Composition, proteolysis, lipolysis, volatile compound profile and sensory characteristics of ripened white cheeses manufactured in different geographical regions of Turkey

Accepted Manuscript Composition, proteolysis, lipolysis, volatile compound profile and sensory characteristics of ripened white cheeses manufactured in different geographical regions of Turkey Pelin Salum, Gokce Govce, Perihan Kendirci, Deniz Bas, Zafer Erbay PII:

S0958-6946(18)30183-3

DOI:

10.1016/j.idairyj.2018.07.011

Reference:

INDA 4365

To appear in:

International Dairy Journal

Received Date: 24 May 2018 Revised Date:

16 July 2018

Accepted Date: 16 July 2018

Please cite this article as: Salum, P., Govce, G., Kendirci, P., Bas, D., Erbay, Z., Composition, proteolysis, lipolysis, volatile compound profile and sensory characteristics of ripened white cheeses manufactured in different geographical regions of Turkey, International Dairy Journal (2018), doi: 10.1016/j.idairyj.2018.07.011. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

ACCEPTED MANUSCRIPT Composition, proteolysis, lipolysis, volatile compound profile and sensory characteristics

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of ripened white cheeses manufactured in different geographical regions of Turkey

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Pelin Salum a, Gokce Govce b, Perihan Kendirci c, Deniz Bas d, Zafer Erbay b*

Department of Food Engineering, Institute of Natural and Applied Sciences,

b

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Cukurova University, 01130 Adana, Turkey.

Department of Food Engineering, Faculty of Engineering, Adana Science and

Technology University, 01250 Adana, Turkey.

Department of Gastronomy and Culinary Arts, Faculty of Tourism, Katip Celebi

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c

University, Izmir, Turkey.

Department of Food Engineering, Faculty of Engineering, Cankiri Karatekin

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d

University, Cankiri, Turkey.

*Corresponding author. Tel.: +90 332 4550000 ext 2080 E-mail address: [email protected] (Z. Erbay)

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ACCEPTED MANUSCRIPT ___________________________________________________________________________ ABSTRACT

Turkish white cheese is the most important cheese variety made in Turkey and significant

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variations are commonly observed in their properties (mainly sensory). White cheese samples were collected from different geographical regions of Turkey and analysed to determine the common characteristics and to investigate the effects of the regional differences arising from

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differential natural conditions and the natural variations in raw milk. Although the samples showed similar compositional properties, there were significant variations in the ripening

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parameters and flavour attributes. Forty-five volatile compounds were identified and a descriptive vocabulary consisting of 15 common attributes were used to describe Turkish white cheese. Cheeses with moderate proteolytic ripening degrees and high lipolytic ripening values caused an increase in the preference score. Moreover, the increase in the volatile acid

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contents positively affected the preference. The most preferred cheeses were manufactured in two neighbouring provinces located in the north-west of the country.

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ACCEPTED MANUSCRIPT 1.

Introduction

Turkish white cheese (original name is “Beyaz peynir”) is one of the most important traditional cheeses manufactured in Turkey. Its annual production represents more than 60%

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of the total cheese production (Hayaloglu, Ozer, & Fox, 2008; USK, 2014). Traditionally, sheep’s or goats’ milk is used in the production of Turkish white cheese. However, due to the insufficient supply of these milks, cows’ milk or the mixture of milks is generally used

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(Hayaloglu, Guven, & Fox, 2002). In 2017, cheese production in Turkey was 688061 tons and 96% of this total corresponded to cheeses made from cows’ milk (TUIK, 2018).

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Turkish white cheese is a brined type cheese and generally defined as white-coloured, rindless, salty, acidic taste, and without eye formation (Kamber, 2008; Özer, Kirmaci, Hayaloglu, Akçelik, & Akkoç, 2011). There is no use of starter cultures in the traditional production technique. While the fresh Turkish white cheese has a soft texture, after a ripening

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period of 3 months, the texture turns into a semi-hard structure (Hayaloglu et al., 2002). In the scientific literature, there are some studies carried out to characterize different properties of Turkish white cheeses found in the market. Gross composition (Güler & Uraz,

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2004; Turantaş, Ünlütürk, & Göktan, 1989), mineral compositions (Mendil, 2006; Merdivan,

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Yilmaz, Hamamci, & Aygun, 2004), microbiological quality (Kivanç, 1989; Turantaş et al., 1989), aflatoxin M1 contents (Ardic, Karakaya, Atasever, & Adiguzel, 2009; Yaroglu, Oruc, & Tayar, 2005), organic acid contents (Akalin, Gönç, & Akbaş, 2002), and free fatty acid profiles (Akalin, Kinik, & Gönç, 1998) have been investigated. However, the variations observed in these properties clearly reveal the differential natural conditions, the use of different types of milk, the natural variations of raw milk, and the diversity of the cheesemaking procedures (Hayaloglu et al., 2002; Kamber, 2008; Kivanç, 1989; Turantaş et al., 1989).

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ACCEPTED MANUSCRIPT On the other hand, there is limited information about the volatile compositions of Turkish white cheeses and no studies, according to the authors’ knowledge, focused on the sensory attributes and their relationship with the volatile profile in the scientific literature (Akalin & Karaman, 2011; Özer et al., 2011; Sahingil, Hayaloglu, Simsek, & Ozer, 2014). In

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this study, nine different Turkish white cheese samples were collected from different

production plants with high capacity located in different provinces of Turkey. Cheeses were manufactured from cows’ milk using the traditional procedure without starter cultures and

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ripened for 6 months. The physico-chemical composition, proteolytic and lipolytic ripening parameters along with free fatty acid and volatile profiles and sensory attributes of the white

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cheeses were analysed. The effects of the raw milk properties connected with regional differences and the common properties of the cheeses have been investigated.

Materials and methods

2.1.

Cheese samples

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2.

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The nine Turkish white cheese samples manufactured with traditional techniques from cows’ milk and ripened for 6 months were collected from different geographical regions of

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Turkey in 2016. The provinces were coded as follows; Çanakkale (1 and 5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9). Minimum collected sample amount was 10 kg for each cheese. They were packed in vacuum and kept at 4 °C until performing colour and sensory analyses. For further analyses, the cheeses were frozen at –18 °C and shredded prior to analyses.

2.2.

Compositional analysis

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ACCEPTED MANUSCRIPT Moisture, fat, and protein (based on total nitrogen, TN) contents of cheeses were determined by the gravimetric (AOAC, 2012d), Gerber (AOAC, 2012c), and Dumas (ISOIDF, 2002) methods, respectively. Ash content was determined by gravimetric method (AOAC, 2012b) and acidity values by titrimetric method (expressed as lactic acid %)

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(AOAC, 2012a). Salt content in the cheeses was determined by potentiometric titration

method (ISO-IDF, 2006). For pH measurements, the samples were diluted with water (1:2), homogenized and the values were obtained by using a pH/ion meter (Mettler Toledo Seven

Colour evaluation

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2.3.

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Compact; Colombus, OH, USA). All compositional analyses were done in triplicate.

The colour of the Turkish white cheese samples was quantified by using a colorimeter (Konica Minolta CM-5, Tokyo, Japan). Colour values, L (lightness), a (redness/greenness),

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and b (yellowness/blueness) for the cheese samples were measured from the inner surfaces of the cheese moulds and the chroma values were calculated as follows: +

(1)

Proteolysis

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2.4.

=√

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ℎ

Total nitrogen (TN), water-soluble nitrogen (WSN), trichloroacetic acid-soluble

nitrogen (TCASN), and phosphotungstic acid-soluble nitrogen (PTASN) contents of samples were measured according to the methods described in Bütikofer, Rüegg, and Ardö (1993). The different parameters of proteolytic ripening were calculated from the values of TN, WSN, TCASN, and PTASN using the following equations (Pereira, Gomes, Gomes, & Malcata, 2008; Pereira, Graça, Ogando, Gomes, & Malcata, 2010):

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ℎ

(

)=

( # ) =

&

× 100

$

(2)

× 100

(%$$ ) = ' $

(3)

× 100

(4)

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%



In addition to PTASN values, the total free amino acid (FAA) contents of cheese

samples were quantified by using 2,4,6-trinitrobenzenesulphonic acid (TNBS) method. TNBS

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reacts with the primary amines and forms a yellow colour, the absorbance of which was

measured at 420 nm (Bouton & Grappin, 1994). By this method, total amino acid content in

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cheeses can be measured more specifically as only free amino groups are quantified (McSweeney & Fox, 1997; Sousa, Ardö, & McSweeney, 2001). In the present study, TNBS method was used with slight modifications. Briefly, 0.5 mL of WSN extract at 5 µL mL-1 concentration was mixed with 0.5 mL borate buffer (0.1 M, pH 9.5) and 1 mL TNBS reagent

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(1 mg mL-1). The mixture incubated in a water bath for 60 min at 37 °C and the incubation stopped with the addition of 2 mL of phosphate buffer (0.1 M). The absorbances of samples were measured at 420 nm with a UV-Vis spectrophotometer (Agilent Technologies, Santa

Lipolysis

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2.5.

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Clara, USA). The results were expressed as mg leucine g-1 cheese.

Lipolysis was evaluated by measuring the acid degree value (ADV) and quantifying

the FFA contents of Turkish white cheese samples. The ADV is a quantitative index for hydrolytic lipolysis in dairy products and indicates the amount of FFA present in fat (Park, 2001). In the present study, samples were de-emulsified and free fat was separated by detergent, heat, and centrifugation. Finally, ADV was determined by titration with alcoholic KOH (Park, 2001).

