research article determination of volatile compounds in four...

Research ArticleDetermination of Volatile Compounds in Four CommercialSamples of Japanese Green Algae Using Solid PhaseMicroextraction Gas Chromatography Mass Spectrometry

Masayoshi Yamamoto1 Susanne Baldermann23 Keisuke Yoshikawa4 Akira Fujita4

Nobuyuki Mase5 and Naoharu Watanabe1

1 Integrated Bioscience Section Graduate School of Science and Technology Shizuoka University 836 Ohya Suruga-kuShizuoka 422-8529 Japan

2 Leibniz-Institute of Vegetable and Ornamental Crops GroszligbeerenErfurt eV Theodor-Echternmeyer-Weg 114979 Groszligbeeren Germany

3 Institute of Nutritional Science University of Potsdam Arthur-Scheunert-Allee 114-116 14558 Nuthetal Germany4Technical Research Institute RampD Center T Hasegawa Company Ltd 29-7 Kariyado Nakahara-ku Kawasaki 211-0022 Japan5 Graduate School of Engineering Shizuoka University 3-5-1 Johoku Naka-ku Hamamatsu 432-8561 Japan

Correspondence should be addressed to Naoharu Watanabe acnwataipcshizuokaacjp

Received 31 August 2013 Accepted 6 October 2013 Published 27 January 2014

Academic Editors M A Strege T Tuzimski and M C Yebra-Biurrun

Copyright copy 2014 Masayoshi Yamamoto et al 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

Green algae are of great economic importance Seaweed is consumed fresh or as seasoning in Japan The commercial value isdetermined by quality color and flavor and is also strongly influenced by the production area Our research based on solid phasemicroextraction gas chromatography mass spectrometry (SPME-GC-MS) has revealed that volatile compounds differ intenselyin the four varieties of commercial green algae Accordingly 41 major volatile compounds were identified Heptadecene wasthe most abundant compound from Okayama (Ulva prolifera) Tokushima (Ulva prolifera) and Ehime prefecture (Ulva linza)Apocarotenoids such as ionones and their derivatives were prominent volatiles in algae from Okayama (Ulva prolifera) andTokushima prefecture (Ulva prolifera) Volatile short chained apocarotenoids are among the most potent flavor components andcontribute to the flavor of fresh processed algae and algae-based products Benzaldehyde was predominant in seaweed fromShizuoka prefecture (Monostroma nitidum) Multivariant statistical analysis (PCA) enabled simple discrimination of the samplesbased on their volatile profiles This work shows the potential of SPME-GC-MS coupled with multivariant analysis to discriminatebetween samples of different geographical and botanical origins and form the basis for development of authentication methods ofgreen algae products including seasonings

1 Introduction

Seaweeds have been traditionally used as medicines foodand seasoning [1 2] Most seaweeds are of commercialimportance in foodstuffs cosmetics and dyes Moreoverthey serve as source for biological active compounds Today75ndash8 million tons of macroalgae are produced per annumand their estimated value is US$ 55ndash6 billion [3] Seaweedsare widely distributed throughout the world and are predom-inantly consumed in Japan China Korea South AustraliaNew Zealand Polynesia and South America [4]

Certain countries or regions have reputations for pro-ducing high-quality green algae including Aonori (a highquality merchandised Japanese green seaweed) and are ableto demand a significantly higher price for their productsIn Japan high-value production areas include the river Shi-manto in Kochi prefecture and river Yoshino in Tokushimaprefecture There is a strong interest in the authenticity ofthese commercial products and up to now consumers haveto rely on package information to provide confirmation thatthe product is from a high-quality region with a traceability

Hindawi Publishing Corporatione Scientific World JournalVolume 2014 Article ID 289780 8 pageshttpdxdoiorg1011552014289780

2 The Scientific World Journal

system such as the Food Marketing Research and Informa-tion Center in Japan [5]

GC-MS techniques coupled with multi-variant analysisare becoming increasingly important tools to monitor thedistribution of counterfeit foodstuffs and have been used inauthenticating many food products such as tea olive oilhoney cheese and wine [6ndash12]

We newly report on nontargeted SPME-GC-MS fol-lowed by multi-variant statistical analysis in Japanese greenmacroalgae Significant differences in the volatile profilesof green algae from four production areas were detectedimplying that markers could potentially be identified usingthis analytical-statistical approach

2 Experimental

21 Commercial Dried Green Algae Samples Ulva proliferaUlva linza and Monostroma nitidum commercially all soldas ldquoAonorirdquo (Japanese green algae) were subjected to sensoryevaluation and used for solid-phase microextraction (SPME)method development Ulva prolifera (2 samples) Ulva linzaand Monostroma nitidum were obtained from OkayamaTokushima Ehime and Shizuoka prefecture respectivelyCommercial Japanese green algae were purchased from awholesale agency in Japan The samples were placed in airtight plastic bags in the dark at room temperature Prior toanalysis the samples were ground to a fine powder usingliquid nitrogen

22 Sensory Evaluation The sensory tests were carried outby nine untrained panelists consuming regularly green algaeHedonic sensory attributes evaluated in this study were colortaste and overall acceptability of four green algae samplesThepanelists (6male 3 female) were between 26- and 40-yearold A nine point hedonic scale (0ndash8) anchored by dislikeextremely to like extremely for the overall score light coloredto strongly colored for the green color and weak to intensefor the taste was employedMore precisely the panelists wereasked to evaluate the following aroma attributes animalicfloral spicy fatty green note marine-like fresh (waterly)powdery and leather-like common flavor attributes of greenalgae Samples were served in glass beakers and providedrandomly to panelists Sensory evaluation procedures wereexplained to the panelists prior the tests

23 Solid Phase Microextraction (SPME) Polydimethylsilox-ane fibers (100 120583m) were mounted in a SPME manual holder(Supelco Bellefonte PA USA) All fibers were conditionedprior to analyses according to the manufacturerrsquos recom-mendations Various parameters (extraction method timeand temperature) influencing SPME analysis were optimizedFor each analysis 10 g of sample was placed in a 40 cm3 vialspiked with 1mg of the internal standard ethyldecanoateand then sealed with an aluminum crimp cap provided witha needle-pierceable polytetrafluoroethylenesilicone septumEach sample vial was placed at 50∘C for 60min to equilibrateand then the septum was pierced with the SPME needleFibers were exposed to the sample headspace for 60min

After the extraction time fibers were retracted into theneedle transferred immediately to the injection port of a GCand desorbed at 230∘C for 1min

24 GCMS Analysis The compounds collected in theheadspace above the green algae samples were analyzed byGC connected to a mass spectrometer (MS) (Shimadzu GC-MSQP5000) which was controlled by a Class-5000 worksta-tion Separation was carried out on a Supelcowax 10 fusedsilica capillary column 30m times 025mm id 025120583m filmthickness (Supelco Bellefonte PA USA) Injection was insplitless mode for 1min at 230∘C with a specific SPME insertTheGC temperaturewas raised from50 to 160∘Cat 3∘Cminminus1and from 160 to 240∘C at 10∘Cminminus1 The mass spectra werescanned at 70 eV over a mass range from mz 50 to 280Nonisothermal linear RI (Kovats type) was calculated afterthe analysis of a C11ndashC30 n-alkane series (GL Science) underthe same conditions Volatile compounds were tentativelyidentified by comparing their mass spectra with those of thereference database (NIST Mass Spectral Data 981015840 edition)Compounds were attributed to molecules through their indi-vidual mass spectrum retention indices (RI) and retentiontime (RT) In addition pure reference compounds were usedto confirm the results Flavor properties were assigned basedon reported odor descriptions [3 13 14]

25 Data Analysis The relative peak areas were calculatedin relation to the peak area of the internal standard ethylde-canoate The identified volatile compounds were classifiedinto different groups of molecules such as alkanes alkenesketones aldehydes sulfur compounds alcohols and estersUnidentified compounds were excluded from this analysisThe relative peak areas of the different compounds in eachcategory were calculated as a ratio of the total peak areaStatistical analyses were conducted using XL-STAT version2013 (Addinsoft New York USA)

