Olive Oil as a Transport Medium for Bioactive Molecules of Plants?-An In Situ Study.

(1) Caries and erosions still remain a challenge for preventive dentistry. Certain plant extracts have shown beneficial effects in preventive dentistry. The aim of this study was to evaluate the antibacterial, anti-adherent and erosion-protective properties of ellagic acid (EA) as a polyphenolic agent. The combination with olive oil was investigated additionally to verify a possible improved bioactive effect of EA. (2) An in situ study was carried out with six subjects. Individual splints were prepared with bovine enamel specimens. The splints were worn for 1 min (pellicle formation time). Thereafter, 10 min rinses were performed with EA in water/in oil. Bacterial adherence was evaluated by fluorescence microscopy (DAPI, ConA, BacLight) after an 8 h oral exposition time. Additionally, the splints were worn for 30 min to quantify demineralization processes. The ultrastructure of the pellicle was investigated after an oral exposure time of 2 h under a transmission electron microscope. Statistical analysis was performed by Kruskal-Wallis tests, Mann-Whitney U tests and Bonferroni-Holm correction. (3) Rinsing with EA led to a significant reduction of adherent vital and dead bacteria. The combination with olive oil did not improve these outcomes. The assessment of glucan structures after rinsing with EA in water showed significant effects. Significant differences were observed for both rinses in calcium release at pH 3.0. After rinsing with EA in oil, significantly less calcium was released compared to rinsing with EA in water (pH = 3.0). (4) Olive oil is not suitable as a transport medium for lipophilic polyphenols. EA has anti-adherent and antibacterial properties in situ. EA also shows erosion-protective effects, which can be enhanced in combination with olive oil depending on the pH value. Ellagic acid has a neutral pH and could be an opportunity in the treatment of specific patient groups (xerostomia or mucositis).

Preventive Applications of Polyphenols in Dentistry-A Review.

Polyphenols are natural substances that have been shown to provide various health benefits. Antioxidant, anti-inflammatory, and anti-carcinogenic effects have been described. At the same time, they inhibit the actions of bacteria, viruses, and fungi. Thus, studies have also examined their effects within the oral cavity. This review provides an overview on the different polyphenols, and their structure and interactions with the tooth surface and the pellicle. In particular, the effects of various tea polyphenols on bioadhesion and erosion have been reviewed. The current research confirms that polyphenols can reduce the growth of cariogenic bacteria. Furthermore, they can decrease the adherence of bacteria to the tooth surface and improve the erosion-protective properties of the acquired enamel pellicle. Tea polyphenols, especially, have the potential to contribute to an oral health-related diet. However, in vitro studies have mainly been conducted. In situ studies and clinical studies need to be extended and supplemented in order to significantly contribute to additive prevention measures in caries prophylaxis.

Identification of New Compounds from Sage Flowers (Salvia officinalis L.) as Markers for Quality Control and the Influence of the Manufacturing Technology on the Chemical Composition and Antibacterial Activity of Sage Flower Extracts.

Parts of Salvia species such as its flowers and leaves are currently used as a culinary herb and for some medicinal applications. To distinguish the different sage extracts it is necessary to analyze their individual chemical compositions. Their characteristic compounds might be established as markers to differentiate between sage flowers and leaf extracts or to determine the manufacturing technology and storage conditions. Tri-p-coumaroylspermidine can be detected only in flowers and has been described here for Salvia and Lavandula species for the first time. Markers for oxidation processes are the novel compounds salviquinone A and B, which were generated from carnosol by exposure to oxygen. Caffeic acid ethyl ester was established as an indirect marker for the usage of ethanol as extraction solvent. The compounds were identified by LC-QTOF-HRESIMS, LC-MS, NMR, IR, and single-crystal X-ray diffraction after isolation by semipreparative HPLC. Furthermore, sage flower resin showed interesting antibacterial in vitro activities against Gram-positive and Gram-negative bacteria.

The chemical composition of the pharmacologically active Thymus species, its antibacterial activity against Streptococcus mutans and the antiadherent effects of T. vulgaris on the bacterial colonization of the in situ pellicle.

