Sodium Dehydroacetate Research Articles

Anticoagulant effect of sodium dehydroacetate (DHA‐S) in rats

Sodium dehydroacetate (DHA-S) is used as a food additive, preservative and antimicrobial agent. Repeated oral administration of DHA-S in rats induced severe hemorrhage in multiple organs and prolongation of blood coagulation factors. To determine the mechanism of hemorrhage, the protective effect of vitamin K (VK) was investigated. Increased VK-dependent blood coagulation parameters, a prolonged prothrombin time (PT) and an activated partial thromboplastin time (APTT) were observed in rats when DHA-S alone was administered, while only a slight change was observed in animals that received a single injection of vitamin K2 following the DHA-S dosing. These results suggest that DHA-S-induced hemorrhage is caused by a deficiency of vitamin K. In addition, the inhibitory effect of DHA-S on vitamin K1 epoxide reductase (VKOR) activity was measured with an in vitro system using liver microsomes of normal male rats. DHA-S concentration-dependently inhibited VKOR activity similar to warfarin, but the inhibitory concentration was high. Therefore, it was concluded that the DHA-S-induced hemorrhage was caused by a depletion of blood VK, associated with any factors including VKOR inhibition.

Effect of addition of sodium dehydroacetate on the fermentation of blue-grass silage

Effect of addition of sodium dehydroacetate on the fermentation of blue-grass silage

Simultaneous determination of sweeteners and preservatives in preserved fruits by micellar electrokinetic capillary chromatography.

A micellar electrokinetic capillary method for the simultaneous determination of the sweeteners dulcin, aspartame, saccharin, and acesulfame-K and the preservatives sorbic acid; benzoic acid; sodium dehydroacetate; and methyl-, ethyl-, propyl-, isopropyl-, butyl-, and isobutyl-p-hydroxybenzoate in preserved fruits is developed. These additives are ion-paired and extracted using sonication followed by solid-phase extraction from the sample. Separation is achieved using a 57-cm fused-silica capillary with a buffer comprised of 0.05 M sodium deoxycholate, 0.02 M borate-phosphate buffer (pH 8.6), and 5% acetonitrile, and the wavelength for detection is 214 nm. The average recovery rate for all sweeteners and preservatives is approximately 90% with good reproducibility, and the detection limits range from 10 to 25 microg/g. Fifty preserved fruit samples are analyzed for the content of sweeteners and preservatives. The sweeteners found in 28 samples was aspartame (0.17-11.59 g/kg) or saccharin (0.09-5.64 g/kg). Benzoic acid (0.02-1.72 g/kg) and sorbic acid (0.27-1.15 g/kg) were found as preservatives in 29 samples.

Cytotoxicity of food preservatives in cultured rat hepatocytes loaded with linolenic acid

We investigated the ability of eight food preservatives to induce lipid peroxidation in normal and α-linolenic acid (LNA)-loaded cultured rat hepatocytes. On the addition of sodium dehydroacetate (DHA-Na), potassium sorbate (SA-K) or thiabendazole (TBZ) to the cell culture, lipid peroxidation, assessed in terms of the production of malondialdehyde (MDA), was induced in LNA-loaded cells, but not in normal cells. At the low concentrations, induction of lipid peroxidation in LNA-loaded cells was highest with TBZ, whereas at high concentrations DHA-Na greatly induced lipid peroxidation. The occurrence of lipid peroxidation in LNA-loaded cells was accompanied by a decrease in cellular GSH levels with the three preservatives and by a decrease in cellular protein-SH levels with DHA-Na and TBZ. Furthermore, cell injury, measured by the release of LDH, was produced in LNA-loaded cells exposed to DHA-Na and SA-K. The addition of TBZ caused substantial cell injury in normal cells, and even greater injury in LNA-loaded cells. The prevention of lipid peroxidation in LNA-loaded hepatocytes by addition of an antioxidant, N, N′-diphenyl- p-phenylenediamine (DPPD) almost completely prevented DHA-Na- and SA-K-induced cell injury, and reduced TBZ-induced cell injury. The addition of diphenyl (DP), o-phenylphenol (OPP) or butyl p-hydroxybenzoate (BHB) caused severe cell injury, in association with a marked decrease in cellular levels of both of GSH and protein-SH in both groups of cells. However, lipid peroxidation was not detectable in either group of cells exposed to these preservatives. Sodium propionate (PA-Na) and sodium benzoate (BA-Na) had little effect on any cytotoxic parameter in either group of cells.

