This work presents the sample extraction methods for solid and liquid sample matrices for simultaneous quantification of oat (Avena sativa L.) and pea (Pisum sativum L.) saponins: avenacoside A, avenacoside B, 26-desglucoavenacoside A, and saponin B and 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) saponin, respectively. The targeted saponins were identified and quantified using a hydrophilic interaction liquid chromatography with mass spectrometric detection (HILIC-MS) method. The simple and high-throughput extraction procedure was developed for solid oat- and pea-based food samples. In addition, a very simple extraction procedure for liquid samples, without the need to use lyophilisation, was also implemented. Oat seed flour (U-¹³C-labelled) and soyasaponin Ba were used as internal standards for avenacoside A and saponin B, respectively. Other saponins were relatively quantified based on avenacoside A and saponin B standard responses. The developed method was tested and successfully validated using oat and pea flours, protein concentrates and isolates, as well as their mixtures, and plant-based drinks. With this method, the saponins from oat- and pea-based products were separated and quantified simultaneously within 6 min. The use of respective internal standards derived from U-¹³C-labelled oat and soyasaponin Ba ensured high accuracy and precision of the proposed method.

The high throughput in genome sequencing and metabolic model (MM) reconstruction has democratised bioinformatics approaches such as flux balance analysis. Fluxes’ prediction accuracy greatly relates to the deepness of the MM curation for a specific organism starting from the cell composition. One component is the cell wall, which is a functional barrier (cell shape, exchanges) with the environment. The bacterial cell wall (BCW), including its thickness, structure, and composition, has been extensively studied in Escherichia coli but poorly described for other organisms. The peptidoglycan (PG) layer composing the BCW is usually thinner in Gram− bacteria than in Gram+ bacteria. In both bacteria groups, PG is a polymeric mesh-like structure of amino acids and sugars, including N-acetylglucosamine, N-acetylmuramic acid, and amino acids. In this study, we propose a high-throughput method to characterise and quantify PG in Gram-positive and Gram-negative bacteria using acidic hydrolysis and hydrophilic interaction liquid chromatography coupled with mass spectrometry (HILIC-MS). The method showed a relatively short time frame (11 min analytical run), low inter- and intraday variability (3.2% and 4%, respectively), and high sensitivity and selectivity (limits of quantification in the sub mg/L range). The method was successfully applied on two Gram-negative bacteria (Escherichia coli K12 MG1655, Bacteroides thetaiotaomicron DSM 2079) and one Gram-positive bacterium (Streptococcus salivarius ssp. thermophilus DSM20259). The PG concentration ranged from 1.6% w/w to 14% w/w of the dry cell weight. The results were in good correlation with previously published results. With further development, the PG concentration provided by this newly developed method could reinforce the curation of MM.

This study investigates the effect of extended rye scald fermentation times (12–48 h) on its biochemical properties and rye bread’s nutritional and sensory qualities. Traditional rye bread production in Latvia involves prolonged fermentation with lactic acid bacteria (LAB), a process that influences the bread’s acidity, sugar content, and concentrations of organic acids, fructans, and phytates. Scald fermentation was analyzed at intervals of 0, 12, 24, 36, and 48 h to monitor microbial activity, particularly LAB population dynamics. Organic acids and sugar profiles were analyzed using HPLC, while phytic acid and fructan concentrations were determined using the Phytic Acid Assay Kit (K-PHYT) and Fructan Assay Kit (K-FRUC). Sensory evaluation assessed attributes including aroma, sour and sweet taste, stickiness, and floury aftertaste. A rapid pH decrease and increased total titratable acidity (TTA) after 12 h confirmed scald’s suitability as a substrate for Lactobacillus delbrueckii metabolism. Lactic acid content increased 13.8-fold after 48 h. Combined scald and dough sourdough fermentation reduced phytic acid by 20% and fructans by 49%, improving mineral bioavailability. Extending fermentation beyond 24 h showed no significant differences in physicochemical parameters, although it improved sensory quality, reduced stickiness, balanced sweet–sour flavors, enhanced aroma, and minimized floury aftertaste.