Exploring the Effect of Milk Fat on Fermented Milk Flavor Based on Gas Chromatography–Ion Mobility Spectrometry (GC-IMS) and Multivariate Statistical Analysis
Abstract
:1. Introduction
2. Results and Discussion
2.1. Effects of Milk Fat on pH and Total Titratable Acidity of Fermented Milk
2.2. Effects of Milk Fat on Water-Holding Capacity, Syneresis, and Color of Fermented Milk
2.3. Effects of Milk Fat on Texture of Fermented Milk
2.4. Results of the Single-Factor Experiment and Response Surface Methodology
2.5. Analysis of E-Nose
2.6. Analysis of GC-IMS
2.6.1. The Volatile Components in Two Fermented Milks Identified by GC-IMS
2.6.2. Multivariate Statistical Analysis by GC-IMS
2.6.3. The Key VOC Analysis by ROAV
2.7. Correlation Analysis of VOCs with an E-Nose
3. Materials and Methods
3.1. Materials and Reagents
3.2. Fermented Milk Preparation
3.3. Physicochemical Determinations
3.4. Texture Profile Analysis
3.5. Experimental Design of Single-Factor Experiments and Response Surface Methodology
3.6. Electronic Nose (E-Nose) Analysis
3.7. Analysis of Volatile Compounds (VOCs) by GC-IMS
3.8. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Xu, H.; Luo, J.; Tian, H.; Li, J.; Zhang, X.; Chen, Z.; Li, M.; Loor, J. Rapid communication: Lipid metabolic gene expression and triacylglycerol accumulation in goat mammary epithelial cells are decreased by inhibition of SREBP-1. J. Anim. Sci. 2018, 96, 2399–2407. [Google Scholar] [CrossRef]
- Aleksandra, W.; Joanna, B.; Olga, B.; Kamila, S.; Jolanta, S.; Katarzyna, K.; Emilia, P.; Monika, A.Z.; Jadwiga, H.; Gabriela, O. Lipid Profile, Lipase Bioactivity, and Lipophilic Antioxidant Content in High Pressure Processed Donor Human Milk. Nutrients 2019, 11, 1972. [Google Scholar] [CrossRef]
- Wang, Y.; Wang, X.; Wang, M.; Zhang, L.; Zan, L.; Yang, W. Bta-miR-34b controls milk fat biosynthesis via the Akt/mTOR signaling pathway by targeting RAI14 in bovine mammary epithelial cells. J. Anim. Sci. Biotechnol. 2021, 12, 83. [Google Scholar] [CrossRef]
- Miles, E.; Calder, P. The influence of the position of palmitate in infant formula triacylglycerols on health outcomes. Nutr. Res. 2017, 44, 1–8. [Google Scholar] [CrossRef]
- Zhao, X.; Yan, H.; Cao, J.; Ye, B.; Zhao, Y.; Liu, L. Effect of milk fat and its main fatty acids on oxidation and glycation level of milk. J. Food Sci. Technol. 2023, 60, 720–731. [Google Scholar] [CrossRef]
- Wang, M.; Fan, M.; Zheng, A.; Wei, C.; Liu, D.; Thaku, K.; Wei, Z. Characterization of a fermented dairy, sour cream: Lipolysis and the release profile of flavor compounds. Food Chem. 2023, 423, 136299. [Google Scholar] [CrossRef] [PubMed]
- Esmaeilzadeh, P.; Ehsani, M.; Mizani, M.; Givianrad, M. Characterization of a traditional ripened cheese, Kurdish Kope: Lipolysis, lactate metabolism, the release profile of volatile compounds, and correlations with sensory characteristics. J. Food Sci. 2021, 86, 3303–3321. [Google Scholar] [CrossRef] [PubMed]
