Free Fatty Acids Quantification in Dairy Products

Quantification of free fatty acids in dairy products is not only important due to their (fatty acids) impact on the flavour and texture of dairy products but also because of their potential impact on nutrition and health, and as anti-microbial agents. This chapter provides an overview of the practical issues associated with existing lipid extraction techniques and quantification procedures using gas chromatography flame-ionization detection. The most widely used methods are compared and recent advancements in the quantification of free fatty acids in dairy products are discussed

Genetic, enzymatic and metabolite profiling of the Lactobacillus casei group reveals strain biodiversity and potential applications for flavour diversification

Aims: The Lactobacillus casei group represents a widely explored group of lactic acid bacteria, characterized by a high level of biodiversity. In this study, the genetic and phenotypic diversity of a collection of more than 300 isolates of the Lact. casei group and their potential to produce volatile metabolites important for flavour development in dairy products, was examined. Methods and Results: Following confirmation of species by 16S rRNA PCR, the diversity of the isolates was determined by pulsed-field gel electrophoresis. The activities of enzymes involved in the proteolytic cascade were assessed and significant differences among the strains were observed. Ten strains were chosen based on the results of their enzymes activities and they were analysed for their ability to produce volatiles in media with increased concentrations of a representative aromatic, branched chain and sulphur amino acid. Volatiles were assessed using gas chromatography coupled with mass spectrometry. Strain-dependent differences in the range and type of volatiles produced were evident. Conclusions: Strains of the Lact. casei group are characterized by genetic and metabolic diversity which supports variability in volatile production. Significance and Impact of the Study: This study provides a screening approach for the knowledge-based selection of strains potentially enabling flavour diversification in fermented dairy products

Comparison and validation of 2 analytical methods for the determination of free fatty acids in dairy products by gas chromatography with flame ionization detection

peer-reviewedAccurate quantification of free fatty acids (FFA) in dairy products is important for quality control, nutritional, antimicrobial, authenticity, legislative, and flavor purposes. In this study, the performance of 2 widely used gas chromatographic flame ionization detection methods for determination of FFA in dairy products differing in lipid content and degree of lipolysis were evaluated. We used a direct on-column approach where the isolated FFA extract was injected directly and a derivatization approach where the FFA were esterified in the injector to methyl esters using tetramethylammonium hydroxide as a catalyst. A comprehensive validation was undertaken to establish method linearity, limits of detection, limits of quantification, accuracy, and precision. Linear calibrations of 3 to 700mg/L (R2>0.999) and 20 to 700mg/L (R2>0.997), and limits of detection and limits of quantification of 0.7 and 3mg/L and 5 and 20mg/L were obtained for the direct injection on-column and the derivatization method, respectively. Intraday precision of 1.5 to 7.2% was obtained for both methods. The direct injection on-column method had the lower levels of limits of detection and quantification, because FFA are directly injected onto the GC as opposed to the split injection used in the derivatization method. However, the direct injection on-column method experienced accumulative column phase deterioration and irreversible FFA absorption because of the acidic nature of the injection extract, which adversely affected method robustness and the quantification of some longer chain FFA. The derivatization method experienced issues with quantification of butyric acid at low concentrations because of coelution with the injection solvent peak, loss of polyunsaturated FFA due to degradation by tetramethylammonium hydroxide, and the periodic emergence of by-product peaks of the tetramethylammonium hydroxide reaction that interfered with the quantification of some short-chain FFA. The derivatization method is more robust, and because the derivatization step can be automated, it is more suitable for routine analysis of FFA in dairy products. However, considerable scope exists to develop an alternative gas chromatography with flame ionization detection method to quantify FFA in dairy products without any limitations that is robust and accurate

