ABSTRACT
Hops and hop extracts are approved and widely used bittering agents in the brewing of beer. During recent years, preisomerized alpha hop acids and reduced preisomerized alpha hop acids have been introduced as effective and economical bittering agents that may be added late in the brewing process. Although hops have been used for centuries, there are few studies in the literature on the safety of this ingredient. The study herein was conducted to determine the effects associated with subchronic oral administration of the reduced preisomerized hop acids, hexahydroisohumulone and tetrahydroisohumulone, in the dog. The results show that these materials are generally well tolerated in the dog. At high dose levels they induce vomiting, and much of the material administered was excreted in the faeces. The no-observed-adverse-effect level (NOAEL) of the compounds were 50 and 100 mg/kg body weight, respectively. Consumption of these ingredients by adult humans drinking 1 litre of beer daily is less than 0.25 mg/kg body weight; their use is thus associated with wide safety margins.
Subject(s)
Feces/chemistry , Flavoring Agents/toxicity , Terpenes/toxicity , Vomiting/chemically induced , Administration, Oral , Animals , Cyclohexenes , Dogs , Dose-Response Relationship, Drug , Female , Flavoring Agents/administration & dosage , Flavoring Agents/chemistry , Isomerism , Male , No-Observed-Adverse-Effect Level , Terpenes/administration & dosage , Terpenes/chemistryABSTRACT
Sucrose acetate isobutyrate (SAIB), a mixture of esters of sucrose with a composition approximating the name sucrose diacetate hexaisobutyrate, has been used for over 30 yr in many countries as a 'weighting' or 'density-adjusting' agent in non-alcoholic carbonated and non-carbonated beverages. As part of the demonstration of safety of SAIB as a direct food additive in human diets, a program of toxicity testing was started in the late 1950s that culminated in extensive studies of SAIB in rodents, monkeys and humans over the last decade. This review summarizes the toxicity data, accrued up until 1988, that precede the safety studies published elsewhere in this issue. SAIB has been shown to have very low acute and chronic toxicities in rats, monkeys, and, except for effects on the liver, in dogs at feeding levels of up to 10% in the diet. Slight effects seen in rats and monkeys at levels of 10% in the diet are unlikely to be directly caused by exposure to SAIB. In dogs, however, SAIB causes decreases in bromosulfophthalein (BSP) and indocyanine green (ICG) elimination from the serum immediately following a single dose, indicative of interference with biliary excretion. On repeated feeding in dogs, SAIB caused increases in serum alkaline phosphatase levels, but enzymes indicative of toxic effects on the liver were unaffected. On prolonged feeding to dogs, SAIB caused changes in liver morphology revealed by electron microscopy. All of these effects were reversed when SAIB was withdrawn from the diet. The no-effect level for these effects in dogs was near 5 mg/kg body weight, but these effects were not seen in rats fed up to 4 g/kg body weight/day, monkeys fed up to 10 g/kg body weight/day, or humans fed up to 20 mg/kg body weight/day. The toxicity and pharmacological studies in dogs, rats and monkeys suggest that the effect of SAIB on biliary excretion and liver morphology in dogs is essentially pharmacological rather than toxicological in nature and that the difference between the effects in dogs at levels as low as 5 mg/kg body weight/day, and the lack of effects in rats or monkeys at levels up to 10 g/kg/day is not merely a quantitative difference between species, but an absolute qualitative difference.
