Phytic Acid Assay Kit

Phytic acid, or myo-inositol hexakisphosphate, is the primary source of inositol and storage phosphorus in plant seeds and has considerable nutritional importance. In this form, phosphorus is unavailable for absorption by monogastric animals, and the strong chelating characteristic of phytic acid reduces the bioavailability of multivalent minerals such as iron, zinc, and calcium. Currently, there is no simple quantitative method for phytic acid; existing methods are complex, and the most commonly accepted method, AOAC Official Method^SM 986.11, has limitations. The aim of this work was to develop and validate a simple, high-throughput method for the measurement of total phosphorus and phytic acid in foods and animal feeds. The method described here involves acid extraction of phytic acid, followed by dephosphorylation with phytase and alkaline phosphatase. The phosphate released from phytic acid is measured using a modified colorimetric molybdenum blue assay and calculated as total phosphorus or phytic acid content of the original sample. The method was validated to a maximum linearity of 3.0 g phytic acid/100 g sample. Accuracy ranged from 98 to 105% using pure phytic acid and from 97 to 115% for spiked samples. Repeatability ranged from 0.81 to 2.32%, and intermediate precision was 2.27%.

The objectives of this study were to investigate whether an increased dietary myo-inositol (MI) supply translates into changes in MI concentrations and endogenous mucosal phosphatase activities in the intestine of laying hens and whether different laying hen strains respond differently to MI supplementation. The diets were corn–soybean meal-based and supplemented without (MI0) or with 1 (MI1), 2 (MI2), or 3 (MI3) g MI/kg feed. Ten hens per strain (Lohmann Brown-classic (LB) and Lohmann LSL-classic (LSL)) and diet were sacrificed at the age of 30 wk following a 4-wk stay in a metabolic unit. The blood plasma, digesta of the duodenum+jejunum and distal ileum, mucosa of the duodenum, and eggs were collected at wk 30. The concentration of MI in the blood plasma was increased by MI supplementation (P < 0.001); however, that of MI3 did not further increase compared with MI2. The concentration of MI in the duodenum+jejunum and ileum increased steadily (P < 0.001). The MI concentration in the duodenum+jejunum was higher in LB than in LSL hens (P = 0.017). The MI concentration in egg yolk was increased by MI supplementation (P < 0.001) and was higher in LB than in LSL hens (P = 0.015). Strain or diet did not affect mucosal phosphatase activity. Myo-inositol flow at the terminal ileum and postileal disappearance increased with each increment in MI supplementation (P < 0.001) and was higher in LB than in LSL hens (P ≤ 0.041). Regression analysis indicated that, on average, 84% of supplemented MI was retained in the body or metabolized and excreted in a different form. Based on the measured MI concentrations in the blood and eggs, dietary MI was not completely absorbed in the small intestine and, to a different extent, in the two laying hen strains. A higher dietary MI supply was followed by higher intestinal absorption or metabolism by microorganisms. The fate of supplemented MI and its relevance to birds warrant further research.

Cereals and pulses are staple foods that provide substantial amounts of dietary protein and essential micronutrients, particularly zinc (Zn) and iron (Fe). The fortification of Zn and Fe in these products has been mandated in 94 countries, making them vital components of a nutritionally balanced diet. However, their bioavailability can be hindered by the anti-nutritional compound phytate. This analysis of cereals and pulses sourced from supermarkets in Adelaide, Australia assessed their phytate content and the bioavailability of Zn and Fe as well as their protein content to evaluate overall quality. Results indicated that pulse products, such as chickpea and lentil, are superior sources of protein and minerals, particularly of Zn and Fe, with higher micronutrient bioavailability compared to cereal products. A calculation based on this survey, 280 g of dried chickpea or lentil provides 100% of the daily Zn intake for women and 57.22% for men. Phytate content and phosphorus nutrition were highly correlated in whole grain products such as brown rice and wheat. In addition, Zn and Fe concentrations in cereal products like wheat and oat flour were lower than other products, suggesting that they may not meet the fortification targets recommended by Food Standards Australia New Zealand.

Legume-based sourdough is gaining momentum. This study aimed to compare the effectiveness of type I sourdough prepared with durum wheat semolina (S1), pea flour (S2) or 50:50 semolina/pea flour (S3) in improving the nutritional quality, antioxidant compounds, in vitro digestibility and aroma of traditional durum wheat focaccia. Six focaccias were prepared: three with 40 % of S1, S2 and S3, and three with the corresponding amount of unfermented flours. Pea sourdough increased the content of phenolic compounds (8.82 ± 0.12 mg GAE/g d.m. in focaccia with 40 % pea flour and 4.92 ± 0.41 mg GAE/g d.m. in unfermented semolina focaccia), and consequently increased the antioxidant activity. Focaccias with pea flour or pea sourdough were “source of protein” and “high fiber”, according to UE Reg. 1924/2006. Pea sourdough slowed down starch in vitro digestibility while enhancing protein digestibility and leading to a more complex volatile profile, with increased content of aldehydes, alcohols and Maillard reaction compounds.