3 Results and Discussion

The results of the sensory analysis revealed that it wasimpossible to differentiate between the samples based on theoverall score (Figure 1) The same applies also to taste andcolor (data not shown) Based on the nine aroma attributes(animalic floral spicy fatty green note marine-like fresh(waterly) powdery and leather-like) it was found that thesamples from Okayama Tokushima and Ehime prefectureare uniform However the sample from Shizuoka prefecturewas differently evaluated in respect of its green-note marine-like fresh and powdery aroma (Figure 2)

As a result it is impossible to distinguish between com-mercially sold ldquoAonorirdquo samples based on sensory evalua-tion Solely in taxonomically different samples OkayamaTokushima and Ehime (Ulva species) versus Shizuoka(Monostroma species) somewhat different scores could beobtained

Volatile compounds are fundamental for algae odor andaromaTherefore it is not surprising that volatile compoundsin algae have been investigated and reports evaluating volatile

The Scientific World Journal 3

Okayama EhimeTokushima0

2

4

6

8

Scor

e

Shizuoka

Figure 1 Overall scores of the sensory evaluation of green algaesamples

001020304050607080Animalic

Floral

Spicy

Fatty

Green noteMarine-like

Fresh

Powderly

Leather-like

OkayamaTokushima

EhimeShizuoka

Figure 2 Sensory characterisation of green algae samples Thefollowing aroma attributes were determined based on panelistrsquosevaluation animalic floral spicy fatty green note marine-likefresh (waterly) powdery and leather-like

metabolites for example in brown and red algae are availablein the current literature [15ndash18]

GC-MS techniques coupled with multi-variant analysishave been used in authenticating many food products andshould be tested in this study for its practicability to dis-tinguish different commercially sold Japanese green algaesamples

Forty-one major volatile compounds classed as alkanesalkenes ketones aldehydes sulfur compounds alcoholsand esters were identified in the four commercial Japanesegreen algae (Table 1) These compounds are derived from

00

200

400

600

800

1000Alkanes

Alkenes

Ketones

AldehydesSulfur compounds

Alcohols

Esters

OkayamaTokushima

EhimeShizuoka

Figure 3 Chemical classification of volatile compounds in relativeconcentrations (119899 = 3) in different commercial green algae samples

different precursors including the amino acids fatty acidsand carotenoids (Table 1)

Hydrocarbons including alkanes and alkenes (eg tetra-decene pentadecane hexadecane and octadecene) wereabundant in three (Okayama Tokushima and Ehime) of thefour samples (Figure 3)

The relative concentrations of alkenes vary among thesamples from different algae of the orderUlvales Green algaeof the Ulva species are rich in alkenes 95 in Okayama923 in Tokushima and 894 in Ehime prefecture Incontrast the green algae of the Monostroma species fromShizuoka prefecture contain only 63 of alkenes An oppo-site trend has been found for the relative content of aldehydesVery low relative contents are present in algae fromOkayama(02) Tokushima (01) and Ehime (01) and about 915in algae from Shizuoka prefecture

Heptadecanes derived from fatty acids were identifiedas major components among all samples 7-Heptadecane hasbeen reported to be one of the major components in marinegreen algae (Ulva pertusa and Chara vulgaris) and is knownto be the character impact compound of edible kelp It alsocontributes to the off-flavor inwater [1 19]There is a seasonalvariation in the concentration of 7-heptadecene and phytolin marine algae It was found that 7-heptadecane was usuallypresent in appreciable amounts throughout the year exceptduring summer [20]

Ketones sulfur compounds and esters occurred in sim-ilar concentration ranges in all of the samples Principalcomponent analysis was performed to identify key volatilescontributing to the flavor characteristic of the four com-mercial algae samples Figure 4 shows the correlation circleof factor 1 against factor 2 and helped us to identify majorvolatiles contributing to the distinct flavor properties of thecommercial samples

4 The Scientific World JournalTa

ble1Vo

latilec

ompo

unds

ofdifferent

greenalgaes

amples

presentedas

relativer

atiosc

omparedto

theinternalstand

ardethyldecanoate

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D1

Dod

ecane

C 10H

22Alkane

Alkane(B)

1185

ND

ND

ND

1200010

2Tridecane

C 13H

28Alkane

Fatty

acids

Alkane(B)

1297

06

000

0306

000

08ND

232

00298

3226-

Trim

ethylcyclohexano

neC 9

H16O

Ketone

Caroteno

ids

Camph

oraceous

tobaccono

tes

thujon

e-lik

e(A)

1308

07

00011

07

00026

ND

ND

4Tetradecane

C 14H

30Alkane

Fatty

acids

Alkane(B)

1397

39

00020

114

000

9771

00158

60

000

655

Tetradecene

C 14H

28Alkene

1444

05

00013

09

000

0521

00025

ND

6Pentadecane

C 15H

32Alkene

Alkane(B)

1502

836

00622

2511

01181

4205

06057

ND

7(EE)-35-Octadien-2-on

eC 8

H12O

Ketone

Fatty

acids

caroteno

ids

Alm

ond(C

)1517

ND

ND

ND

ND

8Be

nzaldehyde

C 7H

5OAldehyde

Aminoacids

Sweetstr

ong

almon

d(A

)almon

dbu

rnt

sugar(B)

1517

1700028

23

000

6809

00015

37773

74504

9Pentadeceneisomer

1C 1

5H30

Alkene

1523

24

00028

36

00025

47

00057

117

00326

10Pentadeceneisomer

2C 1

5H30

Alkene

1546

07

00011

23

00025

1000010

05

00028

11Dim

ethylsulfoxide

C 2H

6OS

Sulfu

rcompo

und

Aminoacids

1557

82

00457

165

00944

80

00500

123

00221

12(EZ)-35-Octadien-2-on

eC 8

H12O

Ketone

Fatty

acids

caroteno

ids

Melo

n-lik

e(C)

1567

27

00100

21

000

6506

00052

03

00029

13Pentadeceneisomer

3C 1

5H30

Alkene

1576

ND

ND

ND

51

00085

14266-Trim

ethyl-2

-hydroxycyclohexano

neC 9

H16O

2Ke

tone

Caroteno

ids

Sweet

tobacco-lik

earom

awith

herbaceous

underton

es(A

)

1594

86

00237

9600223

41

00053

ND

15Hexadecane

C 16H

34Alkane

Alkane(B)

1601

43

00089

56

00122

7700144

282

00563

16120573-C

yclocitral

C 10H

16O

Aldehyde

Caroteno

ids

Fresh(B)

1612

35

00085

35

00074

1400010

ND

17Hexadeceneisomer

1C 1

6H32

Alkene

1619

1200010

25

00027

21

00025

ND

18Hexadeceneisomer

2C 1

6H32

Alkene

1624

09

00024

1100026

22

000

04ND

19Safranal

C 10H

14O

Aldehyde

Caroteno

ids

1637

ND

ND

ND

ND

20Heptadecane

C 17H

36Alkane

Alkane(B)

1707

561

00610

252

00936

255

02227

124

00993

217-Heptadecene

C 17H

34Alkene

1722

29097

10740

40033

33306

39543

55587

1555

03312

22Heptadecene

isomer

1C 1

7H34

Alkene

1733

1800193

ND

120

00030

7000169

23Heptadecadieneisomer

1C 1

7H32

Alkene

1757

1034

006

041392

01109

1487

01725

643

01246


2C 1

7H32

Alkene

1766

1000021

1200018

05

000

4707

00015


3C 1

7H32

Alkene

1769

43

00022

22

00011

08

00074

07

00104


4C 1

7H32

Alkene

1772

38

00037

53

00036

59

000

4111

00198


5C 1

7H32

Alkene

1779

08

00022

ND

03

00059

05

000

9128

Octadecane

C 18H

38Alkane

Alkane(B)

1801

30

00051

16000

4232

00189

21

00059

29Octadecene

C 18H

36Alkene

1817

112

00072

128

00059

181

00085

09

00081

30120572-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

violet-floral(A

)1842

46

00039

56

00139

1800085

ND


Table1Con

tinued

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D31

Non

adecane

C 19H

40Alkane

Alkane(B)

