Influence of milk pasteurization and scalding temperature

HAL Id: hal-00895632https://hal.archives-ouvertes.fr/hal-00895632

Submitted on 1 Jan 2007

HAL is a multi-disciplinary open accessarchive for the deposit and dissemination of sci-entific research documents, whether they are pub-lished or not. The documents may come fromteaching and research institutions in France orabroad, or from public or private research centers.

L’archive ouverte pluridisciplinaire HAL, estdestinée au dépôt et à la diffusion de documentsscientifiques de niveau recherche, publiés ou non,émanant des établissements d’enseignement et derecherche français ou étrangers, des laboratoirespublics ou privés.

Influence of milk pasteurization and scaldingtemperature on the volatile compounds of Malatya, a

farmhouse Halloumi-type cheeseAli A. Hayaloglu, Elizabeth Y. Brechany

To cite this version:Ali A. Hayaloglu, Elizabeth Y. Brechany. Influence of milk pasteurization and scalding temperatureon the volatile compounds of Malatya, a farmhouse Halloumi-type cheese. Le Lait, INRA Editions,2007, 87 (1), pp.39-57. �hal-00895632�

https://hal.archives-ouvertes.fr/hal-00895632

https://hal.archives-ouvertes.fr

39Lait 87 (2007) 39–57© INRA, EDP Sciences, 2007DOI: 10.1051/lait:2006025

Original article

Influence of milk pasteurization and scalding temperature on the volatile compounds of

Malatya, a farmhouse Halloumi-type cheese

Ali A. HAYALOGLUa*, Elizabeth Y. BRECHANYb

a Department of Food Engineering, Inonu University, 44280 Malatya, Turkeyb Food Quality and Safety Group, Hannah Research Institute, Ayr, KA6 5HL, UK

Received 20 July 2006 – Accepted 28 November 2006

Abstract – Malatya cheese, a farmhouse Halloumi-type cheese, was made from raw or pasteurizedmilk and the curds were scalded in hot whey at 60, 70, 80 or 90 oC. After ripening for 30 or 90 d,the cheeses were analyzed for characterization of volatile composition. One hundred and two vola-tile compounds including 11 acids, 13 esters, 15 ketones, 6 aldehydes, 26 alcohols, 2 lactones, 5 sul-fur compounds, 5 terpenes and 19 miscellaneous compounds were identified using solid phasemicro extraction and gas chromatography-mass spectrometry. The use of raw milk in the manufac-ture enriched the volatile profile of the cheese and the majority of volatiles were more abundant inraw milk cheeses than in pasteurized milk cheeses. The cheeses made with raw milk containedhigher levels of acids, esters and lactones and lower levels of aldehydes and sulfur compounds thandid the cheeses made from pasteurized milk. Principal component analysis (PCA) was applied tosimplify interpretation of the GC-MS data and distinguished the raw and pasteurized milk cheeseson the plot. The samples were also classified based on scalding temperature by PCA, but no regulardistribution was observed. The results suggest that the pasteurization of cheese milk had a greatereffect on volatile composition of cheese than scalding temperature of the curd.

pasteurization / raw milk / Malatya cheese / volatile compounds / scalding / aroma

摘要 – 原料奶的巴氏杀菌和热烫温度对一种农家 Halloumi 干酪——Malatya 干酪中挥发性化合物的影响。 Malatya 干酪属于农家 Halloumi 干酪中的一种。由未杀菌奶和巴氏杀菌奶生产的凝块分别在 60、 70、 80 和 90 °C 的乳清中热烫 3 min。干酪成熟 30 d 和 90 d 后分析了其中挥发性化合物的特性。经固相微萃取技术和 GC/MS 分析，共检测出 102 种挥发性化合物成分，其中包括 11 种有机酸、 13 种酯类化合物、 15 种酮类化合物、 6 种醛类化合物、 26 种醇类化合物、 2 种内酯、 5 种硫化物、 5 种萜烯和 19 种其他类化合物。未杀菌奶生产干酪中挥发性化合物成分丰富，与巴氏杀菌奶生产的干酪相比，前者干酪中主要挥发性化合物含量高，其中有机酸、酯、内酯的含量明显高于后者，但是醛类和含硫化合物含量则低于后者。采用主成分分析法 (PCA) 对 GC-MS 测定的数据进行了简化分析，分析了未杀菌奶和巴氏杀菌奶生产的两种干酪的特性。对不同热烫温度下测定的数据采用 PCA 分析法将样品进行了分类，证明他们之间没有规律性的分布。试验结果表明原料奶的巴氏杀菌对干酪中挥发性化合物的影响要显著地高于凝块的热烫温度。

巴氏杀菌 / 原料奶 / Malatya 干酪 / 挥发性成分 / 热烫 / 芳香气味

* Corresponding author (通讯作者): [email protected]

Article published by EDP Sciences and available at http://www.edpsciences.org/lait or http://dx.doi.org/10.1051/lait:2006025

http://www.edpsciences.org/lait

http://dx.doi.org/10.1051/lait:2006025

40 A.A. Hayaloglu, E.Y. Brechany

Résumé – Influence de la pasteurisation du lait et de la température de cuisson sur les compo-sés volatils du Malatya, fromage fermier de type Halloumi. Du Malatya, un fromage fermier detype Halloumi, a été fabriqué à partir de lait cru et de lait pasteurisé, et les caillés obtenus ont étécuits dans du lactosérum chauffé à 60, 70, 80 ou 90 °C. Après 30 ou 90 jours d’affinage, la compo-sition en volatils des fromages a été caractérisée. Cent deux composés volatils incluant 11 acides,13 esters, 6 aldéhydes, 26 alcools, 2 lactones, 5 composés soufrés, 5 terpènes et 19 composés diversont été identifiés par micro-extraction en phase solide et chromatographie gazeuse couplée à la spec-trométrie de masse (CG-MS). L’utilisation du lait cru pour la fabrication a abouti à un profil en vola-tils plus riche et la majorité des volatils étaient présents en quantité plus importante dans les froma-ges au lait cru par rapport aux fromages au lait pasteurisé. Les fromages au lait cru contenaient desteneurs plus élevées en acides, esters et cétones, et des teneurs moins élevées en aldéhydes et com-posés soufrés. Une analyse en composante principale (ACP) a permis de simplifier l’interprétationdes données de CG-MS et de différencier les fromages au lait cru des fromages au lait pasteurisé.Les échantillons ont également été classifiés sur la base de la température de cuisson par ACP, maisaucune distribution régulière n’a pu être observée. Ces résultats suggèrent que la pasteurisation dulait de fabrication a un effet sur la composition en volatils plus marqué que celui de la températurede cuisson du caillé.

pasteurisation / lait cru / Malatya / composé volatil / cuisson du caillé / arôme

1. INTRODUCTION

Cheese ripening is a complex and dy-namic biochemical process that includesprotein breakdown, fat hydrolysis and lac-tose metabolism [25]. Hydrolysis of caseinby proteolytic enzymes produces peptidesand free amino acids. These play a criticalrole in the development of cheese flavour.Many volatile compounds which contributeto the characteristic aroma and flavour oc-cur during the ripening of cheeses. Somefactors affect the composition of volatilefractions in cheese such as animal feeding,the types of milk, coagulant, starter or sec-ondary starters, heat treatment of milk, rip-ening time and temperature [7, 12, 14, 31,39]. Heat treatment of milk alters the fla-vour profile of cheese due to a decrease inthe number of non-starter lactic acid bacte-ria (NSLAB) and possibly inactivation ofindigenous milk enzymes [15, 40]. Heattreatment causes delaying in the flavour de-velopment in cheese. Lau et al. [22] re-ported that a cheese made from pasteurizedmilk took twice as long as that made fromraw milk to develop the same flavour inten-sity and lower levels of soluble nitrogenand free amino acid were obtained in cheesemade from pasteurized milk. The native mi-croorganisms and indigenous milk en-zymes are main contributors for theformation of the characteristic aroma com-ponents in cheese. Heat treatment of milk

prior to cheese making had an effect on mi-crobial flora, proteolysis, free amino acids,free fatty acids, volatile fractions and sen-sory characteristics [6, 12, 17, 26, 31, 39].However, heat treatment of milk prior tocheese making is an essential process inmany cheeses due to the presence of unde-sirable microorganisms which causes somedefects in texture and flavour. In addition,it can ensure the hygienic and standardquality of cheese, the ripening at highertemperature and the reduction of some risksincluding blowing caused by butyric fer-mentation, control over lactic acid produc-tion and off-flavours [30].