含そう用アロプリノール液に使用する保存剤の検討

For the prevention or treatment of stomatitis caused by anticancer agents, allopurinol gargle has been used at Gifu University Hospital. The allopurinol gargle has been prepared by dispersing powdered allopurinol tablets and sodium carboxymethylcellulose into water for injection. As the result of examination of microbial contamination of the allopurinol gargle administered to a patient, Acinetobacter calcoaceticus var. anitratus, Burkholderia pickettii or Candida albicans were isolated. Benzoic acid (BA), sodium dehydroacetate (DA) and ethyl-p-hydroxybenzoate (EP) were used as preservatives for allopurinol gargle. Allopurinol gargles containing 0.05% of BA, 0.1 % of BA, 0.05% of DA, 0.1 % of DA or 0.05% of EP were prepared and antimicrobial activities against strains isolated from allopurinol gargles and those from clinical specimens were assessed based on the criteria for topical preparations in British Pharmacopoeia. Among the alloprinol gargles prepared, only alloprinol gargle containing 0.1% of BA had antimicrobial activity against all strains at storage conditions of 25°C and 37°C. The alloprinol gargle containing 0.1 % of BA did not demonstrate sufficient antimicrobial activity at 4°C. The BA added to allopurinol gargles had no influence on either the stability of allopurinol or on the inhibition of xanthine oxidase by allopurinol. No growth of organisms appeared in the remaining allopurinol gargle containing 0.1 % of BA used by the patient.These findings suggest that the addition of BA at the concentration of 0.1 % is suitable for allopurinol gargle.

Determination of Sodium Dehydro Acetate

Determination of Sodium Dehydro Acetate

Simultaneous determination of preservatives, sweeteners and antioxidants in foods by paired-ion liquid chromatography

A paired-ion, reversed-phase high-performance liquid chromatographic method was developed for the simultaneous determination of sweeteners (dulcin, saccharin-Na and acesulfame-K), preservatives (sodium dehydroacetate, sorbic acid, salicyclic acid, benzoic acid, succinic acid, methyl-para-hydroxybenzoic acid, ethyl-para-hydroxybenzoic acid, n-propyl-para-hydroxybenzoic acid, isopropyl-para-hydroxybenzoic acid, n-butyl-para-hydroxybenzoic acid, and isobutyl-para-hydroxybenzoic acid), and antioxidants (3-tertiary-butyl-4-hydroxyanisole and tertiary-butyl-hydroquinone). A mobile phase of acetonitrile-50 mM aqueous α-hydroxyisobutyric acid solution (pH 4.5) (2.2 : 3.4 or 2.4 : 3.6, v/v) containing 2.5 mM hexadecyltrimethylammonium bromide and a C18 column with a flow rate at 1.0 mL/min and detection at 233 nm were used. This method was found to be very reproducible with detection limits ranged from 0.15 to 3.00 µg. The retention factor (k) of each additive could be affected by concentrations of hexadecyltrimethylammonium bromide and α-hydroxyisobutyric acid, and pH and ratio of mobile phase. The presence of additives in some food samples was determined.

Post-harvest treatment of wild mango ( Irvingia gabonensis) for improved shelf life

The effectiveness of hot water, chemicals, wax and film packaging in extending the post-harvest life of wild mango under tropical ambient conditions was investigated. Dipping wild mangoes in hot (55°C) water or hot 0·1% benomyl, 0·5% sodium dehydroacetate or 0·5% Na 2S 2O 5, followed by waxing or packaging in boxes overwrapped with stretch PVC film, delayed ripening, controlled decay, minimised weight loss and extended the shelf life of the fruits when held at 22–35°C and 70–95% relative humidity without adverse effects on visual and chemical qualities. The best results were obtained when PVC film packaging was preceded by a sulphite dip.

Fungi and decay associated with wild mango fruits (Irvingia gabonensis) under different postharvest conditions

AbstractFungi and percentage decay associated with wild mango fruits during storage at different postharvest conditions were monitored. In addition the effect of chemical treatments (hot water, solutions of benomyl, sodium acetate and sodium metabisulphite) and film packaging on percentage rate of decay was investigated.The characteristic brownish black rot, brownish discolouration and rotting of tissue, and greenish‐black soft rot common in wild mango fruits during storage are associated with fungi of the genera Aspergillus, Botrytis and Penicillium. The use of hot water, hot benomyl, hot sodium dehydroacetate and hot sodium metabisulphite (55 °C each) dips alone, or in combination with film packaging helped in reducing postharvest decay of wild mango fruits. Varietal differences in susceptibility of wild mango fruits to decay was also observed.

