Physicochemical, in vitro digestibility and functional properties of carboxymethyl rice starch cross-linked with epichlorohydrin

Cross-linked carboxymethyl rice starches (CL-CMRSs) were prepared from reactions between native rice starch and varied concentrations (0.1–15% w/w, M-0.1 to M-15) of epichlorohydrin (ECH) in a simultaneous carboxymethylation-crosslinking reaction setup using methanol as the solvent. The degree of carboxymethyl substitution was between 0.24 and 0.28, while apparent amylose contents were lowered due to modification. SEM images showed minor change on the granule surface, while XRD profiles indicated slight loss of crystallinity. DSC thermograms revealed no transition peak in all treated samples. The water uptake (WU), swelling volume (SV) and free swelling capacity (FSC) of CL-CMRSs increased significantly as a result of the modification, while swelling of CMRSs cross-linked with 2% (M-2) and 3% (M-3) ECH yielded FSC values and WU values that were much greater than those of native starches and were comparable to that of Explotab®. All modified starch samples showed increased amount of rapidly digestible starch (RDS), while cross-linking with 5–7.5% ECH raised the resistant starch (RS) content, compared to native starch. M-2 also showed promising results in tablet disintegration test. ECH–CL–CMRSs could potentially be used as an excipient in pharmaceutical and food/food supplement products.     

Physicochemical and nutritional properties of reduced-caloric density high-fibre breads

The influence of some physicochemical properties (chemical and nutritional composition, particle size, colour, dynamic viscosity, viscoelastic moduli) of associated binary mixtures of structuring hydrocolloids (cellulose derivatives, galactomannans) and prebiotic oligosaccharides (fructo- and gluco-oligosaccharides) in diluted hydrated wheat flour matrixes is discussed in terms of nutritional (higher resistant starch, lower digestible starch and lower in vitro Glycaemic Index) and technological functional profiles (higher sensory scores and longer keepability) of reduced-caloric density (−20%) high-fibre (>6 g/100 g) breads. Few technological functional (sensory firmness and overall acceptability) and most nutritional bread properties (protein digestibility, rapidly digestible starch, slowly digestible starch, in vitro expected Glycaemic Index, total digestible starch and resistant starch) were found to depend on dietary fibre molecular characteristics (mean particle diameter (D[4, 3]), storage modulus (G′), loss modulus (G″), complex viscosity (η*), and lactic acid solvent retention capacity. Dietary fibres with larger particle size resulted in highly sensory acceptable breads with higher amounts of resistant starch and slightly lower protein digestibility. Fibres exhibiting high viscoelasticity -G′, G″- and complex viscosity -η*- in concentrated solutions yielded breads with better sensory perception, lower digestible starch and higher resistant starch contents bringing to lower in vitro expected Glycaemic Index.     

Kiwifruit remnants from digestion in vitro have functional attributes of potential importance to health

The physicochemical properties of plant cell walls in the gut are important in modulating processes that influence health. We investigated the physicochemical properties of kiwifruit cell walls digested under gastric and gastroileal conditions in vitro. Soluble and insoluble undigested polymer fractions were measured, the hydration properties of the digested pulp, and the capacity to retard diffusion and mixing in a simulated small intestinal segment. Undigested polymer (dietary fibre) fractions differed little between “Hayward’ (‘Hayward’) and ‘Hort16A’ (gold) kiwifruit cultivars in their relative proportions, although total dietary fibre was greater in ‘Hayward’ than in the ‘Hort16A’. The polysaccharide composition of seed-free digestion-resistant polymer was similar in both cultivars and not affected by in vitro digestion. Indigestible remnants from kiwifruit had strong water retention and swelling capacities, also little affected by digesting, and retarded both glucose diffusion and mixing significantly, especially in the presence of low background viscosity. We conclude that the particulate cell wall remnants of digested kiwifruit retain substantial potential to influence the properties of gut contents.     

