Anti-elastase and anti-hyaluronidase of phenolic substance from Areca catechu as a new anti-ageing agent

Synopsis We have previously screened 150 medicinal plants for the inhibition of elastase and found significant inhibitory effects of the extracts of Areca catechu L. on the ageing and inflammation of skin tissues . To isolate and identify the compounds having biological activity, they were further purified by each fraction of solvents, silica gel column chromatography, preparative TLC and reversed-phase HPLC. The peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as a phenolic substance by using various colorimetric methods, UV and IR. IC₅₀ values of this phenolic substance were 26.9 μg mL⁻¹ for porcine pancreatic elastase (PPE) and 60.8 μg mL⁻¹ for human neutrophil elastase (HNE). This phenolic substance showed more potent activity than that of reference compounds, oleanolic acid (76.5 μg mL⁻¹ for PPE, 219.2 μg mL⁻¹ for HNE) and ursolic acid (31.0 μg mL⁻¹ for PPE, 118.6 μg mL⁻¹ for HNE). According to the Lineweaver–Burk plots, the inhibition against both PPE and HNE by this phenolic substance was competitive inhibition with the substrate. The phenolic substance from A. catechu effectively inhibited hyaluronidase activity (IC₅₀ : 210 μg mL⁻¹ ).
These results suggest that the phenolic substance purified from A. catechu has an anti-ageing effect by protecting connective tissue proteins.     

Selenium content in selected products of animal origin and estimation of the degree of cover daily Se requirement in Poland

The aim of the study was to determine Se concentration in selected products of animal origin (dairy products, pork, beef, chicken, giblets, fish, eggs) and to estimate the degree to which these products cover daily Se requirement in humans. Selenium concentrations were determined using the spectrofluorometric method. Mean Se concentration in the milk, yoghurt, kefir, and probiotic drinks was 0.020 μg mL⁻¹, 0.010 μg mL⁻¹, 0.012 μg mL⁻¹ and 0.012 μg mL⁻¹, respectively. Selenium concentration in cheese ranged 0.022–0.088 μg g⁻¹ wet weight. The average selenium content of meat ranged from 0.064 (beef) to 0.094 (chicken) μg g⁻¹ w.w. The mean Se content of giblets (liver: 0.307–0.401 μg g⁻¹ w.w.) was significantly ( P  < 0.05) higher than in meat. The concentration of Se depends on fish species and in our study it ranged from 0.136 ± 0.023 (flounder) to 0.282 ± 0.024 μg g⁻¹ w.w. (mackerel). The results obtained show that the analysed food provides 22.8% of the daily selenium requirement. Considering that animal products account for 40–45% of the diet daily selenium intake averages 33–37 μg.     

Natural preservative ingredient from marine alga Ulva lactuca L.

The contents of total chlorophyll (T-Chl), carotenoids and phenolics compounds were quantified in the biomasses of Ulva lactuca grown either in normal or artificial sea water under indoor conditions. The antioxidant and antibacterial activities of U. lactuca crude organic extracts ( Ulva- COEs) were determined. Thirty-four compounds in Ulva- COEs were characterised by thin layer chromatography and high-performance liquid chromatography. The major compounds were chlorophyll a (Chl a ) (15.60–30.90%) and b (Chl b ) (12.20–14.89%) , 9-cis β-carotene (13.12–14.47%), α-carotene (11.44–11.47%) and all-trans β-carotene (6.16–29.70%, of total carotenoids).The Ulva- COEs exhibited remarkable antioxidant activity, with an IC₅₀ (concentration which causes a 50% of DPPH radical scavenging activity) values ranged from 16.5 and 18.7 μg mL⁻¹, which could be compared with the synthetic antioxidants: α-tocopherol (14.4 μg mL⁻¹), butylated hydroxyanisol (13.1 μg mL⁻¹) and butylated hydroxyltoluene (13.1 μg mL⁻¹). Also, Ulva- COEs exhibited great potential antibacterial activities against six bacterial strains, with minimal inhibitory concentration values ranged from 0.40 to 0.35 mg mL⁻¹.     