The pharmacological active genus Thymus L. comprises over 200 species. Besides its traditional pharmacological use, thyme may reduce the risk of caries disease, however, there is very little respective literature. The pharmacological effects can be attributed to the secondary plant metabolites. The composition of the essential oil and the polyphenols is important for the evaluation of the pharmacological activity. Nevertheless, there are no studies regarding a comparative analysis of the different pharmacological thyme species. In the present study, four different pharmacology Thymus species were cultivated under comparable conditions, and the volatile compounds as well as the polyphenols were characterized. In addition, the in vitro antibacterial activity against S. mutans, one of the primary cariogenic bacterial species, as well as of the essential oil and of the polyphenols were investigated. Furthermore, the bacterial viability and its effect on the initial bacterial adhesion under oral conditions were evaluated in situ for the essential oil and the polyphenols. By GC-MS, 69 volatile compounds, and by LC-DAD-MS/MS, 46 polyphenols could be identified. The comprehensive examination of the essential oils and the polyphenols revealed that the main compounds were equal. However, the yield of the essential oil and the polyphenol content differed clearly. The essential oils of the four investigated Thymus species exhibited an antibacterial activity against S. mutans in vitro, in contrast to the polyphenols of T. vulgaris. Rinsing with polyphenol-rich infusions reduced the initial bacterial colonization while the essential oil inhibited the bacterial growth on dental enamel in situ.

The Polyphenolic Composition of Cistus incanus Herbal Tea and Its Antibacterial and Anti-adherent Activity against Streptococcus mutans.

The Mediterranean plant Cistus incanus is rich in polyphenols and has shown several pharmacological activities, mainly antibacterial effects. Furthermore, in situ studies revealed that a C. incanus infusion reduces the initial bacterial adhesion in the oral cavity due to the polyphenols, an indication that C. incanus might reduce the risk of caries disease. In the present study, the polyphenols from four different commercial C. incanus herbal teas were extracted by standardized accelerated solvent extraction for in vitro tests and by an infusion for in situ tests. Both extracts were characterized qualitatively and quantitatively by high-performance liquid chromatography and only the polyphenol content differed slightly. By means of diode array detection and mass spectrometry, 29 polyphenols, including ellagitannins, flavanols, and glycosylated flavonols, were identified. Thereby, only quantitative but no qualitative differences between the four samples were detected. Furthermore, the in vitro antibacterial activity of the C. incanus accelerated solvent extracts against Streptococcus mutans, one of the primary cariogenic bacterial species, was examined using a live/dead assay (BacLight®). With this approach, C. incanus yielded antibacterial properties. Additional in situ experiments indicated that rinses with a C. incanus infusion reduced the initial bacterial colonization of enamel samples exposed to oral fluids for over eight hours. Furthermore, it was shown by transmission electron microscopy that the application of a C. incanus infusion modifies the ultrastructure of the acquired enamel pellicle, yielding a more electron-dense morphology. It can be assumed that the polyphenols are responsible for the observed effects.

Protective effects of coffee against induction of DNA damage and pre-neoplastic foci by aflatoxin B1.

SCOPE: Aim of the study was to investigate the protective properties of coffee towards aflatoxin B1 (AFB1) induced formation of pre-neoplastic hepatic foci and the identification of the constituents and molecular mechanisms that account for these effects. MATERIALS AND METHODS: Rats consumed three different brews and were subsequently treated with AFB1 (0.75 mg/kg b.w. intraperitoneally). Ten weeks later, the numbers and areas of hepatic foci were determined. Furthermore, the impact of the brews on AFB1-induced DNA damage was quantified in single cell gel electrophoresis assays and the activities of drug metabolising enzymes and glutathione-related parameters were monitored. Additionally, single cell gel electrophoresis assay experiments were conducted with pure caffeine. CONCLUSION: All brews reduced the frequencies of the hepatic foci. The most pronounced protection (reduction 82%) was seen with the caffeine containing metal and paper filtered brews. DNA migration was reduced between 65 and 75% with the caffeine containing brews. In additional experiments, clear protective effects were found with caffeine at dose levels that corresponded to those contained in the coffee. This observation indicates that the alkaloid accounts partly for the protective effects of coffee. Furthermore, our findings indicate that induction of UDP-glucuronosyltransferase contributes to the chemopreventive effects of coffee since all brews increased the activity of this detoxifying enzyme.