- Su, Y.; Wang, H.; Wu, Z.; Zhao, L.; Huang, W.; Shi, B.; He, J.; Wang, S.; Zhong, K. Sensory Description and Consumer Hedonic Perception of Ultra-High Temperature (UHT) Milk. Foods 2022, 11, 1350. [Google Scholar] [CrossRef]
- Olagunju, A.; Omoba, O.; Enujiugha, V.; Alashi, A.; Aluko, R. Technological Properties of Acetylated Pigeon Pea Starch and Its Stabilized Set-Type Yoghurt. Foods 2020, 9, 957. [Google Scholar] [CrossRef] [PubMed]
- Li, S.; Tang, S.; He, Q.; Hu, J.; Zheng, J. Changes in Proteolysis in Fermented Milk Produced by Streptococcus thermophilus in Co-Culture with Lactobacillus plantarum or Bifidobacterium animalis subsp. lactis during Refrigerated Storage. Molecules 2019, 24, 3699. [Google Scholar] [CrossRef]
- Yadav, V.; Gupta, V.K.; Meena, G.S. Effect of culture levels, ultrafiltered retentate addition, total solid levels and heat treatments on quality improvement of buffalo milk plain set yoghurt. J. Food Sci. Technol. 2018, 55, 1648–1655. [Google Scholar] [CrossRef]
- Trieu, K.; Bhat, S.; Dai, Z.; Leander, K.; Gigante, B.; Qian, F.; Korat, A.; Sun, Q.; Pan, X.; Laguzzi, F.; et al. Biomarkers of dairy fat intake, incident cardiovascular disease, and all-cause mortality: A cohort study, systematic review, and meta-analysis. PLoS Med. 2021, 18, e1003763. [Google Scholar] [CrossRef]
- Zhao, L.; Feng, R.; Ren, F.; Mao, X. Addition of buttermilk improves the flavor and volatile compound profiles of low-fat yogurt. LWT 2018, 98, 9–17. [Google Scholar] [CrossRef]
- Cong, Y.; Yi, H.; Zheng, F. Taste distinguishing of yoghurts based on electronic tongue. Sci. Technol. Food Ind. 2015, 36.04, 49–52+56. [Google Scholar] [CrossRef]
- Christos, S.; Franco, B.; Eugenio, A.; Erna, S.; Luca, C.; Tilmann, D.; Flavia, G. PTR-TOF-MS Analysis for Influence of Milk Base Supplementation on Texture and Headspace Concentration of Endogenous Volatile Compounds in Yogurt. Food Bioprocess Technol. 2012, 5, 2085–2097. [Google Scholar] [CrossRef]
- Chen, L.; Zhang, H.; Shi, H.; Xue, C.; Wang, Q.; Yu, F.; Xue, Y.; Wang, Y.; Li, Z. The flavor profile changes of Pacific oysters (Crassostrea gigas) in response to salinity during depuration. Food Chem. X 2022, 16, 100485. [Google Scholar] [CrossRef]
- He, Z.; Wang, X.; Li, G.; Zhao, Y.; Zhang, J.; Niu, C.; Zhang, L.; Zhang, X.; Ying, D.; Li, S. Antioxidant activity of prebiotic ginseng polysaccharides combined with potential probiotic Lactobacillus plantarum C88. Int. J. Food Sci. Technol. 2015, 50, 1673–1682. [Google Scholar] [CrossRef]
- Xu, X.; Cui, H.; Xu, J.; Yuan, Z.; Liu, X.; Fan, X.; Li, J.; Zhu, D.; Liu, H. Effects of different probiotic fermentations on the quality, soy isoflavone and equol content of soy protein yogurt made from soy whey and soy embryo powder. LWT 2022, 157, 113096. [Google Scholar] [CrossRef]