1898

29

00119

1100039

20

00029

07

00033

32120573-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

dry(A

)1927

125

01087

157

00144

59

000

4007

00031

33Maltol

C 6H

6O3

Ketone

Caramel-

butte

rscotch(A

)caramel(B)

1960

06

00082

ND

ND

1000059

34120573-Io

none

epoxide

C 13H

20O

2Ke

tone

Caroteno

ids

Sweetb

erry

(A)

1980

21

00181

28

00025

09

000

06ND

35Methyltetradecano

ate

C 15H

30O

2Ester

Fainto

nion

ho

ney(A

)orris

(A)(B)

2010

09

00051

00

000

0013

00017

1700036

36Pentadecanal

C 15H

30O

Aldehyde

Fresh(B)

2034

1800103

31

00033

1800103

ND

3761014

-Trim

ethyl-2

-pentadecanon

eC 1

8H36O

Ketone

Caroteno

ids

2139

29

00168

67

00174

7000160

74004

27

38Hexadecanoica

cid

methyleste

rC 1

7H34O

2Ester

Fatty

acids

2217

52

00210

22

00038

9000133

22

00029

39Dihydroactin

idiolid

eC 1

1H16O

2Ke

tone

Caroteno

ids

Musky

orcoum

arin-like

(A)

2347

30

00363

84

00199

42

00022

04

00034

40Methyllinolenate

C 19H

32O

2Ester

Fatty

acids

2566

41

00390

27

000

6373

00268

34

00057

41Ph

ytol

C 20H

40O

Alcoh

olCh

loroph

yll

Faintfl

oral(A

)flo

wer

(B)

2580

28

00191

1200050

09

00023

07

00071

(A)F

oodandAgriculture

Organizationof

theu

nitednatio

ns(http

wwwfaoorg)(B)

Flavornet(httpwwwflavornetorg)and(C

)HuandPan

2000

[14]


DodecaneTridecane

Trimethylcyclohexanone

Tetradecane

TetradecenePentadecane

Benzaldehyde

Pentadecene

Pentadecene

Dimethyl sulfoxide

Pentadecene

hydroxycyclohexanone

Hexadecane

Hexadecene

Hexadecene

Safranal

Heptadecane

Heptadecene

Heptadecene

Heptadecadiene

HeptadecadieneHeptadecadiene

Heptadecadiene

Heptadecadiene

Octadecane

Octadecene

Nonadecane

Maltol

Methyl tetradecanoate

Pentadecanal

pentylidenefuran-2-one

Hexadecanoic acid methylester

Dihydroactinidiolide

Methyl linolenate

Phytol

0

025

05

075

1

025 050 075 1

minus025

minus025

minus075

minus075

minus1

minus1

minus05

minus05

120573-ionone epoxide120573-Ionone

266-Trimethyl-2-

120572-Ionone120573-Cyclocitral

35-Octadien-2-one

61014-Dimethyl

35-Octadiene-2-one

F1 (4813)

226-

Variables (axes F1 and F2 6827)F2

(20

14

)

Figure 4 Correlation circle of volatile compounds from different green algae samples The profiling was carried out based on 41 peaks anddetected corresponding peak area ratios Samples were independently analyzed in triplicate The data was analyzed by PCA using XL-STAT20132

Our analysis revealed benzaldehyde an important com-pound in algae from Shizuoka prefecture originated from theamino acid biosynthesis pathway Benzaldehyde contributesto the sweet almond odor of fruits and flowers and is char-acterized by low odor thresholds (350ndash3500 ppb) Benzalde-hyde can also be liberated from glycosidically bound precur-sors in various foods In addition it has been reported thatconcentrations increase during manufacturing processes forexample production of apple puree or roasting of sesameseeds [21]

Several hydrocarbons such as tridecane pentadecanehexadecane and dodecane were identified as relevant odorcompounds in seaweed from Shizuoka prefecture

It is noticeable that the proportion of volatile apoc-arotenoids is remarkably high in the investigated algaevarieties The importance of carotenoid-derived flavor com-pounds in algae has been previously reported Volatile apoc-arotenoids such as the potent flavor compound120573-ionone areproduced by a diversity of algae taxa for example Ulothrixfimbriata and Asakusa Nori (Porphyra tenera) [22 23] Apartfrom chemical formation pathways [24] it has been demon-strated that functional carotenoid-cleavage like enzymes

contribute to the formation of volatile apocarotenoids in algae[25]

226-Trimethylcyclohexanone (E Z)-35-octadiene-2-one 266-trimethyl-2-hydroxycyclohexa-none 120572-ionone 120573-ionone and 120573-ionone epoxide are abundant in Okayamaand Tokushima Dihydroactinidiolide makes an importantcontribution to the volatile profile of algae from TokushimaIt is also an important component in green tea with a sweetpeach-like flavor [24] In addition 120573-cyclocitral and phytolare characteristic compounds influencing the flavor of algaefrom Okayama and Tokushima prefecture they contributeto fresh and floral odors respectively [18] The ketonessuch as 120573-cyclocitral 120573-ionone and 120573-ionone epoxide andphytol are generated via degradation of carotenoids 120572-Ionone contributes to the violet wood odor 120573-ionone con-tributes to the violet-like and characteristic seaweed odor 120573-ionone epoxide contributes to the sweet berry odor Theseare important aroma impact compounds characterized byvery low odor thresholds for example 120573-ionone has an odorthreshold of 0007 ppm [26] 120573-Ionone accounts for 125157 59 and 07 of the volatiles in the algae harvestedfromOkayama Tokushima Ehime and Shizuoka prefecture


Okayama2Okayama3

Okayama1

1

Tokushima2

Tokushima1

Tokushima3

Ehime3Ehime2

Ehime1

Shizuoka2Shizuoka1

Shizuoka3

0

0

2

2

4

4

6

6

8 10minus8 minus6

minus6

minus4

minus4

minus2

minus2

Observations (axes F1 and F2 6827)

F2

(20

14

)

F1 (4813)

Figure 5 Scoring plots of nontargeted profiling of green algaesamples of different origin by GC-MS-SPME The profiling wascarried out based on 41 peaks and detected corresponding peak arearatios Samples were independently analyzed in triplicate The datawere analyzed by PCA using XL-STAT 20132

respectively Moreover thermal degradation products of120573-carotene including 226-trimethylcyclohexanone benzal-dehyde 266-trimethyl-2-hydroxycyclohexanone 120573-cyclo-citral 56-epoxy-120573-ionone and dihydroactinidiolide werepreviously reported [24]

After the removal of undesirable material green algae aresun dried and packaged It might be speculated that similarreactions occur during the sun drying processing of greenalgae and may have a strong impact on the quality of algaeproducts The occurrence of apocarotenoids is one of themost important indicators of authenticity for Ulva proliferaUlva linza andMonostroma nitidum

Another class of aroma compounds the aliphatic acidmethyl esters including hexadecanoic acid methyl esterand methyl linolenate was abundant in algae from Shizuokaprefecture Methyl linolenate serves as aprecursor of (EZ)-35-octadien-2-one and is one of the most odor-activecompounds [27] It has been found in cod liver oil and Ulvapertusa and contributes to the melon-like odor In this study(E Z)-35-octadien-2-onewas detected in all samples andwasmore abundant in Ulva prolifera

To explain the overall characteristics of the four algaesamples further studies are necessary to elucidate the con-tribution of single components and identify unknown com-pounds

Multi-variant analysis of known components was per-formed on the mass spectral data to explain the overallcharacteristics of the differences among the samples PCAwas performed using a transformed data set consisting ofcorrelation matrix of 41 variables yielding in 11 eigenvectorsFive factors explained 9239 of the total variance indicatingthat a reduced number of volatile compounds could explainthe overall characteristics of the four different samplesFactor 1 explained 4813 of the total variance and factor 2explained 2014 of the total variance Both factors together

enable discrimination of the four algae samples fromdifferentbiological and geographical origins (Figure 5) It is interestingthat there are fewer differences in the volatile profiles ofsamples from close geographic origins such as Tokushimaand Okayama compared with samples which are furtheraway from each other for example Tokushima and EhimeFuture studies should be performed with a higher number ofreplicates and taking into account testing sets