Background: Millets are underutilized grains rich in nutrients. This study aimed to investigate the impact of debranning on the nutritional, cooking, and microstructural properties of five Indian millets namely foxtail, little, kodo, barnyard, and proso millet. Methods: The proximate composition, mineral content, cooking properties (cooking time, solid loss, water uptake, alkali score), Fourier Transform Infra Red (FTIR) spectra, X ray Diffraction (XRD) and microstructural characteristics (Scanning Electron Microscopy) of dehusked and debranned millet samples were examined and analysed. Results: Debranning resulted in decrease in protein (except for little and barnyard millets), dietary fibre, fat, mineral and phytate content in all the millets while enhanced available carbohydrates and amylose content. The cooking times for dehusked millets were significantly higher while, the solid loss and water uptake during cooking of debranned millets were higher. On debranning, Fourier Transform Infra Red (FTIR) spectra showed changes in the pattern with increase in the intensity of amide II (1363 to 1367 cm⁻¹) and amide III (1215 to 1231 cm⁻¹) bands in the debranned foxtail, little, and kodo millets. The X-ray diffractogram (XRD) showed decrease in relative crystallinity on debranning. Scanning Electron Microscopic (SEM) examination revealed that debranning resulted in the loss of seed coat, aleurone layer and partial loss of germ in the millets. Conclusion: Dehusked millets are nutritious and should be promoted in Indian diets to improve diet quality, debranned millets are nutritionally inferior, can increase the glycemic load of Indian diets.

With the rapid development of animal husbandry, the demand for protein feed resources is increasing. Cottonseed meal (CSM) and soybean meal (SBM) are rich sources of protein. However, their application is limited due to the existence of anti-nutrients, which can be harmful to the digestion and absorption. A strain of Bacillus subtilis (Mafic-Y7) was isolated from digesta of intestines of Tibetan pigs. The strain showed high protease activity, which helps in degrading proteinic anti-nutritional factors in grain meal and in vitro degradation of free gossypol. In order to better understand this isolated strain, whole genome of Mafic-Y7 strain was sequenced and analyzed. Different effects on various grain meals were identified.

Jilungin (Terminalia canescens) is a native Australian plant and the Indigenous “Nyul Nyul” people of the  Kimberley region of Western Australia use its leaves to make herbal tea. Due to the rise in the popularity of drinking Jilungin tea among the consumers in Australia and internationally, it is important to study the nutritional and health-beneficial properties as well as safety of Jilungin leaves. This study aims to determine the nutritional composition, anti-nutritional factors, antimicrobial and antidiabetic properties of Jilungin leaves. Also, the phytochemical profiling using UHPLC-MS/MS (Ultra-performance liquid chromatographymass spectrometry) and antioxidant activity of Jilungin methanolic extracts and herbal infusion were investigated. The safety of the leaves and infusion was also investigated by using in vitro mammalian cell lines (Caco2, HT29, and HepG2) through cell viability assays. The leaves are rich in dietary fiber (43.9%) and linoleic acid (30.4% of total fatty acids). Phytochemical profiling revealed ellagic acid, geraniin, pedunculagin, and punicalagin as the major bioactive compounds. The results also demonstrated that Jilungin has strong antioxidant and antidiabetic activities. A significant (p < 0.01) strong positive correlation was observed between the high antioxidant activity of Jilungin infusion with the major bioactive compounds. Jilungin extracts (50 mg/mL) exhibited strong antimicrobial activity against Staphylococcus aureus and Bacillus cereus. Its infusion and methanolic extract were safe on the studied cell lines (Caco-2, HT29, and HepG2) at higher concentrations of 66.6 and 98 mg/mL, respectively. Therefore, Jilungin teas or infusions could be a safe and effective way to promote health and well-being.

Cereal products contribute significantly to dietary intake of essential minerals. In wheat, iron and zinc are stored in specific grain structures including the aleurone, scutellum and embryo. Wheat cell walls are resistant to digestion in the human gastrointestinal tract and therefore this study investigated the hypothesis that physical disruption of the cell walls would increase the bioaccessibility and bioavailability of iron and zinc from wheat-based foods. Flour was micronized using a combination of roller milling and a micro-mill and this reduced median particle size by two-thirds. Hydrothermally processed wheat flour doughs were subjected to in vitro digestion to determine mineral bioaccessibility. Mineral bioavailability from food digests was measured using human intestinal Caco-2 cells. Iron (but not zinc) bioavailability from wheat foods made using the micronized flour (2.5 ± 0.5 nmol/mg cell protein) was increased significantly compared with foods produced from standard milled flour (1.3 ± 0.1 nmol/mg cell protein; P = 0.031). Micronization of wheat flour has the potential to increase the absorption of the endogenous iron present in cereal foods and this might have health benefits for population groups with poor iron status.