- Zeineb, J.; Olfa, O.; Slah, Z.; Touhami, K.; El Halima, H. Co-fermentation process strongly affect the nutritional, texture, syneresis, fatty acids and aromatic compounds of dromedary UF-yogurt. J. Food Sci. Technol. 2020, 58, 1727–1739. [Google Scholar] [CrossRef]
- Ge, X.; Tang, N.; Huang, Y.; Chen, X.; Dong, M.; Rui, X.; Zhang, Q.; Li, W. Fermentative and physicochemical properties of fermented milk supplemented with sea buckthorn (Hippophae eleagnaceae L.). LWT 2022, 153, 112484. [Google Scholar] [CrossRef]
- Molaee, P.; Reza, F.; Mortazavian, A.; Sarem, N.; Ali, M.; Akbar, G.; Khorshidian, N. Comparative effects of probiotic and paraprobiotic addition on microbiological, biochemical and physical properties of yogurt. Food Res. Int. 2020, 140, 110030. [Google Scholar] [CrossRef]
- Zhao, X.; Dun, K.; Zhang, Y. Effect of Inulin on the Texture, Rheological Properties, and Microstructure of Synbiotic Yogurt. Sci. Technol. Food Ind. 2023, 44, 72–77. [Google Scholar] [CrossRef]
- Ayyash, M.; Abdalla, A.; Abu-Jdayil, B.; Huppertz, T.; Bhaskaracharya, R.; Al-Mardeai, S.; Mairpady, A.; Ranasinghe, A.; Al-Nabulsi, A. Rheological properties of fermented milk from heated and high pressure-treated camel milk and bovine milk. LWT 2022, 156, 113029. [Google Scholar] [CrossRef]
- Popescu, L.; Ceșco, T.; Gurev, A.; Ghendov-Mosanu, A.; Sturza, R.; Tarna, R. Impact of Apple Pomace Powder on the Bioactivity, and the Sensory and Textural Characteristics of Yogurt. Foods 2022, 11, 3565. [Google Scholar] [CrossRef] [PubMed]
- Yan, B.; Sadiq, F.; Cai, Y.; Fan, D.; Zhang, H.; Zhao, J.; Chen, W. Identification of Key Aroma Compounds in Type I Sourdough-Based Chinese Steamed Bread: Application of Untargeted Metabolomics Analysisp. Int. J. Mol. Sci. 2019, 20, 818. [Google Scholar] [CrossRef] [PubMed]
- Aljewicz, M.; Majcher, M.; Nalepa, B. A Comprehensive Study of the Impacts of Oat β-Glucan and Bacterial Curdlan on the Activity of Commercial Starter Culture in Yogurt. Molecules 2020, 25, 5411. [Google Scholar] [CrossRef]
- Huang, J.; Kong, X.; Chen, Y.; Chen, J. Assessment of flavor characteristics in snakehead (Ophiocephalus argus Cantor) surimi gels affected by atmospheric cold plasma treatment using GC-IMS. Front. Nutr. 2023, 9, 1086426. [Google Scholar] [CrossRef]
- Li, Y.; Wang, J.; Wang, T.; Lv, Z.; Liu, L.; Wang, Y.; Li, X.; Fan, Z.; Li, B. Differences between Kazak Cheeses Fermented by Single and Mixed Strains Using Untargeted Metabolomics. Foods 2022, 11, 966. [Google Scholar] [CrossRef]
- Zhang, S.; Wang, T.; Zhang, Y.; Song, B.; Pang, X.; Lv, J. Effects of Monascus on Proteolysis, Lipolysis, and Volatile Compounds of Camembert-Type Cheese during Ripening. Foods 2022, 11, 1662. [Google Scholar] [CrossRef]
- Liu, J.; Zhang, W.; Liu, D.; Zhang, W.; Ma, L.; Wang, S. Physicochemical properties of a new structural lipid from the enzymatical incorporation of flaxseed oil into mutton tallow. Heliyon 2022, 8, e09615. [Google Scholar] [CrossRef]
