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International Dairy Journal 18 (2008) 790–800

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Aroma characterisation of Gouda-type cheeses

I. Van Leuven, T. Van Caelenberg, P. Dirinck
Laboratory for Flavour Research, Catholic Technical University St.-Lieven, Gebr. Desmetstraat 1, 9000 Gent, Belgium
Received 30 April 2007; received in revised form 7 January 2008; accepted 8 January 2008

Abstract
The aim of this study was to characterise flavour differences between short-ripened Gouda-type cheeses produced from raw and
pasteurised milk and to follow, in the case of pasteurised milk cheeses, flavour evolution during ripening. The volatile composition of the
cheeses was studied using a combination of simultaneous steam distillation–extraction and gas chromatography–mass spectrometry.
Visualisation of the analytical results was performed by principal component analysis. A clear differentiation between raw milk cheese
and pasteurised milk cheese was observed and the evolution of the volatile composition of the pasteurised cheese samples during ripening
was clearly demonstrated. Partial least squares regression was used to examine possible relationships between the chemical–analytical
and sensory descriptive data.
r 2008 Elsevier Ltd. All rights reserved.

1. Introduction contribution of (ethyl) esters to cheese flavour is concen-

tration-dependent (Liu, Holland, & Crow, 2004). An
Volatiles play an important role in flavour perception of overview of mechanisms of ester biosynthesis in dairy
cheese. Typical cheese aroma is the result of volatiles and non-dairy micro-organisms is given by Liu et al.
formed by lipolysis, proteolysis and metabolism of lactose, (2004).
lactate and citrate (Marilley & Casey, 2004; McSweeney & During ripening of cheese, the enzymatic degradation of
Sousa, 2000; Smit, van Hylckama Vlieg, Smit, Ayad, & amino acids leads to the formation of flavour-impact
Engels, 2002; Urbach, 1997). The fat-derived flavour volatiles (Marilley & Casey, 2004; McSweeney & Sousa,
volatiles in Gouda cheese were clearly described by 2000; Visser, 1993). Caseins are degraded into peptides
Alewijn, Sliwinski, and Wouters (2005). Fatty acids are and amino acids. Methionine, the aromatic and the
the main precursors of the secondary fat-derived com- branched-chain amino acids are the precursors for sulphur
pounds (methyl ketones, free fatty acids (FFAs), components (methional, dimethyl disulphide, dimethyl
aldehydes, lactones and ethyl esters). Alewijn, Smit, trisulphide) (Engels et al., 2002), for aromatic and
Sliwinski, and Wouters (2007) also illustrated that the branched-chain aldehydes (benzaldehyde, phenylacetalde-
formation of lactones in Gouda cheese is a one-step hyde, 3-methylbutanal, 2-methylbutanal, 2-methylpropa-
intramolecular trans-esterification reaction. d-Lactones are nal) and for branched-chain volatile acids and alcohols
produced more rapidly than g-lactones during ripening (3-methylbutyric acid, 2-methylbutyric acid, isobutyric
(Alewijn et al., 2005). Ketones, especially methyl ketones, acid, 3-methylbutanol, 2-methylbutanol).
are produced slightly and do not contribute substantially to Besides lipids and proteins, lactose is also a major milk
the flavour of Gouda-type cheeses (Alewijn et al., 2005). constituent for the formation of cheese aroma (Marilley &
On the contrary, methyl ketones are important flavour Casey, 2004; McSweeney & Sousa, 2000). Lactose, lactate
compounds in Blue-mould-type cheeses (Molimard & and citrate contribute to the formation of diacetyl, acetoine,
Spinnler, 1996). Moulds are responsible for the conversion ethanol and acetate (Cogan & Hill, 1993; Escamilla-
of FFAs into methyl ketones (Gehrig & Knight, 1963). The Hurtado, Tomasini-Campocosio, Valdés-Martinez, &
Soriano-Santos, 1996; Henriksen & Nilsson, 2001; Mel-
Corresponding author. Tel.: +32 92658639; fax: +32 92658638. chiorsen, Jokumsen, Villadsen, Israelsen, & Arnau, 2002;
E-mail address: [email protected] (I. Van Leuven). Syu, 2001).