Micronucleus tests in mice on 39 food additives and eight miscellaneous chemicals

A total of 47 chemical substances including 32 synthetic food additives, seven additives from natural sources, three trihalogenated methanes, two fluoro-compounds for dental use, one insecticide, and two other compounds were subjected to the micronucleus test in mice. Five compounds, i.e. chlorine dioxide, maltol, potassium bromate, sodium chlorite and sodium dehydroacetate, were found to induce micronuclei after a single ip injection. Potassium bromate, sodium chlorite and sodium dehydroacetate were tested further by oral administration, and potassium bromate showed a clearly positive result. Almost all the compounds which were negative with the single treatment were additionally tested by four or five multiple treatments, but none showed any indication of micronucleus induction.

４‐ジメチルアミノアゾベンゼンの発ガン性に及ぼすデヒドロ酢酸ナトリウム同時投与の影響

４‐ジメチルアミノアゾベンゼンの発ガン性に及ぼすデヒドロ酢酸ナトリウム同時投与の影響

4 Final Report on the Safety Assessment of Sodium Dehydroacetate and Dehydroacetic Acid

Sodium Dehydroacetate and Dehydroacetic Acid are used as preservatives in cosmetic formulations at concentrations of 1.0 percent or less. Both compounds are rapidly absorbed when administered orally or on the skin of test animals.Acute toxicity studies indicate that Sodium Dehydroacetate and Dehydroacetic Acid are slightly toxic when administered orally to rats. Neither compound was an irritant when applied to rabbit skin. Sodium Dehydroacetate was found to exhibit minimal eye irritation. Subchronic and chronic studies reveal various toxic effects, primarily due to the incurred lack of appetite and weight loss. No evidence of mutagenicity was reported for either ingredient use. No evidence of tumor induction by Dehydroacetic Acid was detected in a 64-week study. Dehydroacetic Acid had an inhibitory effect on hepatoma induction in rats when fed 4-(dimethylamino)azobenzene. A teratogenicity study in mice revealed no significant findings when compared to untreated controls.Sodium Dehydroacetate, Dehydroacetic Acid, and cosmetics containing these ingredients were found practically nonirritating, nonsensitizing, nonphotosensitizing, and nonphototoxic in numerous clinical tests. On the basis of the available animal and clinical data, it is concluded that Sodium Dehydroacetate and Dehydroacetic Acid are safe as cosmetic ingredients in the present practices of use and concentration.

Absence of Synergistic Toxicity of Two Food Additives, Sodium Dehydroacetate and Dibutyl Hydroxy Toluene, in Rats Treated Orally for 6 Months

Toxicological potentiation of two food additives was investigated in male and female Wistar rats given sodium dehydroacetate (DHA-Na) and dibutyl hydroxy toluene (BHT) in the diet for 6 months. The concentrations of the samples in the diet were 0 (control and pair feeding control), DHA-Na 0.05, 0.15 and 0.5%, BHT 0.1, 0.3 and 1%, DHA-Na 0.05%+BHT 0.1%, DHA-Na 0.5%+BHT 0.1% and DHA-Na 0.05%+BHT 1%. Under the present experimental conditions, no marked potentiation of toxicity was observed in the combination of DHA-Na and BHT.

Effects of Various Preservatives on the Growth and Aflatoxin B1 Production of Aspergillus flavus and Aspergillus parasiticus in Sliced Bread

Various preservatives, such as calcium propionate, sorbic acid, potassium sorbate, dehydroacetic acid, sodium dehydroacetate, benzoic acid, sodium benzoate and isobutyl p-hydroxybenzoate, were examined for their inhibitory effects on the growth and aflatoxin B1 production of A. flavus and A. parasiticus in sliced bread coated with peanut butter.Bread containing each preservative was baked and sliced in our laboratory. The peanut butter containing a preservative and one hundred spores of each fungus was coated on the upper surface of a slice of bread (5.5×6.5×1.0cm), and then the slice was incubated at 28°C in a Petri dish for ten days.Of the eight preservatives, calcium propionate and sorbic acid showed the most inhibitory effects. When bread containing calcium propionate (2.5g/kg) was coated with peanut butter containing sorbic acid (1.5g/kg), the growth was retarded by 48hr (A. flavus) and 72hr (A. parasiticus), while aflatoxin B1 production was retarded by 72hr (both fungal species) in comparison with the control without any preservative.Further experiments were performed using nine slices of the bread with ten spores each, in the same manner as described above. Neither growth of the two fungi nor aflatoxin production was observed in these slices, with one exception, in which the fungal growth of the two species and aflatoxin production were retarded by 48hr as compared with the control.

