RESUMO
The capability of pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) for the direct analysis of endotoxins is demonstrated in this research article using the lipopolysaccharides of Pseudomonas aeruginosa 10. Analytical methods based on evolved gas analysis-MS, single-shot (SS) Py-GC-MS, and multishot heart cut Py-GC-MS were investigated. Among the various methods developed, the SS Py-GC-MS method shows superior potential for identifying bacterial endotoxins effectively through their biomarkers. The results obtained were validated with conventional mass spectral analysis after hydrolysis. The method was also evaluated for its robustness based on quality control criteria indicated by the U.S. EPA Method 8270D. When applied onto endotoxins of different Gram-negative bacteria, this method produced vastly distinct pyrograms. The results show that rapid and sensitive direct detection of endotoxins is possible with the Py-GC-MS method developed.
RESUMO
A significant portion of the scientific effort has been devoted to the detection of endotoxins in pharmaceutical solutions, as they pose major health threats as contaminants even in minute amounts. Conventional methods based on the biological response of endotoxins have been well-established, but as technology advances, many limitations surfaced in the recent years. As a result, information obtained by chemical analytical methods becomes valuable in crossvalidating these results. In addition to providing an overview on the main strategies for detecting the presence of endotoxins, the biological methods are compared with the chemical techniques. The review also investigates future advances aimed toward a more accurate, reliable, and convenient endotoxin test.