Method for determining endotoxin concentration
First Claim
1. In an optical method for determining endotoxin wherein a sample thought to contain endotoxin is contacted with an endotoxin-coagulable protein from Limulus amebocytes to form a reaction product which contains coagulated protein, the improvement comprising contacting the reaction product with a sufficient amount of an ionic surface active suspending agent to dissolve said coagulated protein.
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Abstract
Improvements in the sensitivity and reproducibility of nephelometric endotoxin determinations using endotoxin-activated enzyme from Limulus amebocyte lysate are achieved by contacting endotoxin-coagulated lysate with an ionic surface active suspending agent, generally a mixture of detergent and suspending agent. Instrumentation improvements further enhance results.
25 Citations
31 Claims
- 1. In an optical method for determining endotoxin wherein a sample thought to contain endotoxin is contacted with an endotoxin-coagulable protein from Limulus amebocytes to form a reaction product which contains coagulated protein, the improvement comprising contacting the reaction product with a sufficient amount of an ionic surface active suspending agent to dissolve said coagulated protein.
- 5. In a nephelometric method for determining endotoxin wherein a sample thought to contain endotoxin is contacted with an endotoxin-coagulable protein from Limulus amebocytes to form a reaction product containing coagulated protein, the improvement comprising contacting the reaction product with a sufficient amount of an ionic detergent and a suspending agent to dissolve said coagulated protein.
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30. In a nephelometric method for determining endotoxin wherein a sample thought to contain endotoxin is contacted with an endotoxin-coagulable protein from Limulus amebocytes to form a reaction product containing coagulated protein, the improvement comprising mechanically disrupting said coagulated protein in said product and contacting said disrupted protein with a sufficient amount of a suspending agent to yield a concentration of greater than about 0.01% by weight in said product, with the proviso that said product is not contacted with a detergent.
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31. In an optical method for determining endotoxin wherein a sample thought to contain endotoxin is contacted with an endotoxin-coagulable protein from Limulus amebocytes to form a reaction product which contains coagulated protein, and then the light scattering or absorbence of a suspension of such reaction product determined, the improvement comprising contacting the reaction product with a sufficient amount of ionic surface active suspending agent to dissolve said coagulated protein.
Specification