Determination of a non-volatile N-nitrosamine on a food matrix.
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Biomedical subjects
Publications and source records attributed to C L Walters.
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A procedure is described whereby the nitrosation of sarcosine can be followed spectrophotometrically in the presence of either hydrophilic or lipophilic antioxidants. Using this technique, a number of antioxidants of both classes at a concentration of 2.0 mmol have inhibited strongly the formation of N-nitrosarcosine from 25 mmol-sarcosine and 25 mmol-nitrite.
The concentrations of nitrite and thiocyanate in fasting and pentagastrin stimulated gastric juice and in saliva have been examined. Nitrite was found in all of 17 samples of fasting gastric juice, mean 4-9 +/- 1-1 muM. Stimulation of gastric secretion with pentagastrin caused no significant change in nitrite concentration. Thiocyanate was detected in all of 21 samples of fasting gastric juice and the difference in concentration between smokers and non-smokers probably reflects similar differences in saliva. In contrast to the nitrite data there was a significant drop in thiocyanate concentration of gastric juice after pentagastrin from 0-9 +/- 0-1 mM to 0-3 +/- 0-04 mM, suggesting a salivary origin for the thiocyanate in gastric juice. Thiocyanate is a powerful catalyst of nitrosation, which, together with small amounts of nitrite and naturally occurring amines could lead to the intragastric formation of carcinogenic nitrosamines and in certain circumstances be a factor in the aetiology of gastric cancer.
A case of gustatory sweating after blunt trauma to the left side of the face has been discussed. In addition to a fibular fracture, the patient sustained multiple mandibular fractures as well as paralysis of the left facial nerve, which disappeared within five weeks. Three months after the trauma, the patient had gustatory sweating, which was controlled with topical applications of scopolamine hydrobromide.
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In model system studies simulating the frying of bacon a considerable variation in the production of N-nitrosopyrrolidine (NOPyr) has been observed, as occurs in sliced bacon fried conventionally. The inclusion of ascorbate or erythorbate (isoacorbate) has led consistently to a fall in or an elimination of NOPyr production up to an addition of 300 ppm (1,5 mM). Above this concentration of ascorbate, the elimination of NOPyr formation persisted in some instances but in others an increased production of the nitrosamine occurred. Using alpha-tocopherol, a fall of NOPyr production during the simulated frying of bacon was obtained with increase of concentration of the antioxidant, there being a consistent elimination of the volatile nitrosamine following an additionof 500 ppm (1.16 mM). Whilst ascorbyl palmitate had a similar action in reducing or eliminating NOPyr production, no synergistic effect between it and alpha-tocopherol was noted in an experiment combining the two antioxidants. When bacon was fried in fat containing 400 or 800 ppm alpha-tocopherol, the total production of NOPyr and DMN in the fried bacon, cooked out fat and condensate was markedly reduced but not completely eliminated as in the model system studies. Similarly, the curing of bacon with a brine containing a mixture of alpha-tocopherol, ascorbyl palmitate and citric acid has led to considerable reductions in NOPyr and DMN output on frying when the combined concentrations of the two anti-oxidants were calculated to be 800 ppm.
Nitrite and hydrogen ion concentrations have been measured in the fasting gastric juice of 69 patients undergoing routine gastrointestinal investigations. There was an inverse relationship between nitrite concentration and hydrogen ion concentration, with a highly significant increase in gastric-juice nitrite in hypochlorhydric subjects. Thiocyanate was also found in all specimens in concentrations likely to increase nitrosamine formation, if nitrosation of amines occurs in the fasting stomach. Neutral gastric juice contains metabolically active bacteria capable both of generating nitrite from nitrate and of catalysing nitrosation. In this way an intragastric environment suitable for the formation of carcinogenic nitrosamines exists in the hypochlorhydric and achlorhydric stomach, providing a possible mechanism for the high incidence of gastric cancer in these subjects.
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Higher levels of NPy and NDMA in cooked bacon and its cooked-out fat were associated with those cures which contained higher levels of nitrite, but no correlation between nitrosamine concentrations and initial nitrate levels was observed. There was a higher proportion of nitrosamines in cooked-out fat than in cooked bacon, but by far the greatest amounts were lost in the vapours produced during cooking.
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The velocity of oxidation of exogenous ferrocytochrome c by nitrite under anaerobic conditions in the presence of skeletal muscle mitochondria is dependent upon pH over at least the range 5.6-6.7, increasing markedly as the pH is lowered. A product of the reaction is the complex formed between nitric oxide and ferricytochrome c. At levels up to 20 mM, nitrite inhibits aerobic cytochrome oxidase action; at higher concentrations, however, a partial resuscitation of the oxidation of ferrocytochrome c occurs, the enhancement of reaction velocity being considerably greater at pH 6.0 than at 6.5. Mitochondrial respiration is also inhibited by nitrite but no similar resurgence was, however, observed and thus the oxidation of ferrocytochrome c by high levels of nitrite is considered to be a direct non-enzymic action. Under anaerobic conditions, the rate of increase of the velocity constant of the oxidation of ferrocytochrome c with nitrite concentration in the presence of muscle mitochondria similarly decreased with rise of pH over the same range. The permeability of the muscle mitochondrion to nitrire has been demonstrated by swelling studies and by the rapid conversion of endogenous ferrocytochrome a3 into its nitrosyl-derivative. Over longer periods of anaerobic incubations of mitochondria with nitrite, oxidation of endogenous cytochromes occurs with the formation of nitrosylferricytochrome c. Above a nitrite concentration of 0.3 mM, the mitochondrial enzyme system probably involved is increasingly inhibited but by a concentration of 30 mM a direct non-enzymic oxidation has intervened. Commercial vacuum packed bacons were examined by electron microscopy. Mitochondria were clearly recognisable although they contained fewer cristae than those observed in fresh meat.
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