Pelita Perkebunan 27(1) 2011, 24-35

Analysis of Pyrazine and Volatile Compounds in Cocoa

Beans Using Solid Phase Microextraction

Analisis Pirazin dan Senyawa Volatil pada Biji Kakao

Menggunakan Mikroekstraksi Fase Padat

Misnawi1*) and Ariza Budi Tunjung Sari 1)

Summary

Analysis of pyrazine and volatile compounds in cocoa beans was done by

using solid phase microextraction (SPME), to develop efficient non solvent ex-
traction method. Extraction was carried out in head space technique using
stableflex fiber coated with DVB/Carboxen/PDMS applied on manual sampling
SPME Holder. Five grams of roasted fermented cocoa bean was processed into
butter and placed into 30 ml vial and capped with a rubber septum, then heated
at temperature of 70OC for 30 min for the extraction. The fiber then was placed
in GC header for desorption and separation. Results of the study showed that
the SPME extracted pyrazines were adequate and well detected in a gas chro-
matography system. Peak area resulted from SPME covered 2.83–5.35% peak
area from syringe, however SPME had comparable ability to syringe in extract-
ing volatile compounds. Five most common pyrazines in cocoa bean aroma were
identified, such as 2 methyl pyrazine (2MP); 2.3 and 2.5 dimethyl pyrazine (DMP);
and 2,3,5 trimethyl pyrazine (TrMP) and tetramethylpyrazine (TMP). Other cor-
responding compounds were also detected in cocoa liquor, i.e. alcohols, car-
boxylic acids, aldehydes, ketons, esters, pyrazines, amines and other volatile
compounds and strongly associated to chocolate aroma. The successful extrac-
tion of pyrazine and volatile-semi volatile compounds which contribute to choco-
late aroma indicates SPME is applicable in flavor analysis.

Key words: Cocoa bean, flavour, pyrazine, solid-phase microextraction, head space, extrac-
tion, gas chromatography, maillard.

R ing k a sa n

Analisis pirazin dan senyawa volatil pada biji kakao dilakukan dengan
perangkat mikroekstraksi fase padat (solid phase micro extraction, SPME), untuk
mengembangkan metode ekstraksi tanpa pelarut yang efisien. Perangkat SPME
dilengkapi fiber stableflex dengan polimer DVB/Carboxen/PDMS yang menjerap
senyawa volatil di area headspace. Biji kakao terfermentasi disangrai dan diambil
lemaknya untuk ditempatkan dalam botol bertutup septa. Sampel dipanaskan
pada suhu 70OC dan serat SPME ditusukkan menembus septa untuk mengekstrak
senyawa volatil dari lemak kakao selama 30 menit. Senyawa volatil lemak kakao
akan dijerap oleh serat SPME dan dilepaskan kembali untuk analisis kromatografi
gas. Penelitian menunjukkan pirazin dan senyawa volatil yang diekstrak oleh
serat SPME dapat terdeteksi dengan baik oleh kromatografi gas. Area puncak
yang dihasilkan SPME meliputi 2,83–5,35% dari area puncak yang dihasilkan

Naskah diterima (received) 2 Desember 2010, disetujui (accepted) 24 Maret 2010.

1) Pusat Penelitian Kopi dan Kakao Indonesia, Jl. PB. Sudirman No. 90, Jember, Indonesia.
*) Alamat penulis (Corresponding Author): misnawi@yahoo.com

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 Analysis of pyrazine and volatile compounds of cocoa beans using solid phase microextraction

syringe, kendati demikian kemampuan ekstraksi SPME dapat disetarakan dengan

syringe. Lima jenis pirazin yang sering terdapat di biji kakao telah diidentifikasi,
meliputi metil pirazin (2MP); 2,3 dan 2,5-dimetilpirazin (DMP); dan 2,3,5
trimetilpirazin (TrMP) dan tetrametil pirazin (TMP). Senyawa lainnya juga
terdeteksi meliputi alkohol, asam karboksilat, aldehida, keton, ester, pirazin,
amin dan senyawa volatil lainnya, dan diketahui erat kaitannya dengan aroma
khas cokelat. Keberhasilan SPME dalam ekstraksi pirazin dan senyawa volatil-
semi volatil yang berperan penting dalam pembentukan aroma cokelat
menandakan SPME dapat digunakan lebih lanjut untuk analisis citarasa.