- Martín-Garcia, A.; Comas-Basté, O.; Riu-Aumatell, M.; Latorre-Moratalla, M.; López-Tamames, E. Changes in the Volatile Profile of Wheat Sourdough Produced with the Addition of Cava Lees. Molecules 2022, 27, 3588. [Google Scholar] [CrossRef]
- Yang, L.; Liu, J.; Wang, X.; Wang, R.; Ren, F.; Zhang, Q.; Shan, Y.; Ding, S. Characterization of Volatile Component Changes in Jujube Fruits during Cold Storage by Using Headspace-Gas Chromatography-Ion Mobility Spectrometry. Molecules 2019, 24, 3904. [Google Scholar] [CrossRef]
- Zhou, Y.; Wang, Y.; Pan, Q.; Wang, X.; Li, P.; Cai, K.; Chen, C. Effect of salt mixture on flavor of reduced-sodium restructured bacon with ultrasound treatment. Food Sci. Nutr. 2020, 8, 3857–3871. [Google Scholar] [CrossRef]
- Wang, R.; Zhang, Y.; Lu, H.; Liu, J.; Song, C.; Xu, Z.; Yang, H.; Shang, X.; Feng, T. Comparative Aroma Profile Analysis and Development of a Sensory Aroma Lexicon of Seven Different Varieties of Flammulina velutipes. Front. Nutr. 2022, 9, 827825. [Google Scholar] [CrossRef] [PubMed]
- Chen, C.; Tian, T.; Liu, Z.; Yu, H.; Yuan, H.; Tian, H. Comparative Study on Volatile Flavor of Chinese Acid-curd Cheese Using Sensory Evaluation. Food Sci. 2023, 44, 228–236. [Google Scholar] [CrossRef]
- Feng, T.; Shui, M.; Song, S.; Zhuang, H.; Sun, M.; Yao, L. Characterization of the Key Aroma Compounds in Three Truffle Varieties from China by Flavoromics Approach. Molecules 2019, 24, 3305. [Google Scholar] [CrossRef] [PubMed]
- Sakin-Yilmazer, M.; Dirim, S.; Di, P.; Kaymak-Ertekin, F. Yoghurt with candied chestnut: Freeze drying, physical, and rheological behaviour. J. Food Sci. Technol. 2014, 51, 3949–3955. [Google Scholar] [CrossRef] [PubMed]
- Ye, Y.; Li, P.; Zhou, J.; He, J.; Cai, J. The Improvement of Sensory and Bioactive Properties of Yogurt with the Introduction of Tartary Buckwheat. Foods 2022, 11, 1774. [Google Scholar] [CrossRef] [PubMed]
- Zhang, B.; Sun, Z.; Lin, L.; Zhang, C.; Wei, C. Analysis of the Effect of Mixed Fermentation on the Quality of Distilled Jujube Liquor by Gas Chromatography-Ion Mobility Spectrometry and Flavor Sensory Description. Foods 2023, 12, 795. [Google Scholar] [CrossRef] [PubMed]
- Leng, P.; Hu, H.; Cui, A.; Tang, H.; Liu, Y. HS-GC-IMS with PCA to analyze volatile flavor compounds of honey peach packaged with different preservation methods during storage. LWT 2021, 149, 111963. [Google Scholar] [CrossRef]
- Cao, L.; Jia, P.; Liu, H.; Kang, S.; Jiang, S.; Pang, M. Effects of High-Canolol Phenolic Extracts on Fragrant Rapeseed Oil Quality and Flavor Compounds during Frying. Foods 2023, 12, 827. [Google Scholar] [CrossRef] [PubMed]
Milk Fat Content | Hardness/1/N | Cohesiveness | Springiness/mm | Gumminess/N | Chewiness/mj |
---|---|---|---|---|---|
5% | 0.034 ± 0.006 d | 0.644 ± 0.032 ab | 9.564 ± 0.030 d | 0.023 ± 0.003 d | 0.264 ± 0.096 c |