Malatya, a farmhouse Halloumi-typecheese, is traditionally made from rawewe’s or cow’s milks or their appropriatemixtures and no starter culture is employed.The traditional method is still used in farmsin small scale and villages. Recently, somecheese makers used the pasteurization proc-ess in its manufacture and added a starterculture to standardize the production and toeliminate undesirable microorganisms. Thecharacteristics of this type of cheeses are thescalding process which produces an elasticand compact texture after the pressing ofcurd at 80–90 °C. So, a second heat treat-ment is applied in the manufacture ofMalatya cheese and the method of manu-facture influences the physical, chemicaland sensory characteristics of the cheese.

Volatile compounds in Malatya cheese 41

A number of volatile components be-longing to the chemical groups acids, es-ters, aldehydes, ketones, alcohols, sulfurcompounds and other aromatic hydrocar-bons have been identified in differentcheeses [27–29, 36]. To the authors’ knowl-edge, no previous studies were carried outto identify the volatile compounds contrib-uting to the characteristic volatile profilesof Malatya cheese; consequently, a studywould be useful for the characterization ofthis cheese which is currently manufacturedboth from raw and pasteurized milk. Theobjective of the study was to investigate theinfluence of pasteurization prior to cheesemaking and scalding temperatures on vol-atile composition of Malatya cheese duringripening.

2. MATERIALS AND METHODS

2.1. Cheese-making

Malatya cheese was made in duplicateusing both raw and pasteurized cow’s milkin a local dairy plant (Karlidag Dairy Prod-ucts, Malatya, Turkey). A volume of 200 Lraw cow’s milk was used in the manufac-ture of Malatya cheese with 100 L of themilk used to make raw milk cheese (Ccheeses) and the rest pasteurised at 72 °Cfor 30 s and used for the manufacture of pas-teurized milk cheeses (P cheeses). Afterpasteurization, the milk was cooled to32 °C and the commercial culture consist-ing of selected strains of Lc. lactis ssp. lactisand Streptococcus thermophilus (Sacco,srl, Cadorago, Italy) was added at the man-ufacturer’s recommended dose and held for30–45 min. Both C and P cheeses were co-agulated using commercial calf rennet(>85% chymosin, REN–NA®, Mayasan,Istanbul, Turkey) at the level of 15 g·100 L–1.The coagulation took place at 32 °C for45 min. Following coagulation, the curdwas cut and stirred for about 30 min, andtransferred into cloth bags and then left for30 min to drain its whey with no pressing.The bags which contain approx. 250 g ofcurd were tied up and moulded as a ball, andthen they were pressed between twowooden blocks for 2 h. The cheeses werescalded at 60, 70, 80 or 90 °C for 3 min by

means of dipping their wheys and then thecheese blocks were re-pressed between thesame wooden blocks for 3 min and then im-mediately cooled to room temperature.Then, the cooled blocks were immersed inbrine (10% NaCl) and ripened for 90 d at 6–8 °C.

2.2. Chemical analysis

Cheeses were analyzed in duplicate formoisture, fat, total protein, salt, pH and ti-tratable acidity as described in Hayalogluet al. [16]. Statistical evaluation of thechemical data was analyzed by ANOVAwith significant differences for P < 0.05using SPSS package program version 9.0for Windows (SPSS Inc., Chicago, IL).

2.3. Analysis of volatile components in cheese using Solid Phase Microextraction (SPME)

Cheese samples were sliced, frozen inliquid nitrogen, pulverized into small gran-ules and stored in glass bottles in a freezerat –20 °C. Samples were analyzed withinmax. 14 d. A 3-g portion of sample was thenplaced in a 15-mL vial and allowed to equil-ibrate at 40 °C for 30 min. Essentially,extraction is achieved by injecting a75-µm Carboxen–Polydimethylsiloxane fiber(Sigma-Aldrich, Poole, England) into thevial and exposing to the headspace for30 min at 40 °C. The fiber was positionedat 3.0 scale units in each run. Desorption ofthe extracted volatiles was carried out on anAgilent 5890 Gas Chromatography-5972Mass Spectrometry System (Agilent Tech-nologies, UK Ltd, Cheshire, UK) run insplitless mode. During desorption the fiberremained in the injector for 2 min at a tem-perature of 250 °C, with helium as the car-rier gas at a flow rate of 1.0 mL·min–1. Thecomponents were separated on an AgilentFFAP column, 50 m × 0.2 mm × 0.33 µm(Crawford Scientific, Lanarkshire, Scot-land). The oven was held at 40 °C for 2 min(desorption period), then ramped at 5 °Cper min to 70 °C, which was held for 1 min.The temperature was then raised at 10 °Cper min to 240 °C to give a run time of30 min. The mass spectrometer was set to


record 33–450 atomic mass units, threshold1000, at a sampling rate of 1.11 scans per s.

2.4. Data analysis of SPME

Components were identified using thedata obtained from the mass spectrometerin full scan mode. A database was then setup to quantitate relative amounts of each.The database was constructed using se-lected ion monitoring with specific ions se-lected in order to allow quantitation ofco-eluting peaks. Response factors werenot used in the quantitation of the samplesas these would require the determination ofrate of release from the cheese into theheadspace, and the efficiency of extractionof each component. Thus actual concentra-tions of the components in the cheese arenot known but for the purpose of this projectcomparative values were used to show dif-ferences between the varying treatments.Instead the data was in the form (Area ofpeak/105) and was normalized to a weightof 1 g sample. This comparative data wasthen analyzed using ANOVA whereby themean response of the data, transformed togive the square root (SQRT), was calcu-lated. ANOVA of the SQRT transformeddata was carried out using Minitab Statisti-cal Software version 13 (Minitab Ltd., Cov-entry, England) on the basis of cheese type

and age. To simplify interpretation of the re-sults, principal component analysis (PCA)was performed using the varimax rotationbetween the aroma characters of thecheeses on the ANOVA results. PCA wascarried out using The Unscrambler v9.6free-trial version (CAMO, Software AS,Oslo, Norway).

3. RESULTS AND DISCUSSION

3.1. Chemical composition

The chemical composition and pH ofMalatya cheese at 1st d of ripening areshown in Table I. No significant differenceswere found between cheeses in total solids,salt and fat-in-dry matter. However, ahigher level of titratable acidity and lowerpH values were found in the cheeses madefrom pasteurized milk with a starter culture(P < 0.05). The addition of starter culture tothe pasteurized milk resulted in signifi-cantly lower pH in pasteurized milk cheeses(P) in comparison to raw milk cheeses (C).The gross composition of the cheese sam-ples at the beginning of the ripening periodare in the normal ranges for a salted cheese.These results are in accordance with thoseof Ozer et al. [34] for Urfa cheese andKahyaoglu and Kaya [21] for Gaziantep

Table I. Chemical composition and pH of Malatya cheese made from raw (C) and pasteurized (P)milk after 1 d of ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of the curd (°C).

Cheeses Titratable acidity1

pH Total solid(%)

Salt(%)

FDM2

(%)Total protein

(%)

C60 0.21 ± 0.08ab 6.02 ± 0.5a 39.85 ± 0.55a 2.70 ± 0.0a 42.03 ± 0.05a 14.03 ± 0.32ab

C70 0.14 ± 0.0a 6.03 ± 0.42a 40.20 ± 0.30a 2.81 ± 0.0a 41.64 ± 2.79a 15.04 ± 014bcd

C80 0.22 ± 0.09ab 6.01 ± 049a 38.79 ± 0.70a 2.70 ± 0.0a 45.74 ± 1.10a 16.20 ± 0.28d

C90 0.23 ± 0.07ab 5.96 ± 0.39a 38.90 ± 1.55a 2.70 ± 0.0a 49.48 ± 0.04a 15.68 ± 0.05d

P60 0.37 ± 0.03b 5.62 ± 0.16b 39.22 ± 0.17a 1.93 ± 0.0a 44.60 ± 2.35a 13.74 ± 0.50a

P70 0.39 ± 0.01b 5.57 ± 0.03b 39.20 ± 0.75a 1.76 ± 0.0a 44.68 ± 2.13a 14.94 ± 0.69bcd

P80 0.30 ± 0.02ab 5.81 ± 0.06ab 40.75 ± 2.55a 2.70 ± 0.0a 41.72 ± 0.15a 14.25 ± 0.0abc

P90 0.29 ± 0.04ab 5.81 ± 0.03ab 40.22 ± 0.52a 2.46 ± 0.0a 42.33 ± 5.52a 15.36 ± 0.01cd

Mean ± SD of duplicate determination in two cheesemaking trials. Means in the same column followedby different letters differ (P < 0.05). 1 Titratable acidity expressed as g lactic acid/100 g cheese.2 FDM: fat-in-dry matter.


cheese; these cheeses are scalded and rip-ened in brine like Malatya cheese.

