Fabio Masotti

Liquid infant formulas: technological tools for limiting heat damage.

In a study considering 15 commercial samples of liquid milk-based infant formulas (MBF) from different manufacturers, the levels of selected molecules, that is, furosine (FUR), galactosyl-beta-pyranone (GAP), lactulose (LCT), and lysinoalanine (LAL), have been measured to provide estimation of the heat damage in these products. The ranges of the studied markers were as follows: FUR=153-600 mg 100 g(-1) of protein, GAP=0.5-4.3 mg L(-1), LCT=226-1511 mg L(-1), and LAL=1.0-16.1 mg 100 g(-1) of protein. The highest levels were found in MBF intended for the youngest babies. Experimental samples were produced in an industrial plant to evaluate the relative contribution of individual technological aspects to the final heat damage. About 90% of both GAP and LCT contents was due to the ultrahigh-temperature sterilization process itself. This effect was more than halved when the pH of the ingredient mixture was adjusted from 7.2 to 6.9 before sterilization or when the product recirculated in the plant was discarded. Up to 60 and 20%, respectively, of the FUR and LAL levels in the finished product were already present in protein ingredients (whey powder, whey protein concentrate). Accurate optimization of processing conditions and scrupulous selection of raw materials proved to be effective means to minimize heat damage in such special food products.

Occurrence and fate of ACE-inhibitor peptides in cheeses and in their digestates following in vitro static gastrointestinal digestion

The occurrence of the casein-derived angiotensin converting enzyme-inhibitor (ACE-I) peptides VPP, IPP, RYLGY, RYLG, AYFYPEL, AYFYPE, LHLPLP and HLPLP were investigated in 12 different cheese samples by Ultra Performance Liquid Chromatography/High-Resolution Mass Spectrometry. The total amount of ACE-I peptides was in the range 0.87-331mgkg(-1). VPP and IPP largely prevailed in almost all cheeses. Following in vitro static gastrointestinal digestion of Cheddar, Gorgonzola, Maasdam and Grana Padano cheeses, type and amount of ACE-I peptides changed, and only VPP, IPP, HLPLP and LHLPLP were detected in the intestinal digestates. The results evidenced that the degree of proteolysis itself cannot be regarded as a promoting or hindering factor for ACE-I peptide release during cheese digestion. Moreover, the data indicated that the ACE-I potential of cheeses cannot be inferred based on the type and amount of ACE-I peptides present in undigested samples.

The evolution of chemical and microbiological properties of fresh goat milk cheese during its shelf life

This study investigated the changes in chemical and microbiological properties of fresh goat milk cheese stored in an open deck refrigerated display cabinet (6 ± 2°C) or in a dark cold room (4 ± 1°C). The effects of partial-vacuum packaging and fluorescent lighting were studied during the cheese shelf life (45 d) and 15 d after. Storage conditions did not affect the pH values (4.3), whereas a slight decrease in moisture (ca. 1%) and in water activity (<0.01 units) was recorded. Proteolysis monitored by Kjeldahl determination increased significantly during storage of all samples. The highest increase from 8.5 to 13.0% of soluble nitrogen (expressed as percentage of total nitrogen) was measured in cheese packaged in the presence of air and stored in a lighted cabinet. The proteolytic trend was also studied through capillary zone electrophoresis by monitoring the degradation of the main casein fractions and the formation of new peptides. In particular, 2 indices, based on peak area ratio of new-formed peptides and casein fractions were related to cheese age. Lipolysis, measured by solid-phase microextraction gas chromatography coupled to mass spectrometry of volatile fatty acids, was unaffected by air or light and did not proceed through storage. As expected, hexanal formed mainly in cheeses stored under light and packaged in air. Evaluation of sensorial quality, performed using a hedonic scale, showed significantly lower scores of cheeses kept under light compared with those kept in the dark, both at 45 and 60 d storage. Overall, the microbiological and chemical results suggested that the shelf life of soft goat milk cheese would be extended from 45 to 60 d. Such conclusion was supported also by the sensory quality evaluation.

Nutritional Quality of Milk Proteins

The first part of this chapter outlines general aspects concerning dietary proteins, i.e. protein requirements in human diet, role and nutritional quality of proteins and methods for its evaluation (CS, Protein Digestibility Corrected AA Score [PDCAAS]), protein digestibility and efficiency of protein utilisation.

Rapid determination of sodium in milk and milk products by capillary zone electrophoresis

A capillary zone electrophoresis method for the determination of Na in milk and milk products was developed and compared with an International Organization for Standardization/International Dairy Federation standard method that is based on flame atomic absorption spectrometry. The adoption of a background electrolyte consisting of 10 mM imidazole adjusted to pH 3.75 by the addition of oxalic acid allowed baseline separation of Na from other milk cations and from Li ion, which was adopted as an internal standard. Method validation was performed and the results for linearity, precision, limit of detection, limit of quantification, and recovery are presented. The procedure was tested on commercial milk samples differing in fat content (whole, semiskimmed, and skimmed) and processing conditions (pasteurization, UHT sterilization, and microfiltration). The reliability of the method was confirmed for different varieties of cheese and other milk products. The method enables the routine measurement of Na content by a rapid and accurate capillary zone electrophoresis procedure.