Subject(s)
Food Additives/toxicity , Sucrose/analogs & derivatives , Animals , Carbonated Beverages/analysis , Carbonated Beverages/history , Dogs , Drug Approval , Food Additives/history , Haplorhini , History, 20th Century , Humans , Liver/drug effects , Liver/ultrastructure , No-Observed-Adverse-Effect Level , Product Surveillance, Postmarketing , Rats , Sucrose/history , Sucrose/toxicity , Toxicity TestsABSTRACT
Preliminary short-term toxicity studies of sucrose acetate isobutyrate (SAIB) in the dog demonstrated that addition of this additive to the diet was associated with an increase in liver size and elevated serum alkaline phosphatase activity with no evidence of pathological change by light microscopy. To determine the basis for these changes, a 12-week oral toxicity study of SAIB was conducted in the dog and a similar study was performed in the rat. SAIB was fed in the diet to groups of six beagle dogs of each sex at 0, 0.5, 1.0, 2.0 and 4.0%. SAIB was also fed to groups of 40 Sprague-Dawley rats of each sex at levels of 0, 2.5, 5.0 and 10.0%. In the rat study, in addition to routine toxicology parameters, hepatic microsomal enzyme induction was determined using a zoxazolamine hypnotic test, urinary ascorbic acid excretion and determination of hepatic carboxylesterase activity. Sodium phenobarbital was fed to groups of 20 rats of each sex at a dose of 100 mg/kg body weight/day by gavage as a positive control for hepatic microsomal enzyme induction. In the dog study, routine toxicological tests were supplemented by tests for bromsulfophthalein (BSP) retention, histochemical staining of liver sections for glycogen, phosphorylase, succinate dehydrogenase, and acid and alkaline phosphatases. Levels of liver lipid, protein, glycogen and carboxylesterase activity were also determined. Electron microscopic examinations were made on liver sections from the dog study at the end of the 12-week SAIB feeding period and after a 2-week withdrawal period. Administration of SAIB to rats did not reveal evidence of any effect on hepatobiliary function, and there was no indication of microsomal enzyme induction. Body weight gain of male rats fed SAIB was decreased, probably as the result of decreased palatability of the diet; SAIB did not affect body weight gain in females. The changes observed in the dogs fed SAIB included increased serum alkaline phosphatase activity with no change in serum alanine aminotransferase, aspartate aminotransferase or lactic dehydrogenase activity and no change in serum electrolyte, serum protein, glucose or bilirubin levels. No haematological changes were observed. BSP retention was observed at all SAIB dose levels. There were no SAIB-related pathological changes in any organ when examined by light microscopy. Examination by electron microscope revealed dilatation of bile canaliculi and an increase in smooth endoplasmic reticulum compared with controls. Histochemical studies also indicated increased enzyme activity of the bile canaliculi. The electron-microscope-revealed changes were completely reversed during a 2-week treatment withdrawal period. The dog study did not establish a no-effect level for changes in hepatobiliary function induced by feeding SAIB.
Subject(s)
Food Additives/toxicity , Liver/drug effects , Sucrose/analogs & derivatives , Alkaline Phosphatase/blood , Animal Feed , Animals , Ascorbic Acid/urine , Biomarkers/analysis , Body Weight/drug effects , Carboxylesterase , Carboxylic Ester Hydrolases/analysis , Dogs , Enzyme Induction/drug effects , Female , Food Additives/administration & dosage , Liver/enzymology , Liver/pathology , Male , Microsomes, Liver/drug effects , Microsomes, Liver/enzymology , Organ Size/drug effects , Phenobarbital , Rats , Rats, Sprague-Dawley , Species Specificity , Sucrose/administration & dosage , Sucrose/toxicity , Sulfobromophthalein , Zoxazolamine/pharmacologyABSTRACT
Sucrose acetate isobutyrate (SAIB), a food additive used as a flavour emulsion stabilizer in citrus-based soft drinks, was evaluated for chronic toxicity in B6C3F1 mice and Fischer 344 rats. SAIB dissolved in acetone was blended into NIH07 rodent diet at concentrations that were adjusted weekly during the first 12 to 18 months of the studies so that ingested dose levels per kg body weight were constant. Groups of 20 rats per sex were given dose levels of 0.0, 0.0, 0.5, 1.0 and 2.0 g SAIB/kg body weight for 1 yr, and groups of 50 rats per sex were given dose levels of 0.0, 0.0, 0.5, 1.0 and 2.0 g SAIB/kg body weight for 2 yr. Mice were fed dose levels of 0.0, 0.0, 1.25, 2.5 and 5.0 g SAIB/kg body weight for 2 yr. The highest doses fed, equivalent to dietary concentrations of approximately 5%, were considered to be the maximum concentrations that could be fed without risk of nutritional deficiencies. Depressions in body weight gain were noted, particularly in female rats during the first 12 to 18 months of the studies. Recovery during the last quarter of the 2-yr study suggests that the reduced body weight gain was nutritional rather than SAIB-related. There were no differences in survival between SAIB-treated rats or mice and controls. Decreased body weight gains, primarily in females, but less consistent than those in the rat, were noted in the 2-yr mouse study. No signs of toxicity were observed in clinical chemistry, haematology, organ weights, gross necropsy findings or light microscopy studies in the 1- or 2-yr rat studies. Electron microscopic examinations of liver sections from high dose level rats from the 1-yr study also revealed no effects of SAIB treatment. There were no significant increases in benign or malignant tumours in the long-term rat or mouse carcinogenicity studies. The lowest no-observed-adverse-effect level (NOAEL) was 2 g SAIB/kg body weight derived from the 1- and 2-yr chronic toxicity studies in the rat.
