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Endotoxins

endotoxinstesting

Endotoxins

What is an endotoxin?

  • Found in the outer membrane of Gram-negative bacteria like E. coli.
  • They’re heat-stable and not destroyed by typical sterilization like autoclaving.
  • Even tiny amounts can trigger fever, inflammation, or endotoxin shock in animals or humans.

Why test peptides for endotoxins?

  • During peptide synthesis, handling, or lyophilization, bacterial contamination can occur.
  • For research peptides, endotoxin testing ensures the material is free from these toxic residues, especially if used in in vivo research models.
  • Regulatory labs (like 503B pharmacies) have strict limits—often < 0.25 EU/mL for injectable solutions.

Testing methods

  1. LAL Assay (Limulus Amebocyte Lysate)
    • Uses enzymes from horseshoe crab blood that clot in the presence of endotoxins.
    • Can be gel-clot, turbidimetric, or chromogenic (color-change) formats.
    • Most common in both research and pharmaceutical testing.
  2. Recombinant Factor C (rFC) Assay
    • Synthetic alternative to LAL—no animal harvesting.
    • Fluorescent readout; same sensitivity.

Result format

  • Reported in Endotoxin Units (EU) per milliliter or per milligram of sample.
  • Example: “< 0.05 EU/mg” means the peptide meets extremely low endotoxin limits.