- Research
- Open Access
Lipolysis and antioxidant properties of cow and buffalo cheddar cheese in accelerated ripening
https://doi.org/10.1186/s12944-018-0871-9
© The Author(s). 2018
- Received: 22 February 2018
- Accepted: 21 September 2018
- Published: 2 October 2018
Abstract
Background
Buffalo milk is the second largest source of milk on the globe, it is highly suitable for the preparation of mozzarella cheese, however, it is not suitable for the preparation of cheddar cheese due to high buffering capacity, low acid development, excessive syneresis, lower lipolysis that lead to lower sensory score. Accelerated ripening can enhance lipolysis and improve sensory characteristics of cheddar cheese. Lipolysis and antioxidant capacity of buffalo cheddar cheese in conventional ripening is not previously studied. Optimization of ripening conditions can lead to better utilization of buffalo milk in cheese industry.
Methods
Effect of accelerated ripening on lipolysis and antioxidant properties of cow and buffalo cheddar cheese were investigated. Cheddar cheese prepared from standardized (3.5% fat) cow and buffalo milk was subjected to conventional and accelerated ripening (4 °C and 12 °C) for a period of 120 days. Fatty acid profile, organic acids, free fatty acids, cholesterol, antioxidant activity and sensory characteristics were studied at 0, 40, 80 and 120 days of ripening.
Results
Fatty acid profile of cow and buffalo cheddar in conventional (120 days old) and accelerated ripening were different from each other (p < 0.05). Free fatty acids in 120 days old buffalo and control cheddar, in accelerated ripening were 0.55% and 0.62%. After accelerated ripening, cholesterol in buffalo and control cheddars were 16 and 72 mg/100 g. After accelerated ripening, concentrations of formic, pyruvic, lactic, acetic and citric acids in buffalo cheddar cheese were, 922, 136, 19,200, 468 and 2845 ppm. At the end of accelerated ripening (120 days), concentrations of formic, pyruvic, lactic, acetic and citric acids in cow cheddar cheese were 578, 95, 9600, 347 and 1015 ppm. Total antioxidant capacity of control cow and buffalo cheddar in accelerated ripening was 77.26 and 88.30%. Colour, flavour and texture score of rapid ripened 80 and 120 days old buffalo cheddar was not different from cow cheddar.
Conclusions
Results of this investigations showed that flavour profile buffalo cheddar subjected to accelerate ripening was similar to cow cheddar cheese. Accelerated ripening can be used for better utilization of buffalo milk in cheddar cheese industry.
Keywords
- Buffalo milk
- Cheddar cheese
- Lipolysis
- Fatty acid profile
Background
Buffalo milk contributes 12% of the total milk production in the world [1]. About 80% of total buffalo milk is produced in India and Pakistan [2]. Chemical composition of buffalo milk in terms of major and minor constituents is significantly different from cow milk. Buffalo milk has higher fat, protein, minerals and total solid contents than cow milk [3]. Concentration of cholesterol in buffalo milk ranged from 8 to 10 mg/100 g while, concentration of cholesterol in cow milk ranged from 30 to 35 mg/100 g [4]. Viscosity and calcium content of buffalo milk is also greater than cow milk [5]. As compared to cow milk, buffalo milk has higher levels of vitamin E, C, zinc, selenium and sulphur containing amino acids, these biochemical compounds are responsible for antioxidant activity in milk and milk products [6, 7]. Average size of fat globules in buffalo and cow milk is 8.7 μm and 0.1-5 μm, respectively [8]. Size of milk fat globule has a pronounced effect on manufacturing and ripening of cheese. Camembert cheese prepared from milk with large sized fat globules underwent lower degree of lipolysis [9]. Chemical composition of buffalo milk offers excellent opportunities for the production of various value-added dairy products; however, larger size of fat globules, higher buffering capacity, slower acid development and excessive syneresis makes buffalo milk unfit for making superb quality cheddar cheese. Results of earlier investigations showed that cheddar cheese prepared from buffalo milk underwent excessive syneresis, required long ripening time with lower textural and flavour prospects [10]. Cheddar is recognized as hard cheese and its acceptability mainly depends upon the flavour, which is produced during ripening [11]. Biochemical and