Besides being an excellent source of high-quality nutrients, legumes are deposits of antinutritional factors (ANFs) that serve as natural defences against herbivory and pathogens. This investigation aimed to study the concentration of different ANFs in 13 green gram (Vigna radiata (L.) Wilczek) cultivars and explore how varietal differences affect their levels. Ten improved, and three local cultivars of green gram were evaluated for antinutritional content using different spectrophotometric assays. Results revealed variability across cultivars with 0.45 to 1.14 g/100 g of phytic acid, 1.11 to 1.99 g/100 g of saponins, 0.91 to 1.78 mg CE/g of tannins, 17.17 to 25.55 mg GAE/g of total phenolic content, 4.41 to 10.44 TUI/mg of trypsin inhibitors, 0.15 to 0.28 g/100 g of raffinose, 0.14 to 0.36 g/100 g stachyose and 1.60 to 2.88 g/100 g of verbascose. These significant variations highlight the wide range of ANF composition found among green gram cultivars. Such data could aid in exploring the nutraceutical and pharmacological potential of these compounds.

This study investigated the development of hybrid cheese analogues (HCA) made with fermented brewery side-stream ingredients (spent yeast and malt rootlets) and dairy milk. Different percentages of side-stream flours (3.5%, 5%, and 7.5%) were mixed with pasteurized milk, and the developed HCA were evaluated for their biochemical and textural properties. The addition of a fermentation step improved nutrient availability and led to pH (range 4.79–5.60) and moisture content (range 45.86%–61.29%) similar to traditional animal-based fresh cheeses (control). The inclusion of side-stream flours led to coagulation, even without rennet addition. The higher the concentration of the flour used, the faster the coagulation time, suggesting synergistic effect between the enzymes of the rennet and the enzymes present in the fermented side-stream flours. Nevertheless, textural properties were inferior compared to the control. Selected HCA formulations with added 3.5% flour exhibited increased counts of enterococci and enterobacteria cell densities, ranging from 7.28 ± 0.03 to 7.72 ± 0.09 log.CFU/g and 4.90 ± 0.16 to 5.41 ± 0.01 log CFU/g, respectively. Compared to the control sample, HCA formulations exhibited higher concentrations of organic acids, peptides, and free amino acids (FAAs). Lactic acid reached up to 23.78 ± 0.94 g/kg of dry matter (DM), while the peptide area reached up to 22918.50 ± 2370.93 mL⋅AU. Additionally, the total concentration of individual FAAs reached up to 2809.74 ± 104.85 mg/kg of DM, contrasted with the control, which resulted in lower concentrations (847.65 ± 0.02 mg/kg of DM). The overall findings suggested that despite challenges in microbiological quality and textural properties, HCA produced with the inclusion of up to 3.5% brewery side-stream flours could be a sustainable solution to produce nutritious dairy alternatives.

Edible filamentous fungi, a source of mycoprotein, are one of the sustainable alternative protein. This study compares protein digestibility (DH%) and amino acid and mineral accessibility in Rhizopus oligosporus cultivated in oat flour (OatRO) or glucose media (GluRO) by using the INFOGEST in vitro digestion protocol. Fungal total amino acids was higher in GluRO (39.0 ± 1.1 % dw) than OatRO (21.8 ± 1.3 % dw) which was also the case for calcium and magnesium content. After completed gastrointestinal digestion, there were no significant differences between GluRO and OatRO regarding DH% (27.21 ± 10.4 % and 29.4 ± 0.5 %), however, GluRO provided significantly higher amino acid accessibility compared to OatRO (64.3 ± 1.6 % and 55.1 ± 3.1 %). Mineral accessibility of GluRO was for Ca: 37.9 ± 1.8 %, Zn: 9.3 ± 0.4 %, Fe: 38.2 ± 1.9 %, Mg: 66.5 ± 1.4 % and Cu: 24.7 ± 1.3 % and for OatRO; Ca: −40.2 ± 2.4 %, Zn: −4.13 ± 0.15 %, Fe:14.6 ± 1.6 %, Mg: 74.5 ± 3.1 %, and Cu: 55.95 ± 0.8 %. Despite the low phytic acid content, OatRO thus showed antinutrient properties with respect to calcium, and zinc, suggesting that oat-derived fungi had antinutrients other than phytic acid. This study hereby revealed that the cultivation substrate affect amino acid and mineral accessibility of filamentous fungi and calls for deeper evaluations of antinutrients in oat-derived fungi.