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ACCEPTED MANUSCRIPT The FFAs profiles were obtained according to a gas chromatographic method with some modifications (De Jong & Badings, 1990; Deeth, Gerald, & Snow, 1983). The extraction of FFAs in the samples was made using aminopropyl SPE columns (Agilent Technologies, Santa Clara, CA, USA). Cheese samples of 1 g with 3 g of anhydrous Na2SO4

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were prepared in a glass test tube. Then, 0.3 mL of 2.5 M H2SO4, 1 mL of internal standard solution and 3 mL of solvent (diethyl ether:hexane, 1:1, v/v) added into the samples. Pentanoic, tridecanoic and heptadecanoic acids were used as internal standards at a

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concentration of 0.5 mg mL-1 for each. Afterward, the mixture in the test tubes was mixed with a vortex for 1 min, centrifuged at 700 × g for 2 min at room conditions and the

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supernatant was collected. This extraction procedure was repeated three times and collected supernatant was filtered through an aminopropyl SPE column, which contains activated alumina. Then, alumina was removed, dried and mixed with 3% formic acid in diethyl ether solution. Finally, the mixture was centrifuged for 10 min at 450 × g and 0.5 µL of the liquid

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phase was injected to the GC (Agilent 7890A; Agilent Technologies, Santa Clara, CA, USA) equipped with a flame ionisation detector (FID). A DB-FFAP column (30 m, 0.25 um, 0.25 mm; Agilent Technologies) was used in the separation of the FFAs. The GC oven

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temperature was initially set at 90 °C for 1 min, then raised up to 240 °C with an increase of 7 °C min-1 and maintained at this temperature for 15 min. The analysis was carried out at a

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constant helium flow rate of 2 mL min-1. The injection and FID temperatures were set at 250 and 260 °C, respectively. Analyses were performed using a 1:10 split ratio. Internal and external standard techniques were used to identify and quantify the compounds. Analytical standards obtained from Sigma-Aldrich (Sigma-Aldrich Chemie GmbH, Steinheim, Germany) with higher than 99.5% purities were used. Results were expressed as mg 100 g fat-1; all samples were analysed in triplicate.

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ACCEPTED MANUSCRIPT 2.6.

Volatile compounds

Solid-phase microextraction (SPME) was used for extraction of volatile compounds of Turkish white cheeses. The identification and quantification of volatile compounds were

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carried out by GC-FID (7890B; Agilent Technologies) and mass spectrometry (MS) (5977A MSD; Agilent Technologies). Equilibrium time, extraction and injection steps were

performed by an automatic injection module (GC Injector 80; Agilent). Extractions were

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conducted with Carboxen/Polydimethylsiloxane (CAR/PDMS, Auto Merlin 75 µm,

SU57343U; Agilent) fibre. The extraction and GC conditions were set according to a

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previous optimisation study (Salum, Erbay, Kelebek, & Selli, 2017).

For the identification of the volatile compounds, injections of 59 different standard compounds were performed. The comparison of the mass spectrum for the non-standard compounds was done with the mass spectra in computer memory. NIST11 and Wiley7

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libraries were used for identification of compounds. Retention indices of the compounds were calculated by the retention data of a linear alkane series. The concentrations of aroma compounds were calculated by using the internal standard (9 µg µL-1 2-octanol and 4-

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nonanol) method. Analyses were performed in triplicate.

Sensory analysis

Sensory analysis was conducted by using ranking to determine the preferences and by

descriptive flavour profile analysis (FPA) to predict the sensory attributes of Turkish white cheeses. Descriptive FPA (Shamaila, Skura, Daubeny, & Anderson, 1993) was conducted by the attendance of nine panellists from the Food Engineering Department of Adana Science and Technology University. The panellists who were previously tested for their sensitivity to

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ACCEPTED MANUSCRIPT major tastes, taste thresholds and sensitivity to odours were trained on the flavour profile technique by using reference foods and flavour compounds (Altuğ Onoğur & Elmacı, 2011; Drake, McIngvale, Gerard, Cadwallader, & Civille, 2001; Karagul-Yuceer, Isleten, & UysalPala, 2007). The references and sensory terms are given in Table 1. Three samples were

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given to the panellists in every session and roundtable discussion was conducted. The

intensity of each character was evaluated on a 0–5 scale. Evaluations performed in duplicates and aftertaste impressions of the samples were also defined.

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Preferences of the white cheese samples were determined by using ranking test (ISO, 2006) and the attendance of the same panellists. The panel was conducted in two sessions and

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in both sessions, each panellist evaluated 5 of 9 samples which were served according to balanced incomplete block designs (BIB) (ISO, 2011; Wakeling & Buck, 2001).

Statistical analysis

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2.8.

Data were analysed by analysis of variance (ANOVA) and Duncan Post Hoc Test. Statistical significance was set at P < 0.05 and SPSS statistical package program (SPSS ver.

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13.0 for Windows, SPSS Inc., Chicago, USA) was used. Principle Component Analysis

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(PCA) was performed for the evaluation of the data obtained from volatile analysis and FPA by using XLSTAT 2011 (Addinsoft, New York, NY, USA).

3.

3.1.

Results and discussion

Compositional analysis

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ACCEPTED MANUSCRIPT The chemical compositions of the ripened white cheese samples are given in Table 2. According to the Turkish Food Codex Cheese Communiqué, all examined cheese samples belonged to the full-fat cheese category (Turkish Food Codex, 2015). One of the samples (sample 9) was defined as soft cheese, while the others were classified in the semi-soft group

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(Turkish Food Codex, 2015). It is determined that all samples were suitable according to the moisture content values specified in the relevant legislation. In terms of salt content in dry matter, sample 5 was at the limit value, while samples 7, 8, and 9 had the values slightly

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above the limits.

Titratable acidity and pH values were in line with those reported in the literature for

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ripened white cheeses (Akalin & Karaman, 2010; Biçer, 2014; Çelik & Uysal, 2009; Kamber, 2008). The acidity values of cheeses, in terms of lactic acid, ranged from 0.8% to 1.0% and the pH values were measured in the range 4.8–5.1. Hayaloglu et al. (2002) reported that the salt contents and the titratable acidity values of ripened Turkish white cheeses were in the

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range 3.3–6.9% and 0.7–3.8%, respectively. In an article series about the traditional cheeses manufactured in Turkey, Kamber (2008) stated that the moisture and protein contents of Turkish white cheese varied between 41.5–66.1% and 13.0–38.2%, respectively. In another

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study, the values of moisture, fat, salt, ash, pH and titratable acidity of ripened Turkish white

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cheeses were in the range 48.6–62.6%, 14.6–21.2%, 3.0–8.7%, 6.4–10.0%, 4.5–5.3%, and 0.2–2.4%, respectively (Çelik & Uysal, 2009). The composition results obtained in the present study varied in a much narrower range in accordance with the values reported in the literature.

3.2.

Proteolysis

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ACCEPTED MANUSCRIPT Proteolysis is one of the most important biochemical processes during ripening and dominates the development of the texture and flavour, not only by breaking down of the protein network but also by the formation of flavour and/or off-flavour compounds with secondary catabolic changes of the proteolysis products (McSweeney, 2004; Sousa et al.,

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2001). Although there are many ways to specify the degree of proteolysis, the most common way is to detect the soluble nitrogen fractions in cheeses (McSweeney & Fox, 1997). While water-soluble nitrogen (WSN) is quantified to determine the degree of proteolysis occurs

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mainly by the activity of coagulant and plasmin, the small peptides and amino acids in WSN can be determined by measuring TCASN. On the other hand, while peptides bigger than 600

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Da precipitate with PTA solution and total nitrogen in this solution is determined with PTASN content analysis, specifically free amino groups in water-soluble extract of cheeses are measured by the TNBS method (Bouton & Grappin, 1994; McSweeney & Fox, 1997). The results of the proteolytic ripening parameters of Turkish white cheese samples are

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presented in Table 3. WSN, TCASN, and PTASN values were found to be in the range 0.48– 0.94%, 0.21–0.36%, and 0.03–0.13%, respectively. By using these values, three indices (REI, RDI, and FAAI) were calculated and their values varied between 16.8–32.8%, 7.4–13.4%,

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and 1.0–4.9%, respectively. The total free amino acid (FAA) content of cheese samples

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ranged from 3.5–11.3 mg leucine g cheese-1. Although the proteolytic ripening parameters varied in a wide range, the results obtained from the present study are in accordance with the literature values (Biçer, 2014; Çelik & Uysal, 2009; Hayaloglu et al., 2008; Kamber, 2008). According to the results, the highest REI value was obtained from sample 2 (32.8%)

whereas sample 9 was characterised by reaching the highest values of RDI, FAAI, and total FAA. The results of PTASN content and TNBS analysis showed similar tendencies. The lowest FAAI value was obtained from sample 6. Additionally, the percentage of the TCASN (30.7%) and PTASN (4.1%) contents in WSN for sample 6 were the lowest.

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ACCEPTED MANUSCRIPT 3.3.

Lipolysis

Another important biochemical process during cheese ripening is lipolysis resulting in the release of free fatty acids (FFAs), which directly or indirectly contribute to the cheese

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flavour. ADV analysis, in which the acidity of the cheese fat is measured, is widely used as a rough estimator of the lipolytic ripening degree in cheeses. However, acetic acid, which is basically not a lipolysis end product but produced by lactate fermentation, is included in the

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ADV determination. This may adversely affect the accuracy of the results, as acetic acid may be present in high amounts in the cheese (Kondyli, Katsiari, Masouras, & Voutsinas, 2002).

