Michael R.A. Morgan

Dietary flavonoid and isoflavone glycosides are hydrolysed by the lactase site of lactase phlorizin hydrolase

Lactase phlorizin hydrolase (LPH; EC 3.2.1.62) is a membrane‐bound, family 1 β‐glycosidase found on the brush border of the mammalian small intestine. LPH, purified from sheep small intestine, was capable of hydrolysing a range of flavonol and isoflavone glycosides. The catalytic efficiency (k cat/K m) for the hydrolysis of quercetin‐4′‐glucoside, quercetin‐3‐glucoside, genistein‐7‐glucoside and daidzein‐7‐glucoside was 170, 137, 77 and 14 (mM−1 s−1) respectively. The majority of the activity occurred at the lactase and not phlorizin hydrolase site. The ability of LPH to deglycosylate dietary (iso)flavonoid glycosides suggests a possible role for this enzyme in the metabolism of these biologically active compounds.

Human metabolism of dietary flavonoids: Identification of plasma metabolites of quercetin

The position of conjugation of the flavonoid quercetin dramatically affects biological activity in vitro, therefore it is important to determine the exact nature of the plasma metabolites. In the present study, we have used various methods (HPLC with diode array detection, LCMS, chemical and enzymic synthesis of authentic conjugates and specific enzymic hydrolysis) to show that quercetin glucosides are not present in plasma of human subjects 1.5 h after consumption of onions (a rich source of flavonoid glucosides). All four individuals had similar qualitative profiles of metabolites. The major circulating compounds in the plasma after 1.5 h are identified as quercetin-3-glucuronide, 3′-methyl-quercetin-3-glucuronide and quercetin-3′-sulfate. The existence of substitutions in the B and/or C ring of plasma quercetin metabolites suggests that these conjugates will each have very different biological activities.

Particle-Stabilizing Effects of Flavonoids at the Oil―Water Interface

It has been shown that some common food flavonoids can act as excellent stabilizers of oil-in-water emulsions through their adsorption as water-insoluble particles to the surface of the oil droplets, i.e., Pickering emulsions are formed. Flavonoids covering a wide range of octanol-water partition coefficients (P) were screened for emulsification behavior by low shear mixing of flavonoid+n-tetradecane in a vortex mixer. Most flavonoids with very high or very low P values were not good emulsifiers, although there were exceptions, such as tiliroside, which is very insoluble in water. When a high shear jet homogenizer was used with 20 vol% oil in the presence of 1 mM tiliroside, rutin, or naringin, much finer emulsions were produced: the average droplet sizes (d32) were 16, 6, and 5 μm, respectively. These results may be highly significant with respect to the delivery of such insoluble compounds to the gut, as well as their digestion and absorption.

Effects of pH on the ability of flavonoids to act as Pickering emulsion stabilizers

The flavonoids tiliroside, rutin and naringin have been investigated as stabilizers of Pickering oil-in-water (O/W) emulsions. The mean droplet size of tetradecane emulsions was considerably smaller at higher pH, especially for rutin. The solubility of flavonoids in the aqueous phase was 4-6 times higher at pH 8 compared to pH 2 for tiliroside and rutin, although all absolute solubilities remained low (<1 mM). This agreed with a slight increase in surface activity of tiliroside and rutin at the O-W interface at pH 8 compared to pH 2. However, improved emulsion stabilization at higher pH is better explained by the significant increase in ζ-potential of the flavonoid particles to more negative values at pH 8, which will improve particle dispersion and increase the charge on the droplets stabilized by them. A buckwheat tea extract, rich in rutin, was also shown to be an effective stabilizer of sunflower O/W emulsions.

Comparative chemical and biochemical analysis of extracts of Hibiscus sabdariffa

Hibiscus sabdariffa extracts have attracted attention because of potentially useful bioactivity. However, there have been no systematic studies of extraction efficiencies of H. sabdariffa. The nature of extracts used in different studies has varied considerably, making comparisons difficult. Therefore, a systematic study of extracts of H. sabdariffa made with different solvents was carried out using water, methanol, ethyl acetate and hexane in the presence/absence of formic acid, using different extraction times and temperatures. The extracts were analysed for total polyphenol content, antioxidant capacity using DPPH, FRAP and TEAC assays, and specific anthocyanins were determined using HPLC and LC-MS. The results showed the highest antioxidant capacities were obtained by extracting using water, with or without formic acid, for 10 min at 100°C. These extracts provided the highest concentrations of cyanidin 3-sambubioside and delphinidin 3-sambubioside. It will be important to use extraction conditions giving optimal extraction efficiencies for subsequent bioactivity experiments.

