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We have already reported the effect of ultraviolet-A (UVA)-sensitization of hematoporphyrin (HP) on the production of lipid hydroperoxides in erythrocytes and on their hemolysis. In this report, we investigated these effects under ultraviolet-B (UVB) irradiation and compared the results with those under UVA irradiation. It was found that an increase in lipid hydroperoxide preceded hemolysis, and that the UVB irradiation resulted in an increase in phosphatidylethanolamine hydroperoxide (PEOOH) and 2-thiobarbituric acid reactive substances (TBA-RS) at an earlier stage in comparison with UVA irradiation. Under either UVA or UVB irradiation, hemolysis was inhibited by anti-oxidants such as sodium azide and ascorbic acid (singlet oxygen scavengers), but not by mannitol, sorbitol (hydroxyl radical scavengers) or superoxide disumutase (SOD) (superoxide radical scavenger). These results suggest that singlet oxygen (1O2) produced by UV irradiation peroxidized the lipids of erythrocyte membranes, and therefore, the hemolysis of erythrocytes occured when the amount of hydroperoxides increased to a constant level, but auto-oxidation of lipids did not affect the hemolysis.

To examine biological effects of UVA, we investigated effects of UVA-sensitization with hematoporphyrin (HP) or riboflavin (RF) on the hemolysis and lipid hydroperoxide levels of rabbit erythrocytes. We irradiated UVA (365 nm) at r.t. for certain time, and after the end of irradiation measured hemolysis keeping erythrocytes at r.t. or at 0°C. As the result, though erythrocytes hemolyzed almost 100% at r.t., hemolysis was almost 0% at 0°C. It is evident that HP was a much stronger sensitizer than RF. Moreover, we measured contents of hydroperoxide of phosphatidylcholine (PC) and phosphatidylethanolamine (PE), most of which constructed lipid cellular membrane, after or before UVA irradiation by the chemiluminescence-HPLC method. As the result, before hemolysis, hydroperoxide levels per lipid significantly increased after the end of irradiation, compared with control (1.75→4.57 nmol/mg PC. 6.13→12.9 nmol/mg PE, respectively). These results suggest that celluar membrane lipid hydroperoxide which was formed with singlet oxygen produced by UVA sensitization, might be concerned with hemolysis.

A simple specific determination method was developed for ethylene oxide (EO) and ethylene chlorohydrin (ECH) in cosmetics and surfactants. EO is desorbed from samples by using a nitrogen stream and absorbed into acidic potassium iodide solution, where it is converted to ethylene iodohydrin (EIH). Any remaining ECH in the samples is converted to EO by the addition of sodium hydroxide, and the desorption procedure is repeated with a fresh acidic potassium iodide absorbing solution. EIH is extracted with benzene and determined by gas chromatography with electron capture detection. EO and ECH contents in the samples are calculated from EIH results. Recoveries from water and shampoo samples were 70.3 ± 5.4 and 58.9 ± 1.2%, respectively, for EO and 66.3 ± 4.0 and 64.5 ± 4.6%, respectively, for ECH. Detection limits in 0.2-2.0 g samples were in the 0.005-0.03 μg/g range for EO and 0.01-0.07 μg/g for ECH. High levels of EO (30-394 μg/g as ECH) were found in 5 of 18 polyoxyethylated surfactant samples, but only small amounts (0.07-4.0 μg/g) of ECH were detected in the samples. EO was not detected in cosmetic samples tested, but ECH was present in small quantities (≤1.11 μg/g).

Formaldehyde (FA) donor type preservatives (Donor) Dowicil-200 and Glydant XL-1000 could be analyzed for free FA in them by HPLC-Post Column Acetyl Acetone method (HPLC-PC method) using ODS column1), however, a separation of FA and Germall 115 was not adequate. We developed HPLC-PC analytical method employing carbon column (mobile phase; 6mM Na2HPO4(PH 2.1)) that achieved an excellent separation, and it could also be applied to other Donors. During the stability studies on the Donor using the Acetyl Acetone Colorimetry (AA method)2) and HPLC-PC method, both methods showed relatively good accordance. Therefore, it is concluded that the AA method could be practical, easy and rapid determination method for FA in cosmetics when the shelf life of the cosmetic products is taken into accounts, although the HPLC-PC method is still the most appropriate method for determination of free FA in cosmetics.

Cosmetics are regulated by the pharmaceutical affairs law in our country. In this review, we describe the definition of cosmetics from the standpoint of that law, and also mention the following matters from the view-point of hygienics. 1) Efficacy of cosmetics : evaluation methods by means of dermatological approach, and the problems for further discussion. 2) Safety of cosmetics : test methods, and information in the literature. 3) Microbiology for cosmetics : test methods, and preventive measures for microbiological contamination. 4) Percutaneous absorption for cosmetics : outline, and observations. 5) Analytical chemistry for cosmetics : analytical instruments, and their applications.

The structure of Pigment A obtained in the Voges-Proskauer reaction was determined as 2-(4-methyl-2-morpholino-1 H-5-imidazolylmethylidene)-1, 2-dihydro-1-oxonaphthalene, by the X-ray analysis of Pigment A', a heavy atom derivative of Pigment A. Pigment A' crystallizes in the triclinic system, space group P-1, with unit-cell dimensions a=8.08A, b=14.63A, c=7.43A, α=101.5°, β=97.2°, γ=92.1°, Z=2. The structure was solved by the heavy atom method and refined by the least-squares method to an R value of 0.065.

工場排水中の塩素の定量法を検討した結果,酸性で通気し,塩素を揮散させる方法で良好な結果を得た.定量操作法は試料溶液(50ml中約2mgの塩素を含む)を硫酸でpH1.0にして通気しながら40分間40℃の水浴中で塩素を揮散させ,これを0.00564Nチオ硫酸ナトリウム液に吸収させる.これをヨウ素滴定法によって定量すると,97%の定量値を得た.本操作法によって,有色物質,混濁物質や鉄,マンガン,銅イオンなどの金属イオンおよび有機窒素化合物などの妨害物質に影響されず塩素を定量することができる.なお直接ヨウ素滴定法は,試料がほとんど澄明で,妨害物質があまり含まれていないときに用いる方法であるが,銅イオンは特にその定量に影響を与えるので,この場合には陽イオン交換樹脂,またはcyclohexanediamine tetraacetic acidを用いて銅イオンの妨害を除くことができた.

The structure of Pigment A obtained in the Voges-Proskauer reaction was determined as 2-(4-methyl-2-morpholino-1 H-5-imidazolylmethylidene)-1, 2-dihydro-1-oxonaphthalene, by the X-ray analysis of Pigment A', a heavy atom derivative of Pigment A. Pigment A' crystallizes in the triclinic system, space group P-1, with unit-cell dimensions a=8.08A, b=14.63A, c=7.43A, α=101.5°, β=97.2°, γ=92.1°, Z=2. The structure was solved by the heavy atom method and refined by the least-squares method to an R value of 0.065.