Micronucleus formation in cultured fetal liver blood cells using mitomycin C.

Mouse fetal-liver blood cells were cultured and used to investigate micronucleus formation after exposure to mitomycin C (MMC). The isolated fetal cells were incubated in a medium supplemented with erythropoietin (EPO), and the frequency of micronuclei formation was detected in polychromatic erythrocytes (PCE). The effects of four variables were investigated: (1) MMC exposure dose, (2) MMC exposure time, (3) incubation time, and (4) EPO concentration. PCE were formed by proliferation and differentiation of the erythroid cells in culture. Micronucleated PCE (MNPCE) were observed in a dose-dependent manner after exposing the cultured cells with up to 1.0 microgram/ml MMC. The optimum time of MMC exposure and post-exposure incubation was 3 h and 48 h, respectively, and the optimum EPO concentration was 0.25 U/ml. Mouse fetal-liver PCE are sensitive primordial cell targets that can be obtained in relatively large numbers from a single pregnant animal. The procedure is relatively simple and potentially useful in detecting mutagens and carcinogens capable of causing chromosomal damage.

Determination of volatile and semivolatile mutagens in air using solid absorbents and supercritical fluid extraction.

Volatile toxicants may be present in emissions from mobile and stationary sources as well as in ambient air. Methods for collecting and concentrating volatiles from air samples have been developed. Solid-phase adsorbents were compared in their trapping efficiencies for dichloromethane (DCM), ethylene dibromide (EDB), 4-nitroblphenyl (4-NB), 2-nitrofluorene (2-NF), and fluoranthene (FI). Charcoal and Carbosieve were the most efficient media for retaining DCM, while XAD-4 was the best adsorbent for EDB and the aromatic compounds. Extraction of direct spikes of compounds from adsorbents using supercritical carbon dioxide resulted in greater than 90% recovery of EDB and 60-92% recovery of the aromatics. Integration of trapping and desorption methods with the Salmonella microsuspension bioassay was demonstrated with EDB and 4-NB recoveries from air; chemical analysis and bioassay gave comparable results (within 10%).