metabolic processes lead to the significant changes in flavour and texture of cheese; these processes are glycolysis, lipolysis and proteolysis [12]. Lipolysis plays an important role in the breakdown of triglycerides of milk fat, generation of free fatty acids, organic acids, large number of flavouring compounds such as, thio-esters, alkan-2-ones, alkane-2-ols, ethyl alcohols, lactones etc. [13]. Ripening temperature has great effect on biochemical events which take place during the maturation of cheese, ripening of cheese at relatively higher temperatures can significantly accelerate the metabolic processes in cheese. Ripening of cheddar cheese is mandatory for the development of typical flavour and textural characteristics. Cheddar cheese is usually ripened for 8–12 months at 4-6 °C. Ripening of cheese on such a lower temperature for a fairly long period of time requires massive refrigeration and inventory cost. Accelerated ripening enhanced the concentration of organic acids in cheddar cheese [14]. However, in this study, fatty acid profile, free fatty acids, peroxide value, cholesterol and change in antioxidant properties of buffalo cheddar cheese in accelerated ripening was not investigated. Accelerated ripening may be used to improve the chemical and sensory prospects of buffalo feta cheese [15]. In view of the economic and nutritional importance of buffalo milk and its lack of suitability for the making of hard/ semi-hard cheese, new technologies should be tried or existing techniques should be explored to enhance the suitability of buffalo milk for the manufacturing of cheddar cheese. To our knowledge, fatty acid profile, organic acids, antioxidant and sensory properties of buffalo cheddar Cheese have not been previously investigated. This study aimed to determine the influence of accelerated ripening on lipolysis, antioxidant and sensory characteristics of buffalo cheddar cheese.
Methods
Materials
Cow and buffalo milk were obtained from Dairy Animals Training and Research Centre, University of Veterinary and Animal Sciences Lahore. Starter culture, Streptococcus lactis ssp. lactis and Streptococcus lactis ssp. cremoris were obtained from Christian Hansen, Denmark. Chemicals used in this investigation were HPLC grade and purchased from Sigma Aldrich, St. Louis, MO, USA.
Experimental plan
Cheddar cheese was prepared from standardized buffalo and cow milk (3.5% fat). Cheddar cheese was ripened at 4 °C and 12 °C for a period of 120 days, analysed at 0, 40, 80 and 120 days of ripening period.
Compositional analysis
Fat, protein and moisture content in cheese were determined by following the standard methods [16].
Lipolysis
Peroxide value and iodine value
Peroxide value and iodine value were determined by following the standard methods [19].
HPLC characterization of organic acids
Samples were prepared by adding 0.1 N phosphoric acid to 5 g grated cheese; organic acids were extracted by dispersing in homogenizer (Heidolph Diax 900, Schwabach, Germany) at 1500 rpm for 5 mins, followed by centrifugation at 7000 rpm for 15 min and supernatant was filtered. Organic acids were determined on HPLC (LCM, Waters Corp., Milford, MA, USA) fitted with ion exchange chromatography column (Supelcogel C-610H, 300 × 7.8 mm, Supelco Inc., Bellefonte, PA, USA). The mobile phase was comprised HPLC grade phosphoric acid with a flow rate of 1 ml/minute, injection volume was 40 μl and measurements were performed at 210 nm. Standards of citric, lactic, formic acids (99% pure, Sigma Chemical Company, St Louis, MO, USA) were used for the identification and quantification of organic acids on a calibration curve and reported as mg/kg dry matter [20].
Antioxidant properties of water-soluble extracts
Preparation of water soluble extracts of Cheddar cheese
Water soluble extracts of cow and buffalo cheddar cheese were prepared according to the method prescribed by [21]. Sixty ml double distilled water was mixed with 20 g grated cheese and homogenized (Ultrasonicator, SONICS, VC 750, USA), homogenized stuff was centrifuged at 14000 g for 10 min (Heraeus, Centrifuge, Hanau, Germany). Supernatant fat layer was castoff, extracts were filtered by Whatman No.1 filter paper. For the precipitation of proteins, pH of the extracts was adjusted to 4.1 using 1 N HCl, contents were again filtered by Whatman No.1. Water-soluble extracts were lyophilized in a rotary evaporator (Buchi, Japan).