Meta-analysis of cheese microbiomes highlights contributions to multiple aspects of quality

peer-reviewedA detailed understanding of the cheese microbiome is key to the optimization of flavour, appearance, quality and safety. Accordingly, we conducted a high-resolution meta-analysis of cheese microbiomes and corresponding volatilomes. Using 77 new samples from 55 artisanal cheeses from 27 Irish producers combined with 107 publicly available cheese metagenomes, we recovered 328 metagenome-assembled genomes, including 47 putative new species that could influence taste or colour through the secretion of volatiles or biosynthesis of pigments. Additionally, from a subset of samples, we found that differences in the abundances of strains corresponded with levels of volatiles. Genes encoding bacteriocins and other antimicrobials, such as pseudoalterin, were common, potentially contributing to the control of undesirable microorganisms. Although antibiotic-resistance genes were detected, evidence suggested they are not of major concern with respect to dissemination to other microbiomes. Phages, a potential cause of fermentation failure, were abundant and evidence for phage-mediated gene transfer was detected. The anti-phage defence mechanism CRISPR was widespread and analysis thereof, and of anti-CRISPR proteins, revealed a complex interaction between phages and bacteria. Overall, our results provide new and substantial technological and ecological insights into the cheese microbiome that can be applied to further improve cheese production.Science Foundation Irelan

A chemometric approach to characterize the aroma of selected brown and red edible seaweeds/extracts

Background: Information pertaining to the aromatic profile of seaweeds and seaweed extracts can provide evidence regarding their potential suitability as ingredients in processed foods. To date only limited material has been available on the volatile profiles of some seaweed species. Others in this study have not previously been described. The volatile profiles of dried brown (Himanthalia elongata, Undaria pinnatifida, Alaria esculenta) and red (Porphyra umbilicalis, Palmaria palmata) seaweeds, and a brown seaweed extract (fucoxanthin) from Laminaria japonica were investigated using a chemometric approach to collate data from volatile gas chromatography – mass spectrometry (GC–MS), direct sensory aroma evaluation, and gas‐chromatography – olfactometry (GC‐O) to obtain a better understanding of their volatile profile and sensory perception. Results: More than 100 volatile compounds were identified by static headspace solid phase micro‐extraction (HS‐SPME) and thermal desorption gas chromatography – mass spectrometry (TD GC–MS). Brown seaweeds were characterized by ‘grassy/herbal/floral’, ‘fruity’, and ‘fatty’ aromas, red seaweeds by ‘green/vegetable’, ‘mushroom/earthy’ and ‘sweet/buttery’ aromas, and the fucoxanthin extract by ‘rancid’ and ‘nutty’ aromas with an overall lower intensity. Heptanal appeared to be a major odor‐active compound in all samples. Other volatiles were more characteristic of each individual seaweed: hexanal, (E,Z)‐2,6‐nonadienal and 2‐pentylfuran for H. elongata; ethyl butanoate and 2,3‐butanedione for U. pinnatifida; 6‐dimethylpyrazine, (E,Z)‐2,6‐nonadienal and sulactone for P. palmata; 1‐octen‐3‐ol for P. umbilicalis, heptanone for A. esculenta, and 2‐furanmethanol for fucoxanthin. Conclusion: Brown and red seaweeds had distinct sensory properties with individual seaweeds having differing volatiles and odorants. This study provides additional information that can contribute to the development of products incorporating dried seaweeds / extracts that are more acceptable to the consumer

Evaluation of the Potential of Lactobacillus paracasei Adjuncts for Flavor Compounds Development and Diversification in Short-Aged Cheddar Cheese

peer-reviewedThe non-starter microbiota of Cheddar cheese mostly comprises mesophilic lactobacilli, such as Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus, and Lactobacillus plantarum. These bacteria are recognized for their potential to improve Cheddar cheese flavor when used as adjunct cultures. In this study, three strains of L. paracasei (DPC2071, DPC4206, and DPC4536) were evaluated for their contribution to the enhancement and diversification of flavor in short-aged Cheddar cheese. The strains were selected based on their previously determined genomic diversity, variability in proteolytic enzyme activities and metabolic capability in cheese model systems. The addition of adjunct cultures did not affect the gross composition or levels of lipolysis of the cheeses. The levels of free amino acids (FAA) in cheeses showed a significant increase after 28 days of ripening. However, the concentrations of individual amino acids in the cheeses did not significantly differ except for some amino acids (aspartic acid, threonine, serine, and tryptophan) at Day 14. Volatile profile analysis revealed that the main compounds that differentiated the cheeses were of lipid origin, such as long chain aldehydes, acids, ketones, and lactones. This study demonstrated that the adjunct L. paracasei strains contributed to the development and diversification of compounds related to flavor in short-aged Cheddar cheeses