Lutein-enriched frankfurter-type products: Physicochemical characteristics and lutein in vitro bioaccessibility

This paper reports a study of the physicochemical characteristics of lutein-enriched frankfurter-type products with two fat levels, and the lutein in vitro bioaccessibility in these products. In general, the high-fat products possessed better emulsion stability and greater hardness and chewiness than low-fat samples. For each fat level, the addition of lutein extract in olive oil reduced emulsion stability in meat products. The presence of lutein reduced the lightness and increased the redness of samples. Lutein was highly stable upon in vitro digestion, with overall recovery of over 84% at the end of the duodenal phase and very low isomerisation. Micellarisation was high but depended on the fat content, ranging from 29–34% (for the low-fat sample) up to 73–81% of the amount initially present (for the high-fat sample). Storage (22 days at 4 °C) did not significantly affect lutein content or bioaccessibility. Our results support the suitability of meat products as lutein carriers and as a means to increase the systematic intake of lutein.     

The in vitro and in vivo antioxidant properties of seabuckthorn (Hippophae rhamnoides L.) seed oil

The antioxidant capacity of seabuckthorn (Hippophae rhamnoides L.) seed oil was investigated with a number of established in vitro assays and in an in vivo study of carbon tetrachloride (CCl₄)-induced oxidative stress in mice. The results showed that DPPH radical scavenging activity, ferrous ion chelating activity, reducing power and inhibition of lipid peroxidation activity all increased with increasing concentrations of seabuckthorn seed oil. Moreover, the EC₅₀ values of seabuckthorn seed oil from the hydrogen peroxide, superoxide radical, hydroxyl radical scavenging assays were 2.63, 2.16 and 0.77 mg/ml, respectively. In the in vivo study, seabuckthorn seed oil inhibited the toxicity of CCl₄, as seen from the significantly increased activities of the antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. The GSH content in the liver was also increased, whereas hepatic malondialdehyde was reduced. Taken together, these results clearly indicate that seabuckthorn seed oil has significant potential as a natural antioxidant agent.     

Isolation, solubility and in vitro hydrolysis of chickpea vicilin-like protein

The chickpea vicilin-like globulin was isolated and chromatographed on Sepharose CL-6B and Sephacryl S-300. The native globulin with a molecular weight of 140 kDa was resolved in Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in seven polypeptide bands in the range of 12.4-67 kDa. The solubility profile of the protein in water and NaCl solutions was typical of a legume globulin. The purified vicilin-like globulin, native and heated, was hydrolyzed by pepsin, trypsin and chymotrypsin. The hydrolysis patterns indicated that the native vicilin-like protein was only partially degraded by the enzymes in comparison with casein. Heating increased its susceptibility to hydrolysis relative to the native form, for all the enzymes. However, the results obtained by the pH-drop method revealed that the in vitro digestibility of the vicilin-like protein was not altered by heating, while 11S-like and total globulins suffered a small increase, indicating that the structural characteristics of storage globulins may be important factors limiting the protein digestion.     

Impact of inulin and okara on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product and probiotic survival under in vitro simulated gastrointestinal conditions

The effect of inulin and/or okara flour on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product (FSP) and on probiotic survival under in vitro simulated gastrointestinal conditions were investigated throughout 28 days of storage at 4 °C. Employing a 2² design, four FSP trials were produced from soymilk fermented with ABT-4 culture (La-5, Bb-12, and Streptococcus thermophilus): FSP (control); FSP-I (with inulin, 3 g/100 mL of soymilk); FSP-O (with okara, 5 g/100 mL); FSP-IO (with inulin + okara, ratio 3:5 g/100 mL). Probiotic viabilities ranged from 8 to 9 log cfu/g during the 28 days of storage, and inulin and/or okara flour did not affect the viability of La-5 and Bb-12. Bb-12 resistance to the artificial gastrointestinal juices was higher than for La-5, since the Bb-12 and La-5 populations decreased approximately 0.6 log cfu/g and 3.8 log cfu/g, respectively, throughout storage period. Even though the protective effect of inulin and/or okara flour on probiotic microorganisms was not significant, when compared to a fresh culture, the FSP matrix improved Bb-12 survival on day 1 of storage and may be considered a good vehicle for Bb-12 and could play an important role in probiotic protection against gastrointestinal juices.     

Phenolic profiles of in vivo and in vitro grown Coriandrum sativum L.

