Chromogenic Substrate for Amidolytic Test
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This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition
Issued and maintained by the United States Pharmacopeial Convention (USP)
—Synthetic molecules consisting of tripeptides or tetrapeptides coupled to a chromophore. The terminal Amino acid is specific for the protease utilized. The synthetic peptides mimic the peptide sequence (specific to the activated coagulation factor) of the active site on the natural substrate. The coagulation factor catalyzes the splitting of the chromophore (p-nitroaniline) from the peptide. The amount of release can be measured directly in a spectrophotometer, because the maximum absorbance spectra of the bound and free chromophores differ. The released chromophore is a colored compound; the complete substrate itself is colorless.
Molecular weights of the various substrates range from about 600 to 750 Da. Solubility in aqueous solutions can vary. Not all substrates are of equal sensitivity, and incubation periods may have to be extended.