INTRODUCTION which binds volatiles on a silica fiber coated

by polymer in headspace area of sample
Pyrazines (1,4 diazines) in roasted co- (Berlardi & Pawliszyn, 1989). Exposing fi-
coa bean contribute to a pleasant aroma of ber at headspace area ensures that only
chocolate, and associate with good quality volatile compounds reach and contact to fi-
of roasted cocoa bean perception. Large ber. Extractions can be conducted in room
number of pyrazines have been identified temperature (Vazquez-Landaverde et al.,
in cocoa and are considered extremely im- 2008; Marsili, 2002), but moderate heat
portant in cocoa flavor (Brunetto et al., often increase volatility (Ducki et al., 2008).
2009; Frauendorfer & Schieberle, 2008; Trapped volatile then was released in the
Krings et al., 2006; Bonvehi & Coll, 2002; GC system during injection through des-
Counet et al., 2002; Jinap et al., 1998). De- orption mechanism (Pawliszyn et al., 1997).
tection of volatile compounds including
pyrazine has been developed using gas SPME differs due to its type of fibers;
chromatography which requires extraction polar, non-polar and bipolar fibers. Each
of those compounds from solid or liquid type of fiber adsorbs different type of com-
sample. Schultz et al. (1977) developed a pounds. Polar fiber may be coated by
method of extraction using simultaneous polyacrylate (PA) (Shirey, 1999) or
distillation and extraction (SDE), in which carbowax-divinylbenzene (CW-DVB)
sample is water-steamed resulting evapo- (Shirey & Sidisky, 1999). Nonpolar fiber
ration of volatile compounds which then may be constructed of polydimethyl silox-
being captured by certain solution and con- ane (PDMS). While bipolar fibers are com-
centrated prior to syringe injection into GC bination between polar and nonpolar poly-
system. This SDE-syringe method involves mer. Bipolar fiber could capture larger
high temperature to extract and move the spectrum of compounds and could be de-
volatile compound into trapping solution veloped from materials of poly-
which could detriment the nature of dimethylsiloxane-divinylbenzene (PDMS-
pyrazine. Since pyrazines are product of DVB) or Carboxen-PDMS (Shirey & Sidisky,
heating process, bias in analysis could be 1999).
occurred due to uses of high temperature This research investigated analysis of
during steam distillation. pyrazine and volatile compounds of cocoa
Solid phase microextraction (SPME) is beans extracted by SPME and evaluated to-
developed to allow extraction of volatile ward SDE-syringe extraction. Analysis was
compounds without use of any solvent. brought by gas chromatography-mass
SPME is a solventless extraction technique spectrometry, focusing on pyrazine analysis
introduced by Berlardi & Pawliszyn (1989), especially 2-methylpyrazine (2-MP);

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25
2,5-dimethylpyrazine (2,5DMP); 2,3- and flame ionization detector for pyrazine
dimethylpyrazine (2,3-DMP); 2,3,5-trime- analysis, and mass spectrometry for vola-
thylpyrazine (TrMP) and tetrame- tile compounds analysis. Bipolar fiber
thylpyrazine (TMP). PDMS-DVB SPME from Supelco was used
for extraction.

