Influence of sample preservation on endotoxin measurement in water.
Appropriate preservation of a range of water types prior to analysis for endotoxin was investigated, including sample storage and addition of sodium thiosulphate. Biologically active endotoxin in water samples was assayed using a chromogenic Limulus Amoebocyte Lysate (LAL) assay. Statistical analysis of measured mean endotoxin levels obtained for samples with and without sodium thiosulphate showed no significant difference in results. There was a 44% mean decline in the concentration of detectable endotoxin in water samples stored at -80 degrees C for 4 weeks compared with samples stored at 4 degrees C and analysed within 24 hours. Freezing of water samples at -80 degrees C in pyrogen-free containers for 4 weeks or longer, then thawing may lead to considerable endotoxin loss; however the addition of sodium thiosulphate to water samples interferes minimally with the LAL assay. These results provide methodological information that can be used to assist researchers in future water endotoxin monitoring studies. The validation and standardisation of water sample preservation protocols are necessary, given the likely increase in the quantification of endotoxin levels in a variety of water sources and the use of such results for health effect determinations.
O'Toole
J
J
Department of Epidemiology and Preventive Medicine, Monash University Australia, The Alfred, Commercial Rd, Melbourne 3004, Australia. joanne.o'toole@med.monash.edu.au
Sinclair
M
M
Jeavons
T
T
Leder
K
K
D053000:Analytic Sample Preparation Methods; D000818:Animals; D004731:Endotoxins / Q000032:analysis* / Q000378:metabolism; D005615:Freezing; D006737:Horseshoe Crabs; D008565:Membrane Proteins / Q000378:metabolism; D013696:Temperature; D013885:Thiosulfates / Q000737:chemistry; D013997:Time Factors; D014867:Water / Q000737:chemistry*
D004731:Endotoxins; D008565:Membrane Proteins; D013885:Thiosulfates; C025357:endotoxin binding proteins; D014867:Water; C017717:sodium thiosulfate