547 plusmn 070c

Smell 855 plusmn 050a 595 plusmn 051b 867 plusmn 019a 538 plusmn 078b 873 plusmn 029a 572 plusmn 012b

General impression 899 plusmn 028a 778 plusmn 056b 859 plusmn 057a 658 plusmn 039c 834 plusmn 024ab 528 plusmn 041c

Superscript lowercase letter show the significant differences with each row (119875 lt 005) plusmn standard deviationR and RP fresh and pasteurized juice of Robinson respectively F and FP fresh and pasteurized juice of Fremont respectively S and SP fresh and pasteurizedjuice of Satsuma respectively

and treatment time Xu et al [25] reported that the freefraction of phenolic increased whereas the ester glycosideand ester-bound fractions decreased after heat treatmentMoreover there was a decrease of total phenolic acid contentafter heat treatment and the content of four flavanone gly-cosides (narirutin naringin hesperidin and neohesperidin)declined with heating time and temperature

According to the results of mandarin juices it was foundthat these juices contain a high concentration of bioactivecompounds (ascorbic acid phenolic and carotenoid) andantioxidant activities were determined to be high (7825 plusmn040ndash8513 plusmn 038) as expected Data show that pasteurisa-tion treatment causes an increase in antioxidant activity withan increase of approximately 6 (119875 lt 005)

Xu et al [25] assumed that many antioxidant phe-nolic compounds in plants are usually presented as thecovalently-bound form therefore some processing methodswere employed to liberate them in order to enhance theirantioxidant capacity Jeong et al [26] reported that heattreatment may liberate some low molecular weight phenoliccompounds and increase the antioxidant capacity of citruspeel as a result Hayat et al [24] expressed that aftermicrowave treatment the free fraction of phenolic acidsincreased whereas the bound fractions decreased and antiox-idant activity was increased The increase in the antioxidantcapacities of citrus pomaces was due at least in part to theincreased phenolic content in the free fraction Free phenoliccompounds have been shown to have greater antioxidanteffects than bound forms [27]

Carotenoid Compounds of Mandarin Juices Carotenoid com-pounds of different mandarin juice samples are given inTable 3Themost abundant carotenoid compoundwas foundto be 120573-cryptoxanthin in all samples this compound washigher than others in the Satsuma samples (119875 lt 005) Afterpasteurisation treatments differences between samples werenot found to be important for 120573-cryptoxanthin 120572-caroteneand 120573-carotene but were important for zeaxanthin and lutein(119875 lt 005)

Dhuique-Mayer et al [28] reported that the main provi-tamin A carotenoids were not significantly affected duringconventional thermal processing of citrus juice These resultsagree with previously reported studies [29ndash32] In thesestudies losses of 120573-carotene or 120573-cryptoxanthin were verylow during pasteurisation or the thermal concentration ofdifferent citrus juices Sanchez-Moreno et al [33] reported

that cases of thermal treated orange juice (90∘C for 1min) ledto an increase in 120573-cryptoxanthin (1919) and zeaxanthin(3749) and to a decrease in lutein (2310) whereas nochanges were found in the extraction of 120573-cryptoxanthin orhydrocarbon carotenoids

Phenolic Compounds of Mandarin Juices Phenolic com-pounds of different mandarin juice samples are given inTable 3 As can be seen nine phenolic acids (gallic acidvanillic acid chlorojenic acid caffeic acid protocatechuicacid ethyl ester (PAEE) p-cumaric acid syringic acid sinapicacidand o-cumaric acid) were detected in themandarin juicesamples Ferrulic acid was not detected in any of the samplesThe most abundant phenolic acid was determined as PAEE(418mgL) in the Robinson variety and chlorogenic acid inFremont (532mgL) and Satsuma (789mgL) varieties Inaddition six flavonoids (hesperidin neohesperidin narin-genin isoquercetin luteolin and kamferol) were detectedin mandarin juices but eriocitrin neoeriocitrin naringinquercetin and apijenin were not detected in all mandarinvarieties Also rutin was detected only for the Satsumavariety Our results indicate that themost abundant flavanoneglycoside identified in mandarin juice was hesperidin buthesperidin concentrations were detected for R (20845mgL)F (22623mgL) and S (28295mgL) Naringin hesperidinand neohesperidin are flavanone glycosides that are knownto accumulate specifically in citrus species They have dif-ferent sugar moieties which influence taste naringin andneohesperidin are neohesperidosides with a bitter taste dueto the sugar neohesperidose while the sugar rutinose causeshesperidin to have a neutral taste [34 35]