4 Conclusions

The volatile profiles of different commercial Japanese greenalgae samples originating from Okayama TokushimaEhime and Shizuoka were described for the first timeMultivariate analysis enabled discrimination of the samplesbased on their botanical and geographical origin Hithertothe authenticity of these commercial products is based onpackage information and this study could form the analyticalbasis of authenticity studies of dried green algae samplesusing the SPME-GC-MS-method followed by multivarianteanalysis

Conflict of Interests

The authors declare that there is no conflict of interests in thepaper

Acknowledgments

The authors are grateful for the gift of Monostroma nitidumfrom Kanetsu Sugiura Company Hamamatsu ShizuokaThey acknowledge the valuable discussion about the sensoryevaluation with Dr Bernhard Bruckner and thank all pan-elists for their support

References

[1] H Sugisawa K Nakamura and H Tamura ldquoThe aroma profileof the volatiles in marine green algae (Ulva pertusa)rdquo FoodReviews International vol 6 no 4 pp 573ndash589 1990

[2] S N Boopathy and K Kathiresan ldquoAnticancer drugs frommarine flora an overviewrdquo Journal of Oncology vol 2010Article ID 214186 18 pages 2010

[3] Food amp Agriculture Organization of the United Nations httpwwwfaoorgfoodfood-safety-qualityscientific-advicejecfajecfa-flaven

[4] T Kajiwara K Kodama A Hatanaka and K Matsui ldquoVolatilecompounds from Japanese marine brown algaerdquo in BioactiveVolatile Compounds from Plants vol 525 of ACS SymposiumSeries pp 103ndash120 1993

[5] Food Marketing amp Research Information Center httpwwwfmricorjptracenoriindexhtm

[6] P Poinot J Grua-Priol G Arvisenet et al ldquoOptimisation ofHS-SPME to study representativeness of partially baked breadodorant extractsrdquo Food Research International vol 40 no 9 pp1170ndash1184 2007

[7] D M A M Luykx and S M van Ruth ldquoAn overview of ana-lytical methods for determining the geographical origin of foodproductsrdquo Food Chemistry vol 107 no 2 pp 897ndash911 2008


[8] R Liu K Xiong Y Chao-Luo X Ze-Dai Z Min-Liu and WTong-Xue ldquoChanges in volatile compounds of a native Chinesechilli pepper (Capsicum frutescens var) during ripeningrdquo Inter-national Journal of Food Science amp Technology vol 44 no 12pp 2470ndash2475 2009

[9] I Silva S M Rocha and M A Coimbra ldquoHeadspace solidphase microextraction and gas chromatography-quadrupolemass spectrometry methodology for analysis of volatile com-pounds ofmarine salt as potential origin biomarkersrdquoAnalyticaChimica Acta vol 635 no 2 pp 167ndash174 2009

[10] K Tananuwong and S Lertsiri ldquoChanges in volatile aroma com-pounds of organic fragrant rice during storage under differentconditionsrdquo Journal of the Science of Food and Agriculture vol90 no 10 pp 1590ndash1596 2010

[11] K Kaseleht E Leitner and T Paalme ldquoDetermining aroma-active compounds in Kama flour using SPME-GCMS and GC-olfactometryrdquo Flavour and Fragrance Journal vol 26 no 2 pp122ndash128 2011

[12] A Prades R Assa M Dornier J P Pain and R BoulangerldquoCharacterisation of the volatile profile of coconut water fromfive varieties using an optimized HS-SPME-GC analysisrdquo Jour-nal of the Science of Food and Agriculture vol 92 pp 2471ndash24782012

[13] Flavornet httpwwwflavornetorg[14] S-P Hu and B S Pan ldquoModification of fish oil aroma using a

macroalgal lipoxygenaserdquo Journal of the American Oil ChemistsrsquoSociety vol 77 no 4 pp 343ndash348 2000

[15] Z Kamenarska A Ivanova R Stancheva et al ldquoVolatile com-pounds from some Black Sea red algae and their chemotaxo-nomic applicationrdquo Botanica Marina vol 49 no 1 pp 47ndash562006

[16] M E Hattab G Culioli L Piovetti S E Chitour and R VallsldquoComparison of various extraction methods for identificationand determination of volatile metabolites from the brown algaDictyopteris membranaceardquo Journal of Chromatography A vol1143 no 1-2 pp 1ndash7 2007

[17] V Gressler P Colepicolo and E Pinto ldquoUseful strategies foralgal volatile analysisrdquo Current Analytical Chemistry vol 5 no3 pp 271ndash292 2009

[18] R P Yu L P Wang S F Wu J S Che and Y M Liu ldquoAna-lysis of volatile compounds in Algal bloom water by gaschromatography-mass spectrometry and sniffingrdquo ChineseJournal of Analytical Chemistry vol 38 pp 1349ndash1352 2010

[19] A Rzama B Arreguy-San-Miguel and M Ettalibi ldquoLipidsmetabolites and essential oil from the green alga Chara vulgar-isrdquo Revue Marocaine des Sciences Agronomiques et Veterinairesvol 22 no 2 pp 65ndash70 2002

[20] M Okano and T Aratani ldquoConstituents in marine algae-Iseasonal variation of sterol hydrocarbon fatty acid and phytolfractions in Ulva pertusardquo Bulletin of the Japanese Society ofScientific Fisheries vol 45 pp 389ndash393 1979 (Japanese)

[21] Y H Hui Handbook of Fruit and Vegetable Flavors John Wileyamp Sons Hoboken NJ USA 2010

[22] P Fink E Von Elert and F Juttner ldquoVolatile foraging kair-omones in the littoral zone attraction of an herbivorous fresh-water gastropod to algal odorsrdquo Journal of Chemical Ecology vol32 no 9 pp 1867ndash1881 2006

[23] I Flament and G Ohloff ldquoVolatile constituents of algaeodoriferous constituents of seaweeds and structure of nor-terpenoids identified in Asakusa-Nori flavorrdquo in Proceedings ofthe 4th Weurman Flavour Research Symposium J Adda EdProgress in Flavor Research pp 281ndash300 Elsevier 1985

[24] M Kawakami ldquoIonone series compounds from beta-caroteneby thermal degradation in aqueous mediumrdquo Nippon Nogeika-gaku Kaishi vol 56 no 10 pp 917ndash921 1982 (Japanese)

[25] S Baldermann A N Mulyadi Z Yang et al ldquoApplication ofcentrifugal precipitation chromatography and high-speedcounter-current chromatography equipped with a spiral tubingsupport rotor for the isolation and partial characterization ofcarotenoid cleavage-like enzymes in Enteromorpha compressa(L) Neesrdquo Journal of Separation Science vol 34 no 19 pp2759ndash2764 2011

[26] R G Buttery R Teranishi L C Ling and J G TurnbaughldquoQuantitative and sensory studies on tomato paste volatilesrdquoJournal of Agricultural and Food Chemistry vol 38 no 1 pp336ndash340 1990

[27] F Ullrich and W Grosch ldquoIdentification of the most intenseodor compounds formed during autoxidation of methyl linole-nate at room temperaturerdquo Journal of the AmericanOil ChemistsSociety vol 65 no 8 pp 1313ndash1317 1988

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy


Carbohydrate Chemistry

International Journal of


Journal of

Chemistry


Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of


Chromatography Research International


Applied ChemistryJournal of



Theoretical ChemistryJournal of


Journal of

Spectroscopy

Analytical ChemistryInternational Journal of


Journal of


Quantum Chemistry


Organic Chemistry International

ElectrochemistryInternational Journal of



CatalystsJournal of


system such as the Food Marketing Research and Informa-tion Center in Japan [5]

GC-MS techniques coupled with multi-variant analysisare becoming increasingly important tools to monitor thedistribution of counterfeit foodstuffs and have been used inauthenticating many food products such as tea olive oilhoney cheese and wine [6ndash12]