Storage stability of a stimulant coconut water–acerola fruit juice beverage

The objective of this work was to study the stability of a beverage formulated with acerola fruit juice and green coconut water with added caffeine. The beverage was prepared with 25% acerola pulp, 75% green coconut water and sugar up to 12°Brix, and caffeine (125 mg L⁻¹), heat processed at 90 °C for 30 s and packed in 250-mL glass bottles. Chemical, physicochemical, microbiological and sensory analyses of the beverage were performed just after processing and during 6 months of storage at room temperature (27 °C). The vitamin C content decreased significantly throughout storage, from 399.5 to 189.6 mg 100 mL⁻¹, although it has remained relatively high. The anthocyanins initially present (0.025 mg 100 mL⁻¹) were completely lost during the storage at a mean rate of 4 μg 100 mL⁻¹ month⁻¹. The product was microbiologically stable during storage. Colour changes were also observed with absorbance at 420 nm, with average values ranging from 0.19 to 0.24. However, according to the sensory analyses the product was acceptable during the 6 months of storage, presenting sensory scores (colour, taste and global acceptance) from 6.5 to 5.5, which suggests its potential for market.     

Arbutin determination in medicinal plants and creams

A simple flow injection (FI) manifold with spectrophotometric detection was fabricated and tested for arbutin determination. It is based on the measurement of a red-coloured product at 514 nm formed by the complexation reaction between arbutin and 4-aminoantipyrine (4-AP) in the presence of hexacyanoferrate (III) in an alkaline medium. On injecting 300 μL standard solutions at various concentrations of arbutin into the FI system under optimum conditions, a linear calibration graph over the range of 1.0–30.0 μg mL⁻¹ arbutin was established. It is expressed by the regression equation y  = 0.2188 ± 0.0036 x  + 0.1019 ± 0.0366 ( r ²   = 0.9990, n  = 5). The detection limit (3σ) and the limit of quantitation (10σ) were 0.04 μg mL⁻¹ and 0.13 μg mL⁻¹, respectively. The RSD of intraday and interday precisions were found to be 1.2–1.4% and 1.7–2.7%, respectively. The method was successfully applied in the determination of arbutin in four selected fruits and three commercial whitening cream extracts with the mean recoveries of the added arbutin over the range of 96.2–99.0%. No interference effects from some common excipients used in commercial whitening creams were observed. The method is simple, rapid, selective, accurate, reproducible and relatively inexpensive.     

Efficacy of extract and essential oil of Lantana indica Roxb. against food contaminating moulds and aflatoxin B1 production

The study investigates the antifungal and antiaflatoxigenic efficacy of Lantana indica against Aspergillus flavus , a key storage fungus. The leaf essential oil of L. indica was found more active than leaf extracts. The oil absolutely inhibited the growth of A. flavus at 1.5 mg mL⁻¹ while ethanolic and chloroform extracts of leaf show MIC at 7.5 and 10.0 mg mL⁻¹ concentrations respectively. The oil also showed pronounced antiaflatoxigenic efficacy and completely inhibited the aflatoxin B₁ production at 0.75 mg mL⁻¹. The ethanolic and chloroformic extracts inhibited the aflatoxin B₁ production at 5.0 and 7.5 mg mL⁻¹, respectively while other extracts exhibited poor efficacy. The L. indica essential oil exhibited broad fungitoxic spectrum against twelve different storage moulds. The present findings may recommend the L. indica essential oil and its bioactive leaf extracts as natural preservative would of immense significance in view of the environmental and toxicological implications by indiscriminate use of synthetic pesticides .     

An evaluation of extracts of five traditional medicinal plants from Iran on the inhibition of mushroom tyrosinase activity and scavenging of free radicals

This study aimed to evaluate the free radical scavenging and inhibition properties of five medicinal plants, including Quercus infectoria Olive., Terminalia chebula Retz. , Lavendula stoechas L., Mentha longifolia L., Rheum palmatum L., toward the activity of mushroom tyrosinase using l -tyrosine and l -3,4-dihydroxyphenylalanine ( l -DOPA) as the substrate. The methanol extracts of Q. infectoria and T. chebula showed strong radical scavenging effect in 2,2'-dipheny l -1-picrylhydrazyl (DPPH) assay (IC₅₀ = 15.3 and 82.2 μg mL⁻¹ respectively). These plants also showed inhibitory effects against the activity of mushroom tyrosinase in hydroxylation of l -tyrosine (85.9% and 82.2% inhibition, respectively). These two plants also inhibited the oxidation of l -DOPA similar to kojic acid as positive control (IC₅₀ = 102.8 and 192.6 μg mL⁻¹ respectively). In general Q. infectoria and T. chebula significantly inhibited tyrosinase activity and DPPH radical. Both activities were concentration-dependant but not in linear manner. It is needed to study the cytotoxicity of these plant extracts in pigment cell culture before further evaluation and moving to in vivo conditions.     