0958-6946/$ - see front matter r 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.idairyj.2008.01.001
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I. Van Leuven et al. / International Dairy Journal 18 (2008) 790–800 791

Several parameters determine flavour formation in content of the pasteurised milk cheese ranged from 42.0%
cheese, e.g., starter cultures (Kieronczyk, Skeie, Langsrud, to 44.0% (w/w), 28.0% to 29.0% (w/w) and 1.80% (w/w),
Le Bars, & Yvon, 2004; Smit, Smit, & Engels, 2005), non- respectively. The fat and salt content increased during
starter lactic acid bacteria or NSLAB flora in the cheese ripening (after 10 months: 32.0% (w/w) and 2.20% (w/w)),
plant (Beresford & Cogan, 2000; Østlie, Eliassen, Florvaag, respectively, while the moisture content decreased (after 10
& Skeie, 2005), cheesemaking technology, salt and fat months: 37.0% (w/w)).
content (Banks, Hunter, & Muir, 1993; Mistry & Kasper- Cheese samples were taken in duplicate by cutting 250 g
son, 1998). Flavour formation in cheese is also importantly of sample from the centre to the outer part just before
influenced by the type of milk (raw vs. pasteurised) and the analysis. The cheese samples were stored in a refrigerator at
ripening period. As a result of the destruction of some 4 1C until they were needed for analysis. For sensory
desirable non-starter lactic acid bacteria during pasteurisa- analysis, the samples were held at ambient temperature for
tion, cheese made from pasteurised milk ripens more slowly about 1 h before evaluation.
and develops a less intense flavour than raw milk cheese
(Fox & McSweeney, 1998; Grappin & Beuvier, 1997; 2.2. Simultaneous steam distillation–extraction (SDE)
Johnson, Nelson, & Johnson, 1990; Seifu, Buys, & Donkin,
2004). For the isolation of the cheese volatiles, a sample of 50 g
In this work, the volatile composition of some commer- of blended cheese was suspended in 600 mL of water and
cial Gouda-type cheeses produced from raw and pas- was extracted in a simultaneous SDE apparatus (Alltech
teurised milk were studied using a combination of Associates, Inc., Lokeren, Belgium) using 60 mL of
simultaneous steam distillation–extraction (SDE) and gas dichloromethane. For determination of the semi-quantita-
chromatography–mass spectrometry (GC–MS). In the case tive volatile composition, undecane was used as internal
of the pasteurised milk cheeses also, the evolution of the standard and was spiked into the solvent. Undecane was
volatile composition as a function of ripening was not detected in the samples and was well separated from
followed. Principal component analysis (PCA) was used the other cheese volatiles. After cooling to ambient
to evaluate the semi-quantitative SDE–GC–MS results and temperature, the combined dichloromethane fractions were
to visualise the changes in flavour profile of the cheeses concentrated to a final volume of 0.5 mL in a Kuderna-
during ripening. Additionally, the cheeses were submitted Danish concentrator (Alltech Associated Inc., Deerfield,
to a descriptive sensory test for some selected attributes. IL, USA). For each cheese sample, duplicate isolations of
The relationship between sensory data and analytical volatiles were performed.
GC–MS data was demonstrated by means of partial least
square regression (PLS2). 2.3. Gas chromatography–mass spectrometry (GC–MS)