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Therefore, in addition to the ADV analysis, FFA profiles were also determined (Table 4). According to our results, the total free fatty acid (TFFA) contents of ripened Turkish white cheeses varied from 509.2 to 2015.9 mg 100 g fat-1. Sample 7 had an average of 2015.9 mg 100 g fat-1 with the highest total free fatty acid (TFFA) value, followed by sample 5

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(1567.9 mg 100 g fat-1) and sample 1 (1089.7 mg 100 g fat-1), respectively. FFAs with aliphatic tails from 4 to 10 carbons are generally considered as the volatile FFAs and the effect of them on the flavour of cheese is remarkable because of their low perception

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threshold (Collins, McSweeney, & Wilkinson, 2003). In the Turkish white cheese samples, 4.7–18.5% of TFFA was constituted by the total volatile free fatty acids (TVFFAs). Sample 1

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(201.7 mg 100 g fat-1) had the highest TVFFA content, followed by sample 4 (143.8 mg 100 g fat-1) and sample 7 (126.2 mg 100 g fat-1). Among the samples, sample 6 had the lowest TFFA (509.2 mg 100 g fat-1) and TVFFA (24.2 mg 100 g fat-1). In all samples, the values of palmitic and oleic acids were significantly higher. Sample 1 was distinguished from all other samples for its high butyric acid content (13.3% of TFFAs). It was mentioned in the literature that the main FFAs in the Turkish white cheese are oleic, palmitic, stearic, and myristic acids (Akalin et al., 1998; Güler & Uraz, 2004; Hayaloglu et al., 2008; Özer et al., 2011).

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ACCEPTED MANUSCRIPT There is a general agreement between the results of the ADV and FFA analyses (Table 4). Results showed that ADV of Turkish white cheese samples varied in the range 2.1–6.1. While the lowest ADV was measured in sample 6, the highest values were

3.4.

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determined in samples 1, 4, 5, and 7.

Volatile compounds

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A total of 45 volatile compounds were identified in Turkish white cheese samples including 13 acids, 9 ketones, 7 esters, 6 alcohols, 4 lactones, 2 aldehydes, 2 phenols, 1 furan,

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and 1 terpene. Amount of these compounds (µg kg-1 ± SD) are given in Table 5 with the linear retention index (LRI) values on a DB-Wax column.

Among the samples, sample 5 (41302.3 µg kg-1) had the highest amount of volatile compounds, followed by sample 4 (40045.4 µg kg-1) and sample 1 (38832.1 µg kg-1) whereas

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sample 6 had the lowest amount of volatile compounds (13837.9 µg kg-1) (Table 5). Acids were the dominant group in all samples and their percentages varied from 72.9% to 92.1%. While samples 1, 4, and 5 had the highest contents, the lowest amount of acids was detected

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in sample 6. A significant portion of acids was FFAs which are directly and/or indirectly

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contributing to the flavour of cheeses. Short and medium length chain FFAs (C4:0-C12:0) have low detection thresholds and each gives a characteristic flavor. FFAs are also precursors of methyl ketones, alcohols, lactones and esters (Curioni & Bosset, 2002; Molimard & Spinnler, 1996). It was found that the concentration of hexanoic acid was higher than all other compounds, except in samples 1 and 6. The most abundant volatile in sample 1 was the butanoic acid which located close to sample 1 in PCA biplot (Fig.1). In addition to FFAs, acetic, propanoic, 2-propenoic, and benzoic acids were detected in Turkish white cheeses. In particular, acetic acid was one of the most abundant compounds in the samples, especially

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ACCEPTED MANUSCRIPT sample 6 in which it represented 37.8% of the total volatile compounds. In the PCA biplot, acid compounds other than dodecanoic and acetic acids were positioned on the positive F1 axis (Fig.1). While sample 5 located very close to nonanoic and heptanoic acids, sample 4

close to butanoic, propanoic and benzoic acids.

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was positioned near pentanoic, decanoic, and 9-decenoic acids. Also, sample 1 was quite

The other predominant volatile groups were esters and ketones (Table 5). With respect to the abundance of volatile compounds, acids were followed by ketones in samples 1, 3, 6, 8,

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and 9, and by esters in samples 2, 4, 5, and 7. A total of nine ketone compounds were

detected in Turkish white cheeses, four of which were methyl ketones. Methyl ketones are

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one of the most important compounds influencing the cheese flavour (especially in blue cheeses) and are formed from FFAs by β-oxidation (Collins et al., 2003; Molimard & Spinnler, 1996). Among the analysed cheeses, sample 6 stood out in terms of ketone content as ketones constituted 17.9% of all volatile compounds, being 2-octanone the most abundant

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compound (1045.8 µg kg-1) after acetic and hexanoic acids. For the rest of the samples, ketones percentages varied from 4.0 to 8.7%. In sample 6, 2-octanone was the most abundant compound after acetic and hexanoic acids. Although the highest amount of 2-octanone was

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found in sample 9, it only accounted for 4% of the total volatile compounds. However, its

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percentage was 7.6% in sample 6. Moreover, acetoin was detected only in sample 6. This compound is a product of citrate metabolism or enzymatic catabolism of aspartic acid and this process is known to be triggered by lactic acid bacteria during cheese ripening (Ardö, 2006; McSweeney & Sousa, 2000). Esters are important aroma compounds due to their low threshold values and ability to mask off-odours caused by high amounts of short-chain FFAs, methyl ketones, and amines. However, high levels of ethyl esters of short chain fatty acids may cause fruity flavour defects in cheeses (Curioni & Bosset, 2002; Holland et al., 2005). Ethyl esters can be formed

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ACCEPTED MANUSCRIPT in cheeses by two fundamental enzymatic ways which were esterification and alcoholysis (Holland et al., 2005; Liu, Holland, & Crow, 2004). A total of seven ester compounds were detected and five of them were fatty acid ethyl esters. Ethyl hexanoate came to the front with its high abundance among the ester compounds. It was found in samples 2, 4, and 7 with a

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percentage of 8.8%, 7.3%, and 7.0%, respectively. Additionally, these samples (2, 4, and 7) included high amounts of ester compounds. Sample 4 had the highest ester content (3170.4 µg kg-1), followed by sample 7 (3113.9 µg kg-1) and sample 2 (2862.5 µg kg-1).

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A total of six alcohols were identified in the samples (Table 5). In cheese, alcohol formation can occur via many metabolic pathways such as lactose metabolism, methyl ketone

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reduction, amino acid metabolism as well as through the degradation of linoleic and linolenic acids (Molimard & Spinnler, 1996). While only two alcohol compounds with low total alcohol content were identified in sample 8, it was found that samples 5, 9, and 3 had high alcohol concentrations. 3-Methyl-1-butanol, which is a branched-chain primary alcohol,

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originates from the reduction of the aldehyde produced from leucine (Curioni & Bosset, 2002). This compound was only detected in sample 5 and its concentration was higher than the rest of alcohols. Another prominent alcohol compound identified in all samples was 2-

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heptanol. Secondary alcohols are formed from methyl ketones via the reductase activity

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(Molimard & Spinnler, 1996).

Among the aldehyde group, benzaldehyde and hexanal were detected in Turkish white

cheeses (Table 5). Within these, benzaldehyde was identified in all samples. In sample 9, the level of benzaldehyde was significantly higher than the rest of the aldehydes (P < 0.05). Benzaldehyde, which is characterised by bitter almond odour notes, can be formed phenylpyruvate via alpha-ketoacid degradation (Molimard & Spinnler, 1996; Yvon & Rijnen, 2001). Hexanal was only identified in the samples 1, 2, 6, and 8. Sample 6 had the highest content of hexanal. Hexanal gives green odour note to the cheeses and may result from β-

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ACCEPTED MANUSCRIPT oxidation of unsaturated fatty acids. However, it is known that this compound causes odour defects when its concentration exceeds a certain threshold (Curioni & Bosset, 2002). In the samples, two volatile phenolic compounds were detected (phenol and p-cresol). Phenol was identified in all samples and the highest concentration was found in sample 6

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(276.7 µg kg-1), followed by sample 8 (250.9 µg kg-1) (Table 5). Phenol can give sharp

medical and floral odour notes to cheese. It results from tyrosine amino acid catabolism by tyrosine-phenol lyase enzyme activity (Fox & Wallace, 1997; Molimard & Spinnler, 1996;

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Yvon & Rijnen, 2001). p-Cresol, which also originates from tyrosine, was only identified in samples 2 and 9. This compound was identified as the odour-active compound in Ezine

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cheese and it was characterised with barny odour note (Yuceer et al., 2009). While phenol located near sample 8 in the PCA biplot, p-cresol and sample 9 were positioned very close to each other (Fig.1).

Lactones, which are cyclic esters, can be formed by intramolecular esterification of

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hydroxy acids and they are generally characterized by fruity odour notes. In cheese, FFAs are precursors of lactones (McSweeney, 2004; Molimard & Spinnler, 1996). Four lactones were identified in Turkish white cheese samples and except for samples 5 and 6, the rest of cheeses

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contained 4 lactone compounds (Table 5). According to the PCA biplot, 3 lactone compounds

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(γ-hexalactone, γ-nonalactone, and benzalacetone) were positioned on the positive F1 axis in the PCA biplot like most of the esters and FFAs, which were generally related to each other. While γ-hexalactone and benzalacetone located near samples 4 and 5, respectively, ∆decalactone was positioned very close to samples 2 and 3 (Fig.1). Furthermore, benzofuran, a compound produced by thermal treatments, was detected in the Turkish white cheese samples with exception of samples 1 and 6 (Table 5). In sample 8, the concentration of benzofuran was significantly higher than the others (P < 0.05). Finally, d-limonene, which is a terpene compound, was identified only in sample 4. Terpenes

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ACCEPTED MANUSCRIPT originate from plants are commonly found in milk and dairy products as a result of pasture feeding of grazing animals (Kilcawley, Faulkner, Clarke, O’Sullivan, & Kerry, 2018).

