DETERMINATION OF ENDOTOXIN IN SMALL-VOLUME PARENTERALS (SVPs): A COMPARISON OF LAL GEL-CLOT METHOD WITH U.S.P. RABBIT PYROGEN TEST
Abstract
The simplicity and sensitivity of the Limulus Amebocyte Lysate (LAL) gel-clot method in detection of endotoxin in Small Volume Parenterals (SVPs) were studied and compared with rabbit pyrogen test.
All formulations were artificially spiked with 0.25, 0.5 and 5EU/ml of Escherichia coli 0111:B4 endotoxin (Iambda=0.5EU/ml). The Threshold Pyrogenic Dose (TPD) for E.coli 0111 :B4 endotoxin was found to be more than lng per kg of body weight (0.5EU/ml). However, the sensitivity of LAL test was found to be 0.25 Eu/ml for water for injection and 0.5 EU/ml for other formulations. The observed inhibitory effect of electrolyte containing solutions was deleted by some degree of dilution prior to test. However, none of the samples showed inhibitory at the Maximum Valid Dilution (MVD). The U.S.P. rabbit pyrogen test was insignificant for all the solutions tested. These results suggest, LAL gel-clot test to be useful for determination of bacterial endotoxin as final release test in SVPs. A comparison of the pyrogenicity assays also indicates, that LAL is more sensitive and simple and less time-consuming than the conventional rabbit test.
All formulations were artificially spiked with 0.25, 0.5 and 5EU/ml of Escherichia coli 0111:B4 endotoxin (Iambda=0.5EU/ml). The Threshold Pyrogenic Dose (TPD) for E.coli 0111 :B4 endotoxin was found to be more than lng per kg of body weight (0.5EU/ml). However, the sensitivity of LAL test was found to be 0.25 Eu/ml for water for injection and 0.5 EU/ml for other formulations. The observed inhibitory effect of electrolyte containing solutions was deleted by some degree of dilution prior to test. However, none of the samples showed inhibitory at the Maximum Valid Dilution (MVD). The U.S.P. rabbit pyrogen test was insignificant for all the solutions tested. These results suggest, LAL gel-clot test to be useful for determination of bacterial endotoxin as final release test in SVPs. A comparison of the pyrogenicity assays also indicates, that LAL is more sensitive and simple and less time-consuming than the conventional rabbit test.
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