Radical scavenging and antimicrobial activity of horsetail (Equisetum arvense L.) extracts

A reversed-phase high performance liquid chromatography (RP-HPLC) separation on C₈ column and quantitative method were developed to analyse hydroxyl derivatives of benzoic and cinnamic acid and flavonoids in horsetail ( Equisetum arvense L.) extracts. Total phenolic content of n -butanol, ethyl acetate and water extracts, determined by the Folin-Ciocalteu method, was 96.4, 26.4 and 15.4 mg g⁻¹ of dry extracts, respectively. The antioxidative activity of horsetail extracts was tested by measuring their ability to scavenge stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) and reactive hydroxyl radicals by electron spin resonance spectroscopy. The results demonstrated that the free radical scavenging activity (versus both DPPH and hydroxyl radicals) depended on the type and concentration of applied extracts; the highest DPPH (EC₅₀ = 0.65 mg mL⁻¹) and hydroxyl radical scavenging activities (EC₅₀ = 0.74 mg mL⁻¹) were obtained in the case of n -butanol extract. The radical scavenging activity of extracts significantly correlated with total phenolic content. The antimicrobial tests showed that ethyl acetate and n -butanol extracts inhibited the growth of tested bacteria.     

Rapid and sensitive determination of formaldehyde in some beverages and foods by flow-injection fluorimetric analysis

A simple and sensitive flow-injection (FI) method for rapid determination of formaldehyde in beverage and food products has been developed. Adopting stop-flow technique, the proposed method distinctly improved the sensitivity of FI fluorimetric method for the determination of formaldehyde based on Hantzsch reaction with cyclohexane-1,3-dione. The fluorescent intensity was proportional to formaldehyde concentrations ranging 0.1 ng mL⁻¹ to 1.000 μg mL⁻¹, and the detection limit ( S / N  = 3) was 0.04 ng mL⁻¹ of formaldehyde. The relative standard deviations ( n  = 11) for determination of 0.100 and 0.005 μg mL⁻¹ of formaldehyde were 1.3% and 2.1%, respectively. The analytical frequency was 18 samples per hour. This method was applied directly for the determination of formaldehyde in diluted beverages and extraction solutions of foods, and the results obtained correlated well with those obtained by the standard method in which a sample pretreatment of steam distillation was required.     

Optimisation of the medium composition for production of protease and soybean peptides by Bacillus subtilis SHZ using response surface methodology

Responses surface methodology was employed to enhance the production of protease and soybean peptides by Bacillus subtilis SHZ. For screening of medium composition significantly influencing protease and soybean peptides yield, the two-level Plackett–Burman design was used. Among thirteen variables tested; KH₂PO₄, glucose and defatted soybean flour (DSF) were selected based on their high significant effect on both protease activity and soybean peptides yield. Then, a three-level Box–Behnken design was employed to optimise the medium composition for the production of the protease and soybean peptides in submerged fermentation. Mathematical models were then developed to show the effect of each medium composition and their interactions on the production of protease and soybean peptides. The model estimated that, the maximal protease activity (320 ± 1 U mL⁻¹) could be obtained when the concentrations of glucose, KH₂PO₄, DSF were set at 8–9 g L⁻¹, 2–3 g L⁻¹, 55–65 g L⁻¹, respectively; while a maximal yield of soybean peptides (8.5 ± 0.1 g L⁻¹) could be achieved when the concentrations of glucose, KH₂PO₄, DSF were set at 7–9 g L⁻¹, 3–4 g L⁻¹ and 55–58 g L⁻¹, respectively. These predicted values were also verified by validation experiments.     

A modified colorimetric method for the estimation of β-cyclodextrin using phenolphthalein

The binding equilibrium between β-cyclodextrin and phenolphthalein has been used to develop a method for the estimation of β-cyclodextrin in solution. From logarithmic plots of amounts of β-cyclodextrin against absorbance at 554 nm, a relation log X  = (log A  − log Y )/ B was found, which gave an estimate of β-cyclodextrin in the concentration range 0.0045 mg mL⁻¹ (3.96 μm) to 4.7 mg mL⁻¹ (4.14 mm), where X =  intercept and B  = slope. This method was found to be highly reproducible and reliable.     

Methyl anthranilate content in Italian citrus honeys determined by HS-SPME-GC

An headspace solid phase microextraction-gas chromatography (HS-SPME-GC) method, previously developed and validated, was applied to the determination of the methyl anthranilate (MA) content of 75 Italian citrus honeys (11 of lemon, 44 of orange and 20 of Citrus spp.). Twenty-four samples were purchased on the local market and 51 were provided by CRA-API ( Consiglio Nazionale per la Ricerca e Sperimentazione in Agricoltura-Istituto Nazionale di Apicoltura e Bachicoltura ) (Bologna, Italy). All the samples had a MA content above the limit of detection (LOD) (0.149 μg g⁻¹) of the analytical method. The concentration range was between 0.46 and 2.52 μg g⁻¹ and the overall average MA content was 1.19 μg g⁻¹. The honeys with the highest mean MA content were the orange honeys followed by Citrus spp. and lemon (1.29 ± 0.461, 1.12 ± 0.511 and 0.92 ± 0.39 μg g⁻¹, respectively). No significant differences were measured between commercial and authenticated samples.     

