Densitometry for analysis of protein and peptide hydrolysates: application to luteinizing hormone releasing hormone.
Densitometric analysis has been performed on various standard protein and peptide solutions and on luteinizing hormone releasing hormone hydrolysate. After thin-layer chromatographic separation using two-dimensional separation on cellulose plates, the plates were mapped and each amino acid was assayed individually in order to obtain a specific integration profile of the hydrolysate. Densitometry proved to be a useful method for the screening of protein hydrolysates and is accurate enough for the assay of peptide hydrolysates. The hormone was hydrolysed with hydrochloric acid-trifluoroacetic acid (1:1, v/v) in the presence of thioglycolic acid. This limits the loss of tryptophan and does not interfere with the analysis. Results were sufficiently reliable, accurate and reproducible for routine analysis.
Barthomeuf
C
C
Laboratoire de Pharmacognosie et Biotechnologie, Faculté de Pharmacie, Université de Clermont-I, Clermont-Ferrand, France.
Pourrat
H
H
Regerat
F
F
D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003720:Densitometry; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D010455:Peptides / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*
D010455:Peptides; D011506:Proteins; D007987:Gonadotropin-Releasing Hormone