2. Materials and methods At the inlet of a HP 5890 gas chromatograph coupled to

a HP 5971A MSD mass spectrometer (Agilent Technolo-
2.1. Materials and sampling gies, Santa Clara, CA, USA), 1 mL of a cheese extract was
injected. The GC system was equipped with a DB-1MS
Two commercial Gouda-type cheeses were provided by a capillary column (100% dimethylpolysiloxane, L 60 m 
Belgian raw milk cheese maker and by a Belgian I.D. 0.25 mm  0.25 mm film thickness; J&W Scientific,
pasteurised milk cheese maker. All cheese samples were Folsom, CA, USA). Helium was used as carrier gas with a
produced from cows’ milk. flow rate of 1 mL min 1 and the column temperature was
The milk used for the raw milk cheese samples was only held at 40 1C for 5 min, then programmed at a rate of
heated to 37 1C. The starters consisted of a mixture of 5 1C min 1 to increase to 250 1C, which was maintained for
different strains of Lactococcus lactis subsp. lactis biovar 13 min. A split ratio of 1/10 was used and the injector and
diacetylactis and subsp. cremoris (MPM De Block, detector were maintained at 250 and 280 1C, respectively.
Edegem, Belgium). The raw milk cheese was ripened for The mass spectra were obtained by electron impact of
6 weeks at 85–90% relative humidity and 13 1C. The final 70 eV and the total ion current (TIC) chromatograms were
round-shaped cheeses produced from raw milk had a recorded by monitoring the TIC in a scan range of
weight of 10.0 kg and an initial moisture, fat and salt 40–260 amu (atomic mass units). A solvent delay of
content of 36.6% (w/w), 37.1% (w/w) and 2.0% (w/w), 6.0 min was used to avoid excessive solvent disturbing the
respectively. MS source. Identification of the cheese volatiles was made
For confidentiality reasons, the process parameters and by comparing the mass spectra of the different volatiles
the ripening conditions of the pasteurised milk cheeses were with the mass spectra of a commercial Mass Spectral
not completely revealed. Round-shaped cheeses produced Library, Wiley275 (Wiley, Somerset, NJ, USA). Semi-
from milk of different seasons and ripened for 6 weeks, 4 quantitative data of the isolated aroma compounds were
months and 10 months were delivered at the same moment calculated by relating the peak area of the cheese volatile to
to the laboratory. The cheeses had a weight of 13.0, 12.5 the peak area of undecane. The concentrations of the
and 11.5 kg, respectively. The initial moisture, fat and salt volatiles were expressed as ng g 1 of cheese.
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792 I. Van Leuven et al. / International Dairy Journal 18 (2008) 790–800

2.4. Descriptive sensory analysis buttery’, ‘taste intensity’ and ‘salt’, no differences in scores
by panellists were measured. Although both cheeses had
Descriptive sensory analyses were performed in a similar ripening times, some characteristic differences were
separate panel room using a trained panel of eight assessors observed between cheeses produced from raw and pro-
(three male, five female, aged between 20 and 60 years). cessed milk. The sensory differences between the two types
During training tests, the panellists developed a vocabulary of cheeses probably originate from the used milk (milk pre-
of six flavour attributes and five taste attributes (Table 1). treatment) and from the different fat content of the cheeses.
The selected attributes were based on the sensory Fig. 2 visualises the average sensory scores for
vocabulary used in the experiment of Muir, Hunter, Banks, pasteurised milk cheese of different ripening times. The
and Horne (1995). The assessors were presented cubes of panel scores of most attributes increased as function of
randomly coded cheese samples, equilibrated to room ripening, with exception of the attributes ‘sweet’ and
temperature. They assessed the intensity of the 11 ‘creamy, buttery’, which decreased during ripening. As