Hesperidin concentrations of fresh mandarin juices weredecreased dramatically after pasteurisation treatments (119875 lt005) The amount of this compound was decreased forRobinson 3933 Fremont 3379 and Satsuma 2305after pasteurisation treatments Conversely Dhuique-Mayeret al [28] reported that hesperidin concentration was notsignificantly decreased during thermal treatments (losseslt2 at 90∘C after 240min) in orange juice The same resultswere in agreement with those of Sanchez-Moreno et al [33]who stated that pasteurisation did not modify hesperidincontent In our samples hesperidin was decreased followingpasteurisation of mandarin juices

34 Sensory Properties of Mandarin Juices Sensory prop-erties of mandarin juices are given in Table 4 Colour and


turbidity properties of juice samples ranged between 867ndash894 and 839ndash895 respectively After pasteurisation treat-ment the colour of samples scored higher than fresh samplesbut differences between juice samples were not importantfor colour Also there was no difference between turbidityproperties of samples The best scores for taste and smellwere obtained from fresh juices but the highest scores (737and 595) after pasteurisation treatment were found for theRP sample The general impression is a sensory propertywhich describes the appreciation of a product thereforethis property is very important for food producers Thegeneral impression of fresh mandarin juices scored between834ndash899 After pasteurisation this value was significantlydecreased (119875 lt 005) The general impression of RP FP andSP samples scored 778 658 and 528 respectively As a resultthe most well-liked fresh and pasteurised juice samples werefrom the Robinson variety of mandarin

4 Conclusions

After the pasteurisation process 119871lowast 119886lowast 119887lowast and 119862lowast valuesof all varieties were increased The Huelowast value of Fremont(F) was increased but was decreased for Robinson (R) andSatsuma (S)The highestΔ119864lowast value of samples was calculatedfor the Fremont variety Ascorbic acid concentration wasdetermined to be highest for the Robinson variety and degra-dation was calculated as 220 1686 and 2431 for samplesof R F and S respectively after pasteurisation The highesttotal carotenoid and phenolic contents were determined inS samples In general after the pasteurisation treatmentthe total carotenoid content of juices was slightly increasedbut total phenolic contents were dramatically decreasedPasteurisation treatment caused an increase in antioxidantactivity The most abundant carotenoid compound was 120573-cryptoxanthin in all samples nine phenolic acids and sixflavonoids were detected in the mandarin juice samplesHesperidin was detected at the highest concentration in allmandarin juices but the hesperidin concentration of sampleswas decreased after the pasteurisation treatment In sensoryevaluation colours of pasteurised samples scored higher thanfresh samplesThemost well-liked fresh and pasteurised juicesamples were those of the Robinson variety of mandarinwith regard to taste smell and general impression As aresult these varieties of mandarin are suitable for the fruitjuice processing industry and have rich bioactive componentswhich are important for human health Also by blendingmandarin juices richer bioactive contents and products withhigh consumer appreciation will be obtained

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

This study was supported by Research Project Units ofCukurova University (Project no ZF2007BAP45)

References

[1] FAO ldquoFAOSTAT Statistical Database FAO Romerdquo 2013 httpfaostatfaoorg

[2] A del Caro A Piga V Vacca and M Agabbio ldquoChanges offlavonoids vitamin C and antioxidant capacity in minimallyprocessed citrus segments and juices during storagerdquo FoodChemistry vol 84 no 1 pp 99ndash105 2004

[3] C Dhuique-Mayer C Caris-Veyrat P Ollitrault F Curk andM Amiot ldquoVarietal and interspecific influence on micronutri-ent contents in citrus from the mediterranean areardquo Journal ofAgricultural and Food Chemistry vol 53 no 6 pp 2140ndash21452005

[4] T Wu Y Guan and J Ye ldquoDetermination of flavonoids andascorbic acid in grapefruit peel and juice by capillary elec-trophoresis with electrochemical detectionrdquo Food Chemistryvol 100 no 4 pp 1573ndash1579 2007

[5] A Cano A Medina and A Bermejo ldquoBioactive compoundsin different citrus varieties discrimination among cultivarsrdquoJournal of Food Composition and Analysis vol 21 no 5 pp 377ndash381 2008

[6] M A Anagnostopoulou P Kefalas E Kokkalou A NAssimopoulou and V P Papageorgiou ldquoAnalysis of antioxi-dant compounds in sweet orange peel by HPLC-diode arraydetection-electrospray ionization mass spectrometryrdquo Biomed-ical Chromatography vol 19 no 2 pp 138ndash148 2005