We newly report on nontargeted SPME-GC-MS fol-lowed by multi-variant statistical analysis in Japanese greenmacroalgae Significant differences in the volatile profilesof green algae from four production areas were detectedimplying that markers could potentially be identified usingthis analytical-statistical approach

2 Experimental

21 Commercial Dried Green Algae Samples Ulva proliferaUlva linza and Monostroma nitidum commercially all soldas ldquoAonorirdquo (Japanese green algae) were subjected to sensoryevaluation and used for solid-phase microextraction (SPME)method development Ulva prolifera (2 samples) Ulva linzaand Monostroma nitidum were obtained from OkayamaTokushima Ehime and Shizuoka prefecture respectivelyCommercial Japanese green algae were purchased from awholesale agency in Japan The samples were placed in airtight plastic bags in the dark at room temperature Prior toanalysis the samples were ground to a fine powder usingliquid nitrogen

22 Sensory Evaluation The sensory tests were carried outby nine untrained panelists consuming regularly green algaeHedonic sensory attributes evaluated in this study were colortaste and overall acceptability of four green algae samplesThepanelists (6male 3 female) were between 26- and 40-yearold A nine point hedonic scale (0ndash8) anchored by dislikeextremely to like extremely for the overall score light coloredto strongly colored for the green color and weak to intensefor the taste was employedMore precisely the panelists wereasked to evaluate the following aroma attributes animalicfloral spicy fatty green note marine-like fresh (waterly)powdery and leather-like common flavor attributes of greenalgae Samples were served in glass beakers and providedrandomly to panelists Sensory evaluation procedures wereexplained to the panelists prior the tests

23 Solid Phase Microextraction (SPME) Polydimethylsilox-ane fibers (100 120583m) were mounted in a SPME manual holder(Supelco Bellefonte PA USA) All fibers were conditionedprior to analyses according to the manufacturerrsquos recom-mendations Various parameters (extraction method timeand temperature) influencing SPME analysis were optimizedFor each analysis 10 g of sample was placed in a 40 cm3 vialspiked with 1mg of the internal standard ethyldecanoateand then sealed with an aluminum crimp cap provided witha needle-pierceable polytetrafluoroethylenesilicone septumEach sample vial was placed at 50∘C for 60min to equilibrateand then the septum was pierced with the SPME needleFibers were exposed to the sample headspace for 60min

After the extraction time fibers were retracted into theneedle transferred immediately to the injection port of a GCand desorbed at 230∘C for 1min

24 GCMS Analysis The compounds collected in theheadspace above the green algae samples were analyzed byGC connected to a mass spectrometer (MS) (Shimadzu GC-MSQP5000) which was controlled by a Class-5000 worksta-tion Separation was carried out on a Supelcowax 10 fusedsilica capillary column 30m times 025mm id 025120583m filmthickness (Supelco Bellefonte PA USA) Injection was insplitless mode for 1min at 230∘C with a specific SPME insertTheGC temperaturewas raised from50 to 160∘Cat 3∘Cminminus1and from 160 to 240∘C at 10∘Cminminus1 The mass spectra werescanned at 70 eV over a mass range from mz 50 to 280Nonisothermal linear RI (Kovats type) was calculated afterthe analysis of a C11ndashC30 n-alkane series (GL Science) underthe same conditions Volatile compounds were tentativelyidentified by comparing their mass spectra with those of thereference database (NIST Mass Spectral Data 981015840 edition)Compounds were attributed to molecules through their indi-vidual mass spectrum retention indices (RI) and retentiontime (RT) In addition pure reference compounds were usedto confirm the results Flavor properties were assigned basedon reported odor descriptions [3 13 14]

25 Data Analysis The relative peak areas were calculatedin relation to the peak area of the internal standard ethylde-canoate The identified volatile compounds were classifiedinto different groups of molecules such as alkanes alkenesketones aldehydes sulfur compounds alcohols and estersUnidentified compounds were excluded from this analysisThe relative peak areas of the different compounds in eachcategory were calculated as a ratio of the total peak areaStatistical analyses were conducted using XL-STAT version2013 (Addinsoft New York USA)

3 Results and Discussion

The results of the sensory analysis revealed that it wasimpossible to differentiate between the samples based on theoverall score (Figure 1) The same applies also to taste andcolor (data not shown) Based on the nine aroma attributes(animalic floral spicy fatty green note marine-like fresh(waterly) powdery and leather-like) it was found that thesamples from Okayama Tokushima and Ehime prefectureare uniform However the sample from Shizuoka prefecturewas differently evaluated in respect of its green-note marine-like fresh and powdery aroma (Figure 2)

As a result it is impossible to distinguish between com-mercially sold ldquoAonorirdquo samples based on sensory evalua-tion Solely in taxonomically different samples OkayamaTokushima and Ehime (Ulva species) versus Shizuoka(Monostroma species) somewhat different scores could beobtained

Volatile compounds are fundamental for algae odor andaromaTherefore it is not surprising that volatile compoundsin algae have been investigated and reports evaluating volatile



2

4

6

8

Scor

e

Shizuoka


001020304050607080Animalic

Floral

Spicy

Fatty


Fresh

Powderly

Leather-like

OkayamaTokushima

EhimeShizuoka





00

200

400

600

800

1000Alkanes

Alkenes

Ketones


Alcohols

Esters

OkayamaTokushima

EhimeShizuoka








ble1Vo

latilec

ompo

unds

ofdifferent

greenalgaes

amples

presentedas

relativer

atiosc

omparedto

theinternalstand

ardethyldecanoate

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D1

Dod

ecane

C 10H

22Alkane

Alkane(B)

1185

ND

ND

ND

1200010

2Tridecane

C 13H

28Alkane

Fatty

acids

Alkane(B)

1297

06

000

0306

000

08ND

232

00298

3226-

Trim

ethylcyclohexano

neC 9

H16O

Ketone

Caroteno

ids

Camph

oraceous

tobaccono

tes

thujon

e-lik

e(A)

1308

07

00011

07

00026

ND

ND

4Tetradecane

C 14H

30Alkane

Fatty

acids

Alkane(B)

1397

39

00020

114

000

9771

00158

60

000

655

Tetradecene

C 14H

28Alkene

1444

05

00013

09

000

0521

00025

ND

6Pentadecane

C 15H

32Alkene

Alkane(B)

1502

836

00622

2511

01181

4205

06057

ND


eC 8

H12O

Ketone

Fatty

acids

caroteno

ids

Alm

ond(C

)1517

ND

ND

ND

ND

8Be

nzaldehyde

C 7H

5OAldehyde

Aminoacids

Sweetstr

ong

almon

d(A

)almon

dbu

rnt

sugar(B)

1517

1700028

23

000

6809

00015

37773

74504

9Pentadeceneisomer

1C 1

5H30

Alkene

1523

24

00028

36

00025

47

00057

117

00326

10Pentadeceneisomer

2C 1

5H30

Alkene

1546

07

00011

23

00025

1000010

05

00028

11Dim

ethylsulfoxide

C 2H

6OS

Sulfu

rcompo

und

Aminoacids

1557

82

00457

165

00944

80

00500

123

00221


eC 8

H12O

Ketone

Fatty

acids

caroteno

ids

Melo

n-lik

e(C)

1567

27

00100

21

000

6506

00052

03

00029

13Pentadeceneisomer

3C 1

5H30

Alkene

1576

ND

ND

ND

51

00085

14266-Trim

ethyl-2

-hydroxycyclohexano

neC 9

H16O

2Ke

tone

Caroteno

ids

Sweet

tobacco-lik

earom

awith

herbaceous

underton

es(A

)

1594

86

00237

9600223

41

00053

ND

15Hexadecane

C 16H

34Alkane

Alkane(B)

1601

43

00089

56

00122

7700144

282

00563

16120573-C

yclocitral

C 10H

16O

Aldehyde

Caroteno

ids

Fresh(B)