attributes using an unstructured 100-mm-score line. After expected, ripening seems to have a clear effect on the
sensory analysis, the scores were converted to numerical flavour characteristics of cheese.
values. For data analysis of the sensory profiles of the
cheese samples, mean scores for each attribute were 3.2. SDE–GC–MS profiling of cheese
calculated. The obtained sensory data of the samples were
visualised by spider web diagrams. 3.2.1. Volatile composition
SDE–GC–MS profiling of the Gouda-type cheeses
2.5. Statistical analysis resulted in the identification of 63 volatiles. In accordance
with previous work (Dirinck & De Winne, 1999), 63
PCA was performed on the semi-quantitative aroma volatiles were classified in chemical classes (Table 2)
compositions of the cheese samples using the Unscrambler and were related to their origin and to their sensory
6.1 (Camo, Oslo, Norway). Partial least squares regression characteristics.
(PLS2) (Unscrambler 9.7, Camo) was used to examine Branched-chain aldehydes (3- and 2-methylbutanal),
possible interrelationships between sensory descriptions branched-chain acids (2-methylpropanoic acid, 3- and 2-
and the aroma compounds. All data were weighted with methylbutanoic acid) and branched-chain alcohols (3-
1/SD (standardisation) and a leverage correction was used methylbutanol) are formed by the catabolism of
to validate the models. branched-chain amino acids initiated by an aminotransfer-
ase (Atiles, Dudley, & Steele, 2000; Marilley & Casey,
3. Results and discussion 2004). 3-Methylbutanal and 2-methylbutanal are charac-
terised by a malty flavour (Friedrich & Acree, 1998;
3.1. Descriptive sensory analysis Griffith & Hammond, 1989; Kubickova & Grosch, 1997;
Rychlik & Bosset, 2001b; Thierry & Maillard, 2002). The
Descriptive sensory analyses for six flavour attributes corresponding alcohols, 3-methylbutanol and 2-methylbu-
and five taste attributes were performed on the raw and tanol, have impact on alcoholic and fruity odours (Thierry
pasteurised milk cheese of the same ripening time (6 weeks) & Maillard, 2002). The branched-chain acids (2-methyl-
and on the pasteurised milk cheese of different ripening propanoic acid, 3-methylbutanoic acid and 2-methylbuta-
times (6 weeks, 4 months, 10 months). Fig. 1 illustrates the noic acid) are responsible for sweaty, rancid, faecal,
average sensory scores for raw and pasteurised milk cheese putrid, ester and rotten fruit-like flavours (Brennand,
of 6 weeks of ripening. The raw milk cheese had higher Ha, & Lindsay, 1989; Karagül-Yüceer, Cadwallader,
scores for the attributes ‘sweet’, ‘bitter’, ‘fruity, flowery’, & Drake, 2002; Marilley & Casey, 2004; Moio, Piombino,
‘nutty, chocolate-like’ and ‘animal’. While the pasteurised & Addeo, 2000; Rychlik & Bosset, 2001a; Thierry &
milk cheese had higher scores for the attributes ‘flavour Maillard, 2002).
intensity’, ‘sour’ and ‘pungent’. For the attributes ‘creamy, From the catabolism of aromatic amino acids, com-
pounds such as benzaldehyde, phenylacetaldehyde,
2-phenylethanol, benzenemethanol and indole are formed
Table 1
Descriptive vocabulary of 11 attributes for descriptive sensory analysis of
(Marilley & Casey, 2004; McSweeney & Sousa, 2000).
Gouda-type cheeses Depending on their concentration, those volatiles have an
impact on the cheese aroma. Benzaldehyde is characterised
Flavour attributes Taste attributes by a bitter almond; phenylacetaldehyde by a honey-like,
Flavour intensity Taste intensity floral, rosy and violet-like; 2-phenylethanol by a rosy,
Creamy, buttery Sweet violet-like and floral; indole by a faecal, putrid, musty
Fruity, flowery Sour odour (Curioni & Bosset, 2002; Friedrich & Acree, 1998;
Nutty, chocolate-like Salt Kubickova & Grosch, 1997; Marilley & Casey, 2004; Moio
Pungent Bitter
et al., 2000; Rychlik & Bosset, 2001a; Thierry & Maillard,
Animal
2002).
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flavour intensity
10.0