Coriandrum sativum L. is a source of a variety of polyphenols and other phytochemicals, related to its high antioxidant activity and to its use for indigestion, rheumatism, and prevention of lipid peroxidation damage. Plant cell cultures are a means to study or to produce some active metabolites, such as polyphenols. This technique was applied to the investigation of coriander, and a detailed analysis of individual polyphenols in vivo and in vitro grown samples was performed. The in vivo vegetative parts showed quercetin derivatives as the main flavonoids and quercetin-3-O-rutinoside (3296 mg/kg dw) was the main polyphenol found in this part of coriander. The fruits revealed only phenolic acids and derivatives, caffeoyl N-tryptophan hexoside (45.33 mg/kg dw) being the most abundant phenolic derivative. In vitro samples also gave a high diversity of polyphenols, being C-glycosylated apigenin (2983 mg/kg dw) the main compound. Anthocyanins were only found in clone A, which was certainly related to its purple pigmentation, and peonidin-3-O-feruloylglucoside-5-O-glucoside was the major anthocyanin found (1.70 μg/kg dw). In vitro culture can be used to explore new industrial, pharmaceutical, and medicinal potentialities, such as the production of secondary metabolites like flavonoids.     

In vitro and in vivo protein hydrolysis of beans (Phaseolus vulgaris) genetically modified to express different phaseolin types

Experiments were conducted to study whether phaseolin type could influence proteolysis susceptibility and nutritional value of total bean protein. The DOR-390 bean cultivar was genetically modified to express different phaseolin types (S, T or I). Beans were soaked and autoclaved. A sequential hydrolysis was carried out in vitro with pepsin and pancreatin. Differences in the degree of protein hydrolysis among bean lines started at 30 min and remained until 240 min, with the S bean proteins presenting lower values (P < 0.05). Subsequently, rats were fed with diets containing beans expressing different phaseolin types as the only source of protein for N digestibility and nutritional value determination. No differences (P > 0.05) in ileal protein digestibility and rat growth were observed. In conclusion, the differences in in vitro hydrolysis between bean lines expressing different phaseolin types had no consequences on growth and N retention in rats.     

Physicochemical properties of a fibrous fraction from chia (Salvia hispanica L.)

An evaluation was done of some physicochemical properties of a fiber-rich fraction (FRF) obtained by dry processing of defatted chia (Salvia hispanica) flour. The fiber-rich fraction (FRF) had 29.56 g/100 g crude fiber content and 56.46 g/100 g total dietary fiber (TDF) content, of which 53.45 g/100 g was insoluble dietary fiber (IDF) and 3.01 g/100 g was soluble dietary fiber (SDF). The FRF water-holding capacity was 15.41 g/g, its water absorption capacity 11.73 g/g, and its organic molecule absorption capacity 1.09 g/g. The FRF also had low oil-holding (2.02 g/g) and water adsorption (0.3 g/g) capacities. Emulsifying activity in this fraction was 53.26% and emulsion stability was 94.84%. Its evaluated antioxidant activity was 488.8 μmol/L Trolox equivalents/g FRF, which is higher than for many cereals and similar to drinks such as wine, tea, coffee and orange juice. The chia FRF values for the evaluated properties, particularly for water-holding, oil-holding and organic molecule absorption capacity, suggest it could be a useful ingredient in dietetic products such as baked and fried foods, among others.     

Effects of simulated digestion in vitro on cell wall polysaccharides from kiwifruit (Actinidia spp.)

Cell wall polysaccharides are resistant to digestion and absorption in the human small intestine and are considered to be delivered to the colon in a chemically unaltered state. In this paper, pulp from green and gold kiwifruit was subjected to in vitro upper-intestinal tract digestion and the chemical and physical changes to cell wall polysaccharides (dietary fibre) were investigated. Yields of insoluble fibre decreased slightly with simulated digestion while soluble fibre yields increased. Constituent sugar and glycosyl linkage analysis of the soluble and insoluble fibre fractions revealed that the chemical composition and structure of the non-starch polysaccharides remained largely unchanged. However, the degree of methylesterification of galacturonic acid residues present in the pectin-rich soluble fibre fractions of both fruit decreased with treatment; size-exclusion chromatography detected changes in the molecular weight profiles of these fractions. These changes may affect the physicochemical properties and fermentability of kiwifruit dietary fibre in the large intestine.     