Development and validation of an efficient automated method for the analysis of 300 pesticides in foods using two-dimensional liquid chromatography-tandem mass spectrometry.

In this study, a fully automated system was developed for the determination of more than 300 different pesticides from various food commodities. The samples were extracted with acetonitrile prior to the injection into the two-dimensional LC-system. No manual clean-up was needed. The separation of analytes and matrix compounds was carried out by a YMC-Pack Diol (2.1 mm×100 mm; 5 µm; 120Å) HILIC column in the first dimension. All analytes eluted within one small fraction at the beginning of the run. With a packed loop interface this fraction was transferred to the analytical reversed phase separation performed on an Agilent Poroshell 120 EC-C18 (2.1 mm×100 mm; 2.7 µm; 120Å). Some very polar compounds with a stronger retention on the HILIC column were measured directly. The method was validated for over 300 pesticides in cucumber, lemon, wheat flour, rocket, and black tea. For the large majority of the analytes, the recovery was between 70% and 120% and the relative standard deviation was clearly under 20%. The limits of detection for nearly all the compounds were at least at 0.01 mg/kg. For over 50% of the analytes, good sensitivity was observed even at 0.001 mg/kg. In spite of the injection of a pure sample extract, the method showed robust results even with dirty matrices like hops and tea.

Floral markers of cornflower (Centaurea cyanus) honey and its peroxide antibacterial activity for an alternative treatment of digital dermatitis.

Cornflower (Centaurea cyanus) honey can be characterized by a greenish yellow color and an intense flavor with a bitter aftertaste. Because cornflower honey contains only a limited amount of pollen for the verification of its floral origin, one objective was the characterization of its polyphenol and norisoprenoid contents to assign floral markers. Here, lumichrome (18.8-43.5 mg/kg), 7-carboxylumichrome, (Z/E)-3-oxo-retro-α-ionol, and 3-oxo-α-ionol appeared to be quite suitable for distinguishing cornflower honey from other unifloral honeys. Additionally, due to its comparably high hydrogen peroxide content (0.5-0.9 mM/h) and the associated antibacterial activity, cornflower honey was used as an alternative treatment of digital dermatitis on an organic dairy farm. Cows affected by this hoof disease often show severe lameness and a subsequent decline in milk yield and loss of body condition. The cows' hooves treated with cornflower honey showed significantly faster healing than the control group without any treatment.

Assessment of sample cleanup and matrix effects in the pesticide residue analysis of foods using postcolumn infusion in liquid chromatography-tandem mass spectrometry.

Matrix effect profiles can be used to visualize the effect of the sample matrix to the data signals occurring in a chromatogram. In the present study these profiles were generated by postcolumn infusion of a standard pesticide mix with extracts of different food matrices prepared by the QuEChERS method. Complete raw extracts as well as individual clean-up steps were analyzed. This allowed for a detailed comparison of the interferences caused by the matrix effects from various food samples. It also gave an idea about the efficiency of matrix reduction processes. When analyzing the individual clean-up extracts of the QuEChERS method just a slight reduction of matrix effects could be observed from step to step. Matrices causing strong signal effects in the results of the raw extracts also have strong effects after the final clean-up step. Some of the components responsible for the matrix effects show an extremely high retention time. After the injection of extracts from rocket or different types of tea, significant ion suppressions occurred even after rinsing the analytical column for a long time. The experiments have shown that similar matrices can produce different matrix effect profiles. For example, for black teas and green teas significantly different matrix effect profiles were obtained, while the matrix effects of teas within one of these groups were exactly the same. Analogous results could be found for citrus fruits. In order to overcome interfering matrix effects, analytical systems equipped with different electrospray ion sources were tested. Furthermore, profiles of diluted food extracts were generated. Dilution led to a significant decrease in the matrix effects.

