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The in vitro physicochemical properties of 6 bottled water products (as drinks), advertised as having cluster sizes of H2O molecules smaller than those of regular water, were studied. No significant differences in the boiling temperature (BT, near 100°C) or freezing temperature (FT, near 0°C) were observed between all 8 aqueous solutions examined, including deionized water (DW) and tap water as controls, although DW had the lowest mean BT and highest mean FT values. The specific gravity of all the examined aqueous solutions, including the controls, changed in a temperature-dependently manner, showing the greatest value (near 1.000) at 4°C. Analyses of inorganic components revealed that various mineral components were dissolved in aqueous solutions examined other than DW, suggesting that the trends of the lowest BT and highest FT for the DW control were due to the absence of these solutes. These results did not demonstrate the presence of an abnormality in the hydrogen bonds between H2O molecules in the sample solutions compared to the controls. In other words, the claims by the manufacturers in their advertisements concerning the cluster sizes of H2O molecules for their water products were not confirmed by the findings of this study.

We studied the in vitro antioxidant activities of neutral aqueous solution systems (water products marketed as drinks) containing hydrogen gas (H2), 2-carboxyethyl germanium sesquioxide (Ge-132), and platinum (Pt) nanocolloid as additives. We evaluated the abilities of these aqueous solutions to inhibit the oxidation of biomolecules catalyzed by an enzyme and induced by reactive oxygen species (ROS) and also to scavenge ROS directly using electron spin resonance (ESR) spectrometry. The concentrations of inorganic elements including Ge and Pt were measured by inductively coupled plasma-mass spectrometry (ICP-MS) and inductively coupled plasma-atomic emission spectrometry (ICP-AES). All the water products examined more or less inhibited the oxidation of 3,4-dihydroxyphenylalanine by tyrosinase and that of L-histidine in an L-ascorbic acid/Cu2+ reaction system. The results of ICP-MS and ICP-AES analyses revealed that Ge, Pt, and some major minerals existed in the water products at concentrations approximately equivalent to those reported by their manufacturers. The ESR spectra indicated that the dissolved Ge-132 molecules and the supplemented Pt nanocolloid particles reduced hydroxyl and superoxide anion radicals. However, under the conditions employed, aqueous H2 did not display such a scavenging ability for these ROS. Our results suggest that H2, Ge-132 and Pt nanocolloid dissolved or supplemented in neutral aqueous media exhibited antioxidant activities in vitro due to the direct scavenging of ROS and/or by other mechanisms.

Ketoprofen [(RS)-2-(3-benzoylphenyl)propanoic acid] is widely used for the treatment of inflammatory diseases and musculoskeletal injury. However, there is concern regarding its potential for photosensitization as a side effect. Free radicals and active oxygen species generated from ketoprofen on exposure to ultraviolet (UV) light have been implicated in phototoxicity and photosensitization. In this study, we examined the suppressing ability of some compounds for the free radicals and active oxygen species generated by the photodynamic reaction of ketoprofen, to determine a new resist of photosensitization by ketoprofen. Eight compounds, including six known free radical scavengers were individually mixed with ketoprofen, and the mixtures were exposed to UV. Then, the free radicals and the active oxygen species were determined by the electron spin resonance spectrometry to estimate suppressing and scavenging ability of compounds. The compounds that show promise in suppressing superoxide anion generation from UV-exposed ketoprofen were further evaluated using the on-line photo-irradiated superoxide anion detection system. It was confirmed that quercetin, a flavonoid, strongly suppresses the generation of free radicals and active oxygen species from UV-exposed ketoprofen. The experiments using the experimental formulation of an adhesive skin patch of ketoprofen containing quercetine and the Chemiluminescence analyzer (CLA) indicated that quercetin has high potential for use as an excipient in ketoprofen ointments to suppress phototoxicity and photosensitization by ketoprofen.