In vitro fermentation of 12 dietary fibres by faecal inoculum from pigs and humans

In vitro fermentation of 12 dietary fibres by faecal inocula from pigs and humans were performed. The fibres included homoglucans, mannans, fructans, polyuronides, and complex heteroglycans. Gas production, short chain fatty acid production and fibre degradation products were monitored during fermentation. Human inoculum has more ability to ferment resistant starch and fibres containing uronic acids. In contrast, pig inoculum is able to ferment cellulose, which is hardly fermented by human inoculum. The sugar and linkage composition of the fibres has an important influence on fibre fermentation patterns. Fibres containing uronic acids induced the production of acetate, whereas fibres containing neutral sugars induced the production of propionate or butyrate. Fermentation of the fructans showed that molecular size could be an influential factor, and fermentation of complex heteroglycans showed that the arrangement of sugars in the molecules may also affect the fermentation patterns. This experiment also shows that monitoring of fibre degradation products is important for understanding how fibres are degraded during fermentation.     

In vitro model to correlate viscosity and bile acid-binding capacity of digested water-soluble and insoluble dietary fibres

Binding and excretion of bile acids in the small intestine by water-soluble and insoluble dietary fibres is one of the main mechanisms for their cholesterol-lowering effects. A model for the determination of the bile acid-binding capacity of dietary fibres was developed. The experimental set-up allowed to correlate the bile acid-binding capacities of different fibres with their viscosities after in vitro digestion. For cellulose, native oat fibre and psyllium fibres clear correlations between viscosity and bile acid-binding capacity could be observed, whereas for water-insoluble lupin fibre such a correlation did not exist. Heat-damaged oat fibre also showed bile acid-binding despite of significantly decreased viscosity. The data demonstrated that bile acid-binding of digested dietary fibres may not be solely based on their viscosity but may be influenced by additional binding forces.     

Predicting relative concentrations of bioavailable iron in foods using in vitro digestion: New developments

In vitro methods have been developed for the prediction of iron bioavailability from foods and supplements. The dialyzability method measures dialyzable iron, released during a simulated gastrointestinal digestion, as an index of iron bioavailability. A new setup, that involves six-well plates and a ring insert that holds the dialysis membrane, is proposed for the application of the dialyzability method with the objective to increase efficiency and to allow testing small-volume samples. A series of solutions (water, ascorbic acid, and phytate), liquid foods (fresh milk and condensed milk), and solid foods (bread + meat meal, corn flakes), were tested in the presence or absence of added iron and digested with the new setup and the setup previously described for the dialyzability method. In both cases, percent dialyzable iron in each treatment remained similar (P > 0.05). These results suggest that the new setup can be employed in future applications with similar food matrices of the dialyzability method.     

Antioxidant capacity of four polyphenol-rich Amazonian plant extracts: A correlation study using chemical and biological in vitro assays

Many plants used in Amazonian folk medicine present a high antioxidant activity. In this study, the antioxidant activities of four largely used plants, namely Byrsonima crassifolia, Davilla kunthii, Davilla rugosa and Inga edulis, were evaluated, using methanolic extracts of their leaves, fruits and bark and several different in vitro tests, based either on the capacity to scavenge free radicals (ORAC, TRAP) or on the ability to protect biological structures (LDLs, erythrocytes). The total phenolics (TP), flavanoids (TFA) and flavonols (TFO) were also measured. Almost all extracts performed well in all assays of antioxidative capacity, with best activities found in leaves (compared to fruits and bark). Most antioxidative performance indicators (ORAC, TRAP, LDL protection) correlated well with the TP and TFA content of the extracts. Conversely, correlation was lower between TFO and these indicators, reflecting a lower involvement of these compounds in antioxidant processes. Erythrocyte protection against oxidant-triggered haemolysis showed no correlation with any of the phenolic content indicators, suggesting that most of these compounds have a low ability to protect lipid targets in the erythrocyte membrane. On the other hand, protection of erythrocytes against haemolysis correlated positively with LDL protection. The extract of I. edulis leaves contained average amounts of polyphenols but ranked first in the majority of the tests, indicating the occurrence of particularly efficient compounds with very important antioxidant properties, which could be used for medicinal and other applications.     

