抄録
Separation behavior of polynuclear aromatic hydrocarbons (PAH) in a dual band thin-layer chromatography (TLC) and in a high performance liquid chromatography (HPLC) was studied in order to develop a new analytical method which could also supply the separated PAH samples for a mutation assay or a physico-chemical measurement. PAH was separated on the TLC plate [kieselguhr layer (5×20 cm)-(26% acetylated cellulose+cellulose Avicel SF) (95: 5 w/w) layer (15×20 cm)] with methanol-ether-water (4: 4: 1 v/v) as a developer. Development was continued until the solvent front reach the 10 cm mark on the latter layer of the TLC plate. Developing time was about 60 min. Separation behavior of PAH in HPLC was studied in the following two chromato-systems; Nucleosil 7C18 column (35 mm×4.6 mm, i.d. +250 mm×4.6 mm i.d.)-acetonitrile-water (6: 4 -9: 1 v/v) system, and Nucleosil 5NO2 column (35 mm×4.6 mm i.d. +250 mm×4.6 mm i.d.)-n-hexanebenzene (5: 5 -95: 5 v/v) system.
Separation pattern of PAH was fairly good as compared with that in an ordinary TLC and several carcinogens were easily separated each other. Relative Rf values of PAH were stable and highly reproducible in this TLC. In HPLC, retention time of PAH decreased with the rise of column temperature. There was a linear relation between the logarithm of the retention time and the reciprocal of absolute temperature of the column. Retention time of PAH decreased also logarithmically with the increment of acetonitrile content of the acetonitrilewater mobil phase. Linear relation was observed among the carbon number of PAH and the logarithms oftheir retention times.
Most of 48 PAHs tested could easily separated each other by the combination use of the TLC and the HPLC using Nucleosil 7C18 column.
PAH which were unable to separate each other in this method could be easily identified by spectrofluorometry. It was proved that this method was useful for analyzing PAH in airborne particulates and also for mutagenicity assay of the separated PAH.