We evaluated the properties and real existence of an electrolyzed-reduced water, which we prepared, and three commercially purchased water goods, that are advertised to have antioxidant activities by the action of “active hydrogen,” on the basis of the results of examinations for inhibitory effects on the oxidative reactions of biomolecules, quantitative analyses of the minerals, and the ESR spectral data in measurement of the scavenging ability for reactive oxygen species. The results suggested that all of the examined aqueous solution systems undoubtedly have antioxidant activities in vitro and that such effects are derived from ordinary molecular hydrogen (hydrogen gas) and/or (a) reductive vanadium ion(s). “Active hydrogen” seems to be absent as an effective component of the antioxidant activities of these aqueous solution systems.

dative stress and kidney damage. However, no effect of L-NMMA was observed on the KBrO3-induced reduction of cytoplasmic GPx activity. These results suggest that reduction of cytoplasmic GPx activity resulted from the KBrO3 administration initiates oxidative stress and that NO also participates in the promotion of KBrO3-induced oxidative stress and kidney damage.

Menthol derivatives were synthesized and evaluated for their promoting activity on the percutaneous absorption of ketoprofen and skin irritation in vivo, choosing O-ethylmenthol (MET) as the mother compound. The compound having a C-3 positionned n-butyl group (1-O-ethyl-3-n-buthylcyclohexanol, OEBC) indicated the most promoting activity and caused relatively little skin irritation. In order to understand enhancement mechanism of OEBC an in vitro permeation study of ketoprofen was performed. The time course of the cumulative amounts of drug permeated through the rat skin exhibited a linear relation after an initial lag time. This was analyzed in membrane diffusion model and the diffusion and partition parameters of ketoprofen were estimated. Both parameters were remarkably enhanced when a hydrogel containing a small quantity of OEBC (0.5%) was applied. Furthermore, to clarify the site of action of OEBC, we also investigated in vitro permeation study of ketoprofen employing different skins of state, reversed skin and stratum corneum stripped skin. When OEBC was added to the hydrogels which were applied to the reversed and stripped skins, almost no changes of the flux were observed compared with the control (without OEBC). These results suggested that the site of action of OEBC was stratum corneum. Morphological changes of the stratum corneum surface were microscopically observed with 0-2% OEBC. The spaces between the stratum corneum cells treated with 0.5-2% OEBC became extended and the shape of each cell became clear. This may suggest that the site of action of OEBC was the intercellular of stratum corneum. Furthermore, an electron spin resonance study was performed to investigate the effect of OEBC on the intercellular lipid bilayer fluidity of the stratum corneum and the rotational correlation times were calculated. 2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO) and 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL) were used as the spin label. In use of OEBC, the fluidity of TEMPO labeled the stratum corneum lipid increased as the addition of OEBC. The results suggested that OEBC promote the penetration of drugs by enhancing fluidity of the local lipid bilayers around TEMPO.

Trihalomethanes such as chloroform, dichlorobromomethane, dibromochloromethane and bromoform in food, which might be produced by the treatment of foods with sodium hypochlorite, were determined by the distillation method. The distillation method was modified for the extraction of trihalomethanes from foods containing lipids and/or proteins. Trihalomethanes in the commercially available bean-curd were determined in the range of 19.8 to 53.5 ppb, although they were not detected in ice candy and pickles. The amount of chloroform in the bean-curd increased with an increase in concentration of sodium hypochlorite used for treatment and in storage temperature. The amount of chloroform produced in the Kinu bean-curd was greater than that produced in the Momen bean-curd. In the bean-curd treated with hypochlorite a large amount of trihalomethanes was generated. On the other hand, in the bean-curd treated with chlorine dioxide or chlorite trihalomethanes were hardly generated. This fact recommends the use of chlorine dioxide or chlorite instead of sodium hypochlorite for the disinfection of food. The use of hypochlorite should be prohibited, because the hypochlorite treatment caused the generation of trihalomethanes in food.

Twelve sorts of foods were tested to investigate the changes in the contents of ascorbic acid, vitamin B1 and vitamin B2 by soaking in 0.5 ppm chlorine dioxide solution. These vitamins in food were extracted with ultrasonic generator for 30 min, and the percentages of the loss of ascorbic acid, vitamin B1 and vitamin B2 were determined by the dichlorphenolindophenol method, thiochrome fluorescence method and lumifulavin method, respectively. It was found that there were the loss of 0-5% in the contents of vitamins after soaking for 15 min and 15-30% after 60 min. However, the percentage of the loss induced by the chlorine dioxide solution was less than that induced by 0.5 ppm chlorite, 0.5 ppm hypochlorite and potable water. The existence of Fe3+, Cu2+ in the solution had an obvious effect which accelerates the decomposition of ascorbic acid.