10% | 0.049 ± 0.004 c | 0.621 ± 0.014 b | 11.249 ± 0.200 c | 0.030 ± 0.002 c | 0.371 ± 0.063 c |
15% | 0.060 ± 0.016 bc | 0.636 ± 0.007 ab | 13.794 ± 0.255 b | 0.043 ± 0.003 b | 0.645 ± 0.009 b |
20% | 0.062 ± 0.006 bc | 0.664 ± 0.04 a | 14.067 ± 1.680 ab | 0.045 ± 0.004 b | 0.689 ± 0.069 b |
25% | 0.070 ± 0.004 b | 0.650 ± 0.098 a | 14.995 ± 0.230 ab | 0.045 ± 0.003 b | 0.716 ± 0.212 b |
30% | 0.103 ± 0.023 a | 0.664 ± 0.005 a | 15.432 ± 0.608 a | 0.065 ± 0.004 a | 1.033 ± 0.047 a |
Source | Sum of Squares | df | Mean Square | F Value | p Value | Significance |
---|---|---|---|---|---|---|
Model | 491.74 | 14 | 35.12 | 19.71 | <0.0001 | *** |
A—Content of cultures | 10.08 | 1 | 10.08 | 5.66 | 0.0322 | * |
B—Content of fats | 12.00 | 1 | 12.00 | 6.73 | 0.0212 | * |
C—Fermentation time | 4.08 | 1 | 4.08 | 2.29 | 0.1524 | |
D—Fermentation temperature | 33.33 | 1 | 33.33 | 18.70 | 0.0007 | *** |
AB | 4.00 | 1 | 4.00 | 2.24 | 0.1563 | |
AC | 2.25 | 1 | 2.25 | 1.26 | 0.2801 | |
AD | 0.0000 | 1 | 0.0000 | 0.0000 | 1.0000 | |
BC | 0.2500 | 1 | 0.2500 | 0.1403 | 0.7136 | |
BD | 30.25 | 1 | 30.25 | 16.97 | 0.0010 | *** |
CD | 2.25 | 1 | 2.25 | 1.26 | 0.2801 | |
A2 | 2.96 | 1 | 2.96 | 1.66 | 0.2187 | |
B2 | 182.21 | 1 | 182.21 | 102.24 | <0.0001 | *** |
C2 | 173.71 | 1 | 173.71 | 97.47 | <0.0001 | *** |
D2 | 165.42 | 1 | 165.42 | 92.82 | <0.0001 | *** |
Residual | 24.95 | 14 | 1.78 | |||
Lack of Fit | 21.75 | 10 | 2.17 | 2.72 | 0.1737 | |
Pure Error | 3.20 | 4 | 0.8000 | |||
Cor Total | 516.69 | 28 | ||||
R2 | 0.9517 | Std. Dev. | 1.33 | |||
Adjusted R2 | 0.9034 | Mean | 78.90 | |||
Predicted R2 | 0.7479 | C.V. % | 1.69 | |||
Adeq Precision | 16.4218 |
Count | Compound | CAS# | Formula | MW | RI a | Rt [sec] b | Dt c [RIPrel] | Comment |
---|---|---|---|---|---|---|---|---|
V1 | 2-Nonanone | C821556 | C9H18O | 142.2 | 1091.6 | 481.454 | 1.40809 | |
V2 | 3-Octanone | C106683 | C8H16O | 128.2 | 985.5 | 335.61 | 1.30991 | |
V3 | Benzaldehyde | C100527 | C7H6O | 106.1 | 954.8 | 309.657 | 1.14748 | |
V4 | 2-Heptanone | C110430 | C7H14O | 114.2 | 888.3 | 254.655 | 1.26113 | monomer |
V5 | 2-Heptanone | C110430 | C7H14O | 114.2 | 888.3 | 254.655 | 1.63297 | dimer |
V6 | Ethyl lactate | C97643 | C5H10O3 | 118.1 | 832.7 | 225.752 | 1.15009 | |
V7 | Cyclopentanone | C120923 | C5H8O | 84.1 | 792.5 | 204.866 | 1.1021 | |
V8 | 2,3-Butanediol | C513859 | C4H10O2 | 90.1 | 790.5 | 203.795 | 1.38336 | |
V9 | Methylpyrazine | C109080 | C5H6N2 | 94.1 | 791.5 | 204.331 | 1.07755 | |
V10 | 3-Hydroxy-2-butanone | C513860 | C4H8O2 | 88.1 | 703.4 | 167.648 | 1.06192 | monomer |
V11 | 3-Hydroxy-2-butanone | C513860 | C4H8O2 | 88.1 | 703.4 | 167.648 | 1.32867 | dimer |
V12 | 2-Methylpropanoic acid | C79312 | C4H8O2 | 88.1 | 766.5 | 193.353 | 1.1646 | |
V13 | 4-Methyl-3-penten-2-one | C141797 | C6H10O | 98.1 | 789.4 | 203.26 | 1.43582 | |
V14 | 2-Pentanone | C107879 | C5H10O | 86.1 | 664.8 | 155.331 | 1.11773 | monomer |