Total antioxidant activity
Total antioxidant activity in cheese samples was determined according to the method of [22]. Sample 1 ml was mixed with 3 ml each of 0.6 M sulphuric acid, 28 mM sodium phosphate and 4 mM ammonium molybdate). Test tubes were incubated at 85 °C for 90 min, absorbance was recorded at 695 nm in visible region of spectrum using reagent blank and ascorbic acid as standard. Total antioxidant activity was determined at 0, 40, 80 and 120 days of accelerated ripening and reported as mg Ascorbic Acid Equivalent per gram.
DPPH free radical scavenging activity
DPPH free radical scavenging activity of cow and buffalo cheddar cheese was determined by following the method of [23]. 1 ml sample was mixed with 1 ml DPPH solution (1 mM in methanol) and incubated at room temperature for 30 min. The absorbance was measured on a double beam spectrophotometer (Shimadzu, Japan). DPPH free radical scavenging activity was determined at 0, 40, 80 and 120 days of ripening and expressed as %inhibition.
Sensory evaluation
Sensory evaluation of cheese was performed by a panel of ten trained judges; cheese samples were tempered at 15 °C for 2 h, prior to evaluation. Samples were randomly coded with three-digit random numbers and offered for sensory evaluation in a well illuminated sensory evaluation laboratory at 20 °C, Samples were evaluated for colour, flavour and texture, on 9-point scale at 40, 80 and 120 days of ripening [24].
Statistical analysis
Experiment was planned in a completely randomized design; each treatment was replicated three times. Each sample was analysed three times, results were reported as Mean ± SE, and two-way analysis of variance technique was used to determine the effect of treatments, ripening temperatures and their interaction. T Test was used to determine the significant difference among the treatments (p < 0.05) on SAS 9.1 statistical software [25].
Results and discussion
Chemical composition
Chemical composition of cow and buffalo cheddar cheese ripened at 4 °C and 12 °C
Ripening Temperature | Ripening Days | Cow Cheddar | Buffalo Cheddar | Cow Cheddar | Buffalo Cheddar | Cow Cheddar | Buffalo Cheddar |
---|---|---|---|---|---|---|---|
Moisture % | Moisture % | Fat % | Fat % | Protein % | Protein % | ||
4 °C | 0 | 38.27 ± 1.15a | 35.89 ± 1.28b | 30.55 ± 0.84b | 32.79 ± 0.93a | 24.83 ± 0.55d | 27.18 ± 0.61a |
40 | 38.21 ± 0.99a | 35.62 ± 0.76b | 30.12 ± 0.65b | 32.55 ± 0.65a | 24.64 ± 0.17d | 27.05 ± 0.46a | |
80 | 37.92 ± 0.42a | 35.13 ± 0.48b | 29.41 ± 0.27c | 31.83 ± 0.17b | 24.33 ± 0.36d | 26.88 ± 0.32b | |
120 | 37.73 ± 0.66a | 34.26 ± 0.95c | 29.25 ± 0.52c | 31.27 ± 0.54b | 23.28 ± 0.43e | 26.14 ± 0.29b | |