Optimising the free radical scavenging activity of shrimp protein hydrolysate produced with alcalase using response surface methodology

Response surface methodology (RSM) was used to optimise the hydrolysis parameters of Acetes chinensis by Alcalase 2.4L in order to obtain a hydrolysate with potent radical scavenging activity. The parameters were temperature, pH and enzyme concentration/substrate concentration (E/S) ratio with degree of hydrolysis (DH) being the response. The results showed that the optimum condition was: temperature at 57 °C, pH at 8.0, E/S ratio at 2.6AU 100 g⁻¹ shrimp, hydrolysis time 3 h. The DH was 26.32%, the hydroxyl radical scavenging activity was up to 88.12% and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was 35.61%. The gel column filtration chromatography by a Sephadex G-25 column yielded five fractions. The molecular weight of the most potent free radical scavenging activity fraction ranged from 915 to 207 Da, its IC₅₀ for hydroxyl radical was 0.03 mg mL⁻¹, and IC₅₀ for DPPH radical was 8.86 mg mL⁻¹.     

DPPH free-radical scavenging ability, total phenolic content, and chemical composition analysis of forty-five kinds of essential oils

Forty-five kinds of commonly used essential oils were employed to investigate the DPPH (1,1-diphenyl2-picrylhydrazyl) radical scavenging ability and total phenolic content of major chemical compositions. The free-radical scavenging ability and total phenolic content of cinnamon leaf and clove bud essential oils are the best among these essential oils. One-half milliliter of cinnamon leaf and clove bud essential oils (10 mg mL EtOH) are shown to be 96.74% and 96.12% of the DPPH (2.5ml, 1.52 × 10^-4 M) free-radical scavenging ability, respectively. Their EC50 (effective concentrations) are 53 and 36 (μg mL^-1). One milligram per milliliter of cinnamon leaf, clove bud, and thyme red essential oils were shown to be 420, 480, and 270 (mg g^-1 of GAE) of total phenolic content, respectively. Eugenol in cinnamon leaf and clove bud essential oils (82.87% and 82.32%, respectively) were analyzed by GC-MS. It is clear that the amounts of the phenol compounds in essential oils and the DPPH free-radical scavenging ability are in direct proportion.     

Phenolic content and profiles of selected wild fruits of Zimbabwe: Ximenia caffra, Artobotrys brachypetalus and Syzygium cordatum

The phenolic compound content and profiles of three wild fruits found in Zimbabwe were tentatively identified using the traditional colorimetric methods and high-performance liquid chromatography (HPLC). The fruits assayed were: Ximenia caffra , Artobotrys brachypetalus and Syzygium cordatum . Ximenia caffra fruit peels contained the highest amounts of total phenolics amounting to 1205 μg g⁻¹ in fresh weight, flavonols amounting to 27 μg g⁻¹ and phenolic acids on HPLC tentative identification showed higher concentrations compared with the profiles of the other fruits. Syzygium cordatum fruit peels contained the least amounts of phenolics amounting to 20 μg g⁻¹, flavonols amounting to 8 μg g⁻¹ and phenolic acids' HPLC profiles showed low concentrations. Comparing the peels and pulps of all the fruits, we detected more total phenolics in the peels of X. caffra as high as 1205 μg g⁻¹ and the pulps had 228 μg g⁻¹, more flavonols and phenolic acids while the peels of S. cordatum fruits contained the least with a total phenolic acid content of 20 μg g⁻¹, and had more flavonols in the pulps than the peels, 11 μg g⁻¹ and 8 μg g⁻¹, respectively. Ximenia caffra contained 1.2% and about 1% dry weight condensed tannins in peels and pulps, respectively. In S. cordatum we detected 0.2% and 0.3% dry weight condensed tannins in the peels and pulps, respectively.     