V15 | 2-Pentanone | C107879 | C5H10O | 86.1 | 665.8 | 155.598 | 1.36885 | dimer |
V16 | 2-Butanone | C78933 | C4H8O | 72.1 | 556.3 | 126.68 | 1.24385 | |
V17 | Butanal | C123728 | C4H8O | 72.1 | 597.8 | 137.658 | 1.28961 | |
V18 | 2-Propanone | C67641 | C3H6O | 58.1 | 475.2 | 105.259 | 1.11326 | |
V19 | Ethanol | C64175 | C2H6O | 46.1 | 419.4 | 90.532 | 1.04741 | |
V20 | Propanal | C123386 | C3H6O | 58.1 | 516.7 | 116.237 | 1.04295 | |
V21 | Hydroxyacetone | C116096 | C3H6O2 | 74.1 | 625.2 | 144.888 | 1.04183 | |
V22 | 3-Methyl-2-butenal | C107868 | C5H8O | 84.1 | 773.8 | 196.298 | 1.09094 | |
V23 | Ethyl Acetate | C141786 | C4H8O2 | 88.1 | 598.9 | 137.926 | 1.09429 | |
V24 | Pentanal | C110623 | C5H10O | 86.1 | 687.1 | 161.221 | 1.41684 | |
V25 | Hexanal | C66251 | C6H12O | 100.2 | 784.8 | 200.85 | 1.25612 | |
V26 | Heptanal | C111717 | C7H14O | 114.2 | 898.4 | 261.9 | 1.33202 | |
V27 | (E)-2-Hexenal | C6728263 | C6H10O | 98.1 | 814.7 | 216.38 | 1.17688 | |
V28 | Dimethyl sulfide | C75183 | C2H6S | 62.1 | 510.5 | 114.595 | 0.95577 | |
V29 | 2-Propanol | C67630 | C3H8O | 60.1 | 527.5 | 119.089 | 1.20946 | |
V30 | 1-Pentanol | C71410 | C5H12O | 88.1 | 756.8 | 189.41 | 1.25209 | |
V31 | Isobutanol | C78831 | C4H10O | 74.1 | 601.6 | 138.652 | 1.16475 | |
V32 | 3-Methylbutanal | C590863 | C5H10O | 86.1 | 644.6 | 150.02 | 1.16267 | |
V33 | 1-Hexanol | C111273 | C6H14O | 102.2 | 868.3 | 244.257 | 1.32146 | |
V34 | 2-Hexanone | C591786 | C6H12O | 100.2 | 777.4 | 197.77 | 1.18483 | |
35 | Nonanal | C124196 | C9H18O | 142.2 | 1109.1 | 506.666 | 1.48711 | |
36 | Octanal | C124130 | C8H16O | 128.2 | 1004.2 | 355.979 | 1.41163 | |
37 | 1 | unidentified | nd d | nd | 767.2 | 193.62 | 1.33537 | |
38 | 2 | unidentified | nd | nd | 775.7 | 197.10 | 1.41238 | |
39 | 3 | unidentified | nd | nd | 754.7 | 188.53 | 1.4001 | |
40 | 4 | unidentified | nd | nd | 721.8 | 175.15 | 1.40233 |
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Tan, C.; Tian, Y.; Tao, L.; **e, J.; Wang, M.; Zhang, F.; Yu, Z.; Sheng, J.; Zhao, C. Exploring the Effect of Milk Fat on Fermented Milk Flavor Based on Gas Chromatography–Ion Mobility Spectrometry (GC-IMS) and Multivariate Statistical Analysis. Molecules 2024, 29, 1099. https://doi.org/10.3390/molecules29051099
Tan C, Tian Y, Tao L, **e J, Wang M, Zhang F, Yu Z, Sheng J, Zhao C. Exploring the Effect of Milk Fat on Fermented Milk Flavor Based on Gas Chromatography–Ion Mobility Spectrometry (GC-IMS) and Multivariate Statistical Analysis. Molecules. 2024; 29(5):1099. https://doi.org/10.3390/molecules29051099
Chicago/Turabian StyleTan, Chunlei, Yang Tian, Liang Tao, **g **e, Mingming Wang, Feng Zhang, Zhi** Yu, Jun Sheng, and Cunchao Zhao. 2024. "Exploring the Effect of Milk Fat on Fermented Milk Flavor Based on Gas Chromatography–Ion Mobility Spectrometry (GC-IMS) and Multivariate Statistical Analysis" Molecules 29, no. 5: 1099. https://doi.org/10.3390/molecules29051099