METHOD FOR EVALUATION OF ENDOTOXIN LEVEL IN HEMOGLOBIN SOLUTION

摘要

A procedure for endotoxin level evaluation in the hemoglobin solution by means of application LAL kinetic-turbidimetric method at the wave length of 650 nm and 660 nm by use of adequate filters and adequate modifications of hemoglobin sample of 10%w/v. Hemoglobin solution of 10%w/v is diluted up to the concentracion of 10mg/ml and then incubed in the temperature range of 60-800C/10-20 minutes. Incubed solution is diluted up to the concentracion of 2,5 mg/ml and then tested by application of LAL kinetic-turbidimetric method on the wave lengths of 650 nm and 660 nm with application of the appropriate filters. Endotoxin concentrations are read from the standard curve obtained for LAL kinetic-turbodimetric method on the wave-lengths from 650 nm to 660 nm. The test is valid if the value of the positive check of the product is in range of 50-200% in comparison to the pure sample, and the hemoglobin solution is apyrogenic if the endotoxin level read at the sample is 0,595EU/ml (5,95EU/g hemoglobine).;Postupak za odredjivanje nivoa endotoksina u rastvoru hemoglobina koji obuhvata primenu LAL kinetiko-turbidimetrijske metode na talasnoj duini od 650nm i 660 nm korienjem odgovarajuih filtera i odgovarajuih modifikacija uzorka 10%w/v hemoglobina. 10%w/v rastvor hemoglobina je razblaen do koncentracije 10mg/ml i potom inkubiran u temperaturnom opsegu 60-80В°C/10-20 minuta. Inkubirani rastvor je razblaen dokoncentracije 2,5 mg/ml i potom testiran primenom LAL kinetiko-turbidimetrijske metode na talasnim duinama od 650 nm i 660 nm uz primenu odgovarajuih filtera. Koncentracije endotoksina su oitavane sa standardne krive koja je dobijena za LAL- kinetiko-turbidimetrijsku metodu na talasnim duinama od 650nm i 660nm. Test je validan ako je vrednost pozitivne kontrole proizvoda u opsegu 50-200% u odnosu na ist uzorak, a rastvor hemoglobina je apirogen ako je nivo endotoksina detektovan u uzorku 0,595EU/ml (5,95EU/g hemoglobina).[A procedure for endotoxin level evaluation in the hemoglobin solution by means of application LAL kinetic-turbidimetric method at the wave length of 650 nm and 660 nm by use of adequate filters and adequate modifications of hemoglobin sample of 10%w/v. Hemoglobin solution of 10%w/v is diluted up to the concentracion of 10mg/ml and then incubed in the temperature range of 60-800C/10-20 minutes. Incubed solution is diluted up to the concentracion of 2,5 mg/ml and then tested by application of LAL kinetic-turbidimetric method on the wave lengths of 650 nm and 660 nm with application of the appropriate filters. Endotoxin concentrations are read from the standard curve obtained for LAL kinetic-turbodimetric method on the wave-lengths from 650 nm to 660 nm. The test is valid if the value of the positive check of the product is in range of 50-200% in comparison to the pure sample, and the hemoglobin solution is apyrogenic if the endotoxin level read at the sample is 0,595EU/ml (5,95EU/g hemoglobine).