MATERIAL AND METHODS

Method of extraction
Cocoa beans used were fermented bulk
cocoa beans obtained from Post Harvest Referring to successful pyrazine extrac-
Laboratory of Indonesian Coffee and Co- tion from peanut butter (Supelco, 1998), this
coa Research Institute (ICCRI). The beans study used cocoa butter as the base of
were roasted at temperature of 150OC for pyrazines extraction. Three 30 mL vials,
45 min and manually processed for shell each contained five grams of roasted fer-
removal. A quantity of roasted beans was mented cocoa butter and capped with a rub-
pressed to collect the butter for pyrazine ber septum were immersed in the water-
analysis, while the other was ground to ob- bath. To observe effect of heat to volatile
tain cocoa liquor for volatile compounds compounds extraction, immersion was pre-
pared at temperatures of 50O C, 60OC and
analysis.
70OC respectively. The vial was heated for
Gas chromatography system consisted 30 min for the extraction. The fiber was
of Shimadzu GC-2010 equipped with then placed in GC injector for desorption
Rtx-1 (100% dimethyl polisiloxane) column and separation in capillary column.

Extraction Procedure Desorption Procedure

Prosedur Ekstraksi Prosedur Desorpsi

Retract fiber/remove Retract fiber/remove

Tarik fiber dan Tarik fiber dan
Pierce sample septum Pierce GC inlet septum keluarkan
Suntik septum botol keluarkan
Suntik septum
sampel inlet GC Expose fiber/desorb
Expose fiber/extract keluarkan fiber/desorpsi
Keluarkan fiber/
ekstraksi

Figure 1. Mechanism of extraction and desorption of SPME.

Gambar 1. Mekanisme ekstraksi dan desorpsi SPME.

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 Analysis of pyrazine and volatile compounds of cocoa beans using solid phase microextraction

Table 1. GC conditions applied in Supelco (1998) and Misnawi et al. (2004) pyrazines detection and modified methods
Tabel 1. Parameter kromatografi gas berdasarkan metode Supelco (1998) dan Misnawi et al. (2004)

Properties Supelco (1998) Misnawi et al.(2004) Modified

Parameter

Sample preparation and Headspace SPME- Steam Destillation- Headspace SPME-

injection SPME SPME SPME
Penyiapan contoh dan
injeksi

Injector Parameter (Parameter infektor)

Inlet Liner 0.75 mm 4 mm 0.75 mm
Temperature (Suhu) 270 O C 200 O C splitless/260O C
Pyrazine source (Sumber pirazin) Peanut Cocoa bean Cocoa butter
Column (Kolom) Supelcowax 1030 m x HP-20 M50 m x Rtx-1(Dimethyl
0.25 mm ID x 0.25 µm 0.32 mm ID x 0.3 µm Polysiloxane) 30 m x
I,25 mm ID x 0.25 µm
Oven (Oven)
Initial temperature (Suhu awal) 40O C 60O C 60 OC
Equilibrium (Keseimbangan) 5 min 3 min 3 min
Rate (Laju) 4OC/min 5OC/min 5OC/min
Final temperature (Suhu akhir) 230 O C 180 O C 200 O C
Hold - 5 min -

Detector (Detektor) Ion trap mass spectro- Flame Ionization Flame Ionization
meter, selected ions Detector (FID) Detector (FID)
used for quantification for pyrazine analysis, Mass
Spectrometry (Shimadzu GC-MS 2010)

GC Conditioning pounds for further analysis in GC. Supelco

(1998) recommends using polar fiber for
Gas chromatography adjustment used optimum extraction of polar compounds
in this study was obtained from several pre- and vice versa. Bipolar fiber is expected to
liminary experiments to obtain the best re- provide larger spectrum of adsorption than
sponse of the GC system on pyrazine com- the other two types of fiber. PDMS-DVB bi-
pounds separation and detection. Two polar fiber was employed to obtain most of
methods were used as references i.e.a pyrazine in cocoa butter.
method for peanut pyrazine detection with
SPME extraction from Supelco (1998) and The extraction of both pyrazine and
GC condition for cocoa pyrazine detection volatile compounds in fiber was through
with SDE (simultaneous distillation extrac- adsorption mechanism. There are two pos-
tion) developed by Misnawi et al. (2004). sibilities of how compounds attached on the
Those methods were combined to obtain a fiber, by absorption or adsorption. Absorp-
new detection method as described in tion mechanism is when compounds bind
Table 1. chemically with the matrix, while in ad-
sorption mechanism, compounds only ad-
here on the surface of matrix. After being
RESULTS AND DISCUSSION adsorbed onto fiber surface, volatile com-
pounds subsequently were released by cer-
Performance of SPME tain desorption procedure.
The bipolar fiber PDMS-DVB used in Extraction of analytes will continue
this experiment was able to extract pyrazine running until the fiber reach its equilib-
from cocoa butter and release volatile com- rium state. At equilibrium point, concen-