Cholesterol-raising diterpenes in types of coffee commonly consumed in Singapore, Indonesia and India and associations with blood lipids: a survey and cross sectional study.

BACKGROUND: To measure the content of cholesterol-raising diterpenes in coffee sold at the retailer level in Singapore, Indonesia and India and to determine the relationship of coffee consumption with lipid levels in a population-based study in Singapore. METHODS: Survey and cross-sectional study in local coffee shops in Singapore, Indonesia and India to measure the diterpene content in coffee, and a population-based study in Singapore to examine the relationship of coffee consumption and blood lipid levels. Interviews and coffee samples (n=27) were collected from coffee shops in Singapore, Indonesia and India. In addition, 3000 men and women who were Chinese, Malay, and Indian residents of Singapore participated in a cross-sectional study. RESULTS AND DISCUSSION: The traditional 'sock' method of coffee preparation used in Singapore resulted in cafestol concentrations comparable to European paper drip filtered coffee (mean 0.09±SD 0.064 mg/cup). This amount would result in negligible predicted increases in serum cholesterol and triglyceride concentrations. Similarly low amounts of cafestol were found in Indian 'filter' coffee that used a metal mesh filter (0.05±0.05 mg/cup). Coffee samples from Indonesia using the 'sock' method (0.85±0.41 mg/cup) or a metal mesh filter (0.98 mg/cup) contained higher amounts of cafestol comparable to espresso coffee. Unfiltered coffee from Indonesia contained an amount of cafestol (4.43 mg/cup) similar to Scandinavian boiled, Turkish and French press coffee with substantial predicted increases in serum cholesterol (0.33 mmol/l) and triglycerides (0.20 mmol/l) concentrations for consumption of 5 cups per day. In the Singaporean population, higher coffee consumption was not substantially associated with serum lipid concentrations after adjustment for potential confounders [LDL-cholesterol: 3.07 (95% confidence interval 2.97-3.18) for <1 cup/week versus 3.12 (2.99-3.26) for ≥3 cups/day; p trend 0.12]. CONCLUSIONS: Based on the low levels of diterpenes found in traditionally prepared coffee consumed in Singapore and India, coffee consumption in these countries does not appear to be a risk factor for elevation of serum cholesterol, whereas samples tested from Indonesia showed mixed results depending on the type of preparation method used.

New approaches for determination of glyphosate and aminomethylphosphonic acid from different tea samples--prospects and limits of cleanup with molecularly imprinted polymer and titanium dioxide.

Glyphosate is the most commonly used herbicide worldwide. To identify glyphosate and its metabolite, aminomethylphosphonic acid (AMPA), in the difficult-to-analyze matrix of tea, new methods were developed. The main emphasis was placed on the cleanup procedure. Two different solid-phase extraction methods were tested and evaluated: one with molecularly imprinted polymers, and the other with immobilized titanium dioxide. The optimization was carried out on the basis of aqueous standard solutions and spiked tea extracts. Validated results were presented for pipet tips that contained immobilized titanium dioxide. After the extraction of glyphosate from tea samples using hydrochloric acid (0.1%), glyphosate and AMPA were concentrated on the adsorbent by pipetting up and down several times without changing the tip. The elution was carried out subsequent to a washing step with 5% ammonia. The extract was derivatized with 2,2,2-trifluoroacetic acid and 2,2,2-trifluoroethanol and then analyzed by GC with MS detection in the negative chemical ionization mode. Quantification was carried out in the selected-ion monitoring mode based on the m/z ratio of characteristic ions due to the ionization process. For glyphosate m/z = 370 was applied, and m/z = 351 for AMPA. The detection limit was 0.03 mg/kg tea for glyphosate and 0.006 mg/kg tea for AMPA. The recoveries for the tested working range (0.1 to 2.8 mg/kg tea) were low but constant at 14 and 12%, respectively. Altogether, the method presented here can be carried out easily and quickly, and it gives reliable information on the presence of the analytes in a sample. Quantification is also possible using standard addition.