Analytical methodology optimization to estimate the content of non-flavonoid phenolic compounds in Argentine propolis extracts.

CONTEXT: Traditionally, the content of total phenolics (flavonoid phenolics (FP) and non-flavonoid phenolics (NFP)) and flavonoids (flavone/flavonol and flavonone/dihydroflavonol) in propolis has been determined by different methodologies. Until now, the percentage of total phenolic (TP) compounds that corresponds to FP and NFP, expressed in the same units by a spectrophotometric method, has not been determined. OBJECTIVE: The current study proposes a quick and simple methodology that separates FP and NFP in propolis samples and determines TP, FP, and NFP by the same method. MATERIALS AND METHODS: Propolis samples from five Argentine provinces (Tucumán, Santiago del Estero, Salta, Misiones, and Jujuy) were used. Extraction of TP from the propolis samples was carried out by maceration with 80% ethanol and quantified by Folin-Ciocalteu reagent (FC-R). Then, FP was precipitated with formaldehyde in acid medium. After centrifugation, NFP were determined in the supernatant using FC-R. FP content was calculated as the difference between the content of TP and NFP. The method was also validated using commercial flavonoids and chalcones. RESULTS: FP recovery in all experiments was between 85.95% and 98.29%. Propolis from Tucumán had significantly higher amounts of total phenols than propolis from other provinces. SE5 showed higher content of FP (81.52%) followed by SA1 (74.75%). The propolis from TUC4, SA4, SE3, and MI showed the lowest FP content and highest content of NFP. CONCLUSIONS: This method provides a simple, reliable, and specific spectrophotometric assay to estimate the content of NFP, FP, and TP in propolis samples.

Anti-inflammatory and antioxidant activities, functional properties and mutagenicity studies of protein and protein hydrolysate obtained from Prosopis alba seed flour.

Prosopis species are considered multipurpose trees and shrubs by FAO and their fruit constitute a food source for humans and animals. According to the "Código Alimentario Argentino", "algarrobo flour" is produced by grinding the whole mature pod, but in the traditional process most of the seeds are discarded. In this paper, the flour from seed was obtained. Then, the proteins were extracted and enzymatic hydrolysis was carried out. According to their amino acid profile and chemical score (>100%), the Prosopis alba proteins, are not deficient in essential amino acids considering the amount of amino acid necessary by adults. The protein isolate showed a good solubility (pH 7.4-9), emulsificant capacity, oil binding capacity and water adsorption capacity. The antioxidant ability of proteins was significantly increased with hydrolysis (SC50 values: 50-5µg/mL, respectively). Inhibitory activity of pro-inflammatory enzymes (lipoxygenase and phospholipase) was described. The mutagenicity/antimutagenicity of proteins and protein hydrolysates from seed flour were also analysed. The results suggest that P. alba cotyledon flour could be a new alternative in the formulation of functional foods not only for its high protein content but also by the biological and functional properties of its proteins and protein hydrolysates.

Polyphenolic compounds and anthocyanin content of Prosopis nigra and Prosopis alba pods flour and their antioxidant and anti-inflammatory capacities.

The aim of this study was to determine the content of total free and bound phenolics, free and bound flavonoids, anthocyanins, and alkaloids and the profile of polyphenols of edible ripe pods of Prosopis alba and Prosopis nigra. P. alba flour showed significantly higher total (sum of Free- and Bound) phenolic content and total flavonoid compounds than P. nigra (p<0.05) while P. nigra had higher concentrations of anthocyanins than P. alba (p<0.05). The P. nigra flour shows a pattern characterized by the occurrence of anthocyanins as well as 14 flavonoid glycosides, with higher chemical diversity than P. alba, which shows 8 flavonoid glycosides as relevant constituents. The main compounds were quercetin O-glycosides and apigenin-based C-glycosides. The phenolic composition of two South American algarrobo pod flour is presented for the first time. P. nigra pods having higher content of anthocyanins are darker (purple) than those of P. alba (light brown). Furthermore, the sugar-free polyphenolic extracts of P. nigra and P. alba as well as anthocyanins enriched extracts from P. nigra showed antioxidant activity. P. nigra and P. alba polyphenolic extracts showed activity against a pro-inflammatory enzyme. In conclusion, algarrobo pods meal contained biologically active polyphenols, with a positive impact on human health.

Radical scavenging capacity and antimutagenic properties of purified proteins from Solanum betaceum fruits and Solanum tuberosum tubers.

In this study, antioxidant activities in free-radical-mediated oxidative systems and the genotoxic/antigenotoxic effects of two proteins with molecular mass around 17 kDa, purified from Solanum betaceum fruits (cyphomine) and Solanum tuberosum tubers (solamarine), were investigated. Both proteins inhibited uric acid formation with IC(50) values between 55 and 60 µg/mL, and both proteins were able to reduce oxidative damage by scavenging hydroxyl radicals and superoxide anion in a dose-dependent manner. Furthermore, the DPPH• reduction assay showed SC(50) values of 55-73 µg/mL. Cyphomine and solamarine were able to retain their antioxidant activity after heat treatment at 80 °C for 15 min. Allium cepa and Salmonella /microsome assays showed no genotoxic and mutagenic effects. Solamarine showed an antimutagenic effect against a direct mutagen (4-nitro-o-phenylenediamine). Consequently, the present study showed that the investigated proteins are promising ingredients for the development of functional foods with a beneficial impact on human health and an important source for the production of bioactive peptides.

Autographic assay for the rapid detection of antioxidant capacity of liquid and semi-solid pharmaceutical formulations using ABTS•+ immobilized by gel entrapment.

An autographic assay suitable for the detection of antioxidant compounds in a complex matrix (liquid and semi-solid pharmaceutical formulations) or in isolated compounds was described. The pre-formed radical monocation of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(•+)) was generated by oxidation of ABTS with potassium persulfate and reduced in the presence of hydrogen-donating antioxidants. For a further comparative estimation of its applicability and sensitivity, different medicinal plant extracts, hydrogels and antioxidant compounds were dot seeded or chromatographed on silica gel (TLC) and revealed with ABTS(•+) solution (System I) or ABTS(•+) immobilized by gel entrapment (System II). Both systems were effective and able to detect antioxidant activity in a micromolar range in seconds. System II was more sensitive and reproducible than System I. This micromethod is quick, inexpensive, and particularly helpful whether it works with numerous samples or on a small scale.

Inhibition of hydrolytic enzyme activities and plant pathogen growth by invertase inhibitors.

The invertase inhibitory protein isolated from Cyphomandra betacea Sendt and Solanum tuberosum inhibited the invertase activity from different species, genera and even plant family. Furthermore, proteinaceous inhibitors are not invertase specific; fungal, bacterial and higher plant enzymes including polygalacturonase, pectinase, pectin lyase, alpha-L-arabinofuranosidase and beta-glucosidase are also shown to be inhibited. Both inhibitors exhibited an in vitro antibacterial action against phytopathogenics strains of Xanthomonas campestris pvar vesicatoria CECT 792, Pseudomonas solanacearum CECT 125, Pseudomonas corrugata CECT 124, Pseudomonas syringae and Erwinia carotovora var carotovora.