Subject(s)
Carcinogens/toxicity , Food Additives/toxicity , Neoplasms/chemically induced , Sucrose/analogs & derivatives , Animal Feed , Animals , Dose-Response Relationship, Drug , Female , Food Additives/administration & dosage , Male , Mice , No-Observed-Adverse-Effect Level , Rats , Rats, Inbred F344 , Sex Factors , Species Specificity , Sucrose/administration & dosage , Sucrose/metabolism , Sucrose/toxicity , Time Factors , Weight Gain/drug effectsABSTRACT
Sucrose acetate isobutyrate (SAIB) is used as an emulsion stabilizer in citrus-based soft drinks. A 4-week range-finding study and a 1-year chronic toxicity study were conducted to determine the tolerance and effects of this food additive in cynomolgus monkeys. SAIB was administered by gavage in corn oil solutions to groups of one monkey of each sex in the range-finding study and four monkeys of each sex in the 1-year study. The dose levels employed in both studies were 0, 500, 1450 and 2400 mg/kg body weight. Control monkeys were given corn oil by gavage. Based on the range-finding study, the 2400 mg/kg body weight dose level of SAIB was considered to be the highest dose of SAIB in corn oil that would be tolerated in a long-term study. No differences were observed between treated and control animals in either study with respect to body weight gain, clinical chemistry, haematology, organ weights, gross necropsy or light microscopy findings that could be attributed to SAIB treatment. Results of specific tests of hepatobiliary function in the 1-year study, including serum enzymes, bilirubin and bile acids, bromsulfophthalein retention and electron microscopic studies of the liver, were also negative. It was concluded that the highest dose level fed, 2400 mg/kg body weight, was the no-observed-adverse-effect level (NOAEL).
Subject(s)
Food Additives/toxicity , Liver/drug effects , Sucrose/analogs & derivatives , Animal Feed , Animals , Bile Acids and Salts/blood , Bilirubin/blood , Brain/drug effects , Dose-Response Relationship, Drug , Female , Food Additives/administration & dosage , Liver/enzymology , Liver/ultrastructure , Macaca fascicularis , Male , Microscopy, Electron , No-Observed-Adverse-Effect Level , Organ Size/drug effects , Ovary/drug effects , Sucrose/administration & dosage , Sucrose/toxicity , Time Factors , Weight Gain/drug effectsABSTRACT
A three-generation reproduction study of sucrose acetate isobutyrate (SAIB) in Fischer 344 rats and teratology studies in Fischer 344 rats and New Zealand white rabbits were performed. Dietary SAIB concentrations to provide dose levels of 0, 0.5, 1.0 and 2.0 g/kg body weight were used for the rat studies, and 0, 0.5, 0.85 and 1.2 g/kg body weight doses of SAIB in corn oil were administered by gavage in the rabbit studies. F0 generation male rats were fed SAIB for 10 wk, and female rats were fed SAIB for 2 wk prior to mating. F1 generation rats were raised on the test diets to maturity, mated to produce F2a litters, and remated to produce the F2b litters that were examined for teratology. F2a rats were mated to study fertility indices for the F3 pregnancy. A decrease in female fertility compared with controls was noted at the highest dose of SAIB during breeding of the F1 generation to produce the F2a litters. No difference in fertility rate between controls and treated animals was noted in the results of the other three matings that were performed, and it was concluded that the reduction in female fertility was not related to SAIB treatment. No morphological abnormalities of soft tissue or skeleton were observed in the rat or rabbit teratology studies. The highest dose levels administered, 2.0 g SAIB/kg body weight in the rat and 1.2 g SAIB/kg body weight in the rabbit, were considered to be no-observed-adverse effect levels (NOAEL).