[7] M J Jordan K L Goodner and J Laencina ldquoDeaeration andpasteurization effects on the orange juice aromatic fractionrdquoLWTmdashFood Science and Technology vol 36 no 4 pp 391ndash3962003

[8] C S Chen and M C Wu ldquoKinetic models for thermalinactivation of multiple pectinesterases in citrus juicesrdquo Journalof Food Science vol 63 no 5 pp 747ndash750 1998

[9] A Espachs-Barroso A van LoeyMHendrickx andOMartın-Belloso ldquoInactivation of plant pectinmethylesterase by thermalor high intensity pulsed electric field treatmentsrdquo InnovativeFood Science and Emerging Technologies vol 7 no 1-2 pp 40ndash48 2006

[10] P Schreier and H Idstein ldquoAdvances in the instrumental anal-ysis of food flavoursrdquo Zeitschrift fur Lebensmittel-Untersuchungund -Forschung vol 180 no 1 pp 1ndash14 1985

[11] M O Nisperos-Carriedo and P E Shaw ldquoComparison ofvolatile flavor components in fresh and processed orangejuicesrdquo Journal of Agricultural and Food Chemistry vol 38 no4 pp 1048ndash1052 1990

[12] P J Fellers and R D Carter ldquoEffect of thermal processing andstorage of chilled orange juice onflavor qualityrdquoFruit Processingvol 3 no 12 pp 436ndash441 1993

[13] M G Moshonas P E Shaw and B S Buslig ldquoRetention offresh orange juice flavor and aroma in an aqueous distillate fromValencia orange juicerdquo Journal of Food Quality vol 16 pp 101ndash108 1993

[14] E Agcam A Akyıldız and G A Evrendilek ldquoComparisonof phenolic compounds of orange juice processed by pulsedelectric fields (PEF) and conventional thermal pasteurisationrdquoFood Chemistry vol 143 pp 354ndash361 2014

[15] AOAC Official Method of Analysis of the Association of OfficialAnalytical Chemistry AOAC Arlington Va USA 15th edition1990

[16] S Meydav I Saguy and I J Kopelman ldquoBrowning determi-nation in citrus productsrdquo Journal of Agricultural and FoodChemistry vol 25 no 3 pp 602ndash604 1977


[17] V Gokmen and J Acar ldquoRapid reversed-phase liquid chro-matographic determination of patulin in apple juicerdquo Journalof Chromatography A vol 730 no 1-2 pp 53ndash58 1996

[18] I Klimczak M Małecka M Szlachta and A Gliszczynska-Swigło ldquoEffect of storage on the content of polyphenols vitaminC and the antioxidant activity of orange juicesrdquo Journal of FoodComposition and Analysis vol 20 no 3-4 pp 313ndash322 2007

[19] D A Kimball ldquoJuice cloudrdquo in Citrus Processing QualityControl and Technology pp 117ndash125 Van Nostrand ReinholdNew York NY USA 1991

[20] H S Lee and W S Castle ldquoSeasonal changes of carotenoidpigments and color in Hamlin Earlygold and Budd Bloodorange juicesrdquo Journal of Agricultural and Food Chemistry vol49 no 2 pp 877ndash882 2001

[21] A J Melendez-Martınez I S Vicario and F J HeredialdquoCarotenoids color and ascorbic acid content of a novelfrozen-marketed orange juicerdquo Journal of Agricultural and FoodChemistry vol 55 no 4 pp 1347ndash1355 2007

[22] H S Lee and G A Coates ldquoVitamin C in frozen freshsqueezed unpasteurized polyethylene-bottled orange juice astorage studyrdquo Food Chemistry vol 65 no 2 pp 165ndash168 1999

[23] P Abdullakasim S Songchitsomboon M Techagumpuch NBalee P Swatsitang and P Sungpuag ldquoAntioxidant capacitytotal phenolics and sugar content of selected thai health bever-agesrdquo International Journal of Food Sciences and Nutrition vol58 no 1 pp 77ndash85 2007

[24] K Hayat X Zhang U Farooq et al ldquoEffect of microwavetreatment on phenolic content and antioxidant activity of citrusmandarin pomacerdquo Food Chemistry vol 123 no 2 pp 423ndash4292010

[25] G Xu X Ye J Chen and D Liu ldquoEffect of heat treatment onthe phenolic compounds and antioxidant capacity of citrus peelextractrdquo Journal of Agricultural and Food Chemistry vol 55 no2 pp 330ndash335 2007

