Robert S.U. Baker

Effect of pH on chromium(VI) species in solution.

Published values of equilibrium constants were used to calculate the percentage of each chromium(VI) species (CrO(4)(2-), Cr(2)O(7)(2-), HCrO(4)(-) and H(2)CrO(4)) present in aqueous solution at total chromium(VI) concentrations of 10(-2)-10(-6)M in the pH range 1-8.

Mutagenicity of metal ions in bacteria

The mutagenicity of 24 metal salts was investigated in plate incorporation and fluctuation assays with Salmonella TA strains or Escherichia coli WP2 uvrA pKm 101. Chromate(VI) and selenate(VI) ions were found to be mutagenic in plate incorporation assays employing conventional media. On the other hand, cadmium(II), beryllium(II), chromate(VI), and metavanadate(V) ions were detected in conventional fluctuation assays, indicating the importance of this technique in detection of metal mutagens. Modified culture media, with trimetaphosphate ions in place of orthophosphate as the sole phosphate source for bacterial growth, were also used in this study. The media modifications prevented precipitation of metals such as nickel and cadmium as their insoluble phosphates, and allowed detection of the mutagenicity of metavanadate ions in plate incorporation assays. However, the fluctuation technique using standard media was shown to detect a wider range of mutagenic metal ions than tests with modified media. It is notable that metaarsenite(III), arsenate(V), and nickel(II) ions were not found to be mutagenic in any of the assays although they are known to be carcinogenic and are mutagenic in other test systems. Their lack of mutagenicity in the modified media indicates that precipitation of these ions as orthophosphates is not the reason for their lack of activity in standard bacterial assays.

Application of X-ray photoelectron spectroscopy to the analysis of stainless-steel welding aerosols

Aerosol particles (“fume”) from manual metal arc welding of stainless steel with E316L-16 electrodes were analysed by X-ray photoelectron spectroscopy. The inherent complexity of the particles required the use of a wide range of experimental techniques. These included IR spectrophotometry, TGA/DTA, XRF, XRD, AAS and electron microprobe analysis. The surface of the fume particles comprised ≈ 50 at% NaF and KF, ≈ 8 at.% soluble (probably K) chromate, ≈ 30 at% SiO2 and several at.% transition-metal oxides, hydroxides or silicates. The fluorides and chromates were removed by washing to reveal a surface which was predominantly SiO2 (≈ 60 at%) with the remainder comprising of transition-metal oxides, silicates and fluorides. Approximately 6 at% F remained on the surface of the water-washed particles, presumably as transition-metal fluoro-complexes. The water-soluble fraction of the fume contained K+, Na+, F− and CrO2−4 ions in the mole ratio 5:5:4:3. When aerosol particles are deposited in lung tissues, water-soluble constituents would be expected to dissolve rapidly. In view of the suspected carcinogenicity of stainless steel welding fume, a bio-medical study of the combined effects of F− and CrO2−4 ions on lung tissue is warranted.

Mutagenicity of arylmethane dyes in Salmonella

22 arylmethane dyes which have been used as food colours, commercial dyes, laboratory stains and pH indicators were tested in the Salmonella/mammalian microsome mutagenicity assay. 8 mutagenic dyes were identified, including 5 food colours and 3 common laboratory strains; none of the 11 indicator dyes tested was mutagenic. The commercial and laboratory dyes Methyl Violet 2B C.I. 42535 and Crystal Violet C.I. 42555 were mutagenic in base-pair substitution mutation detector strain TA1535 in the absence of metabolic activation. By contrast, the food colours Benzyl Violet 4B C.I. 42640, Guinea Green B.C.I. 42085, Light Green SF C.I. 42095, Lissamine Green B C.I. 44090 and Violet BNP C.I. 42581 and the bacteriological stain, Basic Fuchsin C.I. 42500-42510, were all mutagenic in frameshift mutation detector strains TA98 and/or TA1538 and required metabolic activation. Most of these compounds gave weak mutagenic responses with Salmonella and were positive only within narrow dose ranges. Since conflicting results were obtained using dyes from different sources, minor dye components may have been responsible for their mutagenicity. This suggests the need to improve knowledge about impurities in arylmethane colours still used in food and to review the toxicological role of such impurities.

Comparison of rat and guinea pig as sources of the S9 fraction in the salmonella/mammalian microsome mutagenicity test

A highly significant enhancement of mutagenicity occurs with 11 polycyclic aromatic hydrocarbons when 3-methylcholanthrene-induced guinea pig liver S9 is substituted for Aroclor-induced rat liver S9 in the Ames test. The use of MC-induced guinea pig liver S9 is particularly valuable for detecting the weak mutagenicity of benz[c]acridine, which is barely positive in a standard Ames assay. However, anthracene and phenanthrene, which are generally considered not to be carcinogens, remain non-mutagenic for strain TA100. This enhancement of mutagenicity does not correlate with arylhydrocarbon hydroxylase activities of the various liver preparations and does not apply to certain other non-PAH mutagens, including beta-naphthylamine, aflatoxin B1 and 4-dimethylaminoazobenzene.

