The Food Sanitation Law established in 1947 was amended in May, 1995. It passed more than 20 years since the last amendment in 1982, and changes of circumstances about foods necessitated the amendment. One of the biggest change was a change of the eating habits and another was increase of quantity and kinds of imported foods. There also was an increase of people's interest in health, and many people have had concern about their foods. Compared with the old law, the most important and remarkable change in the new law was shown in the article concerning the so-called"natural food additives". Namely, it becomes necessary to be permitted by the Ministry of Health and Welfare to use new"natural food additives"after the law becomes effective. According to the newly amended law food additives are divided into the following four categories : "permitted additives"(including both chemical synthetics and natural food additives permitted after the amendment of the law), "existing natural food additives, ""natural perfumes"and"additives usually used as foods."A list for"existing natural food additives"and lists of examples of"origin of natural perfumes"and"additives usually used as foods"have published. All additives included in these categories need labeling, except in the case otherwise specified, when they are used in foods.

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Three chlorhexidine degradation intermediates, named CHDI-B, CHDI-BR and CHDI-D, were purified from the incubation mixture for chlorhexidine degradation by Pseudomonas sp. strain No. A-3. The structures of these compounds were determined by spectroscopic analyses of ¹H-NMR, ¹³ C-NMR and mass spectroscopy. CHDI-B has a structure in which 1 mol of 1-carboxyethyl is substituted at imine in the guanidyl group adjacent to the hexane moiety in chlorhexidine. CHDI-BR has a structure in which the carboxylic-OH in the 1-carboxyethyl moiety in CHDI-B condensed with imine in the adjacent guanidyl group to form a ring. CHDI-D has a structure in which 2 mol of 1-carboxyethyl are substituted at two imine in guanidyl groups adjacent to the hexane moiety in chlorhexidine.

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We examined the effect of N-phenylalkylimidazoles (chain length of 1, 2, 3, 4 and 5 carbon atoms) on the induction of hepatic and renal microsomal cytochrome P450 (P450) content and hepatic P450 species (CYP1A1, CYP1A2, CYP2C6, CYP2E1 and CYP3A2) in male and female rats. Treatment of rats with N-phenylalkylimidazoles resulted in the high magnitude induction of hepatic P450 ; especially, 1-benzylimidazole and 1-(3-phenylpropyl) imidazole were shown to be potent inducers of P450 in male and female rats, respectively. The extent of hepatic P450 induction after treatment with N-phenylalkylimidazoles having an even number of carbon atoms, such as 1-(2-phenylethyl) imidazole, was less than that produced by the compounds with odd carbon numbers (1 or 3 carbon atoms) in both sexes. Thus, the induced levels of hepatic P450 were decreased with increasing methylene bridge except for 1-(2-phenylethyl) imidazole treatment. However, we did not observe such a chain-length differential induction of renal P450 ; 1-(2-phenylethyl) imidazole and 1-benzylimidazole were effective inducers of renal P450 in male and female rats, respectively. Aniline p-hydroxylase activity in male rat liver microsomes was parallel to the increase of P450 content, although N-demethylase activity was not. Immunoblot analysis revealed that the increase in CYP1A2, CYP3A2 and CYP2E1 after treatment with N-phenylalkylimidazoles was parallel to that in total P450 content, but induced levels of CYP2C6 increased in a chain-length dependent manner in both sexes. In addition, induced levels of CYP1A1 after treatment with N-phenylalkylimidazoles were very faint, irrespective of their marked increase in P450 content. These results clearly indicate that phenylalkyl-substituted imidazole compounds cause chain length-and gender-related differential induction of P450 species and hepatic drug-metabolizing enzymes in rats.

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The effect of several antipyretic analgesics on mitomycin C-induced mutagenesis was investigated using the wing spot test in Drosophila melanogaster. Aspirin, ibuprofen and indomethacin showed strong antimutagenicity, while the antimutagenicity of acetaminophen and ethenzamide were weaker. Aspirin inhibited chromosome recombination. A considerable part of the antimutagenicity of aspirin, ibuprofen and indomethacin can be explained by bioantimutagenicity.