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27
 45000
40000
35000
Unit (satuan), mV

30000
25000
20000
15000
10000
5000

45000
40000
35000
Unit (satuan), mV

30000
25000
20000
15000
10000
5000

Figure 2. Chromatogram of volatile compounds extracted from cocoa butter by using SPME (above)
and seven pyrazine standards (below).
Gambar 2. Kromatogram senyawa volatil yang diekstrak dari lemak kakao menggunakan SPME (atas)
dan senyawa pirazin yang dijadikan standar (bawah).

tration of analyte in the fiber is equal with 230OC. Injector temperature above 200O C
concentration of analyte in sampling area, was recommended to accommodate desorp-
Further exposion will result in minor ef- tion of analyte from fiber. This is also con-
fect, since the fiber would meet its limit of firmed by works of Kumazawa et al. (1999),
trapping. Equilibrium state usually accom- Vilchez et al. (2001) and Perraudine et al.
plished in 30 minutes (da Silva, 2008; (2006) that applied injector temperature
Marsili, 2002). over than 200OC. Therefore column con-
The extraction process was followed by structed of dimethylpolisiloxane-crossbond
injection to GC instrument. There are two were applied due to its ability to retain high
methods of setting GC parameters, the first temperature up to 300OC.
developed by Supelco and the second de- The modification allowed better com-
veloped by Misnawi et al. (2004). Supelco ponent separation and took relatively
has successfully analyzed pyrazines from shorter time. Chromatogram of the modi-
peanut butter, while Misnawi et al. (2004) fied method in the volatile compounds
have analysed pyrazine from cocoa bean separation is shown in Figure 2. Standard
by applying SDE extraction. solution consisting of 2-methylpyrazine
Adjustment made to those methods, in- (2MP); 2,5 dimethylpyrazine (2,5 DMP); 2,3
jector temperature was referred to Supelco dimethylpyrazine (2,3 DMP); 2,3,5
methods that apply injector temperature of trimethylpyrazine and 2,3,5,6 tetra-

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 Analysis of pyrazine and volatile compounds of cocoa beans using solid phase microextraction

