Effects of sorbic acid and its salts on chromosome aberrations, sister chromatid exchanges and gene mutations in cultured Chinese hamster cells

The ability of sorbic acid and its potassium and sodium salts to induce chromosome aberrations, sister chromatid exchanges (SCE) and gene mutations in cultured Chinese hamster V79 cells was examined. Sodium sorbate caused significant induction of chromosome aberrations and SCE, and also induced 6-thioguanine-resistant mutations in a dose-dependent manner. The clastogenic potency of sodium sorbate was found to be less than one hundredth of that of the potent clastogen N-methyl-N'-nitro-N-nitrosoguanidine. The induction of SCE by sodium sorbate was twice the control level, whereas that by methyl methanesulphonate, a potent inducer of SCE, was 14 times the control level. The mutagenic potency of sodium sorbate was less than one-tenth that of ethyl methanesulphonate, a potent inducer of mutation, when compared at an equitoxic level. Sorbic acid and its potassium salt induced chromosome aberrations, but only at the highest doses tested. These compounds also induced 1.2 times the control level of SCE, but neither compound induced 6-thioguanine-resistant mutations. The cytogenetic activity of sodium sorbate was concluded not to be due to the effect of osmotic pressure or an impurity. These results indicate that sodium sorbate is a genotoxic agent, although its potency seems to be weak, and that sorbic acid and potassium sorbate are less genotoxic than the sodium salt.     

Evaluation of the genotoxic potential of sorbic acid and potassium sorbate.

The genotoxic potential of sorbic acid and potassium sorbate was investigated in vivo and in vitro. Oral administration of sorbic acid (up to 5000 mg/kg body weight) did not induce sister chromatid exchanges or the formation of micronuclei in bone marrow cells of mice. Intraperitoneal treatment of rats with 400-1200 mg potassium sorbate/kg body weight did not alter the elution profile of DNA from isolated liver cells in the in vivo alkaline elution assay. Sorbic acid did not induce DNA repair in cultured human A549 cells in the unscheduled DNA synthesis (UDS) assay. In vitro incubation of the cells with 1-1000 micrograms potassium sorbate/ml, in the absence or presence of rat liver homogenate, did not result in the formation of DNA single-strand breaks in the alkaline elution assay. These results demonstrate that sorbic acid and its potassium salt are not genotoxic in vivo or in vitro. In contrast to sorbic acid and potassium sorbate, sodium sorbate is very sensitive to oxidative degradation; the main oxidation product was identified to be 4,5-oxohexenoate, which was mutagenic in the Ames test.     

Cytogenetic study in cultured human lymphocytes treated with three commonly used preservatives

Potassium sorbate, sodium benzoate and potassium nitrate have been tested for their genotoxic, cytostatic and cytotoxic potential in human peripheral blood cells in vitro. Potassium nitrate has shown no activity in the test system. When potassium sorbate and sodium benzoate were used at concentrations of 2.0, 0.2 and 0.02 mM no cytostatic activity was detected. However, concentrations of 4 and 8 mM have shown a weak cytostaticity. Additionally, a genotoxic activity using the SCE methodology has been observed at 8 mM of sodium benzoate and at 4 and 8 mM of potassium sorbate. No cytotoxic activity has been induced by the three preservatives. Data demonstrate that the preservatives at low concentrations can be considered as non genotoxic under conditions tested.     

Re-examination of potassium sorbate and sodium sorbate for possible genotoxic potential

Potassium sorbate and sodium sorbate were investigated for possible genotoxic actions using the Salmonella/mammalian-microsome test, HGPRT and sister chromatid exchange (SCE) test with Chinese hamster ovary cells, the micronucleus test on bone marrow cells of mice and Chinese hamsters, and the chromosome aberration and SCE test on Chinese hamsters. In all the in vitro tests no signs of genotoxicity were detected. Whereas no in vivo mutagenicity of potassium sorbate and sodium sorbate with freshly prepared aqueous solutions and with stored potassium sorbate was found, investigations with stored sodium sorbate revealed weak clastogenic activity by increased chromosome aberrations and elevated numbers of micronuclei at doses of 200 mg/kg body weight, but no induction of SCEs.     

