The LAL assay typically used for endotoxin detection depends on the purified portion of the blood of the horseshoe crab to coagulate in the presence of minute amounts of bacterial endotoxins. There are numerous challenges associated with LAL assay and other current endotoxin detection methods such as ; issue of low endotoxin recovery (LER) which results in false negatives, automation of testing among others.
This technology focuses on a method of detecting and quantifying endotoxins using CE-LIF and solves most of the concerns relating to current endotoxin detection methods. The established CE-LIF analytical method provides a simple, efficient and reliable solution for the analysis of endotoxin concentrations in water, especially in cases the water matrix is very complex (e.g. raw water). This method also involves a simple and low cost endotoxin extraction method for reliable, robust analysis and interference removal to overcome the concern of low endotoxin recovery. Sample preparation can be automated to conserve manpower for other critical operations. By coupling CE-LIF analysis and sample extraction method, this invention can be used to monitor the endotoxin concentrations in both simple water matrices (i.e. drinking water and medical solutions) and complex water matrices (i.e. reservoir water and waste water) without using animal resources.
This technology relates to a method of labeling and concentrating endotoxins in water and finally using CE-LIF to detect and quantify endotoxins directly from water samples. The method has been used for the analysis of endotoxins - lipopolysaccharides (LPS) - from 16 different bacterial strains. LPS was labelled with the amino-reactive fluorescent dye, separated by capillary zone electrophoresis (CZE) under the optimized conditions and detected by LIF detector.
To remove possible interference materials in sample matrix and enrich the LPS, solid phase extraction (SPE) method using an extraction kit for LPS extraction from water samples was developed. The limits of detection (LOD) can reach ng/mL level. The method had been applied to the determination of LPS in pond water samples.
This technology can be used to detect and quantify endotoxins in drinking water, reservoirs or waste water or in the detection of endotoxins in the final product of pharmaceutical industries. This technology will be particularly attractive for complex water samples because of the ease in isolating the endotoxins from the rest of the interfering substances.