[26] S Jeong S Kim D Kim et al ldquoEffect of heat treatment onthe antioxidant activity of extracts from citrus peelsrdquo Journal ofAgricultural and Food Chemistry vol 52 no 11 pp 3389ndash33932004

[27] Y Niwa T Kanoh T Kasama and M Negishi ldquoActivation ofantioxidant activity in natural medicinal products by heatingbrewing and lipophilization A new drug delivery systemrdquoDrugs under Experimental and Clinical Research vol 14 no 5pp 361ndash372 1988

[28] C Dhuique-Mayer M Tbatou M Carail C Caris-Veyrat MDornier and M J Amiot ldquoThermal degradation of antioxidantmicronutrients in Citrus juice kinetics and newly formedcompoundsrdquo Journal of Agricultural and Food Chemistry vol55 no 10 pp 4209ndash4216 2007

[29] H S Lee and G A Coates ldquoThermal pasteurization effects oncolor of red grapefruit juicesrdquo Journal of Food Science vol 64no 4 pp 663ndash666 1999

[30] H S Lee and G A Coates ldquoEffect of thermal pasteurization onValencia orange juice color and pigmentsrdquo LWTmdashFood Scienceand Technology vol 36 no 1 pp 153ndash156 2003

[31] J J T Gama and C M Sylos ldquoMajor carotenoid compositionof Brazilian Valencia orange juice identification and quantifi-cation by HPLCrdquo Food Research International vol 38 no 8-9pp 899ndash903 2005

[32] C Cortes F Torregrosa M J Esteve and A Frıgola ldquoCaro-tenoid profile modification during refrigerated storage inuntreated and pasteurized orange juice and orange juice treated

with high-intensity pulsed electric fieldsrdquo Journal of Agriculturaland Food Chemistry vol 54 no 17 pp 6247ndash6254 2006

[33] C Sanchez-Moreno L Plaza P Elez-Martınez B De Ancos OMartın-Belloso and M P Cano ldquoImpact of high pressure andpulsed electric fields on bioactive compounds and antioxidantactivity of orange juice in comparison with traditional thermalprocessingrdquo Journal of Agricultural and Food Chemistry vol 53no 11 pp 4403ndash4409 2005

[34] J J Peterson J T Dwyer G R Beecher et al ldquoFlavanonesin oranges tangerines (mandarins) tangors and tangelosa compilation and review of the data from the analyticalliteraturerdquo Journal of Food Composition and Analysis vol 19 ppS66ndashS73 2006

[35] I Moulehi S Bourgou I Ourghemmi and M S TounsildquoIndustrial crops and products variety and ripening impacton phenolic composition and antioxidant activity of mandarin(Citrus reticulate Blanco) and bitter orange (Citrus aurantiumL) seeds extractsrdquo Industrial Crops and Products vol 39 pp 74ndash80 2012

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


needed Such information will provide a list of varieties thatare more appropriate for the production of juice

The main objective of this study was to compare thechemical composition of three mandarin cultivars (Robin-son Fremont and Satsuma) that are currently grown inTurkey and to determine the chemical changes that occur asa result of pasteurisation

2 Materials and Methods

21 Juice Processing Robinson (R) (Citrus reticulata hybridof Clementine times Orlando) Fremont (F) (Citrus reticulatahybrid of Clementine times Ponkan) and Satsuma (S) (Citrusunshiu) mandarin varieties grown in the Cukurova Regionof Turkey were used in this study After being washed man-darins were cut into two halves and pressed using a bench-scale automatic squeezing machine (CANCAN Turkey)Mandarin juices were passed through 1mm stainless steelsieves to remove seeds and coarse pulp Juicewas immediatelyprocessed by heat pasteurisation The study was carried outwith three replications

22 Heat Pasteurisation The bench-scale system designed inthe Department of Food Engineering of Cukurova University(Adana Turkey) was used for heat pasteurisation applications[14] As a result of the preliminary experiments heat wasapplied at 95∘C for 15 s for pasteurisation Heat-pasteurisedsamples were placed in amber coloured bottles for furtheranalyses Pasteurised samples were called Robinson (RP)Fremont (FP) and Satsuma (SP) in future analyses

23 General Properties of Mandarin Juices Total acidity anddry matter of juice samples were carried out according toAOAC [15] Brix and pH of mandarin juices were analysedwith an Abbe refractometer (Carl Zeiss Jena Germany) andWTW pH-meter (Weilheim Germany) respectively