Sensory and colour properties

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3.5.

The colour properties of the cheese samples were measured and chroma values were calculated (Table 6). The L, a and b values ranged between 86.4–90.7, 0.3–1.8, and 11.0–

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15.4, respectively. These values were in agreement with those found in the literature for Turkish white cheeses (Akalin & Karaman, 2010; Biçer, 2014).

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Sensory descriptors and intensity scores of samples are given in Table 7 and flavour profile diagram is presented in Fig. 2, which was created from common sensory descriptors in samples. Principle component analysis (PCA) was also performed for evaluating the effects of the flavour descriptors on the differentiation of Turkish white cheese samples. Nine

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principle components were found and by using the first two explaining 50.9% of total variations, biplot diagram was conducted (Fig.3). As presented in Table 7 and Fig. 2, salty, bitter, cowy, cooked, sour, fermented,

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oxidized, yeasty, bite, creamy, butter-like, umami, metallic, whey and free fatty acid

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descriptors were detected in all Turkish white cheese samples. Moreover, sulphurous descriptor was determined in all samples except sample 5. Apart from these descriptors, burnt (samples 3, 7, 8, and 9), waxy (samples 3, and 6), milk (sample 6), margarine (sample 6), plastic (samples 3, and 6), off-flavour (sample 6), sweet (sample 6), putrid (sample 7), soapy (sample 4), and ripe fruity (sample 4) notes were detected in the samples. Salty, bitter, sour, fermented, creamy, and butter-like characters stood out with its higher intensity score (except for sample 6). According to FPA, bitter, cooked, sour, oxidised and bite flavours were significantly more intense in sample 8, while sample 7 was

17

ACCEPTED MANUSCRIPT distinguished in terms of fermented and sulphurous characters. Butter-like, metallic, FFA and yeasty characters obtained higher intensity scores in samples 2 and 7, in samples 1 and 9, in sample 5, and in samples 3 and 9, respectively (P < 0.05). Sample 6 clearly distinguished from the other samples (Fig.3). Waxy, milk,

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margarine, plastic, off-flavour, and sweet flavour notes characterised the sample 6 and it was the least preferred sample due to the ranking test results (P < 0.05).

The burnt character, which suggests the importance of the influence of the heat

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treatments on the final product, was found in the samples 3, 7, 8, and 9. Concordantly in these samples, benzofuran was also detected. The ripe fruity character was determined only in

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sample 4, which also had the highest ester content and fruity character may result from ester compounds. Likewise, the putrid character was determined only in sample 7, which had the highest amount of FFAs. Moreover, the results of the ranking analysis were in line with FFA analysis. The lowest TFFA and TVFFA were detected in sample 6, which had the lowest

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preference, whereas the highest three TFFA values obtained from the cheese samples (samples 1, 5, and 7) were the three samples with the highest preference in the ranking test. Furthermore, three cheese samples (samples 1, 4, and 7) with the highest TVFFA were three

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of the four samples that received the highest score in the sensory ranking test. On the other

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hand, the results of the volatile analysis showed that the highest amount of acids and total volatile compounds were detected in four cheese samples (sample 1, 4, 5, and 7) which obtained the highest preference in the ranking test. Additionally, the samples with high preference had generally higher ester and lactone contents whereas the samples with low preference had higher alcohol and aldehyde ratios in total volatile compounds. These results showed that the increase in the abundance of acids in Turkish white cheese caused a rise in sensory preference.

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ACCEPTED MANUSCRIPT 4.

Conclusions

In this study, Turkish white cheese samples manufactured with traditional techniques from cows’ milk and ripened for 6 months were collected from different parts of Turkey and

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analysed. Although the collected samples showed similar properties in terms of

compositional parameters, there were significant variations observed in the ripening and flavour properties of Turkish white cheeses.

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While cheeses with moderate proteolytic ripening degrees were preferred in the

ranking tests, enhanced lipolytic ripening values caused an increase in the preference score.

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The total FAA, REI, RDI, and FAAI values obtained from the most preferred three cheeses were in the range of 3.5–4.9, 16.8–28.5%, 7.6–9.4%, and 1.4–2.0%, respectively. Moreover, the increase in the total number of volatile compounds caused a direct effect on the flavour and preference of cheese. The preference has been positively affected by the increase in the

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abundance of volatile acids, esters, and lactones. Higher concentrations of esters and lactones were thought to mask the sharp flavour resulting from fatty acids and methyl ketones and

cheese.

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lead to a more balanced flavour, which positively influences the quality of Turkish white

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Finally, the most preferred cheeses were the ones manufactured in the western part of Turkey. The most preferred three cheeses were manufactured in the two neighbouring provinces of Çanakkale and Balıkesir, which are located in the north-western part of the country. Further studies should be performed to investigate the relations between the cheese properties and the climate and flora of these provinces. Additionally, researches on the effect of usage of different milk types on the quality of Turkish white cheese and the effect of process conditions with regard to the microbial variations in cheese would provide insights into the causes of alterations in cheese properties.

19

ACCEPTED MANUSCRIPT

Acknowledgements

This work was supported by The Scientific and Technological Research Council of

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Turkey (TUBITAK) [project no: 115O229].

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References

Akalin, A. S., Gönç, S., & Akbaş, Y. (2002). Variation in organic acids content during

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ripening of pickled white cheese. Journal of Dairy Science, 85, 1670–1676. Akalin, A. S., & Karaman, A. D. (2010). Influence of packaging conditions on the textural and sensory characteristics, microstructure and color of industrially produced Turkish white cheese during ripening. Journal of Texture Studies, 41, 549–562.

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Akalin, A. S., & Karaman, A. D. (2011). Influence of packaging systems on the biochemical characteristics and volatile compounds of industrially produced Turkish white cheese. Journal of Food Biochemistry, 35, 663–680.

EP

Akalin, A. S., Kinik, Ö., & Gönç, S. (1998). Research on the determination of fatty acids

AC C

compositions in cheese varieties sold in Izmir local bazaar and market. Gida, 23, 357– 363.

Altuğ Onoğur, T., & Elmacı, Y. (2011). Gıdalarda duyusal değerlendirme. İzmir, Turkey: Sidaş Medya Ltd. Şti.

AOAC. (2012a). Acidity of milk, titrimetric method, method no. 947.05. In G. W. Latimer Jr. (Ed.), Official Methods of Analysis of AOAC International (19th ed.). Gaithersburg, MD, USA: AOAC International. AOAC. (2012b). Ash of milk, gravimetric method, method no. 945.46. In G. W. Latimer Jr.

20

ACCEPTED MANUSCRIPT (Ed.), Official Methods of Analysis of AOAC International (19th ed.). Gaithersburg, MD, USA: AOAC International. AOAC. (2012c). Fat content of raw and pasteurized whole milk, Gerber method by weight, method no. 2000.18. In G. W. Latimer Jr. (Ed.), Official Methods of Analysis of

RI PT

AOAC International (19th ed.). Gaithersburg, MD, USA: AOAC International.

AOAC. (2012d). Loss on drying (moisture) in cheese, method no. 926.08. In G. W. Latimer Jr. (Ed.), Official Methods of Analysis of AOAC International (19th ed.).

SC

Gaithersburg, MD, USA: AOAC International.

Ardic, M., Karakaya, Y., Atasever, M., & Adiguzel, G. (2009). Aflatoxin M1 levels of

M AN U

Turkish white brined cheese. Food Control, 20, 196–199.

Ardö, Y. (2006). Flavour formation by amino acid catabolism. Biotechnology Advances, 24, 238–242.

Biçer, E. B. (2014). The impact of curd cooking temperature and time in traditional white

TE D

cheese produced in the region of Sivas on cheese yield, melting behaviour and textural properties. Sivas, Turkey: Cumhuriyet University. Bouton, Y., & Grappin, R. (1994). Measurement of proteolysis in cheese: Relationship

EP

between phosphotungstic acid-soluble N fraction by Kjeldahl and 2,4,6-

AC C

trinitrobenzenesulphonic acid-reactive groups in water-soluble N. Journal of Dairy Research, 61, 437–440.

Bütikofer, U., Rüegg, M., & Ardö, Y. (1993). Determination of Nitrogen Fractions in Cheese: Evaluation of a Collaborative Study. LWT - Food Science and Technology, 26, 271–

275. Çelik, Ş., & Uysal, Ş. (2009). Composition, quality, microflora and ripening of Beyaz cheese. Atatürk Üniversitesi Ziraat Fakültesi Dergisi, 40, 141–151. Collins, Y. F., McSweeney, P. L. H., & Wilkinson, M. G. (2003). Lipolysis and free fatty

21

ACCEPTED MANUSCRIPT acid catabolism in cheese: A review of current knowledge. International Dairy Journal, 13, 841–866. Curioni, P. M. G., & Bosset, J. O. (2002). Key odorants in various cheese types as

959–984.