Cell-transforming activity of fourteen chemical agents used in dental practice in Syrian hamster embryo cells

Fourteen chemical agents used in dental practice were assessed for their cell-transforming activity using the Syrian hamster embryo (SHE) cell transformation assay system. The cell-transforming activity was quantitatively assessed by the frequency of morphological transformation (MT) in SHE cells induced by these agents. MT was induced by m-cresol, guaiacol, formaldehyde, sodium hypochlorite, hydrogen peroxide, sodium arsenite, acid fuchsin, and basic fuchsin, but not by p-chlorophenol, p-phenolsulfonic acid, glutaraldehyde, and erythrosine B. Iodine and chlorhexidine exhibited positive and pseudopositive responses, respectively. The chemical agents exhibiting a negative or pseudopositive response neither induced nor enhanced MT even in the presence of exogenous metabolic activation.     

Mutagenicity and genotoxicity of sorbic acid-amine reaction products.

Sorbic acid as well as potassium and calcium sorbate (E202 and E203) are legally used as preservatives in numerous processed foods. Owing to its system of conjugated double bonds, sorbic acid is likely to undergo a nucleophilic attack, which may turn it into mutagenic products. The cyclic derivatives resulting from a double addition reaction between sorbic acid and various amines at two different temperatures (50 degrees C and 80 degrees C) have been analysed. A genotoxicity study has been performed with HeLa cells and plasmid DNA. A mutagenesis study has been carried out by using the Ames test. A SOS spot test and a cytotoxicity study have been realised as well. The results showed that the products involved exhibited neither mutagenic nor genotoxic activities.     

Effects of sorbic acid and sorbic acid-nitrite in vivo on bone marrow chromosomes of mice

The effect of sorbic acid alone and in combination with sodium nitrite has been studied on bone marrow chromosomes of mice following 30 days oral treatment. Bone marrow of mice exposed to sorbic acid (15 mg/kg) and sorbic acid nitrite (7.5-1 mg/kg) showed an increase in mitotic index indicating that the drugs had an effect on spindle apparatus. Sorbic acid effected spindle activity but did not damage chromosomes, whereas nitrite itself was clastogenic. However, a combination of half the concentration each of sorbic acid (15 mg/kg) and sodium nitrite (2 mg/kg) together gave synergistic effects, which may be ascribed to the formation of some genotoxic compound in vivo.     

Assessment of genotoxicity of dental antiseptics: ability of phenol, guaiacol, p-phenolsulfonic acid, sodium hypochlorite, p-chlorophenol, m-cresol or formaldehyde to induce unscheduled DNA synthesis in cultured Syrian hamster embryo cells

To assess the genotoxicity of seven dental antiseptics, the ability of these agents to induce unscheduled DNA synthesis (UDS) was examined using Syrian hamster embryo (SHE) cells. Treatment of SHE cells with phenol or formaldehyde induced UDS in a concentration-dependent manner as determined by direct scintillation counting of [3H]thymidine incorporated into DNA during repair synthesis. Guaiacol or m-cresol induced UDS only in the presence of exogenous metabolic activation. p-Phenolsulfonic acid, sodium hypochlorite and p-chlorophenol failed to induce UDS in the presence or absence of exogenous metabolic activation. Our results suggest that phenol, guaiacol, m-cresol and formaldehyde are genotoxic to mammalian cells.     

Morphological cell transformation of Syrian hamster embryo (SHE) cells by the cyanotoxin, cylindrospermopsin

Cylindrospermopsin (CYN) is a cyanotoxin which has been implicated in human intoxication and animal mortality. Genotoxic activity of this hepatotoxin is known but its carcinogenic activity remains to be elucidated. In this work, CYN was assessed for its cell-transforming activity using the Syrian hamster embryo (SHE) cell transformation assay. This in vitro assay is used to evaluate the carcinogenic potential of chemical, physical and biological agents in SHE cells, which are primary, normal, diploid, genetically stable and capable of metabolic activation. We demonstrated that CYN induced a significant increase in morphological cell transformation in SHE cells following a 7-day continuous treatment in the range of non-cytotoxic concentrations 1 × 10⁻⁷-1 × 10⁻² ng/mL.     