3.2. Volatile composition

One hundred and two compounds wereidentified in the volatile fractions ofMalatya cheese including 11 fatty acids, 13esters, 15 ketones, 6 aldehydes, 26 alcohols,2 lactones, 5 sulfur compounds, 5 terpenesand 19 miscellaneous compounds. Thecompounds identified from the cheeseswere listed by chemical group in Tables II–X.Identification of volatile fraction has not

previously been conducted in Malatyacheese or other cheeses where scalding iscarried out on the curd in hot whey. In thepresent study, the use of pasteurised milk inthe manufacture and ripening time has sig-nificantly influenced the volatile fraction inMalatya cheese. Some of the volatile com-pounds (forty) were significantly influ-enced by using pasteurized milk in themanufacture or by scalding the curd in hotwhey and sixty-three compounds were sig-nificantly influenced by cheese age(P < 0.01).

Table II. Concentrations of fatty acids in Malatya cheese made from raw (C) and pasteurized (P)milk during ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of the curd (°C). Theresults were expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Fatty acid Age (d) Cheeses ANOVA

C60 C70 C80 C90 P60 P70 P80 P90 P-type P-age

Formic acid 3090

1.271.37

1.581.51

1.921.38

2.101.15

3.141.70

2.581.25

2.401.13

1.981.32

NS **

Ethanoic acid 3090

16.9323.96

18.8631.37

22.6228.39

23.3826.36

23.1625.00

21.6520.63

22.0418.75

17.6723.11

NS **

Propanoic 3090

1.253.88

1.204.08

1.374.39

1.643.27

1.581.37

0.991.30

0.951.31

0.801.32

*** ***

acid

2-Methyl propanoic acid

3090

1.194.79

1.573.20

1.843.53

1.892.43

1.322.09

1.312.01

1.162.06

1.141.93

** ***

Butanoic acid 3090

12.9921.76

13.6125.03

15.1523.91

14.7722.30

7.169.50

7.25 6.47 5.22 *** ***

7.90 7.66 7.71

2-Methyl butanoic acid

3090

0.712.41

0.611.79

0.712.01

0.801.36

0.631.02

0.580.93

0.500.96

0.480.89

** ***

3-Methyl butanoic acid

3090

2.676.96

2.966.36

3.696.34

3.904.97

3.064.30

3.364.24

3.044.54

2.873.91

* ***

Pentanoic acid

3090

1.732.27

1.852.67

1.832.43

1.812.35

1.201.28

1.201.20

1.081.19

0.871.10

** **

Hexanoic acid 3090

12.4516.08

11.5318.44

11.4016.59

11.9817.49

7.056.86

6.086.03

5.305.76

4.564.52

** **

Octanoic acid 3090

5.555.85

5.526.54

4.746.89

5.427.22

4.022.75

3.342.85

2.712.84

2.611.96

** NS

Decanoic acid 3090

1.962.02

2.032.20

2.122.39

2.362.40

1.841.21

1.521.27

1.341.22

1.220.89

** NS

Total 3090

58.7091.33

61.33103.18

67.3798.25

70.0591.29

54.1457.07

49.8649.60

46.9847.42

38.2148.66

* P < 0.01; ** P < 0.001; *** P < 0.0001; NS: non significant.


3.2.1. Fatty acids

Fatty acids are one of the main chemicalgroups in the volatile fraction of Malatyacheese (Tab. II). Ethanoic and propanoicare produced by the fermentation of lactoseor lactic acid; in contrast, 2-methyl propa-noic, 2- and 3-methyl butanoic acids areproduced by the metabolisms of Val, Leuand Ile amino acids, respectively [29, 40].Use of pasteurized milk in the production ofMalatya cheese and scalding the curd in hotwhey have significantly influenced all fattyacids present in the cheese except formicand ethanoic acids. Concentration of thefatty acids with the exception of octanoicand decanoic acids changed with age. For-mation of the fatty acids in raw or pasteur-ised milk cheeses exhibited a different trendduring ripening. On 90 d of ripening, rawmilk cheeses had the highest concentrationsof ethanoic, propanoic, 2-methyl propa-noic, butanoic, 2-methyl butanoic, 3-methylbutanoic, pentanoic, hexanoic, octanoicand decanoic acids probably due to numer-ous other microflora could be involved incheese lipolysis. The results obtained are inagreement with studies by Buchin et al. [6]and Shakeel-Ur-Rehman et al. [38] whofound that the concentration of acids washigher in raw milk cheese than those of pas-teurised milk cheese during ripening. Pas-teurization of cheese milk had a greatereffect than scalding the curd in hot whey onthe formation of acids in Malatya cheese. Ingeneral, the concentration of the fatty acidsdecreased as scalding temperature increased;however, a strong correlation or relationshipbetween scalding temperature and concen-tration of the fatty acids cannot be found.The concentration of formic and ethanoic ac-ids were not affected by heating of the curdin hot whey. These compounds are pro-duced by several metabolic pathways suchas lactose or butyric acid fermentation, ca-tabolism of some amino acids or hydrolysisof glycerides [29]. Butanoic (butyric) acidhas a strong effect on cheese flavour, sinceit is found only in milk fat and is described ashaving a cheesy and sweaty odour.

3.2.2. Esters

Esters contribute to cheese flavour byminimizing sharpness of fatty acids and bit-

terness of amines [35]. Table III shows themean concentrations of esters identified inMalatya cheeses during ripening. Ethyl es-ters (seven esters) are the principal esters inthe cheese samples and the other esters weremethyl, propyl and butyl (Tab. III). Thehighest level of ethyl esters in the cheesescan be correlated with higher concentra-tions of primary alcohols and fatty acids[11]. Concentration of ethyl acetate, themost abundant of esters, was higher in rawmilk cheese on 30 d of ripening and its con-centration was the same in both raw andpasteurized milk cheeses. Methyl lactate,propyl hexanoate and 3-methylbutyl bu-tanoate were not detected in pasteurizedmilk cheeses, while ethyl propionate wasnot identified in raw milk cheeses. Ethyl bu-tanoate, ethyl hexanoate, ethyl octanoateand ethyl lactate were identified at the high-est concentrations in raw milk cheeses andtheir concentrations increased significantlyduring ripening.

3.2.3. Ketones

A total of 15 ketones were identified inMalatya cheeses made from raw or pasteur-ized milk. Methyl ketones are principalcompounds in blue cheeses and are formedby enzymatic oxidation of free fatty acidsto β-ketoacids and their consequent decar-boxylation to methyl ketones [11, 25], con-tributing to the pungent aroma [31]. Themajority of ketones identified in Malatyacheeses were not different between cheesesmade from raw or pasteurized milk except2-propanone, diacetyl and 2,3-pentanedi-one. Major ketones were 2-propanone, 2-butanone, 2-pentanone, 2-heptanone, 3-hydroxy 2-butanone (acetoin) and diacetyl(Tab. IV) in the cheeses. Diacetyl (2,3-bu-tanedione) is one of the most important ke-tones and it denotes a buttery and nuttyflavours in cheese [29]. Diacetyl is pro-duced by metabolisms of lactose and citrateby cit+ Lactococcus lactis ssp. lactis andLeuconostoc species [11]. Changes in ke-tone concentration showed different trendsduring ripening. That is, some fluctuationswere observed during ripening. Theconcentration of the majority of methyl


ketones decreased during ripening, whilethe concentration of 2-propanone and 2-undecanone increased with ripening time inall cheeses. Interestingly, 1-hydroxy 2-propanone was not identified in pasteurizedmilk cheeses either at d 30 or 90; however,1-hydroxy 2-propanone and 2,3-pentanedi-one were not determined in 90-d old rawmilk cheeses. Concentrations of 3-hydroxy2-butanone and diacetyl were higher in rawmilk cheeses at 30 d but showed a marked de-crease at 90 d as compared to the trend for pas-

teurized cheeses milk. This correlated wellwith studies by Bintis and Robinson [3]who found higher levels of 3-hydroxy 2-butanone and diacetyl in fresh Feta cheesethan those of 60-d old Feta cheeses, proba-bly due to the action of starters, while theydeclined during ripening. Raw milk cheesescontained the highest levels of 2,3-pentane-dione at 30 d of ripening; however, it was notidentified at 90 d of ripening. The concen-tration of 2,3-pentanedione in pasteurizedmilk cheese was half the amount of raw milk