RI PT

determined by gas chromatography-olfactometry. International Dairy Journal, 12,

De Jong, C., & Badings, H. T. (1990). Determination of free fatty acids in milk and cheese procedures for extraction, clean up, and capillary gas chromatographic analysis.

SC

Journal of High Resolution Chromatography, 13, 94–98.

Deeth, H. C., Gerald, F., & Snow, J. (1983). A gas chromatographic method for the

M AN U

quantitative determination of free fatty acids in milk and milk products. New Zealand Journal of Dairy Science and Technology, 18, 13–20.

Drake, M. A., McIngvale, S. C., Gerard, P. D., Cadwallader, K. R., & Civille, G. V. (2001). Development of a descriptive language for Cheddar cheese. Journal of Food Science,

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66, 1422–1427.

Fox, P. F., & Wallace, J. M. (1997). Formation of flavor compounds in cheese. In S. L. Neidleman & A. I. Laskin (Eds.), Advances in applied microbiology (Vol. 45, pp. 17–

EP

75). New York, NY, USA: Academic Press.

AC C

Güler, Z., & Uraz, T. (2004). Relationships between proteolytic and lipolytic activity and sensory properties (taste-odour) of traditional Turkish white cheese. International Journal of Dairy Technology, 57, 237–242.

Hayaloglu, A. A., Guven, M., & Fox, P. F. (2002). Microbiological, biochemical and technological properties of Turkish White cheese “Beyaz Peynir.” International Dairy Journal, 12, 635–648. Hayaloglu, A. A., Ozer, B. H., & Fox, P. F. (2008). Cheeses of Turkey: 2. Varieties ripened under brine. Dairy Science and Technology, 88, 225–244.

22

ACCEPTED MANUSCRIPT Holland, R., Liu, S. Q., Crow, V. L., Delabre, M. L., Lubbers, M., Bennett, M., et al. (2005). Esterases of lactic acid bacteria and cheese flavour: Milk fat hydrolysis, alcoholysis and esterification. International Dairy Journal, 15, 711–718. ISO-IDF. (2002). Milk and milk products – Determination of nitrogen content – Routine

RI PT

method using combustion according to the Dumas principle. Geneva, Switzerland: International Organisation for Standardisation.

ISO-IDF. (2006). Cheese and processed cheese products - Determination of chloride content

SC

- Potentiometric titration method. Geneva, Switzerland: International Organisation for Standardisation.

M AN U

ISO. (2006). Sensory analysis - Methodology - Ranking. Geneva, Switzerland: International Organisation for Standardisation.

ISO. (2011). Sensory analysis -- Methodology -- Balanced incomplete block designs. Geneva, Switzerland: International Organisation for Standardisation.

TE D

Kamber, U. (2008). The traditional cheeses of Turkey: Cheeses common to all regions. Food Reviews International, 24, 1–38.

Karagul-Yuceer, Y., Isleten, M., & Uysal-Pala, C. (2007). Sensory characteristics of Ezine

EP

cheese. Journal of Sensory Studies, 22, 49–65.

AC C

Kilcawley, K., Faulkner, H., Clarke, H., O’Sullivan, M., & Kerry, J. (2018). Factors influencing the flavour of bovine milk and cheese from grass based versus non-grass based milk production systems. Foods, 7, 1–43.

Kivanç, M. (1989). A survey on the microbiological quality of various cheeses in Turkey. International Journal of Food Microbiology, 9, 73–77. Kondyli, E., Katsiari, M. C., Masouras, T., & Voutsinas, L. P. (2002). Free fatty acids and volatile compounds of low-fat Feta-type cheese made with a commercial adjunct culture. Food Chemistry, 79, 199–205.

23

ACCEPTED MANUSCRIPT Liu, S. Q., Holland, R., & Crow, V. L. (2004). Esters and their biosynthesis in fermented dairy products: A review. International Dairy Journal, 14, 923–945. McSweeney, P. L. H. (2004). Biochemistry of cheese ripening. International Journal of Dairy Technology, 57, 127–144.

proteolysis in cheese during ripening. Lait, 77, 41–76.

RI PT

McSweeney, P. L. H., & Fox, P. F. (1997). Chemical methods for the characterization of

McSweeney, P. L. H., & Sousa, M. J. (2000). Biochemical pathways for the production of

SC

flavour compounds in cheeses during ripening: A review. Lait, 80, 293–324.

Food Chemistry, 96, 532–537.

M AN U

Mendil, D. (2006). Mineral and trace metal levels in some cheese collected from Turkey.

Merdivan, M., Yilmaz, E., Hamamci, C., & Aygun, R. S. (2004). Basic nutrients and element contents of white cheese of Diyarbakir in Turkey. Food Chemistry, 87, 163–171. Molimard, P., & Spinnler, H. E. (1996). Compounds involved in the flavor of surface mold-

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ripened cheeses: Origins and properties. Journal of Dairy Science, 79, 169–184. Özer, B., Kirmaci, H. A., Hayaloglu, A. A., Akçelik, M., & Akkoç, N. (2011). The effects of incorporating wild-type strains of Lactococcus lactis into Turkish white-brined cheese

EP

(Beyaz peynir) on the fatty acid and volatile content. International Journal of Dairy

AC C

Technology, 64, 494–501.

Park, Y. W. (2001). Proteolysis and lipolysis of goat milk cheese. Journal of Dairy Science, 84, E84–E92.

Pereira, C. I., Gomes, E. O., Gomes, A. M. P., & Malcata, F. X. (2008). Proteolysis in model Portuguese cheeses: Effects of rennet and starter culture. Food Chemistry, 108, 862– 868. Pereira, C. I., Graça, J. A., Ogando, N. S., Gomes, A. M. P., & Malcata, F. X. (2010). Influence of bacterial dynamics upon the final characteristics of model Portuguese

24

ACCEPTED MANUSCRIPT traditional cheeses. Food Microbiology, 27, 339–346. Sahingil, D., Hayaloglu, A. A., Simsek, O., & Ozer, B. (2014). Changes in volatile composition, proteolysis and textural and sensory properties of white-brined cheese: Effects of ripening temperature and adjunct culture. Dairy Science and Technology,

RI PT

94, 603–623.

Salum, P., Erbay, Z., Kelebek, H., & Selli, S. (2017). Optimization of headspace solid-phase microextraction with different fibers for the analysis of the volatile compounds of

SC

white-brined cheese by using response surface methodology. Food Analytical Methods, 10, 1956–1964.

M AN U

Shamaila, M., Skura, B., Daubeny, H., & Anderson, A. (1993). Sensory, chemical and gas chromatographic evaluation of five raspberry cultivars. Food Research International, 26, 443–449.

Sousa, M. J., Ardö, Y., & McSweeney, P. L. H. (2001). Advances in the study of proteolysis

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during cheese ripening. International Dairy Journal, 11, 327–345. TUIK. (2018). Calendar adjusted milk and milk products production and change ratios, 2017. Ankara, Turkey. Retrieved from

EP

http://www.tuik.gov.tr/PreHaberBultenleri.do?id=27676

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Turantaş, F., Ünlütürk, A., & Göktan, D. (1989). Microbiological and compositional status of Turkish white cheese. International Journal of Food Microbiology, 8, 19–24.

Turkish Food Codex (2015). Turkish food Codex communique on cheese. USK. (2014). Dairy sector statistics: World and Turkey. Ankara, Turkey. Wakeling, I. N., & Buck, D. (2001). Balanced incomplete block designs useful for consumer experimentation. Food Quality and Preference, 12, 265–268. Yaroglu, T., Oruc, H. H., & Tayar, M. (2005). Aflatoxin M1 levels in cheese samples from some provinces of Turkey. Food Control, 16, 883–885.

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ACCEPTED MANUSCRIPT Yuceer, Y. K., Tuncel, B., Guneser, O., Engin, B., Isleten, M., Yasar, K., et al. (2009). Characterization of aroma-active compounds, sensory properties, and proteolysis in Ezine cheese. Journal of Dairy Science, 92, 4146–4157.

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International Dairy Journal, 11, 185–201.

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Yvon, M., & Rijnen, L. (2001). Cheese flavour formation by amino acid catabolism.

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ACCEPTED MANUSCRIPT Figure legends

Fig. 1. PCA biplot diagram for volatile compounds of ripened Turkish white cheese samples. Samples are codified according to the provinces where they were produced: Çanakkale (1 and

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5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9).

Fig 2. Flavour profile diagram with common sensory descriptors of ripened Turkish white

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cheese samples. Samples are codified according to the provinces where they were produced: Çanakkale (1 and 5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8)

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and Edirne (9).

Fig 3. PCA of the flavour descriptors for ripened Turkish white cheese samples. Samples were codified according to the provinces where they were produced: Çanakkale (1 and 5),

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Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9).

ACCEPTED MANUSCRIPT Table 1 Reference samples and descriptive terms used for flavour profiling of Turkish white cheese samples References

Salty

Sodium chloride solution (0.5% in water)

Bitter

Caffeine solution (0.08% in water)

Cowy

Sodium caseinate solution (5% in water)

Cooked

Milk heated to 85 °C for 30 min.

Sour

Citric acid solution (0.08% in water)

Fermented

Fresh yoghurt

Oxidized

Milk powder exposed to UV light

Yeasty

Raw yeast dough (yeast in 3% warm sucrose water)

Bite

Sparkling water

Sulphurous

Smashed boiled egg

Creamy

Cream

Butter-like

Butter

Umami

Monosodium glutamate solution (1% in water)

Metallic

Ferrous (II) sulphate (0.005% in water)

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Capric acid (100 mg mL-1)

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Sweet

20 ppm butyric acid (in 95% ethanol) Milk heated to 121 °C for 25 min.

