Determination of total sulfite in wine vinegar was attempted by the microbial sensor method. The recovery of added sulfite (30 μg/ml) from four samples of wine vinegar was more than 90%. The modified Rankine's titration method of determining sulfite was difficult to apply in wine vinegar because of influence of acetic acid in wine vinegar. The microbial sensor method was hardly affected by acetic acid as the modified Rankine's colorimetric method was. The total sulfite values determined by the microbial sensor method coincided with those obtained by the modified Rankine's colorimetric method. Determination of sulfite by the microbial sensor method was stabler than that of the modified Rankine's colorimetric method. Since the microbial sensor method needs no preparation of reagents just before measurement and the measurement time is about 20 minutes, this method was useful for determining the total sulfite in wine vinegar.

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Red radish (Raphanau sativus L.) pigments were found to contain more than 8 components, Two diacylated anthocyanins were isolated and their structures were determined to be 3-O-(2-O-(β-D-glucopyranosyl))-(6-O-feruyl-β-D-glucopyranosyl)-(5-O-(β-D-glucopyranosyl)) pelargonidin and 3-O-(2-O-(6-O-feruyl-β-D-glucopyranosyl))-(6-O-feruyl-β-D-glucopyranosyl)-(5-O-(β-D-glucopytanosyl)) pelargonidin of the basis of chemical and spectroscopic evidences.

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Anthocyanin colors in food were analyzed by using LC-ESI-MS and 3D-HPLC methods. Calibration curves of each color (red cabbage color, elderberry color, purple com color, purple sweet potato color, boysenberry color, grape juice color, grape skin color, perilla color, strawberry color, and mixed colors of red cabbage color, elderberry color, and strawberry color) using LC-ESI-MS and 3D-HPLC were obtained by the total ion strength of quasi-fragment ion peaks of aglycon and the total peak area, respectively. Recoveries of anthocyanin colors from beverage, candy and jelly were 60.3〜104.6% except for grape skin color (40.9〜45.0% in LC-ESI-MS). Constituents and contents of anthocyanin in commercial foods were also elucidated.

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[in Japanese]

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[in Japanese]

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The effect of antioxidants used concomitantly were studied concerning the fate of antioxidant molecules during the antioxidation process. Phenolic antioxidants are frequently used in combination scince they exhibit marked synergistic antioxidative effects. The fate of antioxidant molecules in the mixture are important to understanding the synergism of the antioxidants. The antioxidative activity of a mixture of butylated hydroxyanisole (BHA) and propyl gallate (PG) in lard, soybean oil and methyl oleate were measured by the active oxygen method (AOM). On the other hand, the chemical changes of substances in oil were analyzed quantitatively by HPLC. A mixture of BHA and PG showed higher antioxidative effect on three oils than BHA or PG alone. The survival rate of antioxidative molecules increased remarkably with a combination of BHA and PG. In particular, BHA molecules were present in oils for many hours. These results are interesting enough to note that they seem to have some relation to the evaluation of synergism of antioxidant when they are concomitantly used.

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[in Japanese]

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[in Japanese]

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Hairy root cultures of Beta vulgaris L., cv. Detroit Dark Red and strain C110 were cultured on liquid 1/2MS medium (half strength of the macro elements was used) and the growth and the contents of betacyanin and betaxiantin were investigated. Hairy roots of cv. Detroit Dark Red and strain C110 proliferated to 1800-fold and 1300-fold, respectively, of the inoculum after 1 month culture. Betacyanin and betaxantin content were increased in a logarithmic way and finally to be maximum at the stationary phase of the cultures. Maximum contents of betacyanin in the hairy roots of the two strains were almost the same, but the maximum content of betaxantin of C110 hairy roots was half of that of Detroit Dark Red hairy root, and did not increase at the early stage of the culture. The increasing rate of both pigments of C110 hairy roots was more moderate comparing with that of Detroit Dark Red hairy roots. Considering both high productivity and the difference in betalain synthesis of Detroit Dark Red and C110 hairy roots, these two hairy roots were useful plant materials in investigation of betalain synthesis.

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Previously, we developed a mouse model of anaphylaxis inducible within 9 days by food samples, and proposed a standard concept to quantify the intensities of the anaphylactic responses reflected by the fall of blood pressure. In the present paper, we describe several improvements over our previous method. In the original paper, because of the intra venous administration for the elicitation of the anaphylaxis, only water soluble fractions were used in the samples. For this reason, samples were extracted by three kinds of buffers, all at different pH levels. In order to obtain more accurate information as to the detection of allergens contained in whole food samples including water-insoluble fractions, preparation of the samples was changed this time to a single physical extraction in saline. Accordingly samples containing water-insoluble matters were administered intra peritoneally in both sensitization and challenge. Freund's incomplete adjuvant (FIA) was found to be usefule in the sensitization. Using the improved method, allergic matters contained in food samples, not only natural, but also processed, could be examined. As a primary experiment, it was demonstrated that mice sensitiged with 0.5 mg of boiled egg-white were elicited anaphylaxis by lysozyme as shown in Fig. 1. This indicated that our method could detect 1.7 μg of lysozyme contained as an allergen in egg-white.

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Among the world today, allergies are one of the most serious health problems. Therefore, it is important to find anti-allergic substances contained in the every day food we eat. Sensitive methods to quickly detect anti-allergic components in food samples should also be established. In our present paper, we modified our previous method of inducing mouse anaphylaxis (mouse anaphylactic hypotension; MAHT), by applying this method to the assay in search of anti-allergic substances. Mice sensitized with Hen Egg-white Lysozyme (HEL) fell in antigen specific anaphylaxis on the seventh day, just after challenged with HEL. This is because of hypotension caused by the released histamine (HEL-specific mouse anaphylactic hypotension; HEL-MAHT). In this system, diphenhydramine (DH), one of the most typical anti-allergic drug was used as the standard anti-allergic aubstance. When administered intraveneously, 1h before being challenged, a dose of 0.1 to 10mg/kg of DH showed effective inhibition to the HEL-MAHT by 3% to 60% (Fig. 1). Administration of a given dose of DH at a different time frame, before being challenged, resulted in a reasonable time dependent inhibition of HEL-MAHT. Several anti-allergic drugs other than DH also showed similar dose and time dependent inhibition of HEL-MAHT. For the primary application, extracts from garlic, welsh onion, Chinese chive and dried mushroom were tested for their anti-allergic activities. They showed inhibition of HEL-MAHT by 9% to 24%, which corresponds to 0.2-0.3mg/kg of DH. Therefore, it can be concluded that our method can be applicable for the detection of anti-allergic substances in food.

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Using fresh vegetables to which Fenitrothion and eleven other agrochemicals had been added, model experiment were carried out to determined the effect of rinsing and cooking processes on their contents. Rinsing removed approximately 20 to 50% of the water soluble aagrochemicals. No significant differences were found among water or detergent rinsing, and sodium chlorite or sodium hypochlorite treatment. With regard to cooking, peeling skins was the most effective means for removing agrochemicals. Boiling reduced the contents of the water soluble agrochemicals by 75% or more while sauteing removed approximately 60% of all types of agent applied. Organic-nitrogen-type agrochemicals were not detectable in the green pepper system. The results obtained in this model experiment show that rinsing and cooking significantly reduced the contents of agrochemicals in fresh vegetables.

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