Subject(s)
Food Additives/toxicity , Reproduction/drug effects , Sucrose/analogs & derivatives , Animal Feed , Animals , Dose-Response Relationship, Drug , Female , Fertility/drug effects , Litter Size/drug effects , Male , No-Observed-Adverse-Effect Level , Pregnancy , Rabbits , Rats , Rats, Inbred F344 , Species Specificity , Sucrose/administration & dosage , Sucrose/toxicityABSTRACT
A study of the effects of sucrose acetate isobutyrate (SAIB) ingestion was conducted in 13 male and 14 female healthy human volunteers. SAIB, in a gum arabic/water emulsion diluted with orange juice, was ingested once daily, at a dose of 20 mg SAIB/kg body weight in a total volume of 1.16 ml/kg body weight, for a period of 2 wk following a 1-week control period. During the control period, the subjects consumed the same preparation without SAIB. The study was performed in a single-blind manner, each subject serving as his or her own control. Haematology and clinical chemistry tests were conducted on blood samples taken on day -6 and day 0 of the control period and at 7 and 15 days during the SAIB dosing period. In addition to routine haematology and clinical chemistry, specific tests of hepatobiliary function included serum alkaline phosphatase, aspartate and alanine aminotransferases, lactic dehydrogenase, gamma-glutamyltransferase, total and direct bilirubin, bile acids and proteins. None of these parameters were affected by ingestion of SAIB. It was concluded that ingestion of 20 mg SAIB/kg body weight daily for 14 days does not affect the hepatobiliary function of human volunteers.
Subject(s)
Bile Acids and Salts/blood , Food Additives/adverse effects , Liver/drug effects , Sucrose/analogs & derivatives , Administration, Oral , Adolescent , Adult , Alanine Transaminase/blood , Alkaline Phosphatase/blood , Aspartate Aminotransferases/blood , Beverages , Bilirubin/blood , Citrus , Emulsions , Female , Food Additives/administration & dosage , Gum Arabic , Humans , L-Lactate Dehydrogenase/blood , Liver/enzymology , Male , Middle Aged , Single-Blind Method , Sucrose/administration & dosage , Sucrose/adverse effects , gamma-Glutamyltransferase/bloodABSTRACT
Concentrated organic residues extracted from 5 blended aliquots of commercial beers were evaluated for their ability to induce sister chromatid exchange (SCE), chromosomal aberrations and forward mutation in Chinese hamster ovary (CHO) cells. Each extract was prepared by blending 4 commercial beers of similar ingredients and brewing method, passing the beer pool over XAD-2 resin, extracting the resin and concentrating the extract. Studies were performed both with and without metabolic activation using variable amounts of reconstituted residues from 225-fold concentrates of the blended samples. CHO cultures were treated with 0.75 microliters/ml through 10.0 microliters/ml of the concentrates in the SCE assays, 1.0 microliters/ml through 10.0 microliters/ml of the extracts in the aberration assays and 2.5 microliters/ml up to 20 microliters/ml for forward mutation assays. In preliminary screening for SCE as an indicator of potential DNA damage, a significant increase was observed for 3 of 5 concentrated samples; however, no increase in SCE was induced by any of the 5 samples when S9 was added as a source of exogenous metabolic activation. More definitive tests for induction of genetic events, i.e., chromosome aberrations and forward HGPRT mutations, were negative for all 5 extracts whether or not S9 mix was present. Since SCE were not induced in tests with metabolic activation and since there was no concordant aberration or point mutation induction, the preliminary indication of potential DNA damage shown by elevated SCE under conditions without metabolic activation appears to have little biological significance.
Subject(s)
Beer/toxicity , Mutagens/toxicity , Sister Chromatid Exchange/drug effects , Animals , Biotransformation/genetics , CHO Cells , Cricetinae , Cricetulus , DNA Damage , Mutagenicity TestsABSTRACT
Dietary sodium saccharin is associated with bladder tumors when fed at high levels to the male rat. Under these conditions urinary pH, sodium concentration, and volume are elevated and proliferative changes are present in the urothelium. Extensive epidemiological studies have shown that saccharin does not increase the risk of bladder cancer in humans and laboratory investigations have shown that sodium saccharin is not mutagenic and does not bind to DNA. Recent research indicates that the urothelium in male rats is damaged under conditions of high urinary pH and sodium levels by a mechanism that involves alpha 2u-globulin and possibly silicate crystalluria. These studies and their implications for human health risk are reviewed.