Table III. Concentrations of esters in Malatya cheese made from raw (C) and pasteurized (P) milkduring ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of the curd (°C). The resultswere expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Ester Age (d) Cheeses ANOVA


Methyl acetate

3090

0.350.37

0.350.34

0.340.39

0.410.31

0.350.50

0.390.53

0.480.55

0.540.63

NS NS

Methyl lactate

3090

ND1.59

ND1.53

ND1.84

ND1.83

NDND

NDND

NDND

NDND

*** ***

Ethyl acetate

3090

11.348.17

10.9311.26

10.9910.66

11.2411.28

7.2210.62

7.2410.42

9.788.81

8.848.92

NS NS

Ethyl propanoate

3090

NDND

NDND

NDND

NDND

ND1.20

ND1.09

ND1.07

ND1.04

*** ***

Ethyl butanoate

3090

4.663.78

4.194.92

4.324.72

4.115.32

1.953.66

2.253.36

2.732.92

2.573.09

* *

Ethyl hexanoate

3090

3.183.78

2.774.62

2.364.24

2.414.93

1.391.27

1.161.52

1.111.13

0.931.06

** **

Ethyl octanoate

3090

1.371.85

1.452.13

0.982.34

1.322.54

1.000.81

0.740.79

0.430.72

0.520.49

** **

Ethyl decanoate

3090

0.550.66

0.590.76

0.520.84

0.580.87

0.480.39

0.690.42

1.000.41

0.780.33

NS NS

Ethyl lactate

3090

1.202.90

1.416.44

1.744.24

1.894.15

1.802.14

1.741.50

1.641.31

1.321.81

** ***

Propyl acetate

3090

1.251.33

1.471.94

1.191.76

1.161.84

1.141.03

1.031.04

1.381.13

1.330.95

NS NS

Propyl hexanoate

3090

0.910.09

0.590.34

0.000.38

NDND

NDND

NDND

NDND

NDND

* NS

3-Methylbutyl acetate

3090

2.072.37

1.402.83

1.313.46

1.772.84

1.261.84

1.221.64

1.371.58

1.231.83

NS *

3-Methylbutyl butanoate

3090

0.68ND

0.49ND

NDND

NDND

NDND

NDND

NDND

NDND

* **

Total 3090

27.5626.89

25.6437.11

23.7534.85

24.8835.89

16.5723.46

16.4622.32

19.9119.63

18.0520.13

* P < 0.01; ** P < 0.001; *** P < 0.0001; NS: non significant; ND: not detected.


cheeses at 30 d of ripening, but their con-centration markedly declined at 90 d. Imhofet al. [19] detected high levels of 2,3-pen-

tanedione in cheeses inoculated with ther-mophilic cultures and suggested that itmight be produced by metabolism of Ile.

Table IV. Concentrations of ketones in Malatya cheese made from raw (C) and pasteurized (P)milk during ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of the curd (°C). Theresults were expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Ketone Age (d) Cheeses ANOVA


2-Propanone 3090

7.029.60

6.717.59

6.408.39

7.637.75

9.3811.92

11.2112.99

10.6414.72

11.5713.25

** **

2-Butanone 3090

10.239.41

10.018.47

9.078.82

11.598.84

15.319.57

9.909.71

10.529.44

10.339.89

NS **

2-Pentanone 3090

8.005.09

6.714.75

6.305.41

5.786.83

5.636.72

9.157.15

8.856.31

7.136.62

NS *

2-Hexanone 3090

1.010.75

0.970.78

0.810.82

0.931.04

0.921.01

1.431.08

1.190.97

1.121.03

NS NS

2-Heptanone 3090

7.976.17

7.665.72

6.146.16

8.079.70

8.446.51

15.258.00

10.965.72

8.826.44

NS *

2-Octanone 3090

0.750.50

0.600.45

0.390.63

0.510.71

0.580.45

1.250.60

0.830.70

0.760.41

NS NS

2-Nonanone 3090

2.942.85

2.922.32

2.082.45

2.773.50

3.252.04

5.322.36

3.622.46

3.181.93

NS *

2-Decanone 3090

0.350.21

0.350.10

0.120.20

0.290.26

0.28ND

0.21ND

NDND

NDND

NS NS

2-Undecanone 3090

0.450.55

0.490.56

0.420.57

0.490.62

0.460.45

0.510.48

0.410.52

0.410.44

NS *

1-Hydroxy 2-propanone

3090

1.49ND

1.57ND

1.68ND

1.78ND

NDND

NDND

NDND

NDND

*** ***

3-Hydroxy 2-butanone

3090

8.182.76

10.572.71

11.122.29

10.662.98

7.656.14

7.614.66

7.655.19

7.975.37

NS ***

3-Hydroxy 2-pentanone

3090

3.090.50

3.140.38

3.70ND

2.260.33

2.281.44

2.730.97

2.621.09

2.661.19

NS ***

2-Hydroxy 3-pentanone

3090

2.920.52

2.920.32

3.42ND

2.17ND

2.081.34

3.040.96

2.521.12

2.601.12

NS ***

Diacetyl 3090

13.804.57

13.725.72

15.625.82

12.297.58

7.746.80

9.196.71

9.507.12

9.306.53

* ***

2,3-Pentane-dione

3090

6.77ND

6.18ND

7.25ND

4.37ND

2.321.48

2.841.19

3.031.72

3.091.45

** ***

Total 3090

74.9643.45

74.5139.88

74.4941.55

71.5750.14

66.3155.87

79.6256.85

72.3357.07

68.9455.67



3.2.4. Aldehydes

Six aldehydes were identified in Malatyacheeses (Tab. V) and are produced by thecatabolism of fatty acids or amino acids viadecarboxylation or deamination. Alde-hydes are transitory compounds and do notaccumulate in cheese because they aretransformed rapidly to alcohols or to cor-responding acids [10, 23]. No significantdifferences in concentrations of 2-methyl-propanal, 2-methylbutanal and 3-methylb-utanal branched-chain aldehydes werefound among the cheeses made from raw orpasteurized milk. These aldehydes are pro-duced from Val, Ile and Leu, respectively,by Strecker degradation or transaminationand are responsible for unclean and harshflavours in Cheddar cheese [10]. The mostabundant aldehyde was 3-methylbutanal in30-d old cheeses; however, its concentra-tion decreased at 90 d and acetaldehyde wasthe highest in 90-d old cheeses. The con-centration of acetaldehyde in the cheesesincreased during ripening as observed inother types of cheese such as Roncal [32],

Emmental [5]. Benzaldehyde is formed incheese by α-oxidation of phenyl acetalde-hyde which is derived from Phe viathe Strecker reaction or β-oxidation ofcinnamic acid [25]. The concentration ofbenzaldehyde was not influenced by pas-teurization and ripening processes.