Analysis of isothiocyanates in newly generated vegetables, Baemuchae (×Brassicoraphanus) as affected by growth

Isothiocyanates in Brassica exhibit anticancer properties related to induction of phase II detoxification enzymes. Currently, variations of isothiocyanates was evaluated in Baemuchae (× Brassicoraphanus ), which was newly generated by inter-specific hybrid crops between radish ( Raphanus sativus L.) and Chinese cabbage ( Brassica campestris L. ssp. pekinensis ). A high-performance liquid chromatography system coupled with an evaporative light scattering detector and gas chromatography–mass spectrometry were utilised to identify and quantify isothiocyanates including sulforaphene and sulforaphane in prepared plants. Sulforaphene constituted a major isothiocyanate in Baemuchae and radish, while sulforaphane was detected in broccoli. Sulforaphane contents varied during broccoli growth, with a constant decrease noted from 144.9 ± 8.69 in sprouts to 8.1 ± 1.07 μg g⁻¹ in young leaves. Sulforaphene contents in Baemuchae and radish sprouts were 172.6 ± 11.23 and 131.3 ± 4.08 μg g⁻¹, respectively, and were rarely affected by plant growth. The result supported that newly generated crop Baemuchae had potential to be utilised as functional material because of presence of sulforaphene.     

Storage stability of a high dietary fibre powder from orange by-products

The goal of this investigation was to evaluate the effects of storage time and conditions on the physicochemical and microbiological properties of a high dietary fibre powder (HDFP). HDFP for this study was manufactured from orange juice industry by-products. The storage period was 11 months and samples were stored under vacuum or air exposure and under dark or light exposure. The HDFP is a stable product (based on chemical, microbiological and physicochemical properties) with large amounts of dietary fibre (71.62 ± 0.24 g 100 g⁻¹ dry sample) and polyphenols content (40.67 ± 0.45 mg g⁻¹ dry sample). Hesperidin is the major compound identified in HDFP (higher than 50% of total phenolic compounds). The packaging material used protects the HDFP only up to 6 months. After this time, the increase in moisture content decreases the quality of the product. The best storage conditions are packaging under vacuum and darkness. Both conditions preserve the colour of the fibres probably by means of avoiding pigment degradation.     

Oxidative stability of brined anchovies (Engraulis encrasicholus) with plant extracts

The effect of brining with plant extracts on the oxidative stability of anchovies was investigated during storage. The brining process was done in 15 g 100 mL⁻¹ of sodium chloride solution with water, and with myrtle, rosemary and nettle extracts. Brined anchovies were stored at 4 ± 1  ° C for 28 days. Brining with plant extracts slowed down the lipid oxidation of anchovies. The highest antioxidant effect was observed in brined anchovies with rosemary and myrtle extracts during storage as indicated by peroxide value (POV), thiobarbituric acid reactive substance and oxidative rancidity (OR) scores. Furthermore, OR scores in brined anchovies were well correlated with thiobarbituric acid reactive substance ( r ² = 0.66, P  < 0.01) and POV ( r ² = 0.87; P  < 0.01). The fatty acid profiles were similar among the brined anchovies with plant extracts. These results suggest that brining with rosemary, myrtle and to a lesser extent, nettle extracts prevents development of oxidation in lipids of anchovies during storage.     

Identification and determination butylmethoxydibenzoylmethane in the presence benzophenone-3 and ethylhexylmethoxycinnamate in suncare preparation

The protection of sun radiation is a problem on global level for all living organisms on Earth. The need of people for the overexposure to the UV radiation led human population towards finding novel ways of protection of this kind of radiation, in form of cosmetic preparations applied on the skin. So far, the high values of protection factors of preparations and total block preparations with sun protection factor of 50+ were achieved. Physical and chemical filters which absorb radiation are constituents of these preparations. European Union has set regulations as which substances and in what amounts could be used as UV absorbers. American FDA (Food and Drug Administration) also gave its list of the most frequently used UV absorbers in the sunscreen products, as well as their declared concentrations. The most frequently used concentrations of UV filters in cosmetics is between 0.1% and 10%. Concentrations of UV filters in sunscreen products have to be monitored in order to ensure that they are not less from the declared levels, on which depends the efficacy and safety of the product.
Butyl methoxydibenzoylmethane (BMDM) is used as a UV-A filter in suncare products. Optimized high performance liquid chromatography method for BMDM determination in the presence of other UV filters in suncare preparations is presented in this paper. Determination was performed on C₈ reversed phase using UV detection at 357 nm and isocratic mobile phase of acetonitrile and 0.5% phosphoric acid (70 : 30 v/v). Proposed method has limit of detection of 0.058 μg mL⁻¹, limit of quantification 0.193 μg mL⁻¹ and linearity correlation coefficient of 0.9989. Commercially available products were analysed using the proposed method. All analysed samples complied with EU directives limit of BMDM content to no more than 5%.