Human platelet 5-hydroxytryptamine concentration as a tool in prediction of solvent induced neurotoxic effects.

Many organic solvents are lipophilic and concentrate in lipid rich tissues e.g. nervous tissue, where they are known to induce toxic effects in humans, especially in the central nervous system. Changes in the presynaptic neurotransmitter metabolism may play a role in these effects. The platelet is proposed as an alternative human model for this complex in the investigation of such changes, especially regarding 5-hydroxytryptamine (5-HT). The platelet 5-HT concentration was measured in platelets isolated from exposed persons with diagnosed neurointoxication caused by exposure to organic solvents and compared to controls. No difference in the concentration was found. Instead of using the platelet 5-HT concentration in the prediction of neurotoxic effects, the platelet 5-HT uptake rate may be an alternative parameter. A specific and sensitive double isotope derivative technique for 5-HT measurements is described. A solvent induced reduction in the number of platelets/ml platelet rich plasma was found and is discussed.

Hypersensitivity reactions to food colours with special reference to the natural colour annatto extract (butter colour).

It is well known that synthetic food colours especially some azo dyes can provoke hypersensitivity reactions such as urticaria, angioneurotic oedema, and astma (Michaëlsson and Juhlin, 1973, Granholt and Thune, 1975). Natural food colours are scarcely investigated with respect to potential allergic properties. Annatto extract, a commonly used food colour in edible fats e.g. butter, has been tested in patients. Among 61 consecutive patients suffereing from chornic urticaria and/or angioneurotic oedema 56 patients were orally provoked by annatto extract during elimination diet. Challenge was performed with a dose equivalent to the amount used in 25 grammes of butter. Twentysix per cent of the patients reacted to this colour 4 hours (SD: 2,6) after intake. Similar challenges with synthetic dyes showed the following results: Tartrazine 11%, Sunset Yellow FCF 17%, Food Red 17 16%, Amaranth 9%, Ponceau 4 R 15%, Erythrosine 12% and Brillant Blue FCF 14%. The present study indicates that natural food colours may induce hypersensitivity reactions as frequent as synthetic dyes.

Evaluation of food safety. National and international aspects.

Evaluation of the safety of chemicals in food have created considerable interest within the public, political, administrative, and scientific sectors in many parts o- the world. Political and administrative approaches may vary from country to country depdneing on traditions and cultural pattern. The political aims and goals of our times may be clearly defined, but they are often rather obscure seen from a scientific point of view. Health authorities are supposed to interpret the political intentions and transcribe them into practical administration. The use, by national and international health authorities, of the available scientific and technological knowledge to evaluate the safety of the food in question is discussed.

Stimulation of DNA synthesis in mouse and rat lung following administration and butylated hydroxytoluene.

Butylated hydroxytoluene (BHT) is a widely used antioxidant in food. It has recently been reported that intraperitoneal injection of BHT to female mice produced, within 3-5 days, a hypertrophy, hyperplasia, and general disorganisation of the cellular components of the lung (Marion and Mitchell, 1972). In male mice it was shown that this effect of BHT was associated with a marked stimulation of DNA synthesis in the lung (Witschi and Saheb, 1974). Measuring the incorporation of thymidine-2-14C into DNA in lung confirmed the results reported in mice. 500 mg of BHT per kg, i.p., resulted in a 20-40 fold increase in thymidine incorporation in both male and female mice after 4 days. In similar experiments in the rat, however, a two-fold increase was observed in female rats, no effect could be demonstrated in male rats. In experiments in male and female mice using p.o. administration of BHT similar results have been obtained as following i.p. administration.

Absorption and excretion of 1-amino-2-naphthol-6-sulphonic acid in rats and rabbits.

1-amino-2-naphthol-6-sulphonic acid (ANSA) and compounds of similar structure are metabolites formed by azo reduction in mammals from several commonly used food colours, e.g. Sunset Yellow FCF and Food Red 17. At our institute it has been shown that ANSA produces bile duct proliferation when given orally to rabbits. Furthermore Orange RN which by azo reduction yields ANSA (and aniline) induce the same effect in pigs (Olsen et al., 1973) but not in rats (Gaunt et al., 1971). Using 35S-labelled ANSA the absorption and excretion as well as the localisation in the liver cell of ANSA was investigated in rats and rabbits. The results after p.o. administration shows that the rabbit absorbs and excretes into the urine a significant larger amount of ANSA (30-40%) than the rat (6-13%). The results after i.v. administration shows that some 60% of the injected dose is excreted in the urine in both species. In the rat 15-20% are recovered from the faeces, while only traces (0.1-0.5%) are found in faeces of rabbits. A significant amount of ANSA was retained in the blood and the liver of both species 24 and 48 hours after administration. After differential centrifugation of liver homogenates the majority of ANSA was found in the 9000 x g supernatant.

Hemiglobin formation in vitro induced by azo dyes and their metabolites.

Azo dyes which after reduction by the intestinal flora yield aniline are known as inducers of Heinz bodies after oral intake (J.J.-P. Drake, 1975). The effect of aniline is thought to depend on its metabolic conversion to the known hemiglobin inducers, phenylhydroxylamine and o- and p-aminophenol (A. de Bruin, 1976). The azo dyes used for colouring of foods are reduced in vivo forming sulphonated primary aromatic amines, many of which are of aminonaphthol structure. Such compounds could be potential hemiglobin inducers. This possibility was investigated in vitro by incubating the following azo dyes and their metabolites, with red cells in Krebs-Ringer phosphate solution: Chrysoin S, Scharlach GN, Azorubine, Sunset Yellow FCF, Food Red 17, Orange GGN, Ponceau 4R, Amaranth, Ponceau 6R and Fast Yellow AB. The concentration in the incubate was 0.5 mmol/l and the hemiglobin formation was measured after 1 h. The effect on red cells from pigs was compared with the effect on human red cells. Only the metabolites containing amino and naphthol groups together were active. The activity depended on the number and position of the sulfonic acid groups.

Studies on the metabolism of Orange Rn in the pig.

Orange RN (monosodium salt of 1-phenylazo-2-naphthol-6-sulphonic acid) was administered to female pigs either by intraveneous injection or by stomach tube. After intravenous injection of Orange RN, 7.8 mg/kg, the following metabolites were indentified in 24-hour urines: Orange RN (31%), 1-(4-hydroxyphenlazo)-2-naphthol-6-sulphonic acid (3%), p-aminophenol and 0-aminophenol (34% and 4% of the theoretical yield, respectively). 1-Amino-2-naphthol-6-sulphonic acid was present, but not determined. When the urine collection was extended to 72 hours the excretion of p-aminophenol accounted for the remainder of the phenylazo moiety. The excretion pattern for p-aminophenol suggest that Orange RN is partly excreted in the bile and thereafter undergoes azo reduction in the gut. After administration of Orange RN, 78 mg/kg, by stomach tube the following metabolites were identified in the urine: Total coloured metabolites (Orange RN and 1-(4-hydroxyphenylazo)-2-naphtholphenol-6-sulphinic acid) (0.4% of the theoretical yield), p-aminophenol and o-aminophenol (52% and 6% of the theoretical yield, respectively) and aniline (0.3% of the theoretical yield). 1-Amino-2-naphthol-6-sulphonic acid was present, but not determined.