山梨酸与山梨酸钾抑菌、抗炎效果比较

目的:测定和比较了山梨酸与山梨酸钾对几种常见食品污染微生物的抑菌活性以及抗炎性能。方法:采用微量肉汤稀释法、Griess试剂法。结果:山梨酸对表皮葡萄球菌、金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌、绿脓杆菌和变形杆菌等受试细菌具有良好的抑制作用,效果强于山梨酸钾;而在抗炎方面,则效果弱于山梨酸钾。     

Genotoxicity testing of four food preservatives and their combinations in the Drosophila wing spot test

In this study, four food preservatives (sodium nitrate, sodium nitrite, potassium nitrate and potassium nitrite) and there five combinations at a concentration of 25 mM have been evaluated for genotoxicity in the somatic mutation and recombination test (SMART) of Drosophila melanogaster. Three-day-old larvae trans-heterozygous including two linked recessive wing hair mutations (multiple wing hairs and flare) were fed at different concentrations of the test compounds (25, 50, 75 and 100 mM) in standard Drosophila Instant Medium. Wings of the emerging adult flies were scored for the presence of spots of mutant cells, which can result from either somatic mutation or mitotic recombination. Also lethal doses of food preservatives used were determined in the experiments. A positive correlation was observed between total mutations and the number of wings having mutation. In addition, the observed mutations in each wing were classified according to the size and type of the mutation. For the evaluation of genotoxic effects, the frequencies of spots per wing in the treated series were compared to the control group, which is distilled water. Chemicals used were ranked as sodium nitrite, potassium nitrite, sodium nitrate and potassium nitrate according to their genotoxic and toxic effects. Moreover, the genotoxic and toxic effects produced by the combined treatments were considerably increased, especially when the four chemicals were mixed. The present study shows that correct administration of food preservatives/additives may have a significant effect on human health.     

Genotoxicity testing of four food preservatives and their combinations in the Drosophila wing spot test

In this study, four food preservatives (sodium nitrate, sodium nitrite, potassium nitrate and potassium nitrite) and there five combinations at a concentration of 25 mM have been evaluated for genotoxicity in the somatic mutation and recombination test (SMART) of Drosophila melanogaster. Three-day-old larvae trans-heterozygous including two linked recessive wing hair mutations (multiple wing hairs and flare) were fed at different concentrations of the test compounds (25, 50, 75 and 100 mM) in standard Drosophila Instant Medium. Wings of the emerging adult flies were scored for the presence of spots of mutant cells, which can result from either somatic mutation or mitotic recombination. Also lethal doses of food preservatives used were determined in the experiments. A positive correlation was observed between total mutations and the number of wings having mutation. In addition, the observed mutations in each wing were classified according to the size and type of the mutation. For the evaluation of genotoxic effects, the frequencies of spots per wing in the treated series were compared to the control group, which is distilled water. Chemicals used were ranked as sodium nitrite, potassium nitrite, sodium nitrate and potassium nitrate according to their genotoxic and toxic effects. Moreover, the genotoxic and toxic effects produced by the combined treatments were considerably increased, especially when the four chemicals were mixed. The present study shows that correct administration of food preservatives/additives may have a significant effect on human health.     

The genotoxicity of sodium saccharin and sodium chloride in relation to their cancer-promoting properties

The literature indicates that sodium saccharin is non-reactive to DNA and inactive as a gene mutagen in vitro. At elevated dose levels it is capable of producing structural disturbances in eukaryotic chromosomes in vitro, and it shows intermittent activity as a very weak germ-cell and somatic-cell mutagen in vivo. Its possible mode of action in these respects is speculated on and related to its ability to promote bladder tumours in rats at elevated dose levels. A review of the toxicology of sodium chloride reveals a profile of genotoxic activities almost identical to that of sodium saccharin. It is suggested that the recorded genotoxic and cancer-promoting activities of these chemicals will only become apparent at elevated dose levels that define them as significant contributors to the biological medium (solvent) rather than as trace xenobiotic toxins (solutes). The possible activity of acid saccharin, or of its potassium, calcium and ammonium salts, as ionic genotoxins requires urgent evaluation.     