Effect of citric acid deamidation on in vitro digestibility and antioxidant properties of wheat gluten

The effects of citric acid deamidation on the physiochemical properties of wheat gluten were investigated. In vitro digestion was carried out to determine changes of molecular weight distribution, amino acids composition and antioxidant efficacy of wheat gluten hydrolysates. Results indicated that citric acid deamidation significantly increased gluten solubility and surface hydrophobicity, at a neutral pH. Deamidation induced molecular weight distribution change of gluten with little proteolysis. Results from FTIR indicated that the α-helix and β-turn of deamidated gluten increased accompanied by a decrease of the β-sheet structure. After deamidation, in vitro pepsin digestibility of wheat gluten decreased, while in vitro pancreatin digestibility increased. The oxygen radical absorbance capacity (ORAC) activity of the in vitro digests decreased with increase of deamidation time. The high Lys and total essential AAs amounts in the final digests suggested that the nutritional values of wheat gluten after deamidation might be enhanced.     

Effect of ultrasound treatments of tomato pulp on microstructure and lycopene in vitro bioaccessibility

The influence of ultrasound treatments of tomato pulp on microstructure and lycopene in vitro bioaccessibility was investigated. To this purpose, samples were subjected to ultrasound at a frequency and amplitude of 24 kHz and 100 μm, respectively, for increasing lengths of time. Results showed that ultrasound was responsible for loss of tomato cell integrity, as well as a decrease in the degree of pectin esterification. In contrast, rheological measurements showed that ultrasonically treated tomato pulp had greater gel-like properties than an untreated sample. It was inferred that ultrasound promoted the formation of a new network due to hydrogen bonding and hydrophobic interactions among the de-esterified pectin molecules. Such a reinforcement of the tomato pulp structure resulted in a decrease in lycopene in vitro bioaccessibility of the ultrasonically treated tomato pulp, probably due to the fact that the presence of a stronger network may make lycopene less available to the digestion process.     

Thermal aggregation,amino acid composition and in vitro digestibility of vicilin-rich protein isolates from three Phaseolus legumes: A comparative study

The heat-induced aggregation and the in vitro digestibility of vicilin-rich protein isolates from three Phaseolus legumes, including kidney bean, red bean and mung bean were investigated and compared, and their amino acid composition and free sulfhydryl (SH) group contents also evaluated. The results showed that the extent in the heat-induced aggregation varied with the type of the protein isolates, and the formation of new disulphide bonds (at the expense of free SH contents) was involved in the formation of the aggregates. The protein isolates with higher levels of hydrophobic and uncharged polar amino acids, and lower basic amino acid contents exhibited lower extent of their heat-induced aggregation. The in vitro pepsin plus trypsin digestibility was different for various native protein isolates. The digestibility was to a varying extent affected by the heat treatment. The influences of heating on the digestibility of these proteins were closely related to the extent of their heat-induced aggregation. The results suggest that the improvement of nutritional property of those vicilin-rich protein isolates by heat treatment is largely dependent upon their amino acid composition as well as the extent of heat-induced aggregation.     

Indigestible fraction of okara from soybean: composition,physicochemical properties and in vitro fermentability by pure cultures of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> and <Emphasis Type="Italic">Bifidobacterium bifidum</Emphasis>

Major non-digestible components of soybean seeds and okara were determined by an in vitro enzymatic-physiological method, alternative to dietary fibre. Total indigestible fraction was higher in okara (41.6%) than in seeds (28.5%), and consisted of soluble and insoluble fractions, mainly composed of non-starch polysaccharides, klason lignin and resistant protein. Total protein was lower in okara (32.29%) than in seeds (46.97%), as were oil (14.72% okara–20.89% seeds) and ash contents (3.18% okara–4.60% seeds). In vitro digestibility of protein was lower for okara (84.3%), than for soybean seed (91.8%). Moreover, okara showed high swelling (10.54 ± 0.14 mL/g d.w.) and water retention capacity (8.87 ± 0.06 g/g d.w.) and was fermented in vitro to a greater extent by Bifidobacterium bifidum (29.8%), than by Lactobacillus acidophilus (8.3%). For its composition, physico-chemical properties and bifidogenic capacity in vitro, okara is a potential candidate to be a prebiotic fibre-rich ingredient of new functional foods. An erratum to this article can be found at