3.2.5. Alcohols

The cheeses made from raw milk con-tained higher levels than those of thecheeses made from pasteurized milk formost of the alcohols, especially after 90 dof ripening (Tab. VI). The concentrations of1-propanol, 2-propen-1-ol, 3-methyl 1-butanol, 3-methyl 2-buten-1-ol, 2-propanol,2-butanol, 2-pentanol, 2-methoxy ethanoland phenethyl alcohol were significantlyhigher in raw milk cheeses, suggesting thepasteurization process negatively influ-enced the production of alcohols in cheeseduring ripening. Primary alcohols such as1-propanol, 1-butanol, 1-pentanol, 1-hexa-nol, 1-heptanol, 1-octanol are producedmainly by the reduction of aldehydes and

Table V. Concentrations of aldehydes in Malatya cheese made from raw (C) and pasteurized (P)milk during ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of the curd (°C). Theresults were expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Aldehyde Age (d) Cheeses ANOVA


Acetaldehyde 3090

1.323.06

1.372.33

1.253.10

1.332.03

2.343.26

2.294.14

2.573.30

2.473.47

* ***

2-Propenal 3090

0.581.20

0.611.35

0.421.21

0.621.00

0.92ND

NDND

NDND

NDND

** *

2-Methyl propanal

3090

0.520.52

0.650.51

0.790.62

0.780.55

0.650.57

0.810.72

1.000.58

0.960.58

NS *

2-Methyl butanal

3090

0.320.39

0.450.37

0.440.52

0.480.41

0.310.43

0.490.53

0.570.54

0.480.34

NS NS

3-Methyl butanal

3090

2.711.06

3.300.95

4.051.35

4.251.01

3.111.20

4.401.12

6.241.05

4.491.36

NS ***

Benzaldehyde 3090

1.531.26

1.501.26

0.871.49

1.111.64

1.381.06

1.551.21

1.271.52

1.281.09

NS NS

Total 3090

6.977.48

7.876.76

7.828.27

8.576.64

8.716.51

9.527.72

11.656.99

9.686.83



Table VI. Concentrations of alcohols in Malatya cheese made from raw (C) and pasteurized (P)milk during ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of the curd (°C). Theresults were expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Alcohol Age (d) Cheeses ANOVA


Ethanol 3090

37.8929.07

36.7334.89

40.0232.20

40.2431.07

38.0339.92

36.7635.81

40.0233.58

38.9942.59

NS **

1-Propanol 3090

3.959.18

4.0410.35

4.488.78

6.148.64

6.433.12

3.473.32

4.113.34

4.203.62

** ***

2-Propen-1-ol 3090

0.572.31

0.853.04

0.702.89

1.353.24

1.31ND

0.57ND

0.55ND

NDND

*** ***

2-Methyl 1-propanol

3090

1.674.55

3.114.39

3.235.62

3.833.97

1.524.49

1.546.35

1.696.60

1.664.56

NS ***

1-Butanol 3090

4.503.35

4.743.57

3.833.54

3.703.31

3.612.58

3.782.79

4.453.00

3.883.15

NS ***

2-Methyl 1-butanol

3090

2.996.55

3.816.56

3.907.44

4.346.04

3.305.51

3.446.13

3.646.20

3.445.36

NS ***

3-Methyl 1-butanol

3090

7.8415.85

10.1716.10

10.6317.21

11.8614.81

8.9612.33

9.3512.94

9.8613.24

9.4412.14

* ***

3-Methyl 3-buten-1-ol

3090

1.202.27

1.333.03

1.382.27

1.392.05

1.351.91

1.351.59

1.321.56

1.221.94

NS **

3-Methyl 2-buten-1-ol

3090

0.901.92

0.882.64

0.872.39

0.982.38

1.221.19

1.011.01

0.941.00

0.801.10

** ***

1-Pentanol 3090

1.591.40

1.671.25

1.551.48

1.561.35

1.451.86

1.631.69

1.791.76

1.611.82

NS NS

1-Hexanol 3090

2.292.27

2.532.34

2.102.11

2.222.31

2.122.05

2.661.96

2.572.06

2.302.06

NS NS

2-Ethyl 1-hexanol

3090

1.782.03

2.062.20

1.682.06

1.972.58

2.232.20

2.472.35

1.762.52

1.442.31

NS *

1-Heptanol 3090

0.570.64

0.570.65

0.530.62

0.580.72

0.540.59

0.540.59

0.490.61

0.470.49

NS *

1-Octanol 3090

0.810.56

0.950.55

1.020.51

1.220.60

0.880.45

0.650.50

0.650.55

0.540.41

NS **

2-Propanol 3090

4.835.46

4.765.73

4.625.76

4.205.14

3.042.59

2.734.24

2.654.28

2.703.64

** **

2-Butanol 3090

4.3720.18

4.5222.76

4.4720.87

7.1319.11

5.504.48

3.474.96

3.865.14

3.615.52

*** ***

2-Pentanol 3090

3.174.68

2.825.66

2.725.12

2.704.86

2.393.06

2.352.75

2.422.96

2.283.56

* ***

2-Hexanol 3090

0.890.88

0.661.03

0.620.94

0.610.93

0.510.61

0.560.62

0.560.63

0.530.65

NS *


methyl ketones [1] and they impart an alco-holic, winey, sweet, fruity and harsh notesin cheese [2]. The concentration of primaryalcohols were higher in the cheeses thanthose of secondary alcohols, probably dueto the higher concentrations of ethanol and3-methyl 1-butanol. The highest amountsof ethanol were found at 30 d of ripening inthe cheeses and its concentration decreasedin 90-d old cheeses. Ethanol is also foundas the principal alcohol in other types ofcheeses such as Feta [3, 18], Roncal [31],Minas [8], Cheddar [1] and Hispanico [33].The presence of the branched-chain pri-mary alcohols including 2-methyl-1-butanol,2-methyl-1-propanol and 3-methyl-1-butanolin the cheeses indicates conversion of alde-hydes produced from Ile, Val and Leu, re-spectively [11]. Secondary alcohols such as2-propanol, 2-butanol, 2-pentanol are formedby enzymatic reduction of methyl ketones[29] and their amounts changed with pas-teurization of milk or ageing. The concen-

tration of 2-butanol was highest withinsecondary alcohols in raw milk cheeses at90 d of ripening. Of the secondary alcohols,the concentrations of 2-heptanol, 2-octanol,2-nonanol which may be produced byreduction of methyl ketones were not influ-enced by pasteurization or ripening proc-esses, suggesting no role of NSLAB orreleased enzymes of starter cultures used inthe production. 2-Methoxy ethanol waspresent in all cheeses at the highest levelsat 30 d of ripening, but its concentration wasinfluenced by pasteurization and ageingprocesses. The formation of 1-methoxy 2-propanol, 2-butoxy ethanol and benzyl al-cohol were not significantly affected bypasteurization or ageing, except 2-butoxyethanol was affected with age. Raw milkcheeses contained a higher concentration ofphenethyl alcohol than pasteurized milkcheeses at each stages of ripening. Phene-thyl alcohol which may be produced bycatabolism of Phe via transamination,

Table VI. (continued).

Alcohol Age (d) Cheeses ANOVA


2-Heptanol 3090

7.254.76

4.415.46

2.845.04

3.025.17

2.642.57

3.352.69

2.682.86

2.442.60

NS NS

2-Octanol 3090

0.470.45

0.680.44

0.470.50

0.570.57

0.550.49

0.620.48

0.620.46

0.640.56

NS NS

2-Nonanol 3090

1.401.28

1.071.32

0.661.41

0.791.53

0.800.63

1.330.66

0.790.68

0.700.54

NS NS

2-Methoxy ethanol

3090

2.542.06

2.552.11

2.432.01

2.302.04

2.231.70

2.231.50

2.141.52

1.991.48

* ***

1-Methoxy 2-propanol

3090

3.142.05

3.852.61

1.432.64

1.862.43

1.941.89

2.222.08

2.202.16

1.982.16

NS NS

2-Butoxy ethanol

3090

1.270.84

1.110.84

0.860.82

0.940.82

0.870.65

0.830.68

0.790.76

0.710.77

NS *

Benzyl alcohol

3090

0.951.05

0.981.06

0.851.10

0.910.93

1.040.51

1.110.67

0.820.87

0.530.60

NS NS

Phenethyl alcohol

3090

1.121.89

0.891.58

0.582.03

0.671.81

0.650.87

0.601.01

0.551.04

0.520.76

* ***

Total 3090

99.91127.51

101.71142.14

98.45137.34

107.04128.40

95.1198.23

90.5899.36

93.9299.36

88.60104.38

* P < 0.01, ** P < 0.001, *** P < 0.0001; NS: non significant; ND: not detected.


decarboxylation or reduction reactions [25]provides a rose like note to cheese and it isone of the main volatiles in Camembertcheese [29]. Correa Lelles Nogueira et al.[8] reported that the floral character of Mi-nas cheese correlated with the availabilityof phenethyl alcohol in the cheese.