Burnt

Milk

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Whey solution (5 g in 100 mL water)

Free fatty acid

Waxy

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Whey

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Descriptive Terms

UHT milk Sucrose solution (5% in water)

Ripe fruity

20 ppm ethyl hexanoate (in 95% ethanol)

Margarine

No reference used

Plastic

No reference used

Off-flavor

No reference used

Putrid

No reference used

Soapy

No reference used

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Table 2

Protein

Salt

Ash

(%)

(%)

(%)

(%)

(%)

1

50.0 ± 0.4a

25.7 ± 0.6d

18.7 ± 0.5c

2.73 ± 0.03a

3.9 ± 0.0b

2

51.2 ± 0.2b

24.5 ± 0.5c

18.3 ± 0.2bc

3.12 ± 0.10bc

4.3 ± 0.0d

3

50.7 ± 0.5b

25.8 ± 0.3de

17.9 ± 0.2b

2.71 ± 0.15a

3.8 ± 0.1a

4

50.0 ± 0.2a

28.2 ± 0.3f

17.1 ± 0.5a

2.60 ± 0.20a

5

52.5 ± 0.4c

22.7 ± 0.3a

18.3 ± 0.3bc

6

54.1 ± 0.3d

22.8 ± 0.3a

7

51.3 ± 0.5b

8 9 a

Fat in

Salt in

Titratable

DM (%)

DM (%)

Acidity (%)

51.3

5.45

0.86 ± 0.02de

5.11 ± 0.10d

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Fat

50.2

6.40

0.79 ± 0.01ab

4.97 ± 0.02bcd

52.4

5.50

1.00 ± 0.01g

4.77 ± 0.09a

3.7 ± 0.1a

56.3

5.20

0.79 ± 0.05ab

5.02 ± 0.18cd

3.09 ± 0.10b

5.3 ± 0.0e

47.7

6.49

0.88 ± 0.00ef

4.81 ± 0.06ab

18.0 ± 0.4bc

2.67 ± 0.07a

3.9 ± 0.0b

49.5

5.81

0.76 ± 0.02a

4.78 ± 0.09a

24.8 ± 0.3c

17.9 ± 0.3b

3.60 ± 0.07d

4.2 ± 0.0c

51.0

7.38

0.81 ± 0.02bc

4.87 ± 0.07abc

50.1 ± 0.3a

26.5 ± 0.5e

18.7 ± 0.1c

3.34 ± 0.07c

4.2 ± 0.1cd

53.1

6.68

0.90 ± 0.02f

4.90 ± 0.07abc

54.6 ± 0.1d

23.5 ± 0.7b

17.1 ± 0.5a

2.98 ± 0.02b

4.1 ± 0.1c

51.8

6.56

0.83 ± 0.04cd

5.10 ± 0.09d

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Moisture

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Sample

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Composition of ripened Turkish white cheeses. a pH

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Abbreviation: DM, dry matter. Samples are codified according to the provinces where they were produced: Çanakkale (1 and 5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9). Values are means ± SD; means within a column with different superscript letters differ significantly (P<0.05).

2

ACCEPTED MANUSCRIPT Table 3 Proteolytic ripening parameters of ripened Turkish white cheeses. a Sample

WSN (%)

TCASN (%)

PTASN (%)

Total FAA

REI

RDI

FAAI

(%)

(%)

(%)

0.837 ± 0.045e

0.231 ± 0.058ab

0.053 ± 0.008bc

4.92 ± 0.02d

28.53

7.87

1.82

2

0.943 ± 0.008f

0.316 ± 0.050cd

0.064 ± 0.008c

5.07 ± 0.10e

32.79

10.98

2.22

3

0.822 ± 0.009e

0.279 ± 0.005bc

0.080 ± 0.004d

4.85 ± 0.10d

29.29

9.93

2.85

4

0.697 ± 0.010

c

0.277 ± 0.005

bc

0.089 ± 0.010

d

5.12 ± 0.26

e

26.07

10.37

3.34

5

0.482 ± 0.009

a

0.271 ± 0.014

bc

0.059 ± 0.006

c

4.61 ± 0.20

c

16.76

9.41

2.04

6

0.677 ± 0.004c

0.208 ± 0.012a

0.028 ± 0.004a

3.98 ± 0.08b

23.93

7.36

0.99

7

0.549 ± 0.006b

0.212 ± 0.026a

0.040 ± 0.004ab

3.49 ± 0.11a

19.59

7.57

1.43

8

0.567 ± 0.027

b

0.348 ± 0.011

d

19.29

11.85

3.63

0.767 ± 0.036

d

0.357 ± 0.023

d

28.66

13.36

4.93

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0.132 ± 0.011

f

5.90 ± 0.10

a

f

11.31 ± 0.17

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9

0.107 ± 0.012

e

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1

g

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Abbreviations are: WSN, water-soluble nitrogen; TCASN, tricholoroacetic acid-soluble nitrogen; PTASN, phosphotungstic acid-soluble nitrogen; FAA, free amino acid (mg leucine g-1 cheese); REI, ripening extension index; RDI, ripening depth index; FAAI, free amino acid index. Samples are codified according to the provinces where they were produced: Çanakkale (1 and 5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9). Values are means ± SD; means within a column with different superscript letters differ significantly (P<0.05).

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Table 4 Acid degree values and free fatty acid values (mg 100 g fat-1) of ripened Turkish white cheese. a Sample 3

4

5

7

8

9

ADV

6.07 ± 0.19f

3.06 ± 0.02b

3.82 ± 0.27c

5.79 ± 0.03e

6.08 ± 0.02f

2.11 ± 0.01a

4.58 ± 0.11d

4.37 ± 0.07d

3.61 ± 0.03c

Butanoic acid (C4:0)

145.3 ± 3.6g

33.4 ± 1.5d

24.0 ±0.3b

56.8 ± 2.3f

24.0 ± 2.1b

6.7 ± 0.1a

40.9 ± 0.2e

28.4 ± 1.0c

41.3 ± 1.7e

Hexanoic acid (C6:0)

18.7± 11.1d

13.1 ± 0.6b

12.5 ±0.5b

22.3 ± 1.6e

11.5 ± 0.7b

3.5 ± 0.1a

22.3 ± 1.6e

15.3 ± 0.8c

20.3 ± 1.3d

Octanoic acid (C8:0)

13.0 ± 0.4cd

11.2 ±0.7bc

10.7 ±0.4b

20.7 ± 1.7f

10.9 ± 0.5b

3.7 ± 0.2a

18.9 ± 2.1e

12.1 ± 0.7bcd

13.2 ± 0.2d

Decanoic acid (C10:0)

24.6 ± 1.2cd

19.9 ± 1.6b

21.9 ± 1.1bc

44.0 ± 3.8e

26.7 ± 0.7d

11.4 ±0.3a

44.2 ± 3.8e

25.0 ± 1.6cd

25.7 ± 0.4d

Dodecanoic acid (C12:0)

40.8 ± 3.0d

26.1 ± 2.2b

32.1 ± 2.5c

58.5 ± 4.6e

40.5 ± 1.3d

17.6 ± 0.4a

68.1 ± 4.2f

37.1 ± 2.2d

39.6 ± 1.1d

Myristic acid (C14:0)

112.3 ± 6.7e

70.2 ± 6.0b

83.3 ± 4.7bc

101.1 ± 8.5de

106.7 ± 7.2de

49.6 ± 2.1a

198.9 ± 17.2f

92.7 ± 5.2cd

99.4 ± 4.7de

Palmitic acid (C16:0)

305.8 ± 13.8d

183.6 ± 9.4a

228.6 ± 16.2b

223.1 ± 16.1b

450.2 ± 41.0e

163.0 ± 3.4a

597.0 ± 33.9f

253.6 ± 13.7bc

266.0 ± 3.3c

Stearic acid (C18:0)

78.8 ± 1.8b

66.0 ± 5.1ab

96.0 ± 8.1c

55.6 ±4.1a

246.5 ± 15.9e

58.4 ± 2.0a

199.3 ± 17.4d

56.5 ± 4.8a

97.1 ± 3.3c

Oleic acid (C18:1)

313.0 ± 3.3c

273.5 ± 16.1b

321.5 ± 25.4c

315.4 ± 23.9c

598.6 ± 33.6d

158.3 ±24.0a

739.7 ± 8.7e

275.7 ± 11.6b

331.2 ± 16.1c

Linoleic acid (C18:2)

32.8 ± 1.2b

18.7 ± 1.6a

31.1 ± 2.6b

41.5 ± 3.8cd

47.6 ± 3.5d

15.1 ± 1.0a

77.5 ± 9.2e

38.1 ± 2.4bc

32.6 ± 2.1b

Linolenic acid (C18:3)

4.4 ± 0.4cd

7.1 ± 0.8e

3.6 ±0.3b

3.7 ± 0.2b

4.7 ± 0.2d

5.0 ± 0.1d

9.1 ± 0.3f

3.9 ± 0.3bc

2.4 ± 0.1a

TVFFA

201.7 ± 4.4g

77.5 ± 4.2bc

69.1 ± 1.4b

143.8 ± 8.9f

73.1 ± 4.0bc

24.2 ± 2.1a

126.2 ±7.3e

80.7 ± 44.0c

100.5 ± 0.5d

TFFA

1089.7 ± 20.7f

722.7 ± 42.0b

865.4 ± 51.1cd

942.6 ± 68.8de

1567.9 ± 92.3g

509.2 ± 32.7a

2015.9 ± 29.1h

838.4 ± 42.8c

968.2 ± 17.3e

EP

a

RI PT

2

SC

1

TE D

6

M AN U

Parameter

AC C

Abbreviations are: ADV, acid degree value; TVFFA, total volatile free fatty acids (C4:0-C10:0); TFFA, total free fatty acids. Samples are codified according to the provinces where they were produced: Çanakkale (1 and 5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9). Values are means ± SD; means within a row with different superscript letters differ significantly (P < 0.05).