Subject(s)
Saccharin/pharmacokinetics , Sodium/urine , Urinary Bladder Diseases/urine , Urinary Bladder Neoplasms/chemically induced , Urinary Bladder/drug effects , Animals , Humans , Hydrogen-Ion Concentration/drug effects , Male , Rats , Risk , Saccharin/chemistry , Urinary Bladder Calculi/chemistry , Urinary Bladder Calculi/complications , Urinary Bladder Diseases/chemically induced , Urinary Bladder Diseases/complicationsABSTRACT
Caramel Colour II is a distinct type of colourant with a pronounced reddish hue. It is made with sulphite reactants but without ammonia. The red colour and a high alcohol solubility provide functional characteristics that are important in foods or beverages containing natural flavour extractives. Caramel Colour II is widely used in ice creams and liqueurs; however, it represents less than 1% of total caramel colour manufacture. The toxicity of Caramel Colour II was evaluated in a 13-wk study in Fischer-344 (F344) rats. The test material was mixed with demineralized water and the solutions were given to the animals ad lib. in the drinking fluid. The concentrations of caramel colour in the drinking fluid were adjusted periodically to achieve the desired caramel colour intake/kg body weight/day. Groups of 20 rats/sex were given Caramel Colour II at levels of 0, 4, 8, 12 or 16 g/kg for at least 13 wk. There were no deaths in any of the groups fed Caramel Colour II. All rats fed caramel colour had soft faeces. All treated groups also had lower fluid consumption that was attributed to poor palatability of the high concentrations of caramel colour that were fed. A number of changes observed (reduced food consumption in all treatment groups except males given 4 g/kg; significantly lower body weights for males given 12 g/kg or more and for females given 8 g/kg or more; lower urine volume and higher specific gravity) were attributed to the reduced water intake and not considered to be toxicologically significant. There were no consistent treatment-related alterations in haematology or blood chemistry variables, and random changes noted were not associated with macroscopic or microscopic pathological alterations. There were no toxicologically important pathological findings. Based on this study, Caramel Colour II was not toxic in F344 rats treated for 13 wk. The highest dose level tested in this study (16 g/kg) was considered to be the no-observed-adverse-effect level.
Subject(s)
Food Coloring Agents/toxicity , Animals , Body Weight/drug effects , Candy , Carbohydrates , Color , Dose-Response Relationship, Drug , Drinking/drug effects , Eating/drug effects , Female , Intestines/drug effects , Kidney/drug effects , Male , Organic Chemicals , Rats , Rats, Inbred F344 , Solubility , Specific Pathogen-Free Organisms , UrineABSTRACT
Caramel colours used in the manufacture of a wide variety of foods and beverages have been an item of commerce for more than one hundred years. The regulatory history of these additives in the US, the UK, the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and the EC is reviewed, and an introduction to the safety studies of caramel colours in this issue of Food and Chemical Toxicology is provided.
Subject(s)
Food Coloring Agents/history , Legislation, Food/history , Candy , Carbohydrates , Food Coloring Agents/toxicity , History, 19th Century , History, 20th Century , Organic Chemicals , United Kingdom , United StatesABSTRACT
Caramel Colour III is used as a colour additive in beers and a variety of foods. Beer is the most important single source of Caramel Colour III in the diet although consumption of dark beers has been decreasing in recent years. The Joint FAO/WHO Expert Committee on Food Additives (JECFA) has established an acceptable daily intake of 200 mg/kg/day for Caramel Colour III. The safety of Caramel Colour III has been questioned during recent years following feeding studies in the rat that were associated with reduced white cell and lymphocyte counts. These effects have been attributed to the presence of 2-acetyl-4(5)-tetrahydroxybutylimidazole (THI) in this class of caramel colour. Short-term oral toxicity studies were conducted on low-THI and high-THI samples of Caramel Colour III (13 wk) and on a sample of THI (28 days). In both studies, the test materials were mixed with demineralized water and the solutions were given to the animals ad lib. in the drinking fluid. In the 13-wk subchronic toxicity study of Caramel Colour III, groups of 20 rats/sex were given concentrations of caramel colour equivalent to intakes of 0, 10, 15 or 20 g low-THI caramel colour/kg body weight/day or 20 g/kg of a high-THI caramel colour. In the 4-wk toxicity study with THI, groups of 20 rats/sex were given 0, 8 or 64 ppm THI (equivalent to approx. 