animal taste intensity

8.0

6.0

nutty, chocolate-like sweet

4.0

2.0

0.0

fruity, flowery sour

creamy, buttery salt

pungent bitter

Fig. 1. Spider web diagram of the average sensorial scores for raw milk (K) and pasteurised milk (~) cheeses of 6-weeks ripening.

flavour intensity
10.0

animal taste intensity

8.0

6.0

nutty, chocolate-like sweet

4.0

2.0

0.0

fruity, flowery sour

creamy, buttery salt

pungent bitter

Fig. 2. Spider web diagram of the average sensorial scores for 6-weeks (~), 4-months (m) and 10-months (’) ripened pasteurised milk cheeses.
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Table 2
Volatile composition of Gouda-type cheeses, obtained by SDE–GC–MS (steam distillation–extraction and gas chromatography–mass spectrometry) and
expressed in ng g 1

Flavour volatiles Average (S.D.)a

Raw 6 weeks Pasteurised 6 weeks Pasteurised 4 months Pasteurised 10 months

Branched-chain and aromatic aldehydes

3-Methylbutanal 12.0 (2.0) 9.32 (2.74) 37.4 (12.0) 45.5 (3.6)
2-Methylbutanal n.q. n.q. 5.11 (0.63) 7.00 (0.37)
Benzaldehyde 12.1 (5.0) 11.4 (0.4) 15.4 (1.1) 18.6 (0.8)
Phenylacetaldehyde 39.4 (3.0) 36.6 (1.8) 118 (17) 138 (9)
Linear aldehydes
Pentanal 5.54 (1.00) 4.00 (0.01) 3.25 (0.34) 2.20 (0.00)
Hexanal 5.00 (0.60) 4.01 (1.22) 3.94 (1.03) 4.48 (1.71)
Heptanal 4.80 (0.42) 4.18 (0.03) n.q. 6.28 (0.47)
Nonanal 5.20 (0.00) 12.4 (2.2) 6.54 (1.71) 7.77 (1.86)
Dodecanal 24.0 (3.0) 23.3 (4.2) 16.0 (1.0) 13.7 (3.1)
Tetradecanal 50.1 (3.6) 47.2 (7.2) 34.0 (4.0) 25.8 (5.3)
Hexadecanal 183 (34) 200 (39) 144 (3) 127 (1)
2-Nonenal 0.00 4.53 (1.00) n.q. 3.46 (1.12)
2-Decenal 0.00 5.17 (0.17) n.q. n.q.
2,4-Heptadienal 12.4 (4.3) 0.00 0.00 0.00
2,4-Decadienal 6.10 (2.00) 0.00 0.00 0.00
Ketones
2-Pentanone 487 (59) 417 (15) 447 (22) 593 (64)
2-Hexanone 17.3 (1.3) 14.5 (8.0) 21.3 (9.0) 32.2 (4.2)
2-Heptanone 2281 (75) 1613 (188) 1770 (53) 2330 (312)
2-Octanone 24.1 (0.1) 17.7 (3.0) 17.1 (2.2) 25.6 (2.3)
2-Nonanone 1833 (1) 1395 (140) 1254 (170) 1878 (16)
2-Decanone 22.4 (0.6) n.q. 11.9 (2.0) 18.0 (4.0)
2-Undecanone 2159 (42) 1465 (221) 1133 (207) 1440 (42)
2-Dodecanone 30.8 (2.0) n.q. n.q. 16.7 (0.0)
2-Tridecanone 2163 (91) 1604 (274) 1182 (196) 1264 (231)
2-Pentadecanone 1745 (113) 1509 (289) 1106 (120) 1405 (234)
3-Hydroxy-2-butanone 1927 (164) 3281 (264) 1395 (511) 355 (177)
Branched-chain volatile acids
2-Methylpropanoic acid 0.00 163 (26) 0.00 0.00
3-Methylbutanoic acid 0.00 1141 (55) 0.00 0.00
2-Methylbutanoic acid 0.00 60.8 (0.0) 0.00 0.00
Free fatty acids
Butanoic acid n.q. 168 (16) n.q. n.q.
Hexanoic acid n.q. 454 (10) 18.0 (0.0) n.q.
Octanoic acid 33.9 (7.3) 1864 (133) 303 (188) 9.13 (0.00)
Nonanoic acid n.q. 35.0 (1.3) n.q. n.q.
Decanoic acid 1524 (39) 6855 (616) 3319 (1174) 720 (343)
Undecanoic acid 21.6 (6.3) 43.0 (4.0) 20.5 (7.0) 7.67 (3.44)
Dodecanoic acid 1384 (110) 3273 (581) 1733 (611) 833 (423)
Tetradecanoic acid 724 (196) 1097 (281) 468 (55) 387 (250)
Hexadecanoic acid 152 (65) 133 (59) 34.0 (6.2) 63.0 (48.0)
Ethyl esters
Ethyl hexanoate 0.00 0.00 10.0 (2.1) 17.3 (0.1)
Ethyl octanoate 0.00 0.00 4.90 (2.50) 20.2 (0.0)
Ethyl decanoate 0.00 0.00 n.q. 16.4 (3.0)
Ethyl dodecanoate 0.00 0.00 12.8 (3.0) 14.6 (0.2)
Ethyl tetradecanoate 0.00 0.00 n.q. 53.0 (14.0)
Ethyl hexadecanoate 0.00 0.00 8.48 (5.10) 22.1 (10.0)
Lactones
g-Nonalactone n.q. n.q. 2.59 (1.10) 9.21 (4.77)
g-Undecalactone 12.0 (0.3) 17.4 (3.6) 8.58 (1.50) 6.83 (0.30)
Cis-g-6-dodecenoic acid lactone 136 (7) 101 (22) 101 (17) 133 (30)
g-Dodecalactone 468 (26) 401 (56) 340 (47) 395 (40)
d-Octalactone 21.0 (0.4) 22.9 (1.9) 9.24 (3.50) 2.42 (0.00)
d-Decalactone 525 (16) 654 (78) 432 (66) 331 (45)
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Table 2 (continued )