Common beans (Phaseolus vulgaris L.) are among the most important legumes for human nutrition. The aim of the present study was to characterize the composition and in vitro bioaccessibility of tocochromanols, carotenoids, and iron from 14 different landraces and 2 commercial common bean varieties. Phytic acid, dietary fiber, and total (poly)phenolic content were determined as factors that can modify the bioaccessibility of the studied compounds. Two carotenoids were identified, namely lutein (4.6-315 ng/g) and zeaxanthin (12.2-363 ng/g), while two tocochromanols were identified, namely γ-tocopherol (2.62–18.01 µg/g), and δ-tocopherol (0.143-1.44 µg/g). The iron content in the studied samples was in the range of 58.7-144.2 µg/g. The contents of carotenoids, tocochromanols, and iron differed significantly among the studied samples but were within the ranges reported for commercial beans. After simulated gastrointestinal digestion, the average bioaccessibility of carotenoids was 30%, for tocochromanols 50%, and 17% for iron. High variability in the bioaccessible content yielded by the bean varieties was observed. Dietary fiber, phytic acid and total (poly)phenol contents were negatively correlated with the bioaccessibility of carotenoids, while iron bioaccessibility was negatively correlated with the total (poly)phenol content. The principal component analysis indicated that the bioaccessibility of lutein was the main variable involved in class separations. The composition of the food matrix plays an important role in the bioaccessibility of carotenoids, tocochromanols and iron from cooked beans.

Mango kernel flour was produced through solid-state fermentation (SSF) by Aspergillus oryzae and Aspergillus awamori for 48 and 96 h at 30°C, followed by a washing step to remove simple carbohydrates (water-soluble fraction, MKWF). The results show that SSF by A. oryzae reduces phytic acid content by more than 60% after only 48 h and increases the protein content in the flour by more than 55% after 96 h. SSF by A. awamori and A. oryzae increases fat content by more than 49 and 25%, respectively. The free sugars recovered in the MKWF are useable by L. plantarum, which achieved at least one log₁₀ CFU/mL of growth. The MKWF also contains a significant amount of malto-oligosaccharides which might be of interest for its prebiotic use. Mango seed kernel can be upcycled through SSF by Aspergillus spp into a functional flour and an MKWF suitable for Lactobacilli biomass production.

Plant protein ingredients are gaining attention for human nutrition, yet they differ significantly from animal proteins in functionality and nutrition. Fungal solid-state fermentation (SSF) can modulate the composition and functionality, increasing their applicability in foods. Quinoa flour (QF) served as a substrate for Aspergillus oryzae and Rhizopus oligosporus, resulting in two fermented ingredients (QFA and QFR) with different nutritional, functional, and aroma characteristics. A higher increase in protein (+35%) and nitrogen (+24%) was observed in the QFA, while fat was predominantly increased in the QFR (+78%). Fermentable oligo-, di-, monosaccharides and polyols (FODMAPs) decreased in the QFR but increased in the QFA due to polyol production. Metabolomic analysis revealed higher lactic acid concentrations in the QFA, and higher citric, malic, and fumaric acid contents in the QFR. The SSF reduced most antinutrients, while R. oligosporus produced saponins. Olfactometry showed the development of fruity ester compounds and a decrease in metallic and cardboard aromas. Both ingredients showed an enhanced water-holding capacity, with the QFA also demonstrating an increased oil-holding capacity. Complex formation increased the particle size, reduced the solubility, and decreased the foaming properties. Mycelium production darkened the ingredients, with the QFR having a higher differential colour index. This study highlights the potential of SSF to produce ingredients with improved nutritional, sensory, and functional properties.

Adzuki bean (Vigna angularis) and mung bean (Vigna radiata) were investigated for their potential as plant-based milk alternatives. Their nutritional, anti-nutrient, stability, and organoleptic characteristics were compared with commercial soymilk and bovine milk. Results showed that both adzuki bean milk (ABM) and mung bean milk (MBM) demonstrated lower protein (ABM: 2.19 ± 0.10 %; MBM: 1.63 ± 0.05 %) and carbohydrate contents compared to soymilk. Additionally, they exhibited magnesium contents similar to bovine milk and iron levels comparable to both soymilk and bovine milk. While total amino acid content was lower in ABM and MBM, their phytic acid and purine contents were significantly lower than in soymilk. Fewer volatile compounds associated with beany flavours were also present in ABM and MBM, and ABM emerged as a palatable choice from the sensory analysis. These findings suggest that ABM and MBM have the potential to offer diverse and palatable plant-based milk options for consumers.