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The efficiency of the equipments on the market for ozone formation and decomposition by ultraviolet rays in the closed room was measured and assessed. There were two stick-type low pressure ultraviolet quartz lamps in the equipment. Lamps were placed in parallel with each other in the box, and the air was discharged after being sucked by a built-in fan. First, the only ozone formation lamp was lit. As the air containing some ozone was absorbed into the testing room (4.5m³) having no ozone, the variation of ozone concentration was measured every one minute. Second, the only ozone decomposition lamp was lit and the air reaching a given ozone concentration was discharged after being sucked in, and then, the variation of the residual ozone concentration was measured. In addition, the equipment was stopped as the ozone concentration reached the plateau level, and then the velocity of the spontaneous ozone decomposition was measured. As a result of the above-mentioned experiment, some equations, figures, and tables concerning the mutual relations among the number of ozone and UV lamps, the initial ozone concentration, the residual ozone concentration, the concentration reaching the steady state level, and the beginning or the finishing time of reactions, etc. were obtained. The above-mentioned data gave us a clue to the control of the ozone concentration in the closed room.

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The efficiency of commercially available equipments for ozonization by ultraviolet rays in the rooms was measured and assessed. The volume of the room was enlarged to 4.5m³ and to 54m³, and then, the tests were held using three types of ozonization equipments. By considering the data including the results of the tests in the small room, the formulas and the line graphs on the relationships between the room volume and the ozone concentration of the steady state, and those between the ozone concentration of the steady state and the input current of a lamp were obtained. As the result of this experiment, a new phenomenon was found out, that is, the apparent velocity of ozone photo-decomposition by an ultraviolet ray lamp having a constant initial current was slower than that of the spontaneous decompositions, when the room volume was enlarged beyond a certain degree. This phenomenon was named an inversion volume. The overall assessments of these three kinds of equipments were as follows : 1) In the case of the ozone formation those equipments are useful, because they are efficient enough to obtain the ozone concentration of the steady state within an hour, though the time consumed to achieve it varies according to its room volume. 2) In the case of the ozone decomposition in the ordinary laboratory, as far as the decomposition velocity is concerned, setting up one ultraviolet ray lamp and one ozone lamp can be less effective than those of the spontaneous decomposition. Therefore, to increase the decomposition velocity, it is necessary to increase the input current of one ultraviolet ray lamp, or to set up the lamps in parallel. 3) In the extremely clean room, such as a bioclean room, the ozone decomposition by ultraviolet ray (using 140W big type of double wall) is thought to be useful. Lastly, the efficiency assessments and the comparisons of all the equipments (including the following system) measured in the past was conducted, and therefore, the one in which the greatest improvement could be made was found.

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A simple, rapid and sensitive screening method and semiquantitative analysis for abused drugs, methamphetamine (MA), morphine (MO), cocaine (COC) and cannabinoid (CN), in the urine sample were investigated usine the Syva Emit st^TM urine assay reagent with Shimadzu CL-770, the temperature-controlled spectrophotometer. A sample was diluted and mixed in an Emit st^TM urine reagent vial with an Emit st^TM dilutor. Thirty seconds after mixing, the mixture was transferred to the flow cell of the spectrophotometer kept at 37°C, and the absorbance change at 349 nm was measured for 30 s (60 s for CN). Calibration curves were examined over a range of 0 to 50 μg/ml for MA, morphine-3-glucuronide (M3G) and benzoylecgonine (BE), 0 to 0.5 μg/ml for Δ⁹-tetrahydrocannabinol-11-oic acid (THC-COOH), and were found to be relatively linear between 0 and 0.5 μg/ml for MA and M3G, 0 and 1 μg/ml for BE, 0 and 0.3 μg/ml for THC-COOH. Cutoff values were able to be set at the concentration of 0.5 μg/ml for MA, 0.5 μg/ml for M3G (corresponding to 0.2 μg/ml), 0.3 μg/ml for BE, 0.1 μg/ml for THC-COOH, and within-run coefficients of variation (C. V.) for the samples of the cutoff levels were less than 2% (n=10). The quantitative results from enzyme multiplied immunoassay technique (EMIT) assays for 100 urine samples were compared with those from GC, GC/MS and LC/MS, and the former showed an excellent correlation with the latter for MA and BE levels. Dihydrocodeine, however, showed a high cross-reactivity with MO, which often caused "false positive results."

