Douwes J, Versloot P, Hollander A, Heederik D, Doekes G
Department of Epidemiology and Public Health, Agricultural University Wageningen, The Netherlands.
Appl Environ Microbiol. 1995 May;61(5):1763-9. doi: 10.1128/aem.61.5.1763-1769.1995.
The influence of various filter types and extraction conditions on the quantitation of airborne endotoxin with the Limulus amebocyte lysate test was studied by using airborne dusts sampled in a potato processing plant. Samples were collected with an apparatus designed to provide parallel samples. Data from the parallel-sampling experiment were statistically evaluated by using analysis of variance. In addition, the influence of storage conditions on the detectable endotoxin concentration was investigated by using commercially available lipopolysaccharides (LPS) and endotoxin-containing house dust extracts. The endotoxin extraction efficiency of 0.05% Tween 20 in pyrogen-free water was seven times higher than that of pyrogen-free water only. Two-times-greater amounts of endotoxin were extracted from glass fiber, Teflon, and polycarbonate filters than from cellulose ester filters. The temperature and shaking intensity during extraction were not related to the extraction efficiency. Repeated freeze (-20 degrees C)-and-thaw cycles with commercial LPS reconstituted in pyrogen-free water had a dramatic effect on the detectable endotoxin level. A 25% loss in endotoxin activity per freeze-thaw cycle was observed. Storage of LPS samples for a period of 1 year at 7 degrees C had no effect on the endotoxin level. House dust extracts showed a decrease of about 20% in the endotoxin level after they had been frozen and thawed for a second time. The use of different container materials (borosilicate glass, "soft" glass, and polypropylene) did not result in different endotoxin levels. This study indicates that the assessment of endotoxin exposure may differ considerably between groups when different sampling, extraction, and storage procedures are employed.
利用在一家马铃薯加工厂采集的空气尘埃,研究了各种滤器类型和提取条件对采用鲎试剂法进行空气中内毒素定量的影响。样品通过一种旨在提供平行样品的装置进行采集。采用方差分析对平行采样实验的数据进行统计学评估。此外,利用市售脂多糖(LPS)和含内毒素的室内灰尘提取物,研究了储存条件对可检测内毒素浓度的影响。无热原水中0.05%吐温20的内毒素提取效率比仅用无热原水时高7倍。从玻璃纤维、聚四氟乙烯和聚碳酸酯滤器中提取的内毒素量是从醋酸纤维素滤器中提取量的两倍。提取过程中的温度和振荡强度与提取效率无关。用无热原水复溶的市售LPS反复冻融(-20℃)对可检测的内毒素水平有显著影响。观察到每冻融循环内毒素活性损失25%。LPS样品在7℃下储存1年对内毒素水平无影响。室内灰尘提取物在第二次冻融后内毒素水平下降约20%。使用不同的容器材料(硼硅酸盐玻璃、“软”玻璃和聚丙烯)不会导致内毒素水平不同。本研究表明,当采用不同的采样、提取和储存程序时,不同组之间内毒素暴露的评估可能存在很大差异。