Flavour volatiles Average (S.D.)a

Raw 6 weeks Pasteurised 6 weeks Pasteurised 4 months Pasteurised 10 months

d-Undecalactone n.q. n.q. n.q. 11.15 (1.61)

d-Dodecalactone 657 (11) 851 (170) 588 (119) 590 (56)
d-Tetradecalactone 248 (11) 353 (75) 246 (26) 234 (8)
d-Hexadecalactone 52.3 (3.1) 68.0 (15.0) 39.1 (1.0) 36.5 (3.5)
Sulphur compounds
Dimethyl disulphide n.q. 3.24 (0.62) 4.78 (0.31) 7.40 (0.39)
Methional 4.30 (2.01) n.q. 10.6 (0.9) 12.1 (2.0)
Dimethyl trisulphide n.q. 1.61 (0.45) 1.83 (0.63) 2.24 (0.19)
Alcohols
1-Butanol n.q. 8.03 (2.55) 18.4 (4.3) 18.9 (1.7)
3-Methyl-1-butanol 6.36 (0.15) 27.4 (0.3) 51.2 (5.8) 27.3 (10.4)
Benzenemethanol 3.86 (1.77) 6.89 (0.47) 5.11 (1.92) 9.52 (0.47)
2-Phenylethanol 13.6 (0.7) 10.9 (1.8) 29.1 (6.0) 59.2 (4.2)
2,3-Butanediol 17.4 (3.2) 11.6 (1.1) 12.7 (0.0) n.q.
Indole 4.28 (0.01) 6.83 (1.22) 3.07 (0.88) 36.6 (2.4)
a
Average of analyses of two different samples (S.D.); n.q.: not quantified.

Sulphur compounds (dimethyl disulphide, dimethyl odour, while unsaturated aldehydes have a fatty, oily,
trisulphide, methional) originating from methionine frying odour (Maarse, 1991). Ethyl esters, originated
(McSweeney & Sousa, 2000; Yvon & Rijnen, 2001) from the enzymatic or chemical esterification of fatty acids
generally have low odour threshold values (Maarse, 1991) with ethanol, possess pleasant sweet and fruity notes
and have powerful odours described as sulphurous or (Bosset & Liardon, 1984; Fernández-Garcı́a, Mariaca, &
cabbage-, onion-, or garlic-like (Maarse, 1991). Nuñez, 1998; Moio & Addeo, 1998; Molimard & Spinnler,
Free fatty acids (even numbered C4–C16, nonanoic and 1996).
undecanoic acid), 2-methyl ketones (C5–C15, major com- From metabolism of lactose, lactate and citrate 2,3-
pounds have odd carbon numbers), d-lactones (even butanedione (diacetyl) and its reduction products
numbered C8–C16 and d-undecalactone) and g-lactones 3-hydroxy-2-butanone (acetoine) and 2,3-butanediol are
(C9, C11, C12 and unsaturated cis-g-6-dodecenoic acid formed (Cogan & Hill, 1993; Marilley & Casey, 2004;
lactone), saturated and unsaturated aldehydes (saturated McSweeney & Sousa, 2000). Unfortunately, because of the
C5, C6, C7, C9, C12, C14, C16, 2,4-heptadienal and high volatility, diacetyl (with a ‘buttery’ note) was not
2,4-decadienal) and ethyl esters (even numbered C6–C16) measured in this method, as the peak was covered by the
are fat-derived flavour volatiles that play a role in the solvent.
overall flavour of cheese (Alewijn et al., 2005). FFAs are
formed by lipid degradation and have a pungent to rancid 3.2.2. Principal component analysis of the semi-quantitative
odour (Gómez-Ruiz, Ballesteros, Gonzalez Vinas, Cabe- SDE–GC–MS data
zas, & Martinez-Castro, 2002). Methyl ketones are present The concentrations of the 63 identified volatile com-
in most cheeses, but in lower levels than in mould-ripened pounds in the Gouda-type cheeses, listed in Table 2, varied
cheeses (Molimard & Spinnler, 1996). These compounds during ripening and were different between the raw and the
are lipid degradation products and are formed by pasteurised cheese samples. To visualise the SDE–GC–MS
b-oxidation and decarboxylation of fatty acids (Bills & results of the cheese samples, PCA was performed on the
Day, 1964; Marilley & Casey, 2004). Methyl ketones complex data (63 volatiles, 4  2 cheese samples). Indivi-
probably contribute to the background aroma of cheese. dual values of the duplicate analyses were used in order to
Lactones are also related to lipid degradation (Alewijn allow visual evaluation of the reproducibility of the
et al., 2007; Collins, McSweeney, & Wilkinson, 2003; SDE–GC–MS technique.
Marilley & Casey, 2004; McSweeney & Sousa, 2000; Fig. 3 illustrates a 2D-PCA biplot of the SDE–GC–MS
Molimard & Spinnler, 1996). The sensory characteristics results of raw and pasteurised milk cheese of 6 weeks of
of lactones are different: buttery, fruity and coconut-like ripening. The biplot explained 64% (PC1: 53%; PC2: 11%)
(Keeney & Patton, 1956; Maarse, 1991). The saturated of the total chemical information. A good classification
aldehydes can originate from fresh milk but they can also between raw and pasteurised milk cheese was obtained.
be formed, together with the unsaturated aldehydes, by The cheese samples produced from raw milk were located
autoxidation of unsaturated fatty acids (Ho & Chen, 1993). on the negative PC1 axis. Also phenylacetaldehyde
Saturated aldehydes have a green-like, hay-like, paper-like and 2-phenylethanol with a ‘flowery’ flavour character
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796 I. Van Leuven et al. / International Dairy Journal 18 (2008) 790–800