0, 0.9 or 7.2 mg/kg/day) and 10 rats/sex were given 1, 2, 4, 16 or 32 ppm THI (equivalent to approx. 0.1, 0.2, 0.5, 1.9 or 3.7 mg/kg/day) for 4 wk followed by a 2-wk recovery phase for 10 rats/sex in the 0, 8 and 64 ppm groups. Rats given Caramel Colour III had soft faeces; there were no other treatment-related clinical observations and no treatment-related deaths occurred. All treated groups given Caramel Colour III had lower food and fluid consumption than controls. Males given 15 or 20 g low-THI caramel colour/kg or 20 g high-THI caramel colour/kg and females given 20 g/kg of either type had lower body weights than controls. In the 4-wk toxicity study with THI, there were no treatment-related ante-mortem observations, and no effects on body weights or food consumption. Fluid consumption by males and females treated with 64 ppm THI was lower than that of controls.(ABSTRACT TRUNCATED AT 400 WORDS)
Subject(s)
Food Coloring Agents/toxicity , Imidazoles/toxicity , Administration, Oral , Animals , Beverages , Body Weight/drug effects , Candy , Carbohydrates , Drinking/drug effects , Drug Combinations , Eating/drug effects , Female , Food Coloring Agents/administration & dosage , Imidazoles/administration & dosage , Leukocyte Count/drug effects , Leukocytes/drug effects , Lymphocytes/drug effects , Male , Organic Chemicals , Rats , Rats, Inbred F344ABSTRACT
Caramel Colour IV, a type of caramel colour used in the manufacture of cola soft drinks, was evaluated for subchronic and chronic toxicity in rats, and carcinogenicity in Fischer-344 (F344) rats and B6C3F1 mice. In each of the studies, Caramel Colour IV was mixed with demineralized water and the solutions given to the animals ad lib. in the drinking fluid. The concentrations of Caramel Colour IV in the drinking fluid were adjusted periodically to achieve the desired caramel colour intake per kg body weight. In the range-finding studies, groups of 30 rats/sex were given Caramel Colour IV at levels of 0, 15, 20, 25 or 30 g/kg for 13 wk, and groups of 10 male rats were given levels of 0, 2.5, 5, 10 or 15 g/kg for 6 wk followed, for some dose groups, by a 2-wk withdrawal period, and then re-initiation of dosing for another 2 wk. In the rat chronic toxicity study, levels of Caramel Colour IV of 0, 2.5, 5, 7.5 or 10 g/kg were given to groups of 25 rats/sex for 12 months. The test groups in the rat and mouse carcinogenicity studies were composed of 50 animals/sex and each species was given the caramel colour at levels of 0, 0, 2.5, 5 or 10 g/kg for 24 months. In each of the studies, treated animals tended to have dose-related lower water consumption than controls. This was attributed to poor palatability of the drinking fluid, and was generally associated with decreased food consumption and body weights. Rats given caramel colour often had soft or liquid malodorous faeces although there were no treatment-related ante-mortem observations in mice. Blood biochemical changes in the rat (i.e. reduced blood urea nitrogen, alkaline phosphatase and total serum protein) appeared to be related to dietary influences and were not considered toxicologically significant. There were no treatment-related alterations in haematological variables or treatment-related differences in survival or in the incidence of benign or malignant tumours among treated and control groups and no toxicologically important pathological findings. On the basis of these studies, Caramel Colour IV was not toxic or carcinogenic in F344 rats or B6C3F1 mice. The highest dose level tested in the long-term studies (10 g/kg) was considered to be the no-observed-adverse-effect level (NOAEL).