24 Colour Measurement Colour (CIE 119871lowast 119886lowast 119887lowast) analysiswas conducted by the Colour Flex HunterLab instrument50mL of juice was transferred to the 20mm Glass OpticalCell Light Path and analysed The results were given accord-ing to the CIELAB colour system In this system 119871lowast defineslightness (0 black 100 white) 119886lowast denotes the redgreen value((+) red (minus) green) and 119887lowast denotes the yellowblue value((+) yellow (minus) blue) In addition the following formulaswere used for the calculations of Huelowast 119862lowast and Δ119864lowast colourvalues

Huelowast = arctan(119887lowast

119886lowast)

119862lowast

= radic(119886lowast

)2

+ (119887lowast

)2

Δ119864lowast

= radic(Δ119871lowast)2

+ (Δ119886lowast)2

+ (Δ119887lowast)2

(1)

25 Determination of Browning Index 5mL of each varietyof mandarin juice was mixed with 5mL ethyl alcohol (95)in teflon tubes and then centrifuged (4000 rpm 10min at

4∘C) The supernatant was passed through a 045 120583m teflonmembrane filter and the absorbance of the supernatant wasobtained at 420 nm in a spectrophotometer (Perkin ElmerLambda 25-UVVIS USA) [16]

26 Determination of Hydroxymethylfurfural (HMF) HMFextractions of samples were carried out according to themethod reported by Gokmen and Acar [17] The bestchromatographic conditions were determined as a result ofpreliminary experiments as follows 20120583L of supernatantwas injected into the C18 ACE (46 times 250mm) columnthe column was maintained at 30∘C with a flow rate of05mLmin and the photodiode array detector was set at285 nmMethanolwateracetic acid (20791 vvv) was usedas mobile phases

27 Antioxidant Activity The antioxidant activity of themandarin juices was evaluated using the DPPHlowast free radical-scavenging method The DPPHlowast free radical-scavengingactivity measurements were carried out according to the pro-cedure of Klimczak et al [18] with some modifications 5mLof mandarin juices was mixed with 5mL of methyl alcohol(80) in teflon tubes and then centrifuged (4000 rpm 10minat 4∘C) Briefly 01mL of supernatant was added to 246mLof11-diphenyl-2-picrylhydrazyl radical (DPPHlowast 0025 gLminus1 in80 methyl alcohol) and mixed by vortex After incubationfor 10min in the dark the absorbance of the sampleswas measured at 515 nm using the spectrophotometer Theantioxidant activity was expressed as the percentage declineof the absorbance as follows

Antioxidant activity () = (119860control minus 119860 sample

119860control) times 100

(2)

where 119860control is the absorbance of the control and 119860 sample isthe absorbance of the sample

28 Determination of PME Activity For the measurementof PME activity 10mL of mandarin juice was mixed with20mL of 1 pectin-salt substrate (01M NaCl) and incu-bated at 30∘C The solution was adjusted to pH 70 with20NNaOH and the pH of the solution was readjusted to77 with 005NNaOH After the pH reached 77 010mL of005NNaOH was added Time was measured (1199051015840) until thepH returned to 77 PME activity () was calculated as follows[19]

PME Activity (119860) =(005NNaOH ) (010mLNaOH)(1199051015840) (10mLsample)

Residual PME Activity () = (119860119905

1198600

) 100

(3)

where 1199051015840 is time in min 1198600is initial PME activity and 119860

119905is

PME activity after pasteurisation

29 Determination of Total Carotenoid Content Totalcarotenoid determination was carried out according to the








2PO4(pH 24)



2CO3solution





















119886lowast













Yellow

Green

Blue

Red

R F S



+alowast +blowast


Croma lowast

Huelowast

minusalowast +a

lowast

minusblowast

+blowast





















































4 Conclusions




Acknowledgment


References













































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








2PO4(pH 24)



2CO3solution





















119886lowast













Yellow

Green

Blue

Red

R F S



+alowast +blowast


Croma lowast

Huelowast

minusalowast +a

lowast

minusblowast

+blowast





















































4 Conclusions




Acknowledgment


References













































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology











119886lowast













Yellow

Green

Blue

Red

R F S



+alowast +blowast


Croma lowast

Huelowast

minusalowast +a

lowast

minusblowast

+blowast





















































4 Conclusions




Acknowledgment


References













































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



















































4 Conclusions




Acknowledgment


References













































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




















4 Conclusions




Acknowledgment


References













































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



4 Conclusions




Acknowledgment


References













































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology





























Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Documents

Research Article A Study on the Quality Criteria of Some Mandarin …downloads.hindawi.com/archive/2014/982721.pdf · 2019. 7. 31. · Research Article A Study on the Quality Criteria