1.0

0.5
PC2

-0.5

-1.0

-1.0 -0.5 0 0.5 1.0

PC1: 53% , PC2: 11% PC1

Fig. 3. Principal component analysis (PCA) of the volatile compositions (SDE–GC–MS) of raw milk (K) and pasteurised milk (~) cheeses of 6-weeks
ripening.

1.0

0.5
PC2

-0.5

-1.0

-1.0 -0.5 0 0.5 1.0

PC1: 37%, PC2: 17% PC1

Fig. 4. Principal component analysis (PCA) of the volatile compositions (SDE–GC–MS) of pasteurised milk cheese of 6-weeks (~), 4-months (m) and
10-months (’) ripening.

(Maarse, 1991) and 2,4-heptadienal and 2,4-decadienal samples. The somewhat higher amount of 3-methylbutanal
with a ‘fried’ impact on the cheese flavour (Maarse, 1991) should be responsible for the higher ‘nutty, chocolate-like’
were located on the negative PC1 axis. It can be observed character (Maarse, 1991) of the raw milk cheese. It is also
that some methyl ketones, linear aldehydes and two clear that the volatile pattern of the pasteurised milk cheese
g-lactones (g-dodecalactone and cis-g-6-dodecenoic acid showed higher levels of sulphur compounds (dimethyl
lactone) were also located near the raw milk cheese disulphide, dimethyl trisulphide) and of the branched-chain
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acids, 2-methylpropanoic acid, 3- and 2-methylbutanoic The evolution of the volatile composition of pasteurised
acid. Together with the higher levels of free fatty acids milk cheese during ripening is illustrated in Fig. 4. The
(octanoic acid, decanoic acid, dodecanoic acid, etc.) these PCA biplot of the semi-quantitative SDE–GC–MS results
compounds (situated on the positive PC1 axis) should be explained 54% (PC1: 37% and PC2: 17%) of the chemical
responsible for the higher ‘flavour intensity’ and the more information. The pasteurised milk cheese of 6 weeks of
‘sour’ and ‘pungent’ character (Maarse, 1991) of the ripening was situated at the negative PC1 axis of the
pasteurised milk cheese. 2D-PCA biplot. It was clear that after 6 weeks of ripening

