Techno-Functional and Sensory Characterization of Commercial Plant Protein Powders.

Many new plant proteins are appearing on the market, but their properties are insufficiently characterized. Hence, we collected 24 commercial proteins from pea, oat, fava bean, chickpea, mung bean, potato, canola, soy, and wheat, including different batches, and assessed their techno-functional and sensory properties. Many powders had yellow, red, and brown color tones, but that of fava bean was the lightest. The native pH ranged from 6.0 to 7.7. The water solubility index was 28% on average, but after heat treatment the solubility typically increased. Soy isolate had by far the best water-holding capacity of 6.3 g (H2O) g-1, and canola had the highest oil-holding capacity of 2.8 g (oil) g-1. The foaming capacity and stability results were highly varied but typical to the raw material. The emulsification properties of all powders were similar. Upon heating, the highest viscosity and storage modulus were found in potato, canola, and mung bean. All powders had raw material flavor, were bitter and astringent, and undissolved particles were perceived in the mouth. Large differences in functionality were found between the batches of one pea powder. In conclusion, we emphasize the need for methodological standardization, but while respecting the conditions found in end applications like meat and dairy analogs.

Development of Extraction Method for Determination of Saponins in Soybean-Based Yoghurt Alternatives: Effect of Sample pH.

The number of plant-based dairy alternative products on the market is growing rapidly. In the case of soybean-based yoghurt alternatives, it is important to trace the content of saponins, the phytomicronutrients with a disputable health effect, which are likely to be responsible for the bitter off-taste of the products. We present a new sample extraction method followed by hydrophilic interaction liquid chromatography with mass spectrometric detection (HILIC-MS) for identifying and quantifying soyasaponins in soybean-based yoghurt alternatives. Soyasaponin Bb, soyasaponin Ba, soyasaponin Aa, and soyasaponin Ab were quantified using commercially available standard compounds and with asperosaponin VI as the internal standard. As the recoveries of soyasaponins were unacceptable in yoghurt alternatives at their natural acidic pH, the adjustment of pH was performed as one of the first steps in the sample extraction procedure to achieve the optimum solubility of soyasaponins. The validation of the method included the assessment of linearity, precision, limit of detection and limit of quantification (LOQ), recovery, and matrix effect. The average concentrations of soyasaponin Bb, soyasaponin Ba, soyasaponin Ab, and soyasaponin Aa in several measured soybean-based yoghurt alternatives utilising the developed method were 12.6 ± 1.2, 3.2 ± 0.7, 6.0 ± 2.4 mg/100 g, and below the LOQ, respectively. This method provides an efficient and relatively simple procedure for extracting soyasaponins from yoghurt alternatives followed by rapid quantification using HILIC-MS and could find a rightful application in the development of healthier and better-tasting dairy alternatives.

Starter culture growth dynamics and sensory properties of fermented oat drink.

In the present study, an oat drink, a plant-based alternative to dairy products, was developed by fermenting the oat base with different vegan starter cultures. The desired pH below 4.2 was achieved in 12 h, regardless of starter culture used. Metagenomic sequencing revealed that S. thermophilus was the dominating species, ranging from 38% to 99% of the total microbial consortia. At lower pH values, population of L. acidophilus, L. plantarum and L. paracasei continued to increase in fermented oat drinks. Lactic acid was produced between 1.6 and 2.8 g/L. The sensory panel showed that all fermented oat drinks had a sour odor and taste. The volatile compounds identified belonged to the ketone, alcohol, aldehyde, acids, and furan classes. The concentration of the most preferred volatile components, such as diacetyl and acetoin, increased during fermentation. However, sensory evaluation showed that all samples were associated with cereals and not dairy in terms of taste and odor. Rheological analysis showed the formation of weak gel-like structures in fermented oat drinks. Overall, fermentation improved flavor and texture of the product. This study provides a broad overview of the oat drink fermentation process from the perspectives of starter culture growth, microbial consortium dynamics, lactic acid bacteria metabolism, and sensory profile formation.

Quantitative Analysis of Oat (Avena sativa L.) and Pea (Pisum sativum L.) Saponins in Plant-Based Food Products by Hydrophilic Interaction Liquid Chromatography Coupled with Mass Spectrometry.