12 °C | 0 | 38.27 ± 1.15a | 35.89 ± 1.28b | 30.51 ± 0.84b | 32.79 ± 0.93a | 24.87 ± 0.55d | 27.18 ± 0.61a |
40 | 38.12 ± 0.75a | 35.42 ± 0.73b | 29.33 ± 1.12c | 32.18 ± 0.72a | 24.54 ± 0.34d | 27.11 ± 0.53a | |
80 | 37.53 ± 0.46a | 34.78 ± 1.05c | 28.18 ± 0.39d | 31.11 ± 0.19b | 23.53 ± 0.49e | 26.29 ± 0.98b | |
120 | 36.19 ± 0.82b | 33.19 ± 0.59d | 26.85 ± 0.77e | 29.23 ± 0.42c | 22.27 ± 0.19f | 25.36 ± 0.62c |
Total antioxidant capacity (TAC)
Antioxidant properties of cow and buffalo milk cheddar cheese in accelerated ripening
Ripening Temperature | Ripening Days | Cow Cheddar | Buffalo Cheddar | Cow Cheddar | Buffalo Cheddar |
---|---|---|---|---|---|
TAC | TAC | DPPH | DPPH | ||
% | % | % | % | ||
4 °C | 0 | 15.42 ± 0.61l | 20.55 ± 1.46k | 9.61 ± 0.62l | 13.82 ± 1.24k |
40 | 27.36 ± 1.13j | 30.72 ± 1.71i | 19.37 ± 0.52j | 29.88 ± 1.97i | |
80 | 40.87 ± 1.59g | 51.66 ± 2.19f | 31.67 ± 1.16h | 48.22 ± 1.73e | |
120 | 53.42 ± 1.38e | 73.91 ± 2.34c | 44.56 ± 1.66f | 63.27 ± 2.81c | |
12 °C | 0 | 15.42 ± 0.88k | 20.55 ± 0.92k | 9.61 ± 0.75l | 13.82 ± 0.62k |
40 | 32.91 ± 1.33h | 37.43 ± 1.55h | 23.46 ± 1.75i | 39.88 ± 2.11g | |
80 | 65.41 ± 1.98d | 77.94 ± 1.28b | 54.22 ± 2.49e | 67.95 ± 1.43b | |
120 | 77.76 ± 1.47b | 88.30 ± 1.47a | 59.67 ± 1.92d | 72.44 ± 3.53a |
DPPH free radical scavenging activity
DPPH free radical scavenging activity is one of the most significant assays for the assessment of antioxidant properties of food systems. In cheese, DPPH free radical scavenging activity indicates characteristics to the presence of peptides. DPPH Free radical scavenging activity of buffalo cheddar in conventional and accelerated ripening was greater than cow cheddar (Table 2). DPPH free radical scavenging activity of cow and buffalo went on increasing in conventional and accelerated ripening up to four months of ripening. DPPH free radical scavenging activity of 120 days old conventionally ripened cow and buffalo cheddar cheese was 44.56% and 63.27%. DPPH free radical scavenging activity of 120 days accelerated ripened cow and buffalo cheddar was 59.67 and 72.44%. DPPH free radical scavenging activity of 80 days rapid ripened old buffalo cheddar cheese was greater than 120 days rapid ripened cow cheddar. A study was performed to assess the antioxidant properties of peptides cheese ripening for six months, HPLC characterization revealed that antioxidant properties of cheese were strongly correlated with water-soluble peptides, DPPH free radicals scavenging activity of matured cheese was more than 95% [34]. Antioxidant characteristics of white brined cheese were investigated, antioxidant activity of water-soluble and fat-soluble fractions increased during the ripening [35].