Use of smear bacteria and yeasts to modify flavour and appearance of Cheddar cheese

The strains Staphylococcus saprophyticus DPC5671 and Corynebacterium casei DPC5298 were applied in combination with Debaryomyces hansenii DPC6258 to the surface of young Cheddar cheese curd to obtain two different smear-ripened cheeses. A surface microbiota developed over the incubation period, comprising of both yeast and bacteria; pulsed field gel electrophoresis confirmed that the inoculated strains of S. saprophyticus DPC5671 or C. casei DPC5298 were the dominant bacterial strains on the surface of the cheese at the end of the ripening period. The smear cultures changed the appearance and aroma, which were significantly different from the control cheese. The approach presented in this study represents a method for the development of new cheese varieties with novel aromas within a short ripening time

The impact of pasture and non-pasture diets on the sensory and volatile properties of whole milk powder

peer-reviewedThis study evaluated the impact of three distinct diets; perennial ryegrass (GRS), perennial ryegrass/white clover (CLV) and total mixed ration (TMR), on the sensory properties and volatile profile of whole milk powder (WMP). The samples were evaluated using a hedonic sensory acceptance test (n = 99 consumers) and by optimised descriptive profiling (ODP) using trained assessors (n = 33). Volatile profiling was achieved by gas chromatography mass spectrometry using three different extraction techniques; headspace solid phase micro-extraction, thermal desorption and high capacity sorptive extraction. Significant differences were evident in both sensory perception and the volatile profiles of the WMP based on the diet, with WMP from GRS and CLV more similar than WMP from TMR. Consumers scored WMP from CLV diets highest for overall acceptability, flavour and quality, and WMP from TMR diets highest for cooked flavour and aftertaste. ODP analysis found that WMP from TMR diets had greater caramelised flavour, sweet aroma and sweet taste, and that WMP from GRS diets had greater cooked aroma and cooked flavour, with WMP derived from CLV diets having greater scores for liking of colour and creamy aroma. Sixty four VOCs were identified, twenty six were found to vary significantly based on diet and seventeen of these were derived from fatty acids; lactones, alcohols, aldehydes, ketones and esters. The abundance of δ-decalactone and δ-dodecalactone was very high in WMP derived from CLV and GRS diets as was γ-dodecalactone derived from a TMR diet. These lactones appeared to influence sweet, creamy, and caramelised attributes in the resultant WMP samples. The differences in these VOC derived from lipids due to diet are probably further exacerbated by the thermal treatments used in WMP manufacture

The impact of sugar particles size and natural substitutes for the replacement of sucrose and fat in chocolate brownies: Sensory and physicochemical analysis

As fat contributes important textural properties such as lubricity and tenderness to cakes, it is plausible to focus on ways to increase the perception of these properties with the aim of creating the illusion of a higher fat. The utilisation of small sugar particles has been shown to increase the moist and soft texture of Chocolate Brownies. The present study assessed three different sugar particle sizes in their ability to create the illusion of fat content and therefore their ability to permit fat replacement (FR) in this product. The unground commercial sugar (200-5181 µm) was used as the control (UC) and two of its sieved sugar separates, Large (L924-1877 µm) and Small (S459-972 µm) were investigated. For each, fat was replaced using pureed black beans. The most accepted sample was used for sucrose replacement (SR) using inulin and Rebaudioside A. (Reb A.). Samples containing the smallest sugar fraction with 25% FR were most significantly associated overall acceptability (OA) (p<0.01). The application of small sugar particles did not significantly negatively affect OA or liking of samples at a level of 75% FR compared to the other two sugar fractions. The utilisation of small sugar particles (459-972 µm) in the preparation of baked goods could aid baking & industry professionals in reducing the fat content of cake-like products