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.

Development of LC-MS-ESI-TOF method for quantification of phytates in food using 13C-labelled maize as internal standard.

In this work, the LC-MS-ESI-TOF method for simultaneous determination of phytates (inositol mono-, bis-, tris-, tetrakis-, pentakis-, and hexakisphosphates, abbreviated to IP1, IP2, IP3, IP4, IP5, and IP6, respectively) in food samples was developed and validated. The suitability of U-13C-labelled maize as a source for labelled internal standards for quantification of phytates was elucidated. The effectiveness of liberating IP1, IP2, IP3, IP4, and IP5 from phytic acid extracted form U-13C-labelled maize was evaluated for a variety of hydrolysis conditions, including enzymatic and acid hydrolysis. Enzymatic degradation of phytic acid using phytase (PHYZYME XP 5000 L) was very effective; phytic acid was degraded to lower phytates, but their distribution was unequal. Chemical hydrolysis was conducted under acidic conditions using hydrochloric acid and elevated temperatures up to 140 °C. The highest yields of IP4, IP5, and IP6 and of IP1, IP2, and IP3 were achieved by chemical hydrolysis at 105 °C for 7 h and 24 h, respectively. Thus, a combination of these two chemical treatments was selected for internal standard production. The developed LC-MS-ESI-TOF method was tested and successfully validated using plant-based food samples with different distribution of phytates. With this method, different forms of phytates in foods were separated and quantified simultaneously within 20 min. The high accuracy and precision of the developed method were guaranteed using respective labelled internal standards derived from U-13C-labelled maize.

Ripening of Hard Cheese Produced from Milk Concentrated by Reverse Osmosis.

The application of reverse osmosis (RO) for preconcentration of milk (RO-milk) on farms can decrease the overall transportation costs of milk, increase the capacity of cheese production, and may be highly attractive from the cheese manufacturer's viewpoint. In this study, an attempt was made to produce a hard cheese from RO-milk with a concentration factor of 1.9 (RO-cheese). Proteolysis, volatile profiles, and sensory properties were evaluated throughout six months of RO-cheese ripening. Moderate primary proteolysis took place during RO-cheese ripening: about 70% of αs1-casein and 45% of ß-casein were hydrolyzed by the end of cheese maturation. The total content of free amino acids (FAA) increased from 4.3 to 149.9 mmol kg-1, with Lys, Pro, Glu, Leu, and γ-aminobutyric acid dominating in ripened cheese. In total, 42 volatile compounds were identified at different stages of maturation of RO-cheese; these compounds have previously been found in traditional Gouda-type and hard-type cheeses of prolonged maturation. Fresh RO-cheese was characterized by a milky and buttery flavor, whereas sweetness, saltiness, and umami flavor increased during ripening. Current results prove the feasibility of RO-milk for the production of hard cheese with acceptable sensory characteristics and may encourage further research and implementation of RO technology in cheese manufacture.

Microcalorimetric study of the growth of Streptococcus thermophilus in renneted milk.

The growth of Streptococcus thermophilus ST12 (ST12) in liquid milk, reconstituted from low-heat skim milk powder reconstituted skim milk (RSM) and in RSM with rennet addition (r-RSM) at 40°C was monitored by microcalorimetry. It was shown that the growth rate of bacteria decreased in renneted samples in comparison with liquid RSM starting from certain sizes of the colonies ("deviation moments"), which depended on the inoculation rates. The hydrolysis of lactose was delayed for about 1 h in the r-RSM in comparison with RSM but otherwise the metabolism of carbohydrates in the renneted and non-renneted milks was similar. The total free amino acids (TFAA) content by the end of fermentations was higher in r-RSM than in RSM presumably due to the enzymatic hydrolytic activity of rennet. The quantitatively dominating amino acids were remarkably different in the r-RSM and RSM indicating that the hydrolysis cascade of caseins and/or metabolism of amino acids by the bacteria functioned differently in the two cases. The data obtained showed potential of microcalorimetry to characterize quantitative differences of growth and metabolism of the bacteria in renneted and liquid samples of milk.