Fatty acid profile
Changes in fatty acid composition of fresh and 120 days ripened cheese
Fatty Acids | 4 °C | 12 °C | ||||||
---|---|---|---|---|---|---|---|---|
Cow | Buffalo | Cow | Buffalo | |||||
Fresh | 120 days old* | Fresh | 120 days old* | Fresh | 120 days old* | Fresh | 120 days old* | |
C4:0 | 3.52 ± 0.11e | 3.84 ± 0.19c | 4.12 ± 0.01b | 4.71 ± 0.21a | 3.52 ± 0.11d | 4.14 ± 0.08b | 4.12 ± 0.01b | 4.62 ± 0.22a |
C6:0 | 2.32 ± 0.05c | 2.57 ± 0.16a | 2.39 ± 0.04c | 2.61 ± 0.05b | 2.32 ± 0.05c | 2.83 ± 0.16a | 2.39 ± 0.04c | 2.88 ± 0.13a |
C8:0 | 1.29 ± 0.09e | 1.65 ± 0.09d | 2.12 ± 0.04c | 2.89 ± 0.07b | 1.29 ± 0.09e | 2.15 ± 0.22c | 2.12 ± 0.04c | 3.12 ± 0.18a |
C10:0 | 3.18 ± 0.04c | 4.39 ± 0.25b | 1.81 ± 0.10e | 2.35 ± 0.13d | 3.18 ± 0.04c | 5.24 ± 0.19a | 1.81 ± 0.10e | 3.24 ± 0.07c |
C12:0 | 4.31 ± 0.13a | 4.12 ± 0.07b | 2.61 ± 0.06d | 2.41 ± 0.09e | 4.31 ± 0.13a | 3.77 ± 0.08c | 2.61 ± 0.06d | 2.24 ± 0.09f |
C14:0 | 11.95 ± 0.19a | 10.42 ± 0.15b | 11.82 ± 0.31a | 11.26 ± 0.34a | 11.95 ± 0.19a | 10.29 ± 0.46 | 11.82 ± 0.31a | 10.19 ± 0.53b |
C16:0 | 32.42 ± 1.17a | 31.17 ± 0.18c | 31.67 ± 0.41c | 30.27 ± 1.59d | 32.42 ± 1.17b | 30.27 ± 1.86d | 31.67 ± 0.41c | 28.79 ± 2.71e |
C18:0 | 11.18 ± 0.71a | 10.61 ± 0.46b | 11.28 ± 0.52a | 11.04 ± 0.76 | 11.18 ± 0.71a | 9.66 ± 1.35b | 11.28 ± 0.52a | 9.80 ± 1.43b |
C18:1 | 21.29 ± 0.82b | 19.52 ± 1.19e | 24.52 ± 0.35a | 22.68 ± 1.28c | 21.29 ± 0.82d | 16.38 ± 0.69g | 24.52 ± 0.35a | 18.32 ± 0.88f |
C18:2 | 1.76 ± 0.06c | 1.34 ± 0.11d | 2.65 ± 0.11a | 2.21 ± 0.04b | 1.762 ± 0.06c | 0.49 ± 0.05e | 2.65 ± 0.11a | 0.28 ± 0.04f |
C18:3 | 0.28 ± 0.04b | 0.17 ± 0.02c | 0.65 ± 0.04a | 0.29 ± 0.03b | 0.28 ± 0.04b | 0.05 ± 0.01e | 0.65 ± 0.04a | 0.08 ± 0.02d |
Organic acids
Effect of Conventional Ripening (4 C) on Concentration of organic acids in buffalo cheddar Cheese
Effect of Conventional Ripening (4 °C) on Concentration of organic acids in cow cheddar Cheese
Fig. 2 Effect of Accelerated Ripening (12 °C) on Concentration of organic acids in Buffalo cheddar Cheese
Effect of Accelerated Ripening (12 °C) on Concentration of organic acids in Cow cheddar Cheese
Lipolysis
Lipolysis of cheddar cheese ripened at 4 °C and 12 °C
Ripening Temperature | Ripening Days | Cow Cheddar | Buffalo Cheddar | Cow Cheddar | Buffalo Cheddar | Cow Cheddar | Buffalo Cheddar | Cow Cheddar | Buffalo Cheddar |
---|---|---|---|---|---|---|---|---|---|
FFA | FFA | PV | PV | IV | IV | Cholesterol | Cholesterol | ||
% | % | (MeqO2/kg) | (MeqO2/kg) | Cg/100 g | Cg/100 g | mg/100 g | mg/100 g | ||
4 °C | 0 | 0.11 ± 0.01h | 0.12 ± 0.02h | 0.25 ± 0.02h | 0.23 ± 0.02h | 36.7 ± 1.62c | 39.4 ± 2.36a | 168 ± 4.53a | 57 ± 1.35g |
40 | 0.14 ± 0.02h | 0.13 ± 0.01h | 0.32 ± 0.06h | 0.28 ± 0.04h | 35.8 ± 1.37e | 38.9 ± 3.15b | 152 ± 3.65b | 58 ± 1.42h | |
80 | 0.19 ± 0.04g | 0.18 ± 0.04g | 0.78 ± 0.11e | 0.65 ± 0.09f | 34.5 ± 1.29f | 37.4 ± 1.52c | 129 ± 3.95d | 38 ± 1.19i | |