Bacteriolytic activity of an enzyme derived from Streptomyces globisporus 1829 on cariogenic microorganisms

An enzyme which possesses lytic activity against cariogenic streptococci was obtained from strains of streptomyces in soil and sewage. The strains of streptomyces which produced a highly active enzyme in the culture supernatant were identified as Streptomyces griseus strain H-402 and Streptomyces gloaisporus strain 1829. The enzyme of S. globisporus strongly lysed the strains of cariogenic streptococci in the tris acid malate-NaOH buffer, pH 7.0.This study is concerned with the estimation of the bacteriolytic activity of the enzyme derived from S. globisporus on cariogenic microorganisms such as the strains of Streptococcus mutans, Lactobacillus casei and Actinomyces viscosus, and also deals with the effect of fluoride ion, sodium N-lauroylsarcosinate and sodium dehydroacetate on the activity of the bacteriolytic enzyme.Twenty seven strains of S. mutans employed in this experiment were all sensitive to the bacteriolytic enzyme. Dergrees of bacteriolysis were compared with each of 5 serological groups of S. mutans. Strains of group a and b were more sensititive to the enzyme than group, c, d and E. Cariogenic L. casei ATCC 4646 and A. viscosus T6 were also extremely sensitive, whereas 4 strains of Streptococcus sanguis were less sensitive to the enzyme.Strains of S. mutans and S. sanguis which showed the least sensitivity to the enzyme were significantly lysed by prolonged incubation with the enzyme.The bacteriolytic activity was apparently enhanced by the presence of 20 μg/ml of lysozyme or 0.3% of sodium N-lauroylsarcosinate. 0.2% of sodium dehydroacetate was also effective in enhanceing of the enzyme activity. The enzyme activity was not influenced by fluoride ions with a concentration of less than 200 ppm, whereas 40% of the enzyme activity was supressed in the presence of fluoride ions with a concentration of more than 400 ppm. The enzyme activity was also supressed by halide ions such as F-, Cl-, Br-, and I-.

Synergistic toxicity of official permissible preservative food additives (author's transl)

In an investigation of the synergistic toxicity of officially permitted preservative food additives, the acute toxicity test and six-months breeding test were used.Part 1. (Acute toxicity test)(1) Ten preservatives were chosen from the officially permitted list: (sodium dehydroacetate, sorbic acid, benzoic acid, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, salicylic acid, sodium propionate, furylfuramide…2%, furylfuramide…pure). Mixtures of two additives each were administered orally to dd mice.(2) Mixtures were made in two ways: eqaul amounts of each ingredient and eqaul LD50 doses of each.(3) In order to evaluate the synergistic toxicity of the mixtures, the theoretical LD50 values of the mixtures were caliculated.(4) The toxicity of the mixtures was found not to exceed the theoretical values; i.e., it was not synergistic.Part 2. (Six-months breeding test)(1) From the preservatives listed in part 1, furylfuramide…2% (F), sorbic acid (S) and ethyl p-hydroxybenzoate (E) were chosen and administered as single doses and in pairs to SD-JCL rats for 25 weeks.(2) The amounts administered corresponded to about 1/5, 1/10 and 1/50 of the LD50. The percentages of preservatives in the diet were: (F) 4.0%, 2.0%, 0.4%; (S) 3.0%, 1.5%, 0.3%; (E) 2.0%, 1.0%, 0.2%.(3) Experimental results.1) During the experimental period, no significant difference was noted in the general appearence, behavior, food consumption, mortality or mean survival times between the experimental and control groups.2) The weight gain was normal except for a slight delay in the F4%-E2% group.3) The red blood cell count hemoglobin, hematocrit and white blood cell count were not affected.4) The results of biochemical serum tests were as follows: the males in groups F4%, F4%-S3%, F2%-S1.5%, and the females in groups F4%, F2%-S1.5%, F0.4%-S0.3%, F4%-E2% and F2%-E1% showed significantly higher total protein and total cholesterol than the control group.5) The F4% and F4%-E2% groups tended to have enlarged livers and kidneys, and F4%-S3%, F2%-S1.5% and F2%-E1% groups tended to have enlarged livers only.6) Pathological examinations showed no abnormal findings either macroscopically or microscopically.(4) These experiments showed no significant synergistic toxicity in mixtures of the above-mentioned food preservatives.

