This article reviews the present status of knowledges with respect to the chemical mutagens in the environment. Literatures on environmental mutagens were surveyed with positive results noted in natural products, food additives, smoking, food contaminants and chemicals resulted from environmental pollutants. Test systems detecting these chemical mutagens are summarized, and the advantage and limitation of its relevant use for the detection method of chemical carcinogens are discussed.

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Sub-nanogram amount of benzo (a) pyrene (B (a) P) was determined in food stuffs by means of high performance liquid chromatography (HPLC) equipped with a fluorescence detector. Food stuffs were extracted and cleaned up by conventional saponification and dimethylsulfoxide partition before subjecting to chromatography. A chromatographic condition was compromised for routine analysis by using octadecyl silate-silica gel micro beads as a support and acetonitrile + water (70 : 30) as a solvent at a flow rate of 2 ml/min. The best set of excitation and emission wavelengths for fluorometry was 384 and 406 nm, respectively. The method provided 0.01 ppb level of detection limit and a mean recovery (70%) with high reproducibility. Examination of Japanese food stuffs revealed that B (a) P was ubiquitously distributed in most of the stuffs and highly present in special food such as dried bonito. Benzo (k) fluoranthene, benzo (b) fluoranthene, benzo (e) pyrene and perylene were also analysed by HPLC with specific excitation and emission wavelengths.

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We have devised a simple and rapid method for analysing polynuclear aromatic hydrocarbons (PAH) in river sediment. This method consists of the following 4 procedures ; drying of a sediment sample, ultrasonic extraction of PAH from the dry sample, separation of PAH into each component by one dimensional dual band thin-layer chromatography (TLC), and identification and determination of PAH by spectrofluorometry. A sediment is passed through a screen of 28 mesh and then dried. PAH in the dry sample are extracted with a mixed solution of ethanol and benzene by ultrasonification. After centrifugation, a known volume of the extracted solution are evaporated just before dryness under a reduced pressure, and dried up by standing it in a room for 10-20 min. The residue is dissolved in benzene, and subjected to TLC. [Thin-layer plate : Kieselguhr G (5×20, cm)-(26% Acetylated cellulose + Avicel SF) (95 : 5, w/w, 15×20, cm). Developer : Ethanol-Methanol-Ether-Water (2 : 2 : 4 : 1, v/v). Detection : Fluorescence under UV ray.]. Each spot on the TLC plate is scrapped off into a small centrifuge tube, and extracted with dimethylsulfoxide by ultrasonification. After centrifugation, PAH in the solution are identified by comparison of the excitation and emission spectra of the solution with those of a PAH reference solution. Quantitative determination of PAH is done spectrofluorometrically with the narrow base line method. In this method, 7 PAH [benzo [a] pyrene, chrysene, benzo [k] fluoranthene, benz [a] anthracene, perylene, pyrene and benzo [ghi] perylene] in a river sediment can be analysed in a short time. Recoveries of PAH in this method were good. Furthermore, it was proved that PAH contents in several sediments determined by this method agreed well with those by soxhlet extraction instead of the ultrasonic extraction.

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Growth of B. megaterium QM B 1551 in Schaeffer's medium was inhibited by more than 5 μM cadmium chloride added to the medium. Cells could not grow in the presence of 50 μM cadmium chloride. The inhibition of growth of cells by cadmium chloride was suppressed by the addition of manganese chloride. Cadmium chloride was incorporated into cells. On the other hand the content of manganese in cells decreased. A large portion of cadmium of cells existed in the soluble fraction of cell free extracts. The greater part of cadmium was bound to proteins whose molecular weights were more than 200000. Activity of phosphoglycerate mutase obtained from cells grown with cadmium chloride was less than that of cells without cadmium chloride. It is discussed that there may be some relationship among a decrease in manganese content of cells, phosphoglycerate mutase activity and inhibition of growth of cells by cadmium chloride.

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The distribution of polychlorinated naphthalenes (PCN) in several organs and tissues of rats after the single oral administration (20 mg/kg) was investigated. The maximum levels of PCN in the liver, kidney, spleen, brain and blood appeared within 3 hr after the administration, and the levels decreased with time remarkably. The organs containing PCN at the highest level was kidney through which PCN might be excreted. At a 24-hr period after the administration, the order of PCN concentration in the tissues was kidney>lung>spleen>brain and liver, and it was kept at least within 5 days. That PCN was detected at the highest concentration in the kidney and the lowest in the liver was markedly different from the distribution of polychlorinated biphenyls reported already. The residue of PCN in adipose tissues at a 15-day period after the administration was detected at extremely low levels, which were higher than those in the organs. The numbers of chlorine substituted in the molecules were not related to the degree of the accumulation to tissues.

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Antisera to optical isomers of methamphetamine were produced in rabbits immunized with d-, dl- or l-N-(4-aminobutyl) methamphetamine conjugated to bovine serum albumin. The specificity of antibodies obtained was shown by cross-reaction studies with the optical isomers of methamphetamine. The results indicated that the antiserum to d- or l-methamphetamine was specific to d- or l-methamphetamine, respectively, although the antiserum to dl-methamphetamine recognized both d- and l-methamphetamine to almost the same degree.

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An anion exchange high speed liquid chromatographic method fof the determination of glycyrrhizinate in various foods is described. The chromatograph was equipped with a Zipax SAX column and an Ultraviolet (254 nm) detector. The mobile phase was 0.05 M NaClO₄-0.05 M phosphate buffer (pH 5.4). Liquid foods were diluted with water. Solid foods were homogenized with water-ethanol (1 : 1) and filtered, and the filtrate were evaporated to approximately 1/4 volume and diluted with water to a suitable volume. Each solution was filtered through a membrane filter and injected into the chromatograph. However, in soy sauce, soybean paste and salted radish ("Takuan"), the aqueous solutions obtained were moreover cleaned up with a polyamide column before injection. The recoveries from these foods fortified at a level of 200 ppm were about 98% by this method.

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A method was described for the quantitative determination of diethylstilbestrol (DES) in cosmetics. The DES was extracted from samples with dichloromethane, the extract was evaporated, the residue was placed on a silicagel column, and the column was washed with benzene to remove impurities. DES was eluted from the silicagel column with ethyl acetate and determined by high-speed liquid chromatography on a zorbax SIL column, by using ethanol-hexane as eluting solvent and a 240 nm UV monitor for detection. A calibration curve showed linearity in the range of 0.05-0.5 μg of DES, and the detection limit was 0.01 μg. Average recoveries of 100 μg DES added to 5 g samples of skin cream and hair tonic were 90.6 % and 92.1 %, respectively.

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Methyl-, ethyl-, n-propyl- and n-butyl p-hydroxybenzoates (parabens) are used as preservatives in pharmaceutical capsules. In this paper, paraben contents in capsules of commercial pharmaceuticals were examined. Aqueous solution of the sample was extracted with ether, then a portion of the evaporated extract was injected into high-performance liquid chromatograph on a column (30 cm×4.0 mm) of μ-Bondapak C₁₈, with water-methanol mixture (1 : 2, v/v) as mobile phase and finally detected at 254 nm. Also a portion of the evaporated extract was qualitatively analized by gas chromatography on a Silicone GE SE-30 column (1.5 m×3 mm). From 50% of hard capsule samples and 93% of soft capsule samples, parabens were detected. Average and standard deviation of total paraben contents were 0.0761% and 0.0827%, respectively in hard capsules. On the other hand, they were 0.0578% and 0.0601%, respectively in soft capsules. From some of the same pharmaceutical products, various contents of parabens were detected. The practical implications of these investigations were discussed.

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