120 | 0.26 ± 0.03e | 0.22 ± 0.05f | 1.19 ± 0.08c | 1.08 ± 0.15d | 32.7 ± 0.81g | 36.5 ± 1.89d | 88 ± 2.39f | 25 ± 1.10j | |
12 °C | 0 | 0.11 ± 0.01 | 0.12 ± 0.02h | 0.25 ± 0.02h | 0.23 ± 0.02h | 36.7 ± 1.62d | 39.4 ± 2.36a | 168 ± 4.53a | 67 ± 1.35g |
40 | 0.27 ± 0.02e | 0.19 ± 0.03g | 0.43 ± 0.03 | 0.52 ± 0.05g | 34.8 ± 0.95f | 38.1 ± 4.34b | 144 ± 2.33c | 53 ± 3.22h | |
80 | 0.47 ± 0.04c | 0.39 ± 0.04d | 1.26 ± 0.15c | 1.18 ± 0.09c | 32.4 ± 1.48g | 35.6 ± 1.49e | 107 ± 1.67e | 31 ± 1.19j | |
120 | 0.62 ± 0.03a | 0.55 ± 0.02b | 2.59 ± 0.08a | 2.38 ± 0.16b | 29.2 ± 1.34h | 33.2 ± 2.74g | 73 ± 3.42g | 16 ± 0.56k |
Sensory evaluation
Sensory characteristics of 120 days old cheddar cheese ripened at 4 °C and 12 °C
Ripening Temperature | Ripening Days | Cow Cheddar | Buffalo Cheddar | Cow Cheddar | Buffalo Cheddar | Cow Cheddar | Buffalo Cheddar |
---|---|---|---|---|---|---|---|
Color | Color | Flavor | Flavor | Texture | Texture | ||
4 °C | 40 | 6.8 ± 0.24a | 6.4 ± 0.35c | 6.5 ± 0.11d | 6.2 ± 0.16d | 6.2 ± 0.34e | 6.0 ± 0.29e |
80 | 7.5 ± 0.15a | 7.1 ± 0.19b | 7.6 ± 0.19b | 7.1 ± 0.10c | 6.9 ± 0.16c | 6.5 ± 0.13d | |
120 | 8.1 ± 0.34a | 7.6 ± 0.16b | 8.0 ± 0.52a | 7.5 ± 0.26b | 8.1 ± 0.26a | 6.8 ± 0.08c | |
12 °C | 40 | 7.1 ± 0.14a | 6.5 ± 0.42c | 6.9 ± 0.15c | 6.3 ± 0.12d | 6.4 ± 0.15d | 6.3 ± 0.12e |
80 | 8.0 ± 0.08a | 8.1 ± 0.05a | 8.2 ± 0.13a | 7.7 ± 0.65a | 8.0 ± 0.14a | 7.9 ± 0.45a | |
120 | 8.2 ± 0.17a | 8.0 ± 0.11a | 8.0 ± 0.16a | 7.9 ± 0.53a | 7.9 ± 0.31a | 8.1 ± 0.27a |
Conclusions
Flavour and texture score of buffalo cheddar ripened at 12 °C for 80 days was not significantly different from cow cheddar. These results suggest that accelerated ripening can be used to improve the sensory and chemical characteristics of buffalo cheddar cheese.
Declarations
Acknowledgements
The authors are highly obliged to the Library Department, University of Veterinary and Animal Sciences (UVAS), Government College University Faisalabad (GCUF) and IT Department, Higher Education Commission (HEC, Islamabad) for access to journals, books and valuable database.
Funding
Financial assistance for this study was provided by Higher Education Commission of Pakistan.
Availability of data and materials
The dataset supporting the conclusions of this article is included within the article.
Authors’ contributions
MB conceptualized and MN provided the technical assistance; ITK, JAB and MA performed the study, guided in the data collection and analyze the data; MI helped for drafting the manuscript. “It’s also confirmed that all the authors read and approved the final manuscript”.
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare that they have no competing interests.
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Authors’ Affiliations
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