1612

35

00085

35

00074

1400010

ND

17Hexadeceneisomer

1C 1

6H32

Alkene

1619

1200010

25

00027

21

00025

ND

18Hexadeceneisomer

2C 1

6H32

Alkene

1624

09

00024

1100026

22

000

04ND

19Safranal

C 10H

14O

Aldehyde

Caroteno

ids

1637

ND

ND

ND

ND

20Heptadecane

C 17H

36Alkane

Alkane(B)

1707

561

00610

252

00936

255

02227

124

00993

217-Heptadecene

C 17H

34Alkene

1722

29097

10740

40033

33306

39543

55587

1555

03312

22Heptadecene

isomer

1C 1

7H34

Alkene

1733

1800193

ND

120

00030

7000169


1C 1

7H32

Alkene

1757

1034

006

041392

01109

1487

01725

643

01246


2C 1

7H32

Alkene

1766

1000021

1200018

05

000

4707

00015


3C 1

7H32

Alkene

1769

43

00022

22

00011

08

00074

07

00104


4C 1

7H32

Alkene

1772

38

00037

53

00036

59

000

4111

00198


5C 1

7H32

Alkene

1779

08

00022

ND

03

00059

05

000

9128

Octadecane

C 18H

38Alkane

Alkane(B)

1801

30

00051

16000

4232

00189

21

00059

29Octadecene

C 18H

36Alkene

1817

112

00072

128

00059

181

00085

09

00081

30120572-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

violet-floral(A

)1842

46

00039

56

00139

1800085

ND


Table1Con

tinued

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D31

Non

adecane

C 19H

40Alkane

Alkane(B)

1898

29

00119

1100039

20

00029

07

00033

32120573-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

dry(A

)1927

125

01087

157

00144

59

000

4007

00031

33Maltol

C 6H

6O3

Ketone

Caramel-

butte

rscotch(A

)caramel(B)

1960

06

00082

ND

ND

1000059

34120573-Io

none

epoxide

C 13H

20O

2Ke

tone

Caroteno

ids

Sweetb

erry

(A)

1980

21

00181

28

00025

09

000

06ND

35Methyltetradecano

ate

C 15H

30O

2Ester

Fainto

nion

ho

ney(A

)orris

(A)(B)

2010

09

00051

00

000

0013

00017

1700036

36Pentadecanal

C 15H

30O

Aldehyde

Fresh(B)

2034

1800103

31

00033

1800103

ND

3761014

-Trim

ethyl-2

-pentadecanon

eC 1

8H36O

Ketone

Caroteno

ids

2139

29

00168

67

00174

7000160

74004

27

38Hexadecanoica

cid

methyleste

rC 1

7H34O

2Ester

Fatty

acids

2217

52

00210

22

00038

9000133

22

00029

39Dihydroactin

idiolid

eC 1

1H16O

2Ke

tone

Caroteno

ids

Musky

orcoum

arin-like

(A)

2347

30

00363

84

00199

42

00022

04

00034

40Methyllinolenate

C 19H

32O

2Ester

Fatty

acids

2566

41

00390

27

000

6373

00268

34

00057

41Ph

ytol

C 20H

40O

Alcoh

olCh

loroph

yll

Faintfl

oral(A

)flo

wer

(B)

2580

28

00191

1200050

09

00023

07

00071

(A)F

oodandAgriculture

Organizationof

theu

nitednatio

ns(http

wwwfaoorg)(B)


)HuandPan

2000

[14]


DodecaneTridecane


Tetradecane


Benzaldehyde

Pentadecene

Pentadecene

Dimethyl sulfoxide

Pentadecene


Hexadecane

Hexadecene

Hexadecene

Safranal

Heptadecane

Heptadecene

Heptadecene

Heptadecadiene


Heptadecadiene

Heptadecadiene

Octadecane

Octadecene

Nonadecane

Maltol


Pentadecanal




Methyl linolenate

Phytol

0

025

05

075

1

025 050 075 1

minus025

minus025

minus075

minus075

minus1

minus1

minus05

minus05


266-Trimethyl-2-


35-Octadien-2-one

61014-Dimethyl

35-Octadiene-2-one

F1 (4813)

226-


(20

14

)








Okayama2Okayama3

Okayama1

1

Tokushima2

Tokushima1

Tokushima3

Ehime3Ehime2

Ehime1

Shizuoka2Shizuoka1

Shizuoka3

0

0

2

2

4

4

6

6

8 10minus8 minus6

minus6

minus4

minus4

minus2

minus2


F2

(20

14

)

F1 (4813)








4 Conclusions




Acknowledgments


References






































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of



2

4

6

8

Scor

e

Shizuoka


001020304050607080Animalic

Floral

Spicy

Fatty


Fresh

Powderly

Leather-like

OkayamaTokushima

EhimeShizuoka





00

200

400

600

800

1000Alkanes

Alkenes

Ketones


Alcohols

Esters

OkayamaTokushima

EhimeShizuoka








ble1Vo

latilec

ompo

unds

ofdifferent

greenalgaes

amples

presentedas

relativer

atiosc

omparedto

theinternalstand

ardethyldecanoate

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D1

Dod

ecane

C 10H

22Alkane

Alkane(B)

1185

ND

ND

ND

1200010

2Tridecane

C 13H

28Alkane

Fatty

acids

Alkane(B)

1297

06

000

0306

000

08ND

232

00298

3226-

Trim

ethylcyclohexano

neC 9

H16O

Ketone

Caroteno

ids

Camph

oraceous

tobaccono

tes

thujon

e-lik

e(A)

1308

07

00011

07

00026

ND

ND

4Tetradecane

C 14H

30Alkane

Fatty

acids

Alkane(B)

1397

39

00020

114

000

9771

00158

60

000

655

Tetradecene

C 14H

28Alkene

1444

05

00013

09

000

0521

00025

ND

6Pentadecane

C 15H

32Alkene

Alkane(B)

1502

836

00622

2511

01181

4205

06057

ND


eC 8

H12O

Ketone

Fatty

acids

caroteno

ids

Alm

ond(C

)1517

ND

ND

ND

ND

8Be

nzaldehyde

C 7H

5OAldehyde

Aminoacids

Sweetstr

ong

almon

d(A

)almon

dbu

rnt

sugar(B)

1517

1700028

23

000

6809

00015

37773

74504

9Pentadeceneisomer

1C 1

5H30

Alkene

1523

24

00028

36

00025

47

00057

117

00326

10Pentadeceneisomer

2C 1

5H30

Alkene

1546

07

00011

23

00025

1000010

05

00028

11Dim

ethylsulfoxide

C 2H

6OS

Sulfu

rcompo

und

Aminoacids

1557

82

00457

165

00944

80

00500

123

00221


eC 8

H12O

Ketone

Fatty

acids

caroteno

ids

Melo

n-lik

e(C)

1567

27

00100

21

000

6506

00052

03

00029

13Pentadeceneisomer

3C 1

5H30

Alkene

1576

ND

ND

ND

51

00085

14266-Trim

ethyl-2

-hydroxycyclohexano

neC 9

H16O

2Ke

tone

Caroteno

ids

Sweet

tobacco-lik

earom

awith

herbaceous

underton

es(A

)

1594

86

00237

9600223

41

00053

ND

15Hexadecane

C 16H

34Alkane

Alkane(B)

1601

43

00089

56

00122

7700144

282

00563

16120573-C

yclocitral

C 10H

16O

Aldehyde

Caroteno

ids

Fresh(B)

1612

35

00085

35

00074

1400010

ND

17Hexadeceneisomer

1C 1

6H32

Alkene

1619

1200010

25

00027

21

00025

ND

18Hexadeceneisomer

2C 1

6H32

Alkene

1624

09

00024

1100026

22

000

04ND

19Safranal

C 10H

14O

Aldehyde

Caroteno

ids

1637

ND

ND

ND

ND

20Heptadecane

C 17H

36Alkane

Alkane(B)