Subject(s)
Food Coloring Agents/toxicity , Neoplasms/chemically induced , Administration, Oral , Animals , Blood Proteins/analysis , Blood Urea Nitrogen , Body Weight/drug effects , Candy , Carbohydrates , Carcinogenicity Tests , Digestive System/drug effects , Drinking/drug effects , Eating/drug effects , Female , Food Coloring Agents/administration & dosage , Kidney/drug effects , Male , Mice , Organic Chemicals , Random Allocation , Rats , Rats, Inbred F344 , Urine/chemistryABSTRACT
Caramel Colour IV prepared from [U-14C]glucose was ultrafiltered in order to isolate the high molecular weight colour fraction (HMCF). The colour fraction that was non-permeable to a 10,000-Da porosity membrane, contained 84% of the colour, 22% of the solids and 24% of the radioactivity of the [14C]Caramel Colour IV. The absorption, distribution and excretion of [14C]HMCF were evaluated in male rats after administration of single or multiple oral doses of the material at a dosage level of 2.5 g/kg body weight. Rats on the multiple oral dosage regimen were given unlabelled HMCF in their drinking water for 13 days before the administration of a bolus dose of [14C]HMCF on day 14. On both dosage regimens, the predominant route of excretion was by way of the faeces. Less than 3% of the administered radioactivity was excreted in the urine and only a negligible amount was found in the expired air. More than 99% of the administered radioactivity was excreted within 96 hr. The principal tissues in which radioactivity was found were the mesenteric lymph nodes, liver, kidney and tissues of the gastro-intestinal tract. No major differences were observed in the absorption, distribution or excretion patterns between the single and multiple oral dose regimens.
Subject(s)
Food Coloring Agents/pharmacokinetics , Absorption , Administration, Oral , Animals , Candy , Carbohydrates , Drinking , Feces/chemistry , Food Coloring Agents/administration & dosage , Food Coloring Agents/analysis , Intestine, Large/metabolism , Kidney/metabolism , Liver/metabolism , Lymph Nodes/metabolism , Male , Mesentery , Organic Chemicals , Rats , Rats, Inbred F344 , Tissue DistributionABSTRACT
Sodium saccharin and sodium ascorbate are known to promote urinary bladder carcinogenesis in rats following initiation with N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) or N-butyl-N-(4-hydroxybutyl) nitrosamine. Sodium salts of other organic acids have also been shown to be bladder tumor promoters. In addition, these substances increase urothelial proliferation in short term assays in rats when fed at high doses. When they have been tested, the acid forms of these salts are without either promoting or cell proliferative inducing activity. The following experiment was designed to compare the tumor promoting activity of various forms of saccharin and to evaluate the role in promotion of urinary sodium, calcium, and pH as well as other factors. Twenty groups of 40 male F344 rats, 5 weeks of age, were fed either FANFT or control diet during a 6-week initiation phase followed by feeding of a test compound for 72 weeks in the second phase. The chemicals were administered to the first 18 groups in Agway Prolab 3200 diet and the last 2 groups were fed NIH-07 diet. The treatments were as follows: (a) FANFT----5% sodium saccharin (NaS); (b) FANFT----3% NaS; (c) FANFT----5.2% calcium saccharin (CaS); (d) FANFT----3.12% CaS; (e) FANFT----4.21% acid saccharin (S); (f) FANFT----2.53% S; (g) FANFT----5% sodium ascorbate; (h) FANFT----4.44% ascorbic acid; (i) FANFT----5% NaS plus 1.15% CaCO3; (j) FANFT----5.2% CaS plus 1.34% NaCl; (k) FANFT----5% NaS plus 1.23% NH4Cl; (l) FANFT----1.15% CaCO3; (m) FANFT----1.34% NaCl; (n) FANFT----control; (o) control----5% NaS; (p) control----5.2% CaS; (q) control----4.21% S; (r) Control----control; (s) FANFT----5% NaS (NIH-07 diet); (t) FANFT----control (NIH-07 diet). NaS, CaS and S without prior FANFT administration were without tumorigenic activity. NaS was found to have tumor promoting activity, showing a positive response at the 5 and 3% dose levels, with significantly greater activity at the higher dose. CaS had slight tumor promoting activity but without a dose response, and S showed no tumor promoting activity. In addition, NaCl showed weak tumor promoting activity, but CaCO3 was without activity. NH4Cl completely inhibited the tumor promoting activity of NaS when concurrently administered with it. NaCl administered with CaS or CaCO3 administered with NaS showed activity similar to that of NaS. Sodium ascorbate was also shown to have tumor promoting activity, with slightly less activity than NaS. Ascorbic acid showed no tumor promoting activity.(ABSTRACT TRUNCATED AT 400 WORDS)
