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.     

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.     

Antioxidant and toxicity tests of roasted noni (Morinda citrifolia) leaf infusion

The antioxidant properties and toxicity profile of roasted noni ( Morinda citrifolia L . ) leaf infusion were evaluated. The 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging activity was greater than green tea infusion (81.6 ± 0.9% vs. 57.5 ± 1.8%, P  < 0.001). The mean quercetin and kaempferol contents of roasted noni leaf infusion, as prepared by the consumer, were 0.24 ± 0.01 and 0.14 ± 0.01 μg mL⁻¹, respectively. Tannic acid content was 10 ± 1 μg mL⁻¹. The infusion was non-mutagenic in the reverse mutation test in Salmonella typhimurium and did not induce primary DNA damage in E. coli PQ37. Further, no significant primary DNA damage was induced by 5,15-dimethylmorindol, which was the only detectable anthraquinone in noni leaves. The infusion was not cytotoxic in the 24 h brine shrimp toxicity test (LC₅₀ > 1 mg mL⁻¹), nor was there any evidence of acute oral toxicity from the freeze–dried infusion in Sprague–Dawley rats (LD₅₀ > 2000 mg kg⁻¹ b.w.).     

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.     

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.     

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.     

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.     

Chemical composition and antimicrobial activities of essential oil from the peel of bingtang sweet orange (Citrus sinensis Osbeck)

The chemical composition of the essential oil obtained from the peel of Bingtang sweet orange ( Citrus sinensis Osbeck) was analysed by gas chromatography and gas chromatography/mass spectrometry (GC/MS). Twenty-seven components were identified. The monoterpenes and sesquiterpene hydrocarbons with 96.03% (w/w) of the total oil were the principal compound groups. Among which, limonene was observed dominant (77.49%), followed by myrcene (6.27%), α-farnesene (3.64%), γ-terpinene (3.34%), α -pinene (1.49%), sabinene (1.29%) and other minor components. Results by disc diffusion method and minimum inhibitory concentration (MIC) determination method showed that the essential oil had a wide spectrum of antimicrobial activities against Staphylococcus aureus , Penicillium chrysogenum , Bacillus subtilis, Escherichia coli and Saccharomyces cerevisiae , with their inhibition zones ranging from 14.57 mm to 23.37 mm and the MIC ranging from 4.66 μL mL⁻¹ to 18.75 μL mL⁻¹.     

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%.     

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.     

J. Cosmet. Sci., 59, 441–448 (September/October 2008)Simultaneous determination of heavy metals in cosmetic products

An extremely small amount of several heavy metals have been detected in cosmetic products as impurities, which can cause skin allergies through percutaneous adsorption on the skin. We present here a fast, accurate, and highly sensitive method for simultaneous determination of Pb²⁺, Fe²⁺, Cu²⁺, Ni²⁺, Zn²⁺, Co²⁺, Cd²⁺ and Mn²⁺ in coloring agents and cosmetic products, to be evaluated by ion chromatography. All of these metals are well separated through a bifunctional ion-exchange column (IonPac CS5A) and detected by post-column reaction and spectrophotometric detection. The calibration graphs are linear ( r ² > 0.999), in the range 0.1–1000 μg ml^-1. Detection limits for a 200-μl sample solution are at the μg L^-1 level, which is sufficient for judging whether the product is safe or not. The relative standard deviations (RSDs) of the retention time and the peak area are less than 0.21 and 1.24%, respectively. The recovery rates are 97–104%. The result shows that the proposed determination method is more sensitive, more accurate, and faster than current methods such as HPLC, ICP-MS and Flame-AAS. The new method was applied to analyse the amount of heavy metals contained in 22 cosmetic products and 11 coloring agents.     

High-performance liquid chromatographic determination of arbutin in skin-whitening creams and medicinal plant extracts

A high-performance liquid chromatographic method was developed for quantitative analysis of arbutin. The arbutin was separated on an ODS Hypersil^® C18 column with a mobile phase of water: methanol: 0.1 M hydrochloric acid (89:10:1, v/v/v). The level of arbutin was measured by means of UV detection at 222 nm. The optimum conditions for arbutin quantitative analysis were investigated. The calibration curve was found to be linear up to 1000 μg/ml^-1 of arbutin concentration, and the working calibration curve for arbutin determination over the range 0.5–30.0 μg/ml^-1 of arbutin ( r ² = 0.9999) was established. The relative standard deviations for intraday and interday were found to be 0.98% and 1.15%, respectively. A detection limit (3σ) and quantitation limit (10σ) of 0.02 μg/ml^-1 and 0.2 μg/ml^-1, respectively, and a mean percentage recovery of the spiked arbutin of 99.88 ± 1.12% were obtained. The proposed method has been applied to the determination of arbutin in commercial skin-whitening creams (Arbuwhite^® cream, Super Whitening^® cream, and Shiseido^® cream) with average contents of 7.60, 5.30, and 57.90 mg/g^-1, respectively. It was also applied to the determination of arbutin in medicinal plant extracts from Betula alnoides Buch. Ham., Clerodendrum petasites S. Moore, Curculigo latifolia Dryand. Var. latifolia, and Hesperethusa crenulata (Roxb.) Roem, levels of which were found to be 3.50, 1.50, 1.10, and 0.12 μg/g^-1, respectively (no article reported in the literature about arbutin analysis). The proposed HPLC method is rapid, simple, and selective for routine analysis.     