methylpyrazine (2,3,5,6 TMP) were ex- peak area of 2,3 DMP > 2,3,5 TrMP > 2
tracted by SPME apparatus. Beside pyrazines, MP > 2,5 DMP.
numbers of volatile and semi-volatile com- The difference might be due to com-
pounds also appeared, such as carboxilyc pound volatility, and also due to difference
acid, hydrocarbons, alcohols, aldehides and in polarity. Interaction between compounds
esters. and SPME polymer was suggested to
This result showed that the developed occurr within polarity variations. Matrix
method was able to detect most of pyrazine effect has been an issue related to the use
compound presented in roasted cocoa bean. of SPME, which affects extraction-desorp-
The 2,5-DMP, 2.3-DMP, 2,3,5-TrMP and tion performance of SPME fiber (Górecki et
tetramethylpyrazine were detected at reten- al., 1999). However by specifying target
tion time of 8.539, 8.733, 11.088 and compound and selecting suitable fiber, this
13.715 minutes, respectively. Due to its mo- matrix effect could be minimized.
lecular weight which affects volatility, 2MP
was among the compounds identified in
early minutes, along with simple carboxylic Effect of extraction temperature
acids, aldehyde and alkane. Detection of
Extraction temperature was set at three
2MP was followed by 2,5 DMP; 2.3 DMP;
levels, 50OC, 60O C and 70O C. GC analy-
2,3,5 TMP and acetylpyrazine.
sis detected 27 compounds, 33 compounds
Compared to SDE-syringe application, and 34 compounds, respectively for extrac-
concentration of extracted compounds us- tion temperature of 50OC, 60OC and 70OC.
ing SPME was much smaller. Peak area re- Temperature of 70OC facilitated liberation
sulted from SPME analysis covered 2.83– of volatile compounds. At this condition,
5.35% of peak area resulted from syringe cocoa butter as the matrix melts down and
(Figure 3). When sampling was carried out provides ways for volatile compounds to
using syringe, all molecules in sample was evaporate (Figure 6).
inhaled into the syringe. While in extrac-
Extraction temperature also affected
tion carried out using SPME, the molecules
peak area performed by volatile compounds.
will be trapped in limited amount.
Compounds of 3-methyl-butyl-acetate;
Adsorbing rate of SPME decreased as the
2,5-dimethyl pyrazine and 2,3-dimethyl-
concentration of analyte increased
pyrazine were found in larger peak area
(Figure 4). This occurred as the fiber became
at temperature 50OC, and decreasing as the
saturated and was unable to trap additional
extraction temperature raised. However
molecules. This saturation stage suggested
benzaldehyde, phenylethyl alcohol and 2-
equilibrium state that is normally accom-
phenylethyl ester showed inclination in
plished in 30 minutes.
peak area at extraction temperature 60O C
The molecules adsorbed in various and 70OC. This result indicated that mod-
quantity, depends on thickness of polymer erate heat facilitated more deliberation of
as well as their own physical properties such compounds in high molecular weight.
as polarity and boiling point. As being
Heat available in the extraction phase
shown in Figure 5, peak area resulted by
also supplies energy to molecules to reach
syringe extraction shows that 2MP has
its boiling point. 2-methyl pyrazine under
larger peak area then 2,3 DMP > 2,5 DMP
76 cmHg, has boiling point of 135OC; 2,3-
> 2,3,5 TrMP. While peak area resulted
dimethyl pyrazine and 2,5-dimethyl
from SPME shows different sequence, where

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29
 25

Luas area puncak, x10 m3, mv

20
Peak area, x10 m3, mv

y=4E+07x-71407
15
R2=0.9869

10

5 y=307135x+552425
R2=0.8779

0
25 25 25 25 25
Pyrazine concentration (Konsentrasi pirazin), ppm

Pyrazine-
Syringe SPME

Figure 3. Comparison of peak area resulted from syringe extraction and SPME
Gambar 3. Perbandingan luas area puncak yang dihasilkan ekstraksi syringe dan SPME.

25
25 22.86

20
20
Adsorped analyte, %
Analit terjerap, %

15
15

10
10

5.35
55 3.3 2.83 2.91
1.62

00
1 2 3 4 5 6
1 2 3 4 5 6 7
Pyrazine concentration, ppm
Konsentrasi pirazin, ppm

Figure 4. Effect of pyrazine concentration on the percentage of adsoped analyte.

Gambar 4. Pengaruh konsentrasi pirazin terhadap persentase analit terjerab.