Heat damage and in vitro starch digestibility of puffed wheat kernels

The effect of processing conditions on heat damage, starch digestibility, release of advanced glycation end products (AGEs) and antioxidant capacity of puffed cereals was studied. The determination of several markers arising from Maillard reaction proved pyrraline (PYR) and hydroxymethylfurfural (HMF) as the most reliable indices of heat load applied during puffing. The considerable heat load was evidenced by the high levels of both PYR (57.6-153.4 mg kg(-1) dry matter) and HMF (13-51.2 mg kg(-1) dry matter). For cost and simplicity, HMF looked like the most appropriate index in puffed cereals. Puffing influenced starch in vitro digestibility, being most of the starch (81-93%) hydrolyzed to maltotriose, maltose and glucose whereas only limited amounts of AGEs were released. The relevant antioxidant capacity revealed by digested puffed kernels can be ascribed to both the new formed Maillard reaction products and the conditions adopted during in vitro digestion.

Protein breakdown and release of β-casomorphins during in vitro gastro-intestinal digestion of sterilised model systems of liquid infant formula.

Protein modifications occurring during sterilisation of infant formulas can affect protein digestibility and release of bioactive peptides. The effect of glycation and cross-linking on protein breakdown and release of β-casomorphins was evaluated during in vitro gastro-intestinal digestion (GID) of six sterilised model systems of infant formula. Protein degradation during in vitro GID was evaluated by SDS-PAGE and by measuring the nitrogen content of ultrafiltration (3kDa) permeates before and after in vitro GID of model IFs. Glycation strongly hindered protein breakdown, whereas cross-linking resulting from β-elimination reactions had a negligible effect. Only β-casomorphin 7 (β-CM7) was detected (0.187-0.858mgL(-1)) at the end of the intestinal digestion in all untreated IF model systems. The level of β-CM7 in the sterilised model systems prepared without addition of sugars ranged from 0.256 to 0.655mgL(-1). The release of this peptide during GID was hindered by protein glycation.

Composition, proteolysis, and volatile profile of Strachitunt cheese.

Strachitunt, a blue-veined Italian cheese, received the Protected Designation of Origin (PDO) label in 2014. Its unique technological feature is represented by the dual-curd method of production. Strachitunt is produced from raw bovine milk with or without the inoculation of natural starter cultures of lactic acid bacteria, and the addition of secondary cultures of mold spores is not permitted by the product specification. Physico-chemical properties, proteolysis, and volatile profile of Strachitunt were investigated in 10 cheese samples (ripened for 75 d) made throughout spring 2015 and provided by the main cheese maker. Overall, composition parameters showed a large variability among samples. Cheese was characterized by an acid paste (pH 5.46) and a lower extent of proteolysis compared with other blue-veined varieties. The main chemical groups of volatile organic compounds were alcohols and esters, whereas ketones represented only a minor component. The erratic adventitious contamination by mold spores of the cheese milk, the unique dual-curd method of cheese-making, and the large time variability between the piercing time and the end of ripening could be highlighted as the main causes of both the distinctive analytical fingerprint and the scarce standardization of this blue-veined cheese.

Detection of milk powder and caseinates in Halloumi cheese

Halloumi cheese is traditionally manufactured from fresh milk. Nevertheless, dried dairy ingredients are sometimes illegally added to increase cheese yield. Lysinoalanine and furosine are newly formed molecules generated by heating and drying milk protein components. The levels of these molecular markers in the finished Halloumi have been investigated to verify their suitability to reveal the addition of skim milk powder and calcium caseinate to cheese milk. Because of the severe heating conditions applied in curd cooking, genuine Halloumi cheeses (n=35), representative of the Cyprus production, were characterized by levels of lysinoalanine (mean value=8.1 mg/100g of protein) and furosine (mean value=123 mg/100g of protein) unusual for natural cheeses. Despite the variability of the values, a good correlation between the 2 parameters (R=0.975) has been found in all cheeses, considering both the fresh and mature cheeses as well as those obtained from curd submitted to a prolonged cooking following a traditional practice adopted by a very small number of manufacturers. Experimental cheeses made by adding as low as 5% of skim milk powder, or calcium caseinate, or both, to cheese milk fell outside the prediction limits at +/-2 standard deviation of the above-reported correlation regardless of curd cooking conditions or ripening length. This correlation may be adopted as a reliable index of Halloumi cheese genuineness.

Functional characterization of Lactobacillus plantarum ITEM 17215: A potential biocontrol agent of fungi with plant growth promoting traits, able to enhance the nutritional value of cereal products

In this work, we explored the potential of 25 Lactobacillus plantarum strains isolated from cereals and milk-based products, testing characteristics related to antifungal activity and to nutritional quality. The tested strains demonstrated interesting beneficial traits, such as the ability to utilize fructo-oligosaccharides, prebiotic substances that help probiotic microorganisms to grow in the human gut, and to reduce phytate, an antinutrient present in cereal sector. Regarding mould inhibition, we highlighted the ability of the strains to inhibit Penicillium roqueforti, Mucor circinelloides and mycotoxinogenic moulds associated with cereal grains as Aspergillus flavus, A. niger, Fusarium verticillioides. Moreover, a moderate reduction of the bioavailability of aflatoxin AFB1 was detected. The selected L. plantarum strain ITEM 17215, showed a strong inhibitory ability towards fungal growth and was able to produce 1,2-dihydroxybenzene, benzoic acid, p-hydroxyphenyllactic acid and 3-phenyllactic acid. The latter compound, already described as efficient antifungal inhibitor, was the most abundant and its concentration was further increased by adding phenylalanine and phenylpyruvic acid in the growth medium. The metabolites produced by strain ITEM 17215 could also be related to the ability of the strain to induce cereal germination and promote plant growth. This aspect, not yet investigated in L. plantarum, could have interesting applications in the agro-food sector.