4

ACCEPTED MANUSCRIPT

Table 5 Volatile acid, ketone and ester compounds and amounts in ripened Turkish white cheese samples. a Compound

LRI

Samples 1

2

3

4

5

6

7

8

9

5229±50b 35.0±2.4a 827.8±40.4a 32.2±3.4a ND 2113±10a ND 1018±66a 28.5±0.8b 433.2±19.8a ND 286.6±1.6e 91.5±6.6a 10094 (72.9%)

7291±613ef 91.0±1.7b 2700±174b 82.0±1.7c 87.5±2.9b 11093±9909de 159.2±6.6c 3733±190c 44.1±1.7e 1013±31c 57.8±1.6b 309.0±28.7e 95.5±2.5a 26755 (81.2%)

6479±474de 54.1±2.7a 3436±122c 62.0±2.0b 101.6±3.6c 9914±711cd 170.6±6.5d 2932±119b 33.9±1.2c 836.9±76.7b 44.0±2.5a 171.9±12.8b 91.1±0.1a 24328 (84.2%)

5798±49bcd 244.1±11.1e 3066±218bc 72.2±0.4c 81.9±2.6b 9773±167cd 157.7±5.6c 3048±24b 27.1±0.2b 909.3±43.5bc 59.1±1.8bc 158.7±9.8ab 117.5±2.0c 23513 (81.1%)

472.3±10.8f 735.2±12.5c ND 803.6±37.1d 57.9±1.0c 111.9±0.3c ND 34.2±1.1d 104.6±1.7c 2320 (5.6%)

331.0±24.1d 1046±17g 58.9±2.4 749.8±61.7d ND 155.2±11.9d ND 16.8±0.8a 119.3±2.7d 2477 (17.9%)

261.5±6.4b 985.2±4.8f ND 918.5±10.5e ND 72.3±1.2a ND 24.3±0.9b 105.7±3.4c 2368 (7.2%)

249.3±12.0b 913.8±42.9de ND 644.8±29.2c 83.5±6.5d 155.0±5.8d ND 22.4±1.6b 80.3±1.6a 2149 (7.4%)

208.7±7a 1158±24h ND 815.4±24.5d ND 107.1±0.7c ND 30.0±0.3c 106.2±0.2c 2452 (8.4%)

1357±81d ND ND 273.6±0.2d 568.7±9.5g 176.9±9.3d 38.8±1.2d 2415 (5.9%)

442.0±16.3a ND ND ND ND 23.4±1.7a 23.5±0.9b 489 (3.5%)

2301±88e ND ND 73.1±0.5b 717.1±10.7h ND 23.0±0.5b 3114 (9.5%)

1193±115cd ND ND 28.4±0.9a 407.2±34.3f 134.7±9.7c 25.7±1.0ab 1789 (6.2%)

1031±22c 38.8±2.2 ND 663.4±38.3e 316.4±4.8e ND 28.3±0.6b 2078 (7.2%)

1471 1556 1640 1658 1755 1857 1949 2051 2154 2252 2315 2429 2455

3330±169a 550.0±51.7f 13697±845e 246.0±4.1g 130.7±1.4e 11831±795e 144.8±6.2b 3983±102c 37.6±0.2d 1359±9d 91.6±2.5e 183.9±7.7bc 188.1±7.7d 35773 (92.1%)

6155±38cd 198.5±6.8d 3644±135c 113.3±2.4d 50.5±3.0a 8251±262b 87.7±0.8a 3117±161b 21.9±0.7a 865.7±17.4b 59.7±1.8bc 220.5±6.5d 114.0±5.4b 22897 (80.4%)

7830±675ef 118.0±2.9c 3146±214bc 82.4±2.3c 87.0±1.1b 8652±200bc ND 2866±170b 26.2±1.2b 856.0±65.5b 62.0±2.0c 135.1±9.2a 92.9±6.1b 23955 (85.1%)

5439±482bc 79.9±1.2b 5855±278d 146.6±3.3e 146.0±9.7f 15337±1056f 213.9±5.1e 5211±17d 62.4±0.6f 1614±157e 94.8±2.1e 209.6±14.0cd 160.7±14.0c 34569 (86.3%)

5766±10bcd 189.0±5.3d 5631±307d 159.4±11.9f 116.5±5.1d 16506±890f 163.3±7.1cd 5283±439d 60.4±4.1f 1462±76d 84.8±2.1d 222.8±11.6d 117.9±4.3c 35761 (86.6%)

Ketones 2-Heptanone 2-Octanone 3-Hydroxy-2-butanone 4-Nonanone* 2-Cyclopenten-1-one* 2-Nonanone 4-Cyclopentene-1,3-dione* 2-Undecanone Acetophenone Total ketones

1195 1281 1295 1309 1389 1407 1599 1612 1648

398.3±4.4e 590.0±35.6a ND 478.6±42.1b ND 89.2±1.5b 32.6±1.3 45.6±5.0f 101.9±5.6c 1736 (4.5%)

301.7±4.3c 892.3±71.3d ND 632.3±26.4c 57.4±2.3c 80.2±3.7a ND 21.3±1.3b 130.5±7.3e 2116 (7.4%)

313.3±16.3cd 957.6±26.9ef ND 889.7±28.5e 23.8±0.9b 108.8±2.1c ND 38.3±1.7e 117.9±2.6d 2450 (8.7%)

260.5±2.9b 663.4±15.7b ND 410.7±13.0a 14.7±0.7a 108.0±3.3c 18.7±0.5 24.3±0.5b 94.1±1.3b 1594 (4.0%)

Esters Ethyl Hexanoate Propyl hexanoate Ethyl heptanoate Ethyl lactate Ethyl octanoate Ethyl decanoate Ethyl dodecanoate Total esters

1243 1298 1321 1369 1451 1648 1850

506.9±35.6ab 47.2±1.2 ND 178.2±11.2c 20.1±0.5a ND 31.8±0.5c 784 (2.0%)

2497±212f ND ND ND 107.6±0.1c 218.3±9.8e 39.5±3.2d 2863 (10.1%)

639.4±45.7b ND ND 55.7±2.2a 255.7±6.2d 101.3±6.9b 28.7±0.8b 1081 (3.8%)

2940±8g ND 106.8±1.8 ND 47.2±3.2b 36.1±0.6a 39.7±2.6d 3170 (%7.9)

SC

M AN U

TE D

EP

AC C

RI PT

Acids Acetic acid Propanoic acid Butanoic acid 2-Propenoic acid* Pentanoic acid Hexanoic acid Heptanoic acid Octanoic acid Nonanoic acid Decanoic acid 9-Decenoic acid Benzoic acid Dodecanoic acid Total acids

5

ACCEPTED MANUSCRIPT

1248 1290 1325 1378 1416 1571

ND 34.4±0.8b 62.5±1.3cd ND ND 30.9±1.2a 128 (0.3%)

ND 28.4±1.8b 48.4±2.7b 35.5±3.5b ND ND 112 (0.4%)

ND 76.3±6.1c 51.4±0.7b 55.7±1.4d 52.5±2.7b 36.5±2.2c 272 (1.0%)

ND 20.6±2.1a 66.9±0.3d ND ND 35.0±0.4bc 123 (0.3%)

168.5±1.7 ND 83.1±7e 28.2±1.5a ND 32.7±2.6ab 312 (0.8%)

ND 32.5±1.3b 117.8±1.5f ND 78.3±2.0c ND 229 (1.7%)

ND ND 38.7±2.8a 44.1±4.1c 50.1±0.6ab 58.2±1.2d 191 (0.6%)

ND 27.6±1.1b 59.6±2.5c ND ND ND 87 (0.3%)

ND 86.1±2.2d 79.0±2.7e 35.9±2.2b 47.8±0.4a 34.3±1.5abc 283 (1.0%)

Aldehydes Hexanal Benzaldehyde Total aldehydes

1102 1542

75.9±2.2c 95.3±3.5c 171 (0.4%)

51.1±2.5b 95.2±0.4c 146 (0.5%)

ND 73.8±2.3b 74 (0.3%)

ND 113.2±2d 113 (0.3%)

ND 112.7±2.7d 113 (0.3%)

103.8±0.1d 78.3±0.4b 182 (1.3%)

ND 117.4±1.2d 117 (0.4%)

24.8±0.8a 59.9±2.3a 85 (0.3%)

ND 302.3±26.8e 302 (1.0%)

Phenolic compounds Phenol p-Cresol Total phenolic compounds

1998 2142

132.6±0.1a ND 133 (0.3%)

150.9±6.4b 22.6±0.7 174 (0.6%)

146.5±0.9ab ND 147 (0.5%)

235.7±6.1e ND 236 (0.6%)

203.4±9.1cd ND 203 (0.5%)

276.7±1.0g ND 277 (2.0%)

213.9±13.2d ND 214 (0.7%)