Evaluation of metals in in vitro assays, interpretation of data and possible mechanisms of action†

Metal compounds have been investigated in a wide range of in vitro assays for their potential genotoxicity. While inter‐laboratory variability could be responsible for some lack of agreement between different laboratories using the same test methods, there appears to be consistency of results where compounds have been examined with a number of different assay methods, particularly those employing eukaryotic cells. A major barrier for proper evaluation of metals in vitro has been the lack of agreement between eukaryotic and prokaryotic assays. Compounds of nickel, cadmium and beryllium have failed repeatedly to be detected in the widely used Salmonella assay, and yet are active in diverse mammalian cell assay systems. From previous studies with organic chemical mutagens, these major differences in sensitivity of test systems are entirely unexpected, and may be due to a combination of factors: different sensitivity of the various genetic end‐points; variations in physiological state of target cells; variati...

Sensitivity of two Chinese hamster cell lines to SCE induction by a variety of chemical mutagens

SCE induction in Chinese hamster Don (lung) cells was compared with that in CHO (ovary) cells exposed under identical conditions to 14 known mutagens. Test protocols used for comparison were selected following a study of Don and CHO cell responses to aflatoxin B1 and benzo[a]pyrene. In the absence of added metabolizing enzymes 9-aminoacridine, 4-nitroquinoline 1-oxide, N-methyl-N-nitrosourea, dimethylcarbamoyl chloride, beta-propiolactone, daunomycin, aflatoxin B1 and 2-aminoanthracene were directly active in both cell lines; every substance positive in CHO cells was also positive in Don cells. However, the latter detected cyclophosphamide, hydrazine sulphate, benz[c]acridine, 3-methylcholanthrene and benzo[a]pyrene without addition of S9. CHO cells did not respond equivalently to these mutagens, either in the presence or absence of S9. Other differences between the cell lines depended on chemical exposure time, S9 pre-incubation or co-incubation conditions. For example, the ability of CHO cells to detect SCEs due to 2-aminoanthracene was acutely dependent on exposure time. In addition, Don cells exhibited lower background SCE values which were less variable than those of CHO cells under the same culture conditions. Although incapable of detecting 4-dimethylaminoazobenzene (butter yellow) and not as sensitive to cyclophosphamide as certain cell lines of liver origin, the pseudodiploid Don cell line possesses other desirable characteristics required for in vitro SCE assays, particularly with regard to intrinsic metabolic activation of polycyclic aromatic hydrocarbons and related substances.

Metabolism of the carcinogen 7-methylbenz[c]-acridine in the rat

The biliary excretion of radioactivity by adult Wistar rats given i.v. 7-methyl-[7-14C]benz[c]acridine(14C-7-MBAC) and [methyl-3H]-7-methylbenz[c]acridine (3H-7-MBAC) (2 mg/kg) was 61% and 48%, respectively, in males in six hours. Females excreted 33% of a 2 mg/kg dose of 3H-7-MBAC in the same time-period. For male rats, the urinary and faecal excretions were about 10% and 61% of the dose of 14C-7-MBAC, respectively, in seven days. No enterohepatic circulation could be demonstrated in control male rats. The biliary excretion of radioactivity by phenobarbital- and 3-methylcholanthrene-induced male rats given 14C-7-MBAC was similar to or greater than that of control male rats. The organo-soluble biliary metabolites after beta-glucuronidase/arylsulphatase hydrolysis were separated by h.p.l.c., and quantitative metabolite distributions were obtained for induced and control rats by comparison with metabolite standards. The mutagenicity of bile from carcinogen-dosed control rats was greater than that of equivalent bile from carcinogen-dosed 3-methylcholanthrene-pretreated animals.

Mutagenic activity of heated potato/oil systems

Mutagens detected with Salmonella typhimurium strain TA 98 in the presence of liver S9 mix were extracted from potato slices, but not pure potato starch, after frying in oil. No mutagenic activity was detected using strain TA 100, in the presence or absence of S9 mix with either fried potato slices or potato starch. Mutagenic activity was detected at frying temperatures of 140 degrees C and above. The mutagenic activity was limited to the outer portion of the fried potato slices and increased with frying time and temperature. Mutagenic activity ratios for extraction with both (NH4)2SO4/NH4OH and Na2SO4/NaOH were similar.

Effect of induction by DDT on metabolism and mutagenicity of benzo[a]pyrene

Liver microsomal enzymes are essential for the detection of benzo[a]pyrene (B[a]P)-mediated mutagenesis in the Salmonella/mammalian microsome mutagenicity test and, furthermore, this mutagenicity is considerably enhanced by induction of hepatic enzymes involved with drug metabolism. Although Aroclor 1254 is most commonly used for induction of S9 enzymes, DDT is also capable of this induction. This paper reports a comparison of liver S9 fraction induced by the two agents: there is a marked difference in their concentration optima for metabolism of B[a]P; greater numbers of revertant colonies are seen with Aroclor-induced S9, which is optimal at a concentration of 10% (v/v), whereas DDT-induced S9 is optimal at 2.5% (v/v); Aroclor induces aryl hydrocarbon hydroxylase (AHH), cytochrome P-450 and epoxide hydrase while DDT induces only AHH, to about half the level detected in the Aroclor-induced S9 fraction. A comparison of metabolite distribution for Aroclor- and DDT-induced hepatic microsomes reveals quantitative differences only. DDT-induced microsomes yield a greater proportion of B[a]P-4,5-oxide and its metabolic product B[a]P-4,5-dihydrodiol than do Aroclor-induced microsomes. Time course studies on the mutagen half-life measured on the agar plate provides good evidence that metabolites responsible for mutagenicity were different for each inducer.