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We investigated an extracting method of morphine having rapidity, low cost, simplicity and high recovery. The samples containing morphine were dehydrated by adding anhydrous sodium sulfate and extracted by dichloromethane with shaking in the presence of anhydrous sodium sulfate. Morphine could be extracted by the dehydrating action of anhydrous sodium sulfate more efficiently as compared with the liquid-liquid extraction method using dichloromethane. After derivatization with pentafluoropropionic anhydride, morphine was determined with gas chromatography/mass spectrometry (GC/MS) by the selected ion monitoring (SIM) mode. Ethylmorphine was used as an internal standard. The recoveries from 3 ml of urine and hydrolyzed urine containing 830.1ng of morphine were 93.6% and 94.7%, respectively. The recoveries from 3 ml of blood and hydrolyzed blood containing 207.6 ng of morphine were 76.9% and 93.9%, respectively. The standard deviations (n=5) of the recoveries from urine, hydrolyzed urine, blood and hydrolyzed blood were 3.6%, 4.1%, 6.6% and 2.3%, respectively.

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We performed in vitro screening of human and bovine placenta extracts (PE) used as an active component in cosmetics. Evaluation of the cell proliferative action and determination of the sialic acid content and alkaline-phosphatase (ALP) activity showed marked differences among PE samples, but no association among the results was observed. The stability of L-ascorbic acid phosphate magnesium salt (VC-PMg) used for whitening in cosmetics was also examined by PE. VC-PMg was hydrolyzed to L-ascorbic acid following the pseudo-first-order reaction by PE showing ALP activity. PE with high ALP activity required only a short time to degrade VC-PMg by 50%. Each PE sample differed in its efficacy as a cosmetic material. Attention should be given paid to its combination with VC-PMg as a whitening agent.

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To investigate if K-ras point mutations could occur at an early stage of urethane-induced lung tumorigenesis in mice, we applied a sensitive mutation specific PCR assay technique (MSPA) to DNA from lung at the early stage of this condition. At 7 or 14 d after urethane injection (750 mg/kg, i. p.), pulmonary DNA was isolated and the existence of activated K-ras gene with 61st codon A→T mutation was checked according to the MSPA. This technique can detect mutant alleles in the presence of a very large excess of normal ras alleles ; however, the K-ras point mutation could not be detected by the present MSPA. These results suggest that the K-ras point mutations did not occur at the stage of lung tumorigenesis in mice or that the number of mutant alleles was below the detectable range of the technique.

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A long-term (1970-1994) study of atmospheric aerosol has been conducted in Kitakyushu City. The annual mean concentrations of aerosol had gradually decreased until 1983 and they have been stable since then. To investigate the change of the aerosol quality, the total mass and constituents of aerosols collected for 1972-1974 (period I : heavily polluted period) and 1984-1988 (period II : improved period) were compared. The decrease rate (%) of all constituents (inorganic ions : SO₄^2-, NO₃^-, F^-, Cl^-, Ca²⁺ and Mg²⁺ ; metals : Fe, Mn, Zn, V and Pb) except SO₄^2- was much larger than that of the aerosol amount. The concentration of NO₃^- in the aerosol in period II was 54% of that in period I. It was considered that this result was based on the decrease (period II/period I : 45%) of atmospheric NO_x, which was supposed to be a precursor. In contrast, there was a nonproportional relationship between the decrease (88%) of SO₄^2- in the aerosol and the decrease (20%) of atmospheric SO₂. These results show the characteristic of the change of aerozol in Kitakyushu City for 24 years (1970-1994).

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A total of 471 samples of livestock and fishery products were analyzed for antibacterial residues. Samples were collected in Osaka from 1990 to 1994. Livestock products were analyzed for five kinds of sulfa drugs, oxolinic acid, furazolidone, thiamphenicol, decoquinate and amprolium. Cultured fishes were analyzed for three kinds of sulfa drugs, oxolinic acid, sodium nifurstyrenate and furazolidone. Antibacterial residues were detected in two out of 471 examined samples. Sulfadimidine was detected in one (0.04 mg/kg) of 44 samples of imported pork. Oxolinic acid was detected in one (0.03 mg/kg) of 16 samples of imported eel. Antibacterial agents were not detected in pork (domestic), beef, chicken, duck, chicken egg, sweet fish, rainbow trout, yellow tail, sea breams, bastard halibut, flatfish, horse mackerel, puffer and prawn.

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