1707

561

00610

252

00936

255

02227

124

00993

217-Heptadecene

C 17H

34Alkene

1722

29097

10740

40033

33306

39543

55587

1555

03312

22Heptadecene

isomer

1C 1

7H34

Alkene

1733

1800193

ND

120

00030

7000169


1C 1

7H32

Alkene

1757

1034

006

041392

01109

1487

01725

643

01246


2C 1

7H32

Alkene

1766

1000021

1200018

05

000

4707

00015


3C 1

7H32

Alkene

1769

43

00022

22

00011

08

00074

07

00104


4C 1

7H32

Alkene

1772

38

00037

53

00036

59

000

4111

00198


5C 1

7H32

Alkene

1779

08

00022

ND

03

00059

05

000

9128

Octadecane

C 18H

38Alkane

Alkane(B)

1801

30

00051

16000

4232

00189

21

00059

29Octadecene

C 18H

36Alkene

1817

112

00072

128

00059

181

00085

09

00081

30120572-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

violet-floral(A

)1842

46

00039

56

00139

1800085

ND


Table1Con

tinued

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D31

Non

adecane

C 19H

40Alkane

Alkane(B)

1898

29

00119

1100039

20

00029

07

00033

32120573-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

dry(A

)1927

125

01087

157

00144

59

000

4007

00031

33Maltol

C 6H

6O3

Ketone

Caramel-

butte

rscotch(A

)caramel(B)

1960

06

00082

ND

ND

1000059

34120573-Io

none

epoxide

C 13H

20O

2Ke

tone

Caroteno

ids

Sweetb

erry

(A)

1980

21

00181

28

00025

09

000

06ND

35Methyltetradecano

ate

C 15H

30O

2Ester

Fainto

nion

ho

ney(A

)orris

(A)(B)

2010

09

00051

00

000

0013

00017

1700036

36Pentadecanal

C 15H

30O

Aldehyde

Fresh(B)

2034

1800103

31

00033

1800103

ND

3761014

-Trim

ethyl-2

-pentadecanon

eC 1

8H36O

Ketone

Caroteno

ids

2139

29

00168

67

00174

7000160

74004

27

38Hexadecanoica

cid

methyleste

rC 1

7H34O

2Ester

Fatty

acids

2217

52

00210

22

00038

9000133

22

00029

39Dihydroactin

idiolid

eC 1

1H16O

2Ke

tone

Caroteno

ids

Musky

orcoum

arin-like

(A)

2347

30

00363

84

00199

42

00022

04

00034

40Methyllinolenate

C 19H

32O

2Ester

Fatty

acids

2566

41

00390

27

000

6373

00268

34

00057

41Ph

ytol

C 20H

40O

Alcoh

olCh

loroph

yll

Faintfl

oral(A

)flo

wer

(B)

2580

28

00191

1200050

09

00023

07

00071

(A)F

oodandAgriculture

Organizationof

theu

nitednatio

ns(http

wwwfaoorg)(B)


)HuandPan

2000

[14]


DodecaneTridecane


Tetradecane


Benzaldehyde

Pentadecene

Pentadecene

Dimethyl sulfoxide

Pentadecene


Hexadecane

Hexadecene

Hexadecene

Safranal

Heptadecane

Heptadecene

Heptadecene

Heptadecadiene


Heptadecadiene

Heptadecadiene

Octadecane

Octadecene

Nonadecane

Maltol


Pentadecanal




Methyl linolenate

Phytol

0

025

05

075

1

025 050 075 1

minus025

minus025

minus075

minus075

minus1

minus1

minus05

minus05


266-Trimethyl-2-


35-Octadien-2-one

61014-Dimethyl

35-Octadiene-2-one

F1 (4813)

226-


(20

14

)








Okayama2Okayama3

Okayama1

1

Tokushima2

Tokushima1

Tokushima3

Ehime3Ehime2

Ehime1

Shizuoka2Shizuoka1

Shizuoka3

0

0

2

2

4

4

6

6

8 10minus8 minus6

minus6

minus4

minus4

minus2

minus2


F2

(20

14

)

F1 (4813)








4 Conclusions




Acknowledgments


References






































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


ble1Vo

latilec

ompo

unds

ofdifferent

greenalgaes

amples

presentedas

relativer

atiosc

omparedto

theinternalstand

ardethyldecanoate

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D1

Dod

ecane

C 10H

22Alkane

Alkane(B)

1185

ND

ND

ND

1200010

2Tridecane

C 13H

28Alkane

Fatty

acids

Alkane(B)

1297

06

000

0306

000

08ND

232

00298

3226-

Trim

ethylcyclohexano

neC 9

H16O

Ketone

Caroteno

ids

Camph

oraceous

tobaccono

tes

thujon

e-lik

e(A)

1308

07

00011

07

00026

ND

ND

4Tetradecane

C 14H

30Alkane

Fatty

acids

Alkane(B)

1397

39

00020

114

000

9771

00158

60

000

655

Tetradecene

C 14H

28Alkene

1444

05

00013

09

000

0521

00025

ND

6Pentadecane

C 15H

32Alkene

Alkane(B)

1502

836

00622

2511

01181

4205

06057

ND


eC 8

H12O

Ketone

Fatty

acids

caroteno

ids

Alm

ond(C

)1517

ND

ND

ND

ND

8Be

nzaldehyde

C 7H

5OAldehyde

Aminoacids

Sweetstr

ong

almon

d(A

)almon

dbu

rnt

sugar(B)

1517

1700028

23

000

6809

00015

37773

74504

9Pentadeceneisomer

1C 1

5H30

Alkene

1523

24

00028

36

00025

47

00057

117

00326

10Pentadeceneisomer

2C 1

5H30

Alkene

1546

07

00011

23

00025

1000010

05

00028

11Dim

ethylsulfoxide

C 2H

6OS

Sulfu

rcompo

und

Aminoacids

1557

82

00457

165

00944

80

00500

123

00221


eC 8

H12O

Ketone

Fatty

acids

caroteno

ids

Melo

n-lik

e(C)

1567

27

00100

21

000

6506

00052

03

00029

13Pentadeceneisomer

3C 1

5H30

Alkene

1576

ND

ND

ND

51

00085

14266-Trim

ethyl-2

-hydroxycyclohexano

neC 9

H16O

2Ke

tone

Caroteno

ids

Sweet

tobacco-lik

earom

awith

herbaceous

underton

es(A

)

1594

86

00237

9600223

41

00053

ND

15Hexadecane

C 16H

34Alkane

Alkane(B)

1601

43

00089

56

00122

7700144

282

00563

16120573-C

yclocitral

C 10H

16O

Aldehyde

Caroteno

ids

Fresh(B)

1612

35

00085

35

00074

1400010

ND

17Hexadeceneisomer

1C 1

6H32

Alkene

1619

1200010

25

00027

21

00025

ND

18Hexadeceneisomer

2C 1

6H32

Alkene

1624

09

00024

1100026

22

000

04ND

19Safranal

C 10H

14O

Aldehyde

Caroteno

ids

1637

ND

ND

ND

ND

20Heptadecane

C 17H

36Alkane

Alkane(B)

1707

561

00610

252

00936

255

02227

124

00993

217-Heptadecene

C 17H

34Alkene

1722

29097

10740

40033

33306

39543

55587

1555

03312

22Heptadecene

isomer

1C 1

7H34

Alkene

1733

1800193

ND

120

00030

7000169


1C 1

7H32

Alkene

1757

1034

006

041392

01109

1487

01725

643

01246


2C 1

7H32

Alkene

1766

1000021

1200018

05

000

4707

00015


3C 1

7H32

Alkene

1769

43

00022

22

00011

08

00074

07

00104


4C 1

7H32

Alkene

1772

38

00037

53

00036

59

000

4111

00198


5C 1

7H32

Alkene

1779

08

00022

ND

03

00059

05

000

9128

Octadecane

C 18H

38Alkane

Alkane(B)

1801

30

00051

16000

4232

00189

21

00059

29Octadecene

C 18H

36Alkene

1817

112

00072

128

00059

181

00085

09

00081

30120572-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

violet-floral(A

)1842

46

00039

56

00139

1800085

ND


Table1Con

tinued

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D31

Non

adecane

C 19H

40Alkane

Alkane(B)