Subject(s)
Ascorbic Acid/toxicity , Saccharin/toxicity , Urinary Bladder Neoplasms/chemically induced , Animals , Body Weight/drug effects , Butylhydroxybutylnitrosamine , Calcium Carbonate/toxicity , Diet , Drinking/drug effects , Drug Synergism , FANFT , Hydrogen-Ion Concentration , Kidney/drug effects , Male , Rats , Rats, Inbred F344 , Urinary Bladder/drug effects , Urinary Bladder/pathology , UrineABSTRACT
Three methods used to detect proliferative changes in the rat urothelium, light microscopy, scanning electron microscopy, and autoradiography, were compared for their sensitivity in detecting changes produced by administration of sodium saccharin. Weanling male F344 rats were fed sodium saccharin as 0, 3, 5, or 7.5% of the diet, and the bladders were evaluated after 4, 7, and 10 wks of feeding. Light microscopic changes and an increase in labeling index were seen at all time points in rats fed 7.5% sodium saccharin, but not at the lower doses. A slight increase in labeling index was also observed at 10 wks in the 5.0% group. Scanning electron microscopic changes were evident as early as 4 wks with increasing severity at the 3, 5, and 7.5% doses. This study demonstrates that the hyperplastic response of the urothelium to sodium saccharin administration varies with dose and time, and that observation by scanning electron microscopy is the most sensitive of the three methods evaluated for detecting these changes.
Subject(s)
Saccharin/pharmacology , Sodium/analysis , Urinary Bladder/cytology , Animals , Autoradiography , Cell Division/drug effects , Diet , Dose-Response Relationship, Drug , Male , Microscopy, Electron, Scanning , Rats , Rats, Inbred F344 , Saccharin/analysis , Urinary Bladder/drug effects , Urinary Bladder/ultrastructureABSTRACT
Commercial beer was subjected to an investigation in order to establish standard conditions for preparing organic solvent extracts to be used in short-term genetic screening assays. Test samples for use in the evaluation were prepared by mixing several brands of commercially available beer into a composite pool which was then spiked with the mutagen, 2-nitrofluorene. The composite sample was then concentrated using varying ratios of beer to XAD-2 resin in a 1.5 cm X 30 cm column. Dry-weight analyses indicated that significant amounts of residue could be trapped by XAD-2 resin. Columns were sequentially eluted by methylene chloride, acetone and methanol followed by evaporation of the solvents under nitrogen gas. Residues from commercial products were not mutagenic, but mutagenic activity could be detected in residues from spiked beer, yielding nearly 90% of the expected biological activity in S. typhimurium TA98. A standard method amenable to processing large volumes of beer products was devised for application to other projects.
Subject(s)
Beer/analysis , Mutagens/analysis , Fluorenes/isolation & purification , Fluorenes/pharmacology , Mutagenicity Tests , Salmonella typhimurium/drug effects , SolventsABSTRACT
5 concentrated extracts of commercial beers were prepared using XAD-2 resin. The residues were subjected to evaluation for mutagenic activity in Salmonella typhimurium strains TA98, TA100 and TA102. The tests were conducted using preincubation protocols including provisions for S9 metabolic activation. Although the extracts did produce moderate toxicity to the Salmonella organisms used in the assays, none of the residues were found to induce mutation up to their maximum testable concentrations.
Subject(s)
Beer/toxicity , Mutagens , Biotransformation , Mutagenicity Tests/methods , Salmonella typhimurium/drug effectsABSTRACT
The effect of vitamin E, folic acid, pyridoxine (vitamin B6) and choline on the reduction in circulating lymphocytes in the blood of rats fed Caramel Colour (III) also known as ammonia caramel colour (beer type; AC) has been examined. It was found that the reduction in the number of circulating lymphocytes in rats fed AC could be prevented by the addition of pyridoxine to the diet. The activity of AC in reducing the number of circulating lymphocytes also closely resembled that of the pyridoxine antagonist 4'-deoxypyridoxine. After the isolation and identification of 2-acetyl-4(5)-tetrahydroxybutylimidazole (THI), comparative studies indicated that THI was the component of AC responsible for reducing the number of circulating lymphocytes. Although the effect of AC was reduced or prevented by increasing dietary pyridoxine, the lymphocyte reduction associated with the administration of THI was not materially affected by the dietary level of pyridoxine.