Determination of caffeine and potassium sorbate in a neonatal oral solution by HPLC

A reversed-phase HPLC method was developed and validated for the determination of caffeine and potassium sorbate in a neonatal oral solution. Chromatography was performed with a 100 × 4.5 mm Spherisorb 5 μm hexyl column with a mobile phase of 12% acetonitrile in 0.1 M sodium acetate buffer pH 4.5 and ultraviolet detection at 258 nm. The method is stability indicating and there was no interference from a number of common formultion ingredients, although benzoic acid did interfere.     

影响盐酸金刚烷胺糖浆性状稳定性的因素及改进方法

目的:研究使盐酸金刚烷胺糖浆出现结晶现象的因素。方法:通过冷藏5℃、恒温15℃、加速实验、室内等条件按《中国药典》2005年版工艺处方生产的盐酸金刚烷糖浆和按改变工艺处方生产的盐酸金刚烷胺糖浆性状进行稳定性观察。结果:用苯甲酸钠、苯甲酸、山梨酸钾配制的盐酸金刚烷胺糖浆在低温条件5℃(冬天)容易出现结晶现象。结论:盐酸金刚烷胺糖浆使用羟苯乙酯作为防腐剂或在15℃以上储存质量比较稳定。     

关于饮料中山梨酸钾的测定方法的研究

目的:采用HPLC方法测定饮料中防腐剂山梨酸钾的含量。方法:分别将相同对照品溶液、供试品溶液注入Agilent1100型高效液相色谱仪,在不同的流动相比例条件下,分别记录色谱图,进行比较分析。结果:在流动相:甲醇:乙酸铵溶液(0.02mol/L)(10:90)条件下,样品中山梨酸钾的色谱峰与干扰峰的分离度明显高于流动相为甲醇:乙酸铵溶液(0.02mol/L)(5:95)时样品中山梨酸钾的色谱峰与干扰峰的分离度。结论:本方法灵敏、简便、准确。     

Mutagenicity and DNA-damaging activity caused by decomposed products of potassium sorbate reacting with ascorbic acid in the presence of Fe salt.

Although potassium sorbate (PS), ascorbic acid and ferric or ferrous salts (Fe-salts) are used widely in combination as food additives, the strong reactivity of PS and oxidative potency of ascorbic acid in the presence of Fe-salts might form toxic compounds in food during its deposit and distribution. In the present paper, the reaction mixture of PS, ascorbic acid and Fe-salts was evaluated for mutagenicity and DNA-damaging activity by means of the Ames test and rec-assay. Effective lethality was observed in the rec-assay. No mutagenicity was induced in either Salmonella typhimurium strains TA98 (with or without S-9 mix) or TA100 (with S-9 mix). In contrast, a dose-dependent mutagenic effect was obtained when applied to strain TA100 without S-9 mix. The mutagenic activity became stronger increasing with the reaction period. Furthermore, the reaction products obtained in a nitrogen atmosphere did not show any mutagenic and DNA-damaging activity. PS, ascorbic acid and Fe-salts were inactive when they were used separately. Omission of one component from the mixture of PS, ascorbic acid and Fe-salt turned the reaction system inactive. These results demonstrate that ascorbic acid and Fe-salt oxidized PS and the oxidative products caused mutagenicity and DNA-damaging activity.     

AROMATIC l-AMINO ACID DECARBOXYLASE: HISTOCHEMICAL LOCALIZATION IN RAT KIDNEY AND LACK OF EFFECT OF DIETARY POTASSIUM OR SODIUM LOADING ON ENZYME DISTRIBUTION

Utilizing a mono-specific antiserum produced in rabbits to hog kidney aromatic L-amino acid decarboxylase (AADC), the enzyme was localized in rat kidney by immunoperoxidase staining. AADC was located predominantly in the proximal convoluted tubules; there was also weak staining in the distal convoluted tubules and collecting ducts. An increase in dietary potassium or sodium intake produced no change in density or distribution of AADC staining in kidney. An assay of AADC enzyme activity showed no difference in cortex or medulla with chronic potassium loading. A change in distribution or activity of renal AADC does not explain the postulated dopaminergic modulation of renal function that occurs with potassium or sodium loading.     

Comparative toxicity of physiological and biochemical parameters in Euglena gracilis to short-term exposure to potassium sorbate

Ecotoxicology - Potassium sorbate is the potassium salt of sorbic acid, is a widespread and efficient antioxidant that has multiple functions in plants, traditionally associated with the reactions...