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 Analysis of pyrazine and volatile compounds of cocoa beans using solid phase microextraction

1200000

1000000
Luas area puncak, x 10m3, mL
Peak area, x 10m3, mL

800000

600000

400000

200000

0
2 MP 2,5 DMP 2,3 DMP 2,3,5 TrMP

Syringe SPME

Figure 5. Comparison of peak area of four pyrazines extracted by syringe dan SPME.
Gambar5. Perbandingan luas area empat pirazin yang diekstrak dengan syringe dan SPME.

pyrazine have boiling point of 155OC, while Extraction of Cocoa Volatile

2,3,5-trimethyl pyrazine has boiling point Compounds by using SPME
of 171OC (Burdock, 2005).
Detection of other volatile compounds
Most of pyrazines underwent loss of in cocoa bean extracted by SPME was per-
quantity during heat treatment, as occurred formed using GC-MS. Complete separation
in pyrazine, 2-MP, 2,5-DMP, 2,3-DMP and of cocoa aroma chromatograms were ob-
acetylpyrazine (Figure 7). Significant loss tained during running time for total 36
was found in 2,5-DMP and ACP, where the min, yet the chromatogram at 30 min run-
compounds were almost eliminated. Slight ning time showed small peak areas. Iden-
reductions were found in pyrazine and tification by using GC-MS library showed
2-MP, which the final amount was is some- that the major peaks were acetic acid,
what less than it was. tetramethyl pyrazine, 3-methyl pentanoic
Instead of heat loss, TMP and TrMP un- acid and 2,3-dimethyl oxirane exposed at
derwent increasing quantity at higher tem- retention time (RT) of 17.54, 18.26, 22.52
perature. TrMP was suggested being syn- and 25.99 min, respectively. Other vola-
thesized during heat treatment, as being tile compound such as dodecanoic acid (RT
showed by gradual supplementation. Ad- 26.12), benzene-acetaldehyde (RT 28.59)
ditional TMP was detected in 70OC, after mi- and 1,4-bis (morpholinoacetyl)piperazine
nor reduction in 60OC. (RT 32.70) were detected in small peak
areas.

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31
Unit (satuan), mV

Figure 6. Chromatogram of cocoa butter, headspace extraction in 70OC.

Gambar 6. Kromatogram senyawa volatil lemak kakao, ekstraksi headspace pada 70OC .

10
9
8
Pyrazine (Pirazin), ppm

7
6
5
4
3
2
1
0
pyrazine 2MP 2,5DMP 2,3DMP 2,3,5TrMP AcP TMP

50°C 60°C 70°C

Figure 7. Effect of extraction temperature on seven pyrazine compounds.

Gambar 7. Pengaruh suhu ekstraksi terhadap konsentrasi 7 senyawa pirazin.

Analysis showed that 36 compounds volatile compounds associated in fermented

were detected in fermented cocoa liquor cocoa aroma. Frauendorfer and Schieberle
volatile compounds which representing (2006) identified 35 most active compounds
alcohols, carboxylic acids, aldehydes, ketons, from cocoa powder extract off however
esters, pyrazines, amines and other vola- Jinap et al. (1998) stated that the main
tile compounds (Table 2). They were most aroma compounds contribute to chocolate

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32
 Analysis of pyrazine and volatile compounds of cocoa beans using solid phase microextraction

