Yoikabin C, one of food additives submitted to be authorized was analyzed by means of spectrophotometry, nonaqueous titration, and some other methods. It was found that it had the structure of lauryltrimethylammonium-2, 4, 5-trichlorophenoxide in which lauryltrimethylammonium was bound with 2, 4, 5-trichlorophenol by ionic bond, and had one or half a mol of water of crystallization.

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It has been already known that dehydroacetic acid (DHA) produced a chelate compound with ferric salts. The colour developed by this reaction was stable, and the reaction was so sensitive that could be utilized as a colorimetric method for determining DHA.
A aqueous solution containing 0.5-3.5mg of DHA was transferred into a 25-ml flask, added 5ml of 2% iron alum solution, and diluted to the mark with water. The resulting solution was mixed thoroughly, the absorbances of the solution was determined at the wavelength of 440mμ.
Sorbic acid, salicylic acid, methylnaphthoquinone, nitrofurazone, nitrofurylacrylamide, acids, alkalis or colouring matters interfered with the method. Benzoic acid and esters of p-hydroxybenzoic acid did not interfere with this method in the concentration below 50mg per 100ml. However, these interferring substances could be eliminated by the steam distillation method described in the second report.

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It has been reported already that dehydroacetic acid reacted with ferric iron to produce a chelate compound. The reaction was so sensitive and quantitative that it was utilized for the colorimetric determination of dehydroacetic acid.
Suitable amount of the distillate from foods, which was made by the steam distillation method reported on Shokuhin Eiseigaku Zashi, 5, 148 (1965), was transferred into a 25-ml flask, added with 5ml of iron alum solution and diluted to the mark with water. The absorbance of the solution was measured at 440mμ. More than 10μg/ml of dehydroacetic acid in the distillate could be determined by this method. Sorbic acid and salicylic acid interfered with this method, but other food preservatives did not interfere within the regular amount. Nitrofurazone, nitrofurylacrylamide, artificial sweetners and coloured substances did not appear in the distillate.

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Benzoic acid in foodstuffs has been detected by various chromatography, and the identification of small amounts of benzoic acid is possible. The acid has been determined ultraviolet spectrophotometrically, or volumetrically. However, the determination of the acid is disturbed by other acidic substances.
In the present report, studies on the infrared spectrometric analysis of the acid are described. The results obtained are as follows:
1. Characteristic absorption of benzoic acid in carbon tetrachloride in IR-region provides an infrared spectrometric method of analysis of the acid.
2. 1690cm^-1 are useful as key band for the quantitative analysis of the acid.
3. The interfering effects of other food additives on this method have been investigated, and the analysis of benzoic acid was found to be possible, except sorbic acid.
4. This method and ultraviolet spectrophotometry were applied to the market foods, and both measured values were found nearly equal.

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In general, dehydroacetic acid in foodstuffs has been determined by ultraviolet spectrophotometry, or colorimetry with salicylaldehyde.
In the present report, studies on the infrared spectrometric analysis of the acid are described.
The results obtained are as follows:
1. Characteristic absorption of dehydroacetic acid in carbon tetrachloride in IR-region provides an infrared spectrometric method of analysis of the acid.
2. Carbonyl band 1, 750cm^-1 are useful as key band for the quantitative analysis of the acid.
3. The interfering effects of other food additives on this method have been investigated, and the detection of the acid was found to be possible, but its determination was found to be difficult.

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The toxic effects of nitrofurazone have been investigated in rat testis histopathologically.
Marked atrophy of the testis, severe degeneration and necrosis of the seminiferous epithelium were selectively caused by continued high oral administration of nitrofurazone. The testis weight of treated animals was reduced to half of its of controls by 10 days feeding of 0.1% dietary level. Histologically, it was observed that mature spermatozoa, spermatid, and secondary spermatocyte were primarily affected, and primary spermatocyte and spermatogonia were delayed in the destruction, but no changes were noted in the Sertoli cells, interstitial cells, and basement membrane of the tubules by the one month's treatment. Three months after the first administration, however, moderate hyperplasia of Leydig cells and slight thickening of the basement membrane were recognized.
Testicular degenerative changes were arised from 3 days feeding of 0.1% dietary level, and minimal toxic dose to the testicular epithelium was at the 0.06% level of nitrofurazone in the diet.
Almost complete anatomical restoration of the seminiferous epithelium recognized in 60 days after return to the basal diet followed 32 days' administration of the drug. It was shown that regenerative activity was considerably well preserved even in the long duration of the treatment.
It was proved that maximal dose of the nothing of the testicular changes in the rat was about 35mg/kg of nitrofurazone a day, and this dose was corresponding to 2, 500 pieces of fish sausage (net weight 140g) in a man of the body weight of 50kg. From these results, it may be considered that the use of nitrofurazone as a food preservative is practically non-toxic.

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[in Japanese]

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[in Japanese]

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