抗菌性物質の作用機作に関する研究(第2報) : デハイドロ醋酸ナトリウムのD-アミノ酸酸化酵素に対する阻害作用

Sodium dehydroacetate has been found to inhibit one of flavin enzymes, D-amino acid oxidase. This inhibition was analyzed by enzyme reaction kinetics using the enzyme purified from hog kidney. Sodium dehydroacetate was found to inhibit D-amino acid oxidase by two inhibitive factors, (1) the one factor based on direct combination of sodium dehydroacetate and coenzyme flavin adenine dinucleotide (FAD), and (2) the factor based on competitive reaction between sodium dehydroacetate and FAD, in saturated system of the substrate. The dissociationconstant of FAD-sodium dehydroacetate complex was measured by the enzyme reaction kinetic method as Kif=2.7×10-2M/liter, and this value agreed approximately with Kif. FAD=3.85×10-2M/liter measured earlier by the physicochemical method (fluorescence method) for the same complex. The dissociation constant of the apoenzymesodium dehydroacetate complex formed by competition between sodium dehydroacetate and FAD was measured as Kip=5.9×10-2M/liter. The flavin-sodium dehydroacetate interaction, previously proved physicochemically, was also proved by enzyme reaction kinetics through inhibition of sodium dehydroacetate against D-amino acid oxidase.

Metabolic Activities of Lactic Acid Bacteria for Dairy Use when Affected by Added Preservatives

The inhibitory effects of sodium dehydroacetate (DHA-Na) and sodium sorbate (SA-Na), added at 0.02-0.10% level, on acid production, proteolysis and aroma production by five strains of lactic acid bacteria, were investigated in skimmilk and some other milk media. Acid production of some lactobacilli including L. bulgaricus when incubated at 37°C for 24 and 48hr, was inhibited remarkably by the addition of 0.10% DHA-Na to skimmilk media. The similar inhibition occurred with 0.02-0.10% level of SA-Na, especially in the case of S. lactis. Proteolysis by the cultures when incubated for two days was retarded significantly by the addition of the preservatives at 0.08-0.10% level, whereas at the same level of preservatives the inhibition of the cultures when incubated for four days was not so significant. The presence of 0.02% level of preservatives had little effect on proteolysis. On the aroma (diacetyl and acetoin) production by S. lactis and some lactobacilli incubated for two and four days, some inhibitory effect took place when 0.04% DHA-Na or SA-Na was present, and it became nearly complete at the 0.08-0.10% level. Conclusively, some critical problems concerning reasonability of the addition of preservatives to commercial fermented milk beverage were discussed.

Studies on mode of action of antibacterial substances. I. Interaction between sodium dehydroacetate and flavins

An evidence was provided that a kind of a complex is formed by interaction between sodium dehydroacetate with flavin in aqueous solution from the result of physicochemical examination proving the following four points. In the presence of sodium dehydroacetate, (1) quenching of fluorescence characteristic to flavin occurs, (2) a change occurs in the ultraviolet absorption spectrum of flavin, the absorption maximum at 445-450 mμ shifing to a longer wavelength region, with decrease in its intensity, (3) photodecomposition of flavin is markedly inhibited, and (4) solubility of sparingly soluble riboflavin increases markedly. Dissociation constant of flavin-sodium dehydroacetate complex was measured by the fluorescence method, absorption spectroscopy, and by the solubilization method and the value was found to be approximately 2 to 6×10-2 M/L (pH 5.8-8.6, 20°). The site of the dehydroacetic acid molecule interacting with flavin seems to be the enolic hydroxyl in 4-position of its pyrone ring.

Results of Using Sodium Dehydroacetate Applications to Reduce Discoloration of Snapbeans Damaged by Machine Harvesting1

Abstract Sodium dehydroacetate solutions in concentrations up to 0.8% were found to be effective in reducing discoloration of cut and bruised surfaces of mechanically harvested snapbeans and in reducing some mold and bacterial damage. Techniques for objectively measuring color differences using the Hunterlab Meter D 25 are described.