Contamination of nanoparticles by endotoxin: evaluation of different test methods

纳米粒子的内毒素污染:不同测试方法的评估

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作者:Stijn Smulders, Jean-Pierre Kaiser, Stefano Zuin, Kirsten L Van Landuyt, Luana Golanski, Jeroen Vanoirbeek, Peter Wick, Peter Hm Hoet

Background

Nanomaterials can be contaminated with endotoxin (lipopolysaccharides, LPS) during production or handling. In this study, we searched for a convenient in vitro method to evaluate endotoxin contamination in nanoparticle samples. We assessed the reliability of the commonly used limulus amebocyte lysate (LAL) assay and an alternative method based on toll-like receptor (TLR) 4 reporter cells when applied with particles (TiO(2), Ag, CaCO(3) and SiO(2)), or after extraction of the endotoxin as described in the ISO norm 29701.

Conclusion

We demonstrated that nanoparticles can interfere with endotoxin detection systems indicating that a convenient test method must be chosen before assessing endotoxin contamination in nanoparticle samples.

Results

Our results indicate that the gel clot LAL assay is easily disturbed in the presence of nanoparticles; and that the endotoxin extraction protocol is not suitable at high particle concentrations. The chromogenic-based LAL endotoxin detection systems (chromogenic LAL assay and Endosafe-PTS), and the TLR4 reporter cells were not significantly perturbed.

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