3.2.6. Lactones

Only two lactones, butyrolactone and γ -hexanolactone were identified in Malatyacheeses (Tab. VII) and no significant dif-ferences were seen between cheeses for bu-tyrolactone; its concentration was at ahigher level at 30 d of ripening and then de-clined. γ -Hexanolactone was present inonly P60 cheeses within pasteurized milkcheeses at 30 d of ripening and its concen-tration in raw milk cheeses did not changewith age. Lactones contribute to cheese fla-vour giving fruity notes (peach, apricot, andcoconut) [29] and may be formed by in-tramolecular esterification of hydroxy fattyacids by action of microorganisms; that is,they form spontaneously once the fatty acidis released by lipolysis [25]. This suggeststhat the lactones contribute to the creamycoconut note of the flavour of Malatya cheese.

3.2.7. Sulfur compounds

The concentrations of sulfur compoundsin Malatya cheese declined with age(Tab. VIII). The most abundant compoundwas carbon disulfide at 30 d of ripening inthe cheeses. Carbon disulfide was also the

most abundant sulfur compound in Man-chego cheese made from raw or pasteurizedmilk and no significant differences werefound between cheeses [12]. The concen-trations of sulfur compounds were not in-fluenced by milk pasteurization exceptdimethyl disulfide. Dimethyl trisulfide wasnot identified in raw milk cheeses at any rip-ening period. Methanethiol was not presentin any of the cheeses probably due to the ex-traction method of the volatiles; this com-pound is considered a key compound forCheddar flavour [40, 41]. The lower levelsof dimethyl trisulfide in cheeses (absent inraw milk cheeses) can be linked to the ab-sence of methanethiol which is a precursorof dimethyl disulfide and dimethyl tri-sulfide [11].

3.2.8. Terpenes

Five terpenes were identified in Malatyacheese and α-pinene is the principal terpenein the cheese (Tab. IX). α-Pinene was alsoidentified in Cheddar [9], Minas [8] andManchego [12] cheeses. These compoundsoriginate from plants which represent thefeed mixture of pasture and are then trans-ferred to milk and milk product via grazinganimals [8]. Camphene was not identifiedin raw milk cheeses and it was found in pas-teurized milk cheeses only at 90 d of ripen-ing. β-Pinene was not identified at d 30 forall cheeses, while its concentration washigher in pasteurized milk cheeses than rawmilk cheese at 90 d of ripening. Limoneneand p-cymene were present in all cheeses

Table VII. Concentrations of lactones in Malatya cheese made from raw (C) and pasteurized (P)milk during ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of the curd (°C). Theresults were expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Lactone Age (d) Cheeses ANOVA


Butyrolactone 3090

1.080.81

1.080.87

0.910.80

0.940.72

0.890.79

0.900.74

0.850.72

0.810.71

NS *

γ -Hexanolactone 3090

0.430.26

0.350.34

0.290.36

0.310.37

0.24ND

NDND

NDND

NDND

* NS

Total 3090

1.511.07

1.431.21

1.201.15

1.241.10

1.130.79

0.900.74

0.850.72

0.810.71

* P < 0.01; NS: non significant; ND: not detected.


and there were no significant differencesamong cheeses, suggesting no role of pas-teurization and ripening process to the for-mation of limonene and p-cymene.

3.2.9. Miscellaneous compounds

A number of miscellaneous compoundswere identified in Malatya cheese (Tab. X).

Table VIII. Concentrations of sulfur compounds in Malatya cheese made from raw (C) andpasteurized (P) milk during ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of thecurd (°C). The results were expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Sulfur compound Age (d) Cheeses ANOVA


Carbon disulphide 3090

2.190.84

1.660.89

2.110.93

1.771.19

1.780.91

2.770.88

1.810.69

3.080.80

NS ***

Dimethyl sulphide 3090

0.830.77

0.700.77

0.800.84

0.701.27

0.730.93

0.900.99

0.650.61

0.820.96

NS *

Dimethyl disulphide 3090

0.570.38

0.550.54

0.400.46

0.430.56

0.590.83

0.850.76

0.820.69

0.920.75

* NS

Dimethyl trisulphide 3090

NDND

NDND

NDND

NDND

ND0.28

0.420.32

0.39ND

0.460.21

NS NS

Dimethyl sulphone 3090

0.450.37

0.510.41

0.620.44

0.620.39

0.680.39

0.630.31

0.610.35

0.490.32

NS *

Total 3090

4.042.35

3.422.61

3.922.68

3.523.41

3.793.34

5.553.25

4.272.34

5.773.03

* P < 0.01; *** P < 0.0001; NS: non significant; ND: not detected.

Table IX. Concentrations of terpenes in Malatya cheese made from raw (C) and pasteurized (P)milk during ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of the curd (°C). Theresults were expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Terpene Age (d) Cheeses ANOVA


α-Pinene 3090

0.661.96

0.722.17

0.612.20

0.562.27

0.484.80

0.574.87

0.574.25

0.604.65

** ***

Camphene 3090

NDND

NDND

NDND

NDND

ND0.55

ND0.54

ND0.50

ND0.50

*** ***

β-Pinene 3090

ND0.34

ND0.21

ND0.33

ND 0.37

ND0.70

ND0.73

ND0.62

ND0.72

NS ***

Limonene 3090

1.260.64

1.250.70

0.770.66

0.750.86

0.720.59

0.700.77

0.800.70

0.830.66

NS NS

p-Cymene 3090

1.540.83

1.460.68

0.660.72

0.950.84

0.901.06

0.731.29

0.621.25

0.700.99

NS NS

Total 3090

3.463.77

3.433.75

2.033.90

2.264.34

2.097.70

2.018.20

1.997.31

2.137.52

** P < 0.001; *** P < 0.0001; NS: non significant; ND: not detected.


2,5-Dimethyl pyrazine was detected in 30-dold cheeses at very low levels, but not de-termined in C80 and P60 cheeses. Thecompound is produced in cheese bycondensation of amino ketones, which areformed by Maillard and Strecker degrada-tion reactions [13] and imparts roasted andhazelnut notes [27]. Phenolic compoundswhich are mainly present in sheep’s milk asconjugates of phosphate and sulphate con-tribute to cheese aroma at about thresholdconcentration [40]. Phenol is an importantcompound for surface ripened cheese andits contribution to cheese aroma is per-ceived as sharp and medicinal notes [29].The concentrations of phenol and acetophe-none were not different among the cheesesmade from raw or pasteurized milk. Theirconcentration increased significantly withage. Small differences, but not significant,were seen between acetophenone levels ofraw or pasteurized milk cheeses at 90 d ofripening. Acetophenone is produced incheese via β-oxidation of phenylpropionicacid followed by decarboxylation of the β-ketoacid [37]. 4-Methyl phenol (p-cresol) isformed via catabolism of aromatic aminoacids and contributes to cheese flavour asunclean or off-flavours [25]. In Malatyacheese, p-cresol was not found in pasteur-ized milk cheeses and found only in 90-dold raw milk cheeses. Methyl salicylate andp-methyl anisole were present in all cheesesat similar abundances and their concentra-tions declined with age. Hydrocarbons aresecondary products of lipid autoxidationand are precursors for the formation of otheraromatic compounds [5]. Benzene, tolueneand ethyl benzene were found in the volatilefraction of Malatya cheese and their con-centrations were not influenced by pasteur-ization or ripening processes. Of thesehydrocarbons, toluene was the most abun-dant compound and it was also identified athigh levels in Feta-type [3] and a Spanishewe’s milk semi-hard [24] cheeses. In ad-dition, benzyl nitrile which is a benzyl com-pound was identified only in 90-d old rawmilk cheeses. The concentrations of carbondioxide was similar in all cheeses, this com-pound is produced by catabolism of lactate,citrate and fatty acids [25]. Its concentrationin raw milk cheeses did not change during

ripening, but decreased in pasteurized milkcheeses. Styrene which has a strong plasticodour was found in Malatya cheeses madefrom raw or pasteurized milk and differ-ences between cheeses were not significant.Styrene was also detected at trace levels inProvola dei Nebrodi cheese [42], Parmi-giano [4], Roncal [20] and Camembert [29]cheeses. Diethyl ether, chloroform and bro-moform which are likely external origincompounds were also identified in Malatyacheeses. Diethyl ether and bromoform werefound in the cheeses at similar concentra-tions; however, chloroform had higher lev-els in pasteurized milk cheeses. Chloroformwas also identified in Roncal [32], Feta-type [3] and Provola dei Nebrodi [42]cheeses.