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.     

In vitro evaluation of the cutaneous penetration of sprayable sunscreen emulsions with high concentrations of UV filters

The aim of this study was to evaluate the possible penetration through human skin of organic and inorganic filters contained in sunscreen emulsions packaged in aerosol cans, using an in vitro method. Experiments were carried out on two different types of emulsion: W/Si and W/O. This study was conducted using static diffusion cells (Franz cells). The determination of organic UV filters [Methylene Bis Benzotriazolyl Tetramethylbutylphenol (MBBT); Bis-Ethylhexyloxyphenol Methoxyphenyl Triazine (BEMT); Diethylamino Hydroxybenzoyl Hexyl Benzoate (DHHB); Ethylhexyl Methoxycinnamate (EMC); and 2-Ethylhexyl Dimethyl PABA (ED-PABA)] was performed by High Performance Liquid Chromatography (HPLC). Therefore, it was important to develop a single analytical method for the quantification of the five organic filters with the aim of facilitating the experiment. The determination of inorganic filters [titanium dioxide (TiO₂) and zinc oxide (ZnO)] was performed using an emission spectrometric analysis method (ICP-OES). The HPLC and ICP-OES methods were validated. After a penetration test of 24 h duration, the results showed very low penetration only for two of the organic filters (maximum penetration of 1.21 μg cm⁻² h⁻¹ for EMC and 0.14 μg cm⁻² h⁻¹ for MBBT) and no penetration for the inorganic filters. Moreover, more than 50% of each sunscreen agent stayed on the surface on the skin. These results are consistent with those in the literature that presents similar experiments. This study showed that the sprayable sunscreen products developed, which contained high concentrations of UV filters, presented a low level of skin penetration.     

Influence of particle size fractions on the physicochemical properties of maize flour and textural characteristics of a maize-based nonfermented food gel

The physicochemical properties of fractionated maize flour and the textural characteristics of a maize-based nonfermented food gel (maize tuwo ) prepared from the respective fractionated flours were evaluated. The maize flour was fractionated into four fractions: <75 μm, 75–150 μm, 150–300 μm, 300–425 μm and whole meal (<425 μm). There were variations in the selected chemical constituents of fractionated maize flour including protein (2.9–4%), ash (0.80–0.97%), crude fibre (0.73–0.91%) and damaged starch (10.1–17.4%). The fractionated maize flour gave variable bulk density (0.80–0.93 g cm⁻³), water absorption capacity (1.9–2.1 g g⁻¹) and oil absorption capacity (1.7–2.1 g g⁻¹). The colour characteristics of the fractionated maize flour and the pasting properties were all affected by the fractionation. The cohesiveness index (strain at peak compressive force) of the food gel from the flour fractions ranged between 15% and 19.5% while the softness index of the food gel ranged between 16.7 and 17.5 mm. The relative high cohesiveness and softness indexes (i.e. 19.5% and 17.4 mm respectively) of maize tuwo prepared from the flour fraction of 75–150 μm can predispose the food gel towards easier hand-mouldability and swallowability respectively; being important quality indicators for its acceptability.     

The level of polyaromatic hydrocarbons in kajal and surma of major Indian brands

Kajal and surma are eye cosmetics extensively used in Indian subcontinent. Kajal is prepared by burning of vegetable oil and butter oil while surma by grinding of the stones. High performance liquid chromatography and gas chromatography–mass spectrometry instruments were used for quantification and confirmation of 16 polyaromatic hydrocarbons (PAHs). Significant concentration of PAH was found in all the samples examined. The median concentration of PAH ranged from 0.14 (lowest, anthracene) to 31.18 μg g⁻¹ [dibenz(a,h)anthracene] in kajal sample and from not detectable concentration (naphthalene) to 197.47 μg g⁻¹ of benzo(a)pyrene in surma sample. Fifteen PAHs were detected in all the samples. Therefore the use of kajal and surma in eye should be strictly restricted.