250.9±3.8f ND 251 (0.9%)

198.1±6.4c 26.0±0.8 224 (0.8%)

Lactones γ-Hexalactone Benzalacetone γ-Nonalactone ∆-Decalactone Total lactones

1723 1959 1245 2197

24.9±0.5d 11.0±0.6b 22.9±0.3bc 47.6±1.4a 106 (0.3%)

16.9±0.7b 9.2±0.3a 21.9±0.7bc 46.0±3.5a 94 (0.3%)

16.1±0.3b 11.9±0.5c 19.1±0.5a 48.9±2.7ab 96 (0.3%)

21.9±0.4c 19.3±0.4d 28.2±0.1d 58.7±1.4de 128 (0.3%)

24.3±0.1d ND 30.5±2.4e 55.4±5.4cd 110 (0.3%)

ND ND 21.7±0.3b 62.6±1.8d 84 (0.6%)

22.6±0.1c 10.0±0.3a 26.7±0.3d 53.2±2.3bc 112 (0.3%)

10.0±0.4a 12.5±0.4c 23.6±0.9c 49.4±2.1ab 95 (0.3%)

31.5±0.9e 12.2±0.3c 19.2±0.8a 54.8±2.0cd 118 (0.4%)

Furans Benzofuran Total furans

1519

ND ND

65.1±0.1a 65 (%)

66.4±2.5a 96 (%)

62.4±2.2a 62 (%)

62.3±3.1a 62 (%)

ND ND

72.6±0.8b 73 (%)

111.4±1.7c 111 (%)

63.1±5.0a 63 (%)

Terpenes d-Limonene Total terpenes

1205

ND ND

ND ND

ND ND

45.4±3.3 45 (%)

ND ND

ND ND

ND ND

ND ND

ND ND

38831

28467

28140

40041

41297

13832

32944

28895

29007

a

SC

M AN U

TE D

EP

AC C

Total volatile compounds

RI PT

Alcohols 3-Methyl-1-butanol 4-Heptanol* 2-Heptanol 1-Hexanol 3-Octanol 1-Octanol Total alcohols

Abbreviation: LRI, Linear retention index calculated on DB-Wax capillary column. Samples are codified according to the provinces where they were produced: Çanakkale (1 and 5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9). An asterisk indicates that only mass spectrometry (MS) and LRI is available for the identification of these compounds; therefore, it must be considered as an attempt of identification; in the identification of other compounds, chemical standards were used in conjunction with LRI and MS. Values (in (µg kg-1) and means ± SD; means within a row with different superscript letters differ significantly (P < 0.05). 6

ACCEPTED MANUSCRIPT Table 6 Colour properties of ripened Turkish white cheeses. a a

b

Chroma

1

87.38 ± 0.25c

1.54 ± 0.12e

14.18 ± 0.40e

14.27 ± 0.40e

2

86.40 ± 0.25a

1.77 ± 0.08f

15.36 ± 0.48f

15.46 ± 0.47f

3

88.30 ± 0.24e

1.06 ± 0.05c

12.76 ± 0.21d

12.80 ± 0.21d

4

88.83 ± 0.41f

1.04 ± 0.09c

12.85 ± 0.33d

12.90 ±0.33d

5

88.86 ± 0.47f

1.03 ± 0.10c

11.68 ± 0.34b

11.72 ± 0.33b

6

87.02 ± 0.27b

1.04 ± 0.03c

15.25 ± 0.08f

15.28 ± 0.08f

7

90.55 ± 0.30g

1.38 ± 0.05d

11.01 ± 0.21a

11.10 ± 0.21a

8

90.70 ± 0.46g

0.61 ± 0.04b

12.26 ± 0.34c

9

87.79 ± 0.28d

0.27 ± 0.07a

12.96 ± 0.29d

SC

RI PT

L

12.27 ± 0.34c

12.96 ± 0.29d

M AN U

Sample

a

AC C

EP

TE D

Samples are codified according to the provinces where they were produced: Çanakkale (1 and 5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9). Values are means ± SD; means within a column with different superscript letters differ significantly (P < 0.05).

7

ACCEPTED MANUSCRIPT

Table 7 Flavour descriptors, intensity scores and result of ranking analysis of ripened Turkish white cheese samples. a

28

28

a

6 2.37±0.29a 1.65±0.21b 0.30±0.09a 1.28±0.17ab 1.77±0.14a 1.70±0.13b 0.42±0.10ab 0.80±0.13c 0.25±0.08a 0.62±0.10bc 2.25±0.15d 2.58±0.24abc 0.73±0.12e 0.68±0.15c 1.02±0.13b 0.28±0.04a ND 0.88±0.13 1.17±0.19 3.73±0.16 2.42±0.17 3.37±0.15 0.60±0.15 ND ND ND

7 3.45±0.12c 1.63±0.25b 0.50±0.22ab 1.85±0.12d 2.17±0.28b 3.67±0.15g 1.03±0.26d 0.42±0.23a 1.23±0.23cd 1.30±0.29d 1.38±0.34b 3.15±0.27d 0.90±0.13f 0.68±0.20c 1.25±0.16bc 0.55±0.23ab 0.42±0.08a ND ND ND ND ND ND 2.30±0.23 ND ND

8 3.33±0.26c 2.96±0.29e 2.17±0.15e 2.52±0.37e 3.12±0.28c 3.05±0.39e 2.33±0.41e 0.72±0.23bc 1.48±0.27d 0.48±0.18ab 2.27±0.29d 2.54±0.10ab 0.42±0.10bc 0.47±0.08ab 1.22±0.25bc 1.08±0.20de 0.55±0.27ab ND ND ND ND ND ND ND ND ND

9 2.95±0.23b 2.27±0.16d 0.87±0.38c 1.60±0.38bcd 2.18±0.26b 3.37±0.15f 1.20±0.25d 1.22±0.27e 1.42±0.49cd 0.72±0.18c 3.00±0.28e 2.85±0.18c 1.17±0.15g 1.28±0.25d 0.58±0.10a 0.52±0.16ab 0.33±0.15a ND ND ND ND ND ND ND ND ND

32

33

9

34

30

28

RI PT

34

5 4.78±0.17d 1.60±0.11b 0.55±0.21ab 1.32±0.52abc 2.25±0.37b 1.10±0.28a 0.62±0.12bc 0.93±0.16cd 1.12±0.37c ND* 0.68±0.13a 2.37±0.20ab 0.25±0.14a 0.35±0.12a 1.02±0.26b 1.35±0.12e ND ND ND ND ND ND ND ND ND ND

SC

Ranking Score

4 2.85±0.16b 0.8±0.13a 0.65±0.08bc 1.48±0.29bcd 1.53±0.29a 2.20±0.19c 0.33±0.15a 0.45±0.21a 0.35±0.16ab 0.33±0.10a 2.40±0.40d 2.53±0.21ab 0.33±0.08ab 0.33±0.10a 1.50±0.17c 0.58±0.25ab ND ND ND ND ND ND ND ND 0.47±0.21 0.58±0.20

M AN U

3 2.42±0.31a 2.07±0.51cd 0.50±0.23ab 1.33±0.32abc 2.20±0.17 b 2.33±0.16c 1.10±0.14d 1.10±0.22de 0.53±0.05ab 0.47±0.21ab 2.03±0.28cd 2.65±0.37bc 0.58±0.10de 0.57±0.20bc 1.33±0.52 bc 0.97±0.31cd 0.72±0.25b 0.40±0.11 ND ND 1.33±0.20 ND ND ND ND ND

TE D

2 2.70±0.32b 2.08±0.34cd 1.20±0.18d 1.03±0.19a 1.60±0.20a 1.82±0.37b 1.17±0.24d 0.55±0.14ab 0.32±0.12ab 0.75±0.19c 2.40±0.55d 3.28±0.23d 0.50±0.06cd 0.52±0.17bc 0.67±0.21 a 1.18±0.42de ND ND ND ND ND ND ND ND ND ND

EP

Salty Bitter Cowy Cooked Sour Fermented Oxidized Yeasty Bite Sulphurous Creamy Butter-like Umami Metallic Whey FFA1 Burnt Waxy Milk Margarine Plastic Off-flavour Sweet Putrid Soapy Ripe fruity

Sample 1 2.75±0.18b 1.78±0.25bc 1.95±0.19e 1.72±0.23cd 2.38±0.32b 2.68±0.26d 0.77±0.23c 0.48±0.15ab 0.63±0.12b 0.45±0.12ab 1.67±0.19bc 2.32±0.28a 1.33±0.25h 1.17±0.08d 1.23±0.23bc 0.75±0.29bc ND ND ND ND ND ND ND ND ND ND

AC C

Descriptor

Abbreviation: FFA, free fatty acid. Samples are codified according to the provinces where they were produced: Çanakkale (1 and 5), Kars (2), Antalya (3), Denizli (4), Mersin (6), Balıkesir (7), Aksaray (8) and Edirne (9). Values are means ± SD (ND, not detected); means within a row with different superscript letters differ significantly (P < 0.05).

8

ACCEPTED MANUSCRIPT

TE D

M AN U

SC

RI PT

1

2 3

EP

4

AC C

5 6 7 8 9

Figure 1.

TE D

M AN U

SC

RI PT

ACCEPTED MANUSCRIPT

10 11

EP

12

14 15 16 17 18

AC C

13

Figure 2.

TE D

M AN U

SC

RI PT

ACCEPTED MANUSCRIPT

19 20

22 23 24 25 26 27

Figure 3

AC C

EP

21