Correlation Loadings
X: flavour volatiles and Y: sensory descriptors

1.0

0.5
PC2

-0.5

-1.0

1.0 0.5 0 -0.5 -1.0

X-expl: 43%, 24%; Y-expl: 69%, 23% PC1

2
PC2

-2

-4

-8 -6 -4 -2 0 2 4 6 8
X-expl: 43%, 24% Y-expl: 69%, 23% PC1

Fig. 5. (A) Partial least squares regression (PLS2) correlation loadings plot (X-variables: flavour volatiles; Y-variables: sensory descriptors) of Gouda-type
cheeses (raw milk cheese of 6 weeks, pasteurised milk cheese of 6 weeks, 4 months and 10 months). (B) Partial least squares regression (PLS2) scores plot
(raw milk cheese of 6 weeks, pasteurised milk cheese of 6 weeks, 4 months and 10 months) of Gouda-type cheeses, made of raw and pasteurised milk.
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higher concentrations of a lot of fat-derived flavour have low odour threshold values and can have a great
volatiles (FFAs, d-lactones and linear aldehydes) were impact on the cheese aroma. It can be observed that most
determinated, but also 3- and 2-methylbutanoic acid and volatile acids (FFAs and branched-chain acids) have
2-methylpropanoic acid (amino acid-related) had the negative correlations with most sensory attributes. How-
highest levels in the pasteurised milk cheese of 6 weeks of ever, they should be important for the flavour of the
ripening. Also 3-hydroxy-2-butanone (acetoine), that pasteurised milk cheese after 6 weeks of ripening. By
should contribute to the ‘buttery’ note (Maarse, 1991), comparing Fig. 5A (correlation X and Y loadings plot)
was found at a higher concentration in the 6-weeks-ripened with Fig. 5B (scores plot), the raw milk cheese seems to
pasteurised milk cheese. have a good correlation with the attributes ‘fruity, flowery’,
The cheeses ripened for 4 and 10 months were located at ‘sweet’ and ‘creamy, buttery’, while the more ripened
the positive PC1 axis and the PC2 axis differentiated the pasteurised milk cheeses yielded good correlation with the
samples of respectively 4 and 10 months of ripening. There descriptors ‘taste intensity’, ‘pungent’ and ‘flavour inten-
seemed to be no clear effect of ripening on the sensory sity’. The attributes ‘animal’, ‘bitter’, ‘salt’ and ‘sour’
important g- and d-lactones which were situated at both cannot directly be correlated with flavour volatiles. Those
sides of the PC1 axis. From the biplot, it was clear that attributes are more related to less volatile high-mass
during ripening there was an increase of the concentration molecules.
of 3- and 2-methylbutanal, sulphur compounds (methional,
dimethyl disulphide, dimethyl trisulphide), phenylacetalde- 4. Conclusions
hyde, 2-phenylethanol, benzaldehyde and indole. These
strong odour active volatiles should have a major impact In this study, the flavour differences between the selected
on the ‘aged’ cheese flavour. From Fig. 4 and Table 2 it was Gouda-type raw and pasteurised milk cheeses and the
also clear that ethyl esters (ethyl hexanoate, ethyl flavour evolution during ripening of the investigated
octanoate, etc.) were only formed after long ripening pasteurised milk cheeses were evaluated. The differences
periods and increased as a function of ripening time. Ethyl in the composition of volatile flavour compounds between
esters contributed to the ‘floral’ and ‘fruity’ note (Liu et al., short-ripened raw and pasteurised milk cheeses were
2004; Maarse, 1991) of cheeses. demonstrated. During ripening of pasteurised Gouda-type
cheeses, there was an increase of more amino acid-related
3.3. Correlation between sensory and chemical–analytical flavour volatiles and of ethyl esters, while the amount of
analysis linear aldehydes and volatile acids (FFAs and branched-
chain acids) decreased. Correlation analysis suggested that
Partial least squares regression (PLS2) was performed on ‘sweet’ and ‘fruity’ notes correlated with the raw milk
the data from raw and pasteurised milk cheeses of different cheese of 6 weeks ripening, while sulphur compounds
ripening times to examine possible relationships between correlated strongly with the descriptor ‘flavour intensity’ of
sensory attributes and chemical aroma compounds. The the more ripened pasteurised milk cheeses.
X-matrix contained the volatile compounds and the In further research, these techniques will be used for
Y-matrix contained the sensory scores for each attribute. objective evaluation of a whole range of flavour influencing
The ellipses indicate 50% and 100% of explained variance. parameters (e.g., fat and salt reduction, influence of starter
Fig. 5A shows the correlation between the X and cultures, influence of slicing and packaging). An ob-
Y variables. The ‘fruity, flowery’ note can be correlated jective analytical approach can contribute substantially to
with some methyl ketones (2-undecanone and 2-trideca- product and process innovation for the cheese producing
none). Methyl ketones give Blue cheeses a ‘fruity’, ‘spicy’ companies.
odour (Heath, 1978). The attributes ‘sweet’ and ‘creamy,
buttery’ have a positive correlation with some lactones Acknowledgement
(g-dodecalactone and d-octalactone) and with 3-hydroxy-2-
butanone. Lactones can have ‘buttery’ and ‘fruity’ flavour This work was supported by the Institute for the
notes, while 3-hydroxy-2-butanone is known for its Promotion of Innovation by Science and Technology in
‘buttery’ character. It was also clear that most amino Flanders (IWT-Vlaanderen).
acid-related volatiles (2-phenylethanol, benzaldehyde,
3-methylbutanal, phenylacetaldehyde) correlated with the References
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