Table 2. Volatile compounds of cocoa beans extracted by SPME

Tabel 2. Senyawa volatile dari biji kakao yang diekstrak menggunakan SPME
Volatile compounds Coresponding odour Volatile compounds Coresponding odour
Senyawa volatil Senyawa volatil
Alcohol (Alkohol) Ester
2,3-Butanediol sweet, creamy 1-Butanol, 3-methyl-, lemon-like, flowery,
acetate sweet, fruity
Phenylethyl Alcohol caramel-like, alcohol-like, Acetic acid, 2-phenylethyl cereal-like, roasted cocoa
sweet ester
Cyclobutanol roasted cocoa Propanol, methoxy-, acetate flowery, green
2-Nonanol no smell Benzeneacetic acid, ethyl nutty, bean-like
ester
2-Heptanol no smell Decanoic acid, ethyl ester nutty
1,6-Octadien-3-ol, 3,7- bean-like Dodecanoic acid, ethyl ester sweet, creamy
dimethyl-
Carboxylic acid Propanol, methoxy-, acetate no smell
Acetic acid sour, nutty Octanoic acid, ethyl ester alcohol-like
Pentanoic acid, 3- sweet, rancid Pyrazines
methyl-
Octanoic Acid sweet, chocolate-like Pyrazine, trimethyl- sweet,nutty, bean-like,
smoky
n-Hexadecanoic acid no smell Pyrazine, tetramethyl- bean-like, chocolate,
rancid
Butanoic acid, 3-methyl- sour 2,3- Dimethy l-5- roasted cocoa
Aldehyde ethylpyrazine no smell
2,3,5-Trimethyl-6-
Benzaldehyde bean-like ethylpyrazine
Benzeneacetaldehyde, caramel-like, smooky, Amines caramel-like, sweet
alpha ethylidene- nutty 1,4-
Bis(morpholinoacetyl)
5-Methyl-2-phenyl-2- roasted cocoa piperazine caramel-like
hexenal 7H-Pyrro lo(2,3-
Keton d)pyrimidin- citrus-like
Ethanone, 1-(1H-pyrrol- sweet, caramel-like, 4-amine
2-yl)- honey- like, nutty 1,2-Propanediamine
Cyclobutanone, 2-ethyl- no smell Other volatiles smoky

2-Butanone, 3-hydroxy- no smell Oxirane, 2,3-dimethyl-, caramel-like

2-Nonanone no smell trans- roasted cocoa
Propane, 2-(ethenyloxy)-
Hydrazine, 1,1-dimethyl-

flavor are pyrazine, carbonyl, ester, alco- CONCLUSION

hol, hydrocarbon and phenol.
SPME offers accurate, easier extraction
This result also implies that cocoa bean
technique, shorter extraction time and to
aroma is characterized by presence of sweet,
extract specific compounds with lower ex-
caramel-like, nutty and bean-like odour.
traction temperature. Selection can be made
Those odours are expressed by pyrazines,
by choosing the type and fiber specification.
ethyl ester and alcoholic compounds, par-
SPME extracted pyrazine was adequate and
ticularly trimethylpyrazine, tetramethyl-
well detected in a gas chromatography sys-
pyrazine, 2,3-butanediol, dodecanoic acid,
tem equipped either with FID or Mass Spec-
phenylethyl alcohol, ethanone, benzene-
trometry detector. Over thirty compounds
acetal-dehyde and 1,4-bis (morpho-
were detected as the most representative
linoacetyl) piperazine. Few unusual odours
volatile-semi volatile compounds from
might also present, for instance rancid and
roasted cocoa beans including 2 methyl
lemon-like that came with 1-Butanol, 3-
pyrazine (2MP); 2,3 and 2,5 di-methyl
methyl-, acetate and 3-methyl-pentanoic
pyrazine (DMP); and 2,3,5 trimethyl
acid.

PELITA PERKEBUNAN, Volume 27, Nomor 1, Edisi April 2011

33
Pyrazine (TrMP) were identified. Alcohols, Da Silva, G.A.; F. Augusto & R.J. Poppi (2008).
carboxylic acids, aldehydes, ketons, esters, Exploratory analysis of the volatile
pyrazines, amines and other volatile com- profile of beers by HS-SPME. Food
Chemistry, 111, 1057—1063.
pounds were also extracted and associated
to chocolate aroma. The presence of other Ducki, S.; J. Miralles-Garcia; A. Zumbé; A.
volatile compounds which are the key con- Tornero & D.M. Storey (2008). Evalu-
tributor to chocolate aroma indicates SPME ation of solid-phase micro-extraction
coupled to gas chromatography-mass
extraction is applicable in aroma analysis.
spectrometry for the headspace analy-
Limiting factor for SPME is in quantity of sis of volatile compounds in cocoa pro-
compound trapped from the extraction ducts. Talanta, 74, 1166—1174.
which is lower than that of resulted from
Frauendorfer, F. & P. Schieberle (2008).
syringe injection. Changes in key aroma compounds of
criollo cocoa beans during roasting.
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