Determination of amygdalin in apple seeds, fresh apples and processed apple juices

Cyanogenic glycosides are natural plant toxicants. Action by endogenous plant enzymes can release hydrogen cyanide causing potential toxicity issues for animals including humans. We have quantified amygdalin in seeds from different apple varieties, determined the effects of processing on the amygdalin content of apple juice and quantified amygdalin in commercially-available apple juices. Amygdalin contents of seeds from fifteen varieties of apples ranged from 1 mg g(-1) to 4 mg g(-1). The amygdalin content of commercially-available apple juice was low, ranging from 0.01 to 0.04 mg ml(-1) for pressed apple juice and 0.001-0.007 mg ml(-1) for long-life apple juice. Processing led to juice with low amygdalin content, ranging from 0.01 mg ml(-1) to 0.08 mg ml(-1). The results presented show that the amygdalin contents of commercially-available apple juices are unlikely to present health problems to consumers.

Transport of trans-tiliroside (kaempferol-3-β-D-(6"-p-coumaroyl-glucopyranoside) and related flavonoids across Caco-2 cells, as a model of absorption and metabolism in the small intestine.

Abstract 1. Absorption and metabolism of tiliroside (kaempferol 3-β-D-(6″-p-coumaroyl)-glucopyranoside) and its related compounds kaempferol, kaempferol-3-glucoside and p-coumaric acid were investigated in the small intestinal Caco-2 cell model. Apparent permeation (Papp) was determined as 0.62 × 10−6 cm/s, 3.1 × 10−6 cm/s, 0 and 22.8 × 10−6 cm/s, respectively. 2. Mechanistic study showed that the transportation of tiliroside, kaempferol-3-glucoside and p-coumaric acid in Caco-2 model were transporter(s) involved, while transportation of kaempferol was solely by passive diffusion mechanism. 3. Efflux transporters, multi-drug-resistance-associated protein-2 (MRP2), were shown to play a role in limiting the uptake of tiliroside. Inhibitors of MRP2, (MK571 and rifampicin) and co-incubation with kaempferol (10 μM), increased transfer from the apical to the basolateral side by three to five fold. 4. Metabolites of kaempferol-3-glucoside and p-coumaric acid were not detected in the current Caco-2 model, while tiliroside was metabolised to a limited extent, with two tiliroside mono-glucuronides identified; and kaempferol was metabolised to a higher extent, with three mono-glucuronides and two mono-sulfates identified. 5. In conclusion, tiliroside was metabolised and transported across Caco-2 cell membrane to a limited extent. Transportation could be increased by applying MRP2 inhibitors or co-incubation with kaempferol. It is proposed that tiliroside can be absorbed by human; future pharmacokinetics studies are warranted in order to determine the usefulness of tiliroside as a bioactive agent.

IgE binding to proteins from sesame and assessment of allergenicity: implications for biotechnology?

Successful prediction of the potential allergenicity of a protein may be a key factor in the development of novel, genetically modified foods. The use of the decision tree approach for the prediction of allergenicity is discussed. The methods currently used for identifying allergenic proteins (including use of IgE from patient sera for recognition of proteins) are reviewed. Finally, a specific review of the literature concerning identification of allergens from sesame leads to the conclusion that in the absence of validated animal models, identification of allergenicity (and, consequently, prediction of allergenicity) may be problematic.

Effect of acid pretreatment and the germination period on the composition and antioxidant activity of rice bean (Vigna umbellata)

This research evaluated effect of germination period and acid pretreatment on chemical composition and antioxidant activity of rice bean sprouts. Moisture, total phenolics, reducing sugar and B vitamins (thiamine, riboflavin, and niacin) content of steamed sprouts increased with increasing germination time (p⩽0.05). Pretreatment with 1% (w/v) citric acid for 6h significantly increased the total phenolic content. The 18-h-germinated rice beans showed the highest crude protein content, as determined using the Kjeldahl method. During germination, acid pretreatment led to a significant decrease in the intensity of the 76-kDa band. Germination caused a significant increase in radical scavenging activity and ferric reducing antioxidant power, especially in sprouts from citric acid-treated seeds. The antioxidant activities of the ethanolic extracts from both pretreated beans and the control were 1.3-1.6 times higher than those obtained from the water extracts. Major phenolics found in both 0-h and 18-h-germinated rice beans were catechin and rutin.