Importance of sampling, extraction and preservation for the quantitation of biologically active endoto

The influence of filter media, extraction solution and preservation method on detection of biologically active endotoxin in the LAL assay was studied with air samples collected from wastewater treatment plants. The four most common types of filters were used as collection media. The extraction solutions compared were nonpyrogenic water, KH2PO4-triethylamine and Trizma buffers. The effect of preservation on endotoxin air samples was ascertained by storing both the filters without extraction, and samples extracted in the collection day for a few weeks at various temperatures. Samples collected on glass fibre filters showed the highest amounts of detectable endotoxin, while the concentrations of endotoxin were significantly lower when cellulose-mixed esters, polycarbonate or polyvinyl chloride membrane filters were used for air sampling. After collection, the best efficiency for glass fibre filters was attained by extraction with nonpyrogenic water within 8 hours after sampling and storage of the extracts at 4 degrees C until they were analysed. If the filters were stored without extraction, the reduction in endotoxin levels of the sample was about 30% after 1 week preservation and about 70% after 2 weeks. The study shows that the effect of the filter material and preservation practice was significant. These factors play critical roles in assessing exposure to bacterial endotoxins within wastewater aerosols.