3.2.10. Principal component analysis

Principal component analysis (PCA)was applied to those variables which pre-sented significant (P < 0.01) differencesamong the cheeses to clarify separation ofthe cheese samples and interpretation of theresults. Significant differences among thesamples on the first two principal com-ponents on the PCA were determined byANOVA. The results of PCA indicated thatthe cheeses were distinguished according totheir GC-MS profiles. Two bi-plots of thesample scores and variable loadings forPC1 and PC2 is shown in Figures 1a and 1bfor 30 or 90 d of ripening, respectively. PC1and PC2 explained 88 and 6% of the var-iation between the volatiles of the cheeses,respectively, at 30 d. For 30 d, PC1 sep-arated the cheeses on the basis of milk typeused (raw or pasteurized) in the productionof Malatya cheese. Samples were dis-tributed into two main groups: raw milkcheeses located on the positive side on PC1and pasteurized milk cheeses located on thenegative side of PC1. The cheeses madewith raw milk contained higher levels of ac-ids, esters and lactones and lower levels ofaldehydes and sulfur compounds than didthe cheeses made from pasteurized milk. At30 d, the cheeses made from raw milk weredescribed as higher abundance of diacetyl,octane, pentanoic acid (5,0), 2-propenal,2-propanol, 2-propen-1-ol and 1-hydroxy


Table X. Concentrations of miscellaneous compounds in Malatya cheese made from raw (C) andpasteurized (P) milk during ripening. Codes 60, 70, 80 or 90 refer to scalding temperature of thecurd (°C). The results were expressed as SQRT [Area/105] from triplicate analysis of each cheese.

Miscellaneous Age (d) Cheeses ANOVA


2,5-Dimethyl pyrazine

3090

0.26ND

0.28ND

NDND

0.210.26

NDND

0.25ND

0.25ND

0.10ND

NS NS

Phenol 3090

0.700.86

0.680.90

0.550.89

0.640.89

0.650.79

0.600.89

0.590.87

0.550.85

NS **

2-Methyl phenol 3090

ND0.47

ND0.47

ND0.52

ND0.55

NDND

NDND

NDND

NDND

** ***

4-Methyl phenol 3090

0.060.53

ND0.51

ND0.61

ND0.61

NDND

NDND

NDND

NDND

** ***

Acetophenone 3090

0.911.23

1.201.18

0.761.40

0.911.47

0.980.99

1.211.08

0.861.32

0.890.96

NS *

p-Methyl anisole 3090

0.520.69

0.720.55

0.470.54

0.730.71

0.720.37

0.920.52

0.800.51

0.790.35

NS **

Methyl salicylate 3090

0.980.61

1.020.53

0.650.54

0.780.56

0.770.32

0.690.39

0.550.38

0.570.23

NS **

Benzene 3090

1.010.79

1.210.86

0.990.84

0.990.88

0.751.13

0.871.18

1.141.25

1.081.05

NS NS

Toluene 3090

5.433.18

5.943.52

4.333.69

4.253.91

3.494.71

4.254.89

4.664.88

4.454.75

NS NS

Ethyl benzene 3090

3.672.50

3.802.70

2.882.68

2.683.08

2.522.91

2.933.03

2.822.93

3.07 NS NS

2.85

Carbon dioxide 3090

9.329.79

9.6510.60

10.6011.28

11.059.50

12.379.29

11.088.27

10.967.78

9.518.28

NS *

Styrene 3090

3.663.04

4.582.69

2.933.07

3.173.43

3.013.00

3.373.51

3.083.79

3.293.21

NS NS

Benzyl nitrile 3090

ND0.37

ND0.46

ND0.50

ND0.48

NDND

NDND

NDND

NDND

*** ***

Pentane 3090

5.113.49

6.023.89

4.344.10

4.253.68

4.134.97

3.554.79

4.555.25

4.454.11

NS NS

Hexane 3090

3.541.61

3.561.91

2.332.12

3.061.40

1.771.73

1.771.54

3.901.83

1.741.47

NS **

Octane 3090

0.590.60

0.380.53

0.630.60

ND0.72

NDND

ND0.56

NDND

ND0.49

* *

Diethyl ether 3090

6.364.24

5.974.94

5.635.59

5.634.15

4.316.74

4.977.07

7.336.02

7.416.51

NS NS

Chloroform 3090

5.173.41

5.573.59

5.003.72

4.463.07

3.664.37

4.135.07

6.195.56

7.344.78

* **

Bromoform 3090

0.690.33

0.750.40

0.580.33

0.670.40

0.450.37

0.660.41

0.810.40

0.570.43

NS **

Total 3090

47.9737.74

51.3340.23

42.6643.03

43.4639.75

39.6041.68

41.2543.20

48.5042.75

45.8040.31



Figure 1. Bi-plots of principal components 1 and 2 showing the cheese sample scores and variableloadings from principal component analysis of the volatile data for Malatya cheeses made from raw(C) and pasteurized (P) milk and scalded at 60, 70, 80, or 90 °C after 30 (a) and 90 (b) d of ripening.


2-propanone than those of pasteurized milkcheeses. PCA was also applied to the GC-MS data obtained from the 90-d oldcheeses. Samples were distributed into twogroups (raw and pasteurized milk cheeses)as in 30-d old cheeses. PC1 and PC2 ac-counted 98% and 1% of the variance, re-spectively. PC1 separated the cheeses onthe basis of use of raw or pasteurized milkin the cheese manufacture, while no regulardistribution was observed by PC2 based onscalding temperature of the curd. Raw milkcheeses were described as containinghigher levels of pentanoic acid, 2-methylpropanoic acid, 2-propan-1-ol, 2-propanal,3-methylbutyl butanoate and 1-hydroxy 2-propanone than those of pasteurized milkcheeses. The similarity of the plots obtainedfor each ripening period showed that thepasteurization of milk had a greater effecton the volatile composition of the cheesethan scalding temperature of the curd.

4. CONCLUSION

The results of this study showed that theuse of raw milk in the manufacture en-hanced the volatiles in cheese, especiallyacids, alcohols and esters. Also, it can beconcluded that the pasteurization of themilk has a decreasing and/or delaying ef-fects on the development of the volatilecompounds of cheese. Scalding the curd inhot whey at different temperatures has aslight effect on the volatile compounds inMalatya cheese and pasteurization ofcheese milk had a greater effect on the vol-atile composition of cheese than the scald-ing temperature of the curd. Many volatilecompounds are responsible for the charac-teristic flavour of Malatya cheese.

Acknowledgements: The authors gratefullyexpress their gratitude to Karlidag Dairy Prod-ucts (Malatya, Turkey) for using their facilityduring cheese making and Prof. A.H. Pripp(Norway) for advice on multivariate statisticalanalysis.

REFERENCES

[1] Arora G., Cormier F., Lee B., Analysis ofodor active volatiles in Cheddar cheeseheadspace by multidimensional GC/MS/Sniffing, J. Agric. Food Chem. 43 (1995)748–752.

[2] Barron L.J.R., Redondo Y., Aramburu M.,Pérez-Elortondo F.J., Albisu M., NajeraA.I., de Renobales M., Variations in volatilecompounds and flavour in Idiazabal cheesemanufactured from ewe’s milk in farmhouseand factory, J. Sci. Food Agric. 85 (2005)1660–1671.

[3] Bintis T., Robinson R.K., A study of theeffects of adjunct cultures on the aromacompounds of Feta-type cheese, FoodChem. 88 (2004) 435–441.

[4] Bosset J.O., Gauch R., Comparison of thevolatile flavour compounds of six European“AOC” cheeses by using a new DynamicHeadspace GC-MS method, Int. Dairy J. 3(1993) 359–377.

[5] Bosset J.O., Bütikofer U., Gauch R., Sieber R.,Ripening of Emmental cheese wrapped infoil with and without addition of Lactobacil-lus casei subsp. casei. II. Gas chromato-graphic investigation of some volatile neu-tral compounds using dynamic headspace,Lebensm.-Wiss. Technol. 30 (1997) 464–470.

[6] Buchin S., Delague V., Duboz G., BerdaguéJ.L., Beuvier E., Pochet S., Grappin R.,Influence of pasteurization and fat composi-tion of milk on the volatile compounds andflavor characteristics of a semi-hard cheese,J. Dairy Sci. 81 (1998) 3097–3108.