1898

29

00119

1100039

20

00029

07

00033

32120573-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

dry(A

)1927

125

01087

157

00144

59

000

4007

00031

33Maltol

C 6H

6O3

Ketone

Caramel-

butte

rscotch(A

)caramel(B)

1960

06

00082

ND

ND

1000059

34120573-Io

none

epoxide

C 13H

20O

2Ke

tone

Caroteno

ids

Sweetb

erry

(A)

1980

21

00181

28

00025

09

000

06ND

35Methyltetradecano

ate

C 15H

30O

2Ester

Fainto

nion

ho

ney(A

)orris

(A)(B)

2010

09

00051

00

000

0013

00017

1700036

36Pentadecanal

C 15H

30O

Aldehyde

Fresh(B)

2034

1800103

31

00033

1800103

ND

3761014

-Trim

ethyl-2

-pentadecanon

eC 1

8H36O

Ketone

Caroteno

ids

2139

29

00168

67

00174

7000160

74004

27

38Hexadecanoica

cid

methyleste

rC 1

7H34O

2Ester

Fatty

acids

2217

52

00210

22

00038

9000133

22

00029

39Dihydroactin

idiolid

eC 1

1H16O

2Ke

tone

Caroteno

ids

Musky

orcoum

arin-like

(A)

2347

30

00363

84

00199

42

00022

04

00034

40Methyllinolenate

C 19H

32O

2Ester

Fatty

acids

2566

41

00390

27

000

6373

00268

34

00057

41Ph

ytol

C 20H

40O

Alcoh

olCh

loroph

yll

Faintfl

oral(A

)flo

wer

(B)

2580

28

00191

1200050

09

00023

07

00071

(A)F

oodandAgriculture

Organizationof

theu

nitednatio

ns(http

wwwfaoorg)(B)


)HuandPan

2000

[14]


DodecaneTridecane


Tetradecane


Benzaldehyde

Pentadecene

Pentadecene

Dimethyl sulfoxide

Pentadecene


Hexadecane

Hexadecene

Hexadecene

Safranal

Heptadecane

Heptadecene

Heptadecene

Heptadecadiene


Heptadecadiene

Heptadecadiene

Octadecane

Octadecene

Nonadecane

Maltol


Pentadecanal




Methyl linolenate

Phytol

0

025

05

075

1

025 050 075 1

minus025

minus025

minus075

minus075

minus1

minus1

minus05

minus05


266-Trimethyl-2-


35-Octadien-2-one

61014-Dimethyl

35-Octadiene-2-one

F1 (4813)

226-


(20

14

)








Okayama2Okayama3

Okayama1

1

Tokushima2

Tokushima1

Tokushima3

Ehime3Ehime2

Ehime1

Shizuoka2Shizuoka1

Shizuoka3

0

0

2

2

4

4

6

6

8 10minus8 minus6

minus6

minus4

minus4

minus2

minus2


F2

(20

14

)

F1 (4813)








4 Conclusions




Acknowledgments


References






































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


Table1Con

tinued

No

Volatilec

ompo

unds

Form

ula

Class

Orig

inOdo

rdescrip

tion

RIRe

lativec

ontents(averaged

GCpeak

area

ratio

(119899=3))

Okayama

STD

Toku

shim

aST

DEh

ime

STD

Shizuo

kaST

D31

Non

adecane

C 19H

40Alkane

Alkane(B)

1898

29

00119

1100039

20

00029

07

00033

32120573-Io

none

C 13H

20O

Ketone

Caroteno

ids

Warmw

oody

dry(A

)1927

125

01087

157

00144

59

000

4007

00031

33Maltol

C 6H

6O3

Ketone

Caramel-

butte

rscotch(A

)caramel(B)

1960

06

00082

ND

ND

1000059

34120573-Io

none

epoxide

C 13H

20O

2Ke

tone

Caroteno

ids

Sweetb

erry

(A)

1980

21

00181

28

00025

09

000

06ND

35Methyltetradecano

ate

C 15H

30O

2Ester

Fainto

nion

ho

ney(A

)orris

(A)(B)

2010

09

00051

00

000

0013

00017

1700036

36Pentadecanal

C 15H

30O

Aldehyde

Fresh(B)

2034

1800103

31

00033

1800103

ND

3761014

-Trim

ethyl-2

-pentadecanon

eC 1

8H36O

Ketone

Caroteno

ids

2139

29

00168

67

00174

7000160

74004

27

38Hexadecanoica

cid

methyleste

rC 1

7H34O

2Ester

Fatty

acids

2217

52

00210

22

00038

9000133

22

00029

39Dihydroactin

idiolid

eC 1

1H16O

2Ke

tone

Caroteno

ids

Musky

orcoum

arin-like

(A)

2347

30

00363

84

00199

42

00022

04

00034

40Methyllinolenate

C 19H

32O

2Ester

Fatty

acids

2566

41

00390

27

000

6373

00268

34

00057

41Ph

ytol

C 20H

40O

Alcoh

olCh

loroph

yll

Faintfl

oral(A

)flo

wer

(B)

2580

28

00191

1200050

09

00023

07

00071

(A)F

oodandAgriculture

Organizationof

theu

nitednatio

ns(http

wwwfaoorg)(B)


)HuandPan

2000

[14]


DodecaneTridecane


Tetradecane


Benzaldehyde

Pentadecene

Pentadecene

Dimethyl sulfoxide

Pentadecene


Hexadecane

Hexadecene

Hexadecene

Safranal

Heptadecane

Heptadecene

Heptadecene

Heptadecadiene


Heptadecadiene

Heptadecadiene

Octadecane

Octadecene

Nonadecane

Maltol


Pentadecanal




Methyl linolenate

Phytol

0

025

05

075

1

025 050 075 1

minus025

minus025

minus075

minus075

minus1

minus1

minus05

minus05


266-Trimethyl-2-


35-Octadien-2-one

61014-Dimethyl

35-Octadiene-2-one

F1 (4813)

226-


(20

14

)








Okayama2Okayama3

Okayama1

1

Tokushima2

Tokushima1

Tokushima3

Ehime3Ehime2

Ehime1

Shizuoka2Shizuoka1

Shizuoka3

0

0

2

2

4

4

6

6

8 10minus8 minus6

minus6

minus4

minus4

minus2

minus2


F2

(20

14

)

F1 (4813)








4 Conclusions




Acknowledgments


References






































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


DodecaneTridecane


Tetradecane


Benzaldehyde

Pentadecene

Pentadecene

Dimethyl sulfoxide

Pentadecene


Hexadecane

Hexadecene

Hexadecene

Safranal

Heptadecane

Heptadecene

Heptadecene

Heptadecadiene


Heptadecadiene

Heptadecadiene

Octadecane

Octadecene

Nonadecane

Maltol


Pentadecanal




Methyl linolenate

Phytol

0

025

05

075

1

025 050 075 1

minus025

minus025

minus075

minus075

minus1

minus1

minus05

minus05


266-Trimethyl-2-


35-Octadien-2-one

61014-Dimethyl

35-Octadiene-2-one

F1 (4813)

226-


(20

14

)








Okayama2Okayama3

Okayama1

1

Tokushima2

Tokushima1

Tokushima3

Ehime3Ehime2

Ehime1

Shizuoka2Shizuoka1

Shizuoka3

0

0

2

2

4

4

6

6

8 10minus8 minus6

minus6

minus4

minus4

minus2

minus2


F2

(20

14

)

F1 (4813)








4 Conclusions




Acknowledgments


References






































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of


Okayama2Okayama3

Okayama1

1

Tokushima2

Tokushima1

Tokushima3

Ehime3Ehime2

Ehime1

Shizuoka2Shizuoka1

Shizuoka3

0

0

2

2

4

4

6

6

8 10minus8 minus6

minus6

minus4

minus4

minus2

minus2


F2

(20

14

)

F1 (4813)








4 Conclusions




Acknowledgments


References






































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of































Journal of

Chemistry


Advances in

Physical Chemistry



Journal of

Volume 2014














Journal of

Spectroscopy



Journal of


Quantum Chemistry






CatalystsJournal of

research article determination of volatile compounds in four...

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