[7] Bugaud C., Buchin S., Hauwuy A., CoulonJ.B., Relationships between flavour andchemical composition of Abondance cheesederived from different types of pastures, Lait81 (2001) 757–773.

[8] Correa Lelles Nogueira M., Luachevsky G.,Rankin S.A., A study of volatile composi-tion of Minas cheese, Lebensm.-Wiss.Technol. 38 (2005) 555–563.

[9] Curioni P.M.G., Bosset J.O., Key odorantsin various cheese types as determined by gaschromatography-olfactometry, Int. Dairy J.12 (2002) 959–984.

[10] Dunn H.C., Lindsay R.C., Evaluation of therole of microbial Strecker-derivated aromacompounds in unclean-type flavours ofCheddar cheese, J. Dairy Sci. 68 (1985)2859–2874.


[11] Engels W.J.M., Dekker R., de Jong C.,Neeter R., Visser S.A., A comparative studyof volatile compounds in the water-solublefraction of various types of ripened cheese,Int. Dairy J. 7 (1997) 255–263.

[12] Fernandez-Garcia E., Carbonell M., NunezM., Volatile fraction and sensory character-istics of Manchego cheese. 1. Comparison ofraw and pasteurized milk cheese, J. DairyRes. 69 (2002) 579–593.

[13] Frank D.C., Owen C.M., Patterson J., Solidphase microextraction (SPME) combinedwith gas-chromatography and olfactometry-mass spectrometry for characterization ofcheese aroma compounds, Lebensm.-Wiss.Technol. 37 (2004) 139–154.

[14] Gardiner G.E., Ross R.P., Wallace J.M.,Scanlan F.P., Jagers P.P.J.M., FitzgeraldG.F., Collins J.K., Stanton C., Influence of aprobiotic adjunct culture of Enterococcusfaecium on the quality of Cheddar cheese, J.Agric. Food Chem. 47 (1999) 4907–4916.

[15] Grappin R., Beuvier E., Possible implica-tions of milk pasteurization on the manufac-ture and sensory quality of ripened cheese,Int. Dairy J. 7 (1997) 751–761.

[16] Hayaloglu A.A., Guven M., Fox P.F.,McSweeney P.L.H., Influence of starters onchemical, biochemical, and sensory changesin Turkish White-brined cheese during rip-ening, J. Dairy Sci. 88 (2005) 3460–3474.

[17] Horne J., Carpino S., Tuinello L., Rapisarda T.,Corallo L., Licitra G., Differences in vola-tiles, and chemical, microbial and sensorycharacteristics between artisanal and indus-trial Piacentinu Ennese cheeses, Int. Dairy J.15 (2005) 605–617.

[18] Horwood J.F., Lloyd G.T., Stark W., Someflavour compounds of Feta cheese, Aust. J.Dairy Technol. 36 (1981) 34–37.

[19] Imhof R., Glattli H., Bosset J.O., Volatileorganic aroma compounds produced by ther-mophilic and mesophilic mixed strain dairystarter cultures, Lebensm.-Wiss. Technol.27 (1994) 442–449.

[20] Izco J.M., Torre P., Characterization of Ron-cal cheese volatile flavour compoundsextracted by the purge and trap method andanalysed by GC-MS, Food Chem. 70 (2000)409–417.

[21] Kahyaoglu T., Kaya S., Effect of heat treat-ment and fat reduction on the rheologicaland functional properties of Gaziantepcheese, Int. Dairy J. 13 (2003) 867–875.

[22] Lau K.Y., Barbano D.M., Rasmussen R.R.,Influence of pasteurization of milk on pro-

tein breakdown in Cheddar cheese duringaging, J. Dairy Sci. 74 (1991) 727–740.

[23] Le Quere J.L., Molimard P., Cheese flavour,in: Roginski H., Fuquay J.W., Fox P.F.(Eds.), Encyclopedia of Dairy Sciences,vol. 1, Academic Press, London, UK, 2002,pp. 330–340.

[24] Mariaca R.G., Fernandez-Garcia E.,Mohedano A.F., Nunez M., Volatile fractionof ewe’s milk semi-hard cheese manufac-tured with and without the addition ofcysteine proteinase, Food Sci. Technol. Int.7 (2001) 131–139.

[25] McSweeney P.L.H., Sousa M.J., Biochemi-cal pathways for the production of flavourcompounds in cheese during ripening, Lait80 (2000) 293–324.

[26] McSweeney P.L.H., Fox P.F., Lucey J.A.,Jordan K.N., Cogan T.M., Contribution ofthe indigenous microflora to the maturationof Cheddar cheese, Int. Dairy J. 3 (1993)613–634.

[27] Moio L., Addeo F., Grana Padano cheesearoma, J. Dairy Res. 65 (1998) 317–333.

[28] Moio L., Piombino P., Addeo F., Odour-impact compounds in Gorgonzola cheese, J.Dairy Res. 67 (1999) 273–285.

[29] Molimard P., Spinnler H., Compoundsinvolved in the flavor of surface mold-rip-ened cheese: origins and properties, J. DairySci. 79 (1996) 169–184.

[30] Ordonez A.I., Ibanez F.C., Torre P., Barcina Y.,Effect of ewe’s milk pasteurization on thefree amino acids in Idiazabal cheese, Int.Dairy J. 9 (1999) 135–141.

[31] Ortigosa M., Torre P., Izco J.M., Effect ofpasteurization of ewe’s milk and use of anative starter culture on the volatile compo-nents and sensory characteristics of Roncalcheese, J. Dairy Sci. 84 (2001) 1320–1330.

[32] Ortigosa M., Arizcun C., Torre P., Izco J.M.,Use of wild Lactobacillus strains in anadjunct culture for a Roncal-type cheese, J.Dairy Res. 72 (2005) 168–178.

[33] Oumer A., Gaya P., Fernandez-Garcia E.,Mariaca R., Garde S., Medina M., Nunez M.,Proteolysis and formation of volatile com-pounds in cheese manufactured with a bacte-riocin-producing adjunct culture, J. DairyRes. 68 (2001) 117–129.

[34] Ozer B.H., Robinson R.K., Grandison A.S.,Textural and microstructural properties ofurfa cheese (a white-brined Turkish cheese),Int. J. Dairy Technol. 56 (2003) 171–176.


[35] Pinho O., Peres C., Ferreira I.M.P.L.V.O.,Solid-phase microextraction of volatilecompounds in “Terrincho” ewe cheese.Comparison of different fibers, J. Chroma-togr. A 1011 (2003) 1–9.

[36] Sable S., Cottenceau G., Current knowledgeof soft cheeses flavor and related com-pounds, J. Agric. Food Chem. 47 (1999)4825–4836.

[37] Seitz E.W., Microbial and enzyme-inducedflavours in dairy foods, J. Dairy Sci. 73(1990) 3664–3691.

[38] Shakeel-Ur-Rehman, McSweeney P.L.H.,Banks J.M., Brechany E.Y., Muir D.D., FoxP.F., Ripening of Cheddar cheese made fromblends of raw and pasteurized milk, Int.Dairy J. 10 (2000) 33–44.

[39] Shakeel-Ur-Rehman, Banks J.M., BrechanyE.Y., Muir D.D., McSweeney P.L.H., Fox

P.F., Influence of ripening temperature onthe volatiles profile and flavour of Cheddarcheese made from raw or pasteurized milk,Int. Dairy J. 10 (2000) 55–65.

[40] Urbach G., The chemical and biochemicalbasis of cheese and milk aroma, in: LawB.A. (Ed.), Microbiology and Biochemistryof Cheese and Fermented Milk, 2nd edn.,Blackie Academic and Professional, Lon-don, UK, 1997, pp. 253–298.

[41] Weimer B., Seefeldt K., Dias B., Sulfurmetabolism in bacteria associated withcheese, Anton. Leeuwenhoek 76 (1999)247–261.

[42] Ziino M., Condurso C., Romeo V., GiuffridaD., Verzera A., Characterization of “Provoladei Nebrodi”, a typical Sicilian cheese, byvolatiles analysis using SPME-GC/MS, Int.Dairy J. 15 (2005) 585–593.

To access this journal online:www.edpsciences.org

Documents

Influence of milk pasteurization and scalding temperature