Document Detail

Determination of tryptophan as the reduced derivative by acid hydrolysis and chromatography.
MedLine Citation:
PMID:  6528998     Owner:  NLM     Status:  MEDLINE    
A new procedure for the analyses of tryptophan and the total amino acid composition of proteins was based on the observations that pyridine borane reduces tryptophan in trifluoroacetic acid, while other amino acids remain intact [M. Kurata, Y. Kikugawa, T. Kuwae, I. Koyama, and T. Takagi (1980) Chem. Pharm. Bull. 28, 2274-2275; W.S.D. Wong, D.T. Osuga, and R.E. Feeney (1984) Anal. Biochem. 139, 58-67]. Concentrated HCl was used instead of trifluoroacetic acid for analytical purposes. The products were stable to hydrolysis in 6 N HCl, and the reduction did not interfere with hydrolysis and subsequent analyses. Quantitative recovery was achieved with most proteins when they were subjected to acid reduction in ice-cooled concentrated HCl with two incremental additions of pyridine borane. The reaction was terminated after 10 min by dilution with an equal volume of H2O, vacuum sealing, and hydrolyzing at 110 degrees C for 22 h. The yields of the expected values for cytochrome c, catalase, bovine serum albumin, subtilisin BPN', trypsin, chymotrypsin, beta-lactoglobulin, lysozyme, and pepsin were obtained. Ovotransferrin and ovalbumin, however, yielded values for tryptophan lower than literature values. With two different ion-exchange methods, the recoveries of all other amino acids were comparable to those obtained by acid hydrolysis with 6 N HCl. Since the same hydrolysate can be analyzed for both tryptophan and all the other amino acids, the procedure is a more convenient method than those requiring separate determinations. Initial results indicate that the method may be applied to high-performance liquid chromatographic procedures with adaptations of the protocols if necessary.
W S Wong; D T Osuga; T S Burcham; R E Feeney
Related Documents :
2391248 - A kit for labelling erythrocytes or leukocytes with technetium-99m.
21535788 - Effect of salicylic acid treatment on postharvest quality, antioxidant activities, and ...
7348198 - Laboratory method for measuring small amounts of tissue glycogen.
15501668 - The use of smoke acid as an alternative coagulating agent for natural rubber sheets' pr...
15952798 - Site-directed mutagenesis and use of bile acid-mts conjugates to probe the role of cyst...
15284388 - Conjugated linoleic acid isomers and trans fatty acids inhibit fatty acid transport in ...
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Analytical biochemistry     Volume:  143     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1984 Nov 
Date Detail:
Created Date:  1985-03-27     Completed Date:  1985-03-27     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  62-70     Citation Subset:  IM    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Amino Acids / analysis*
Chemical Phenomena
Chromatography, High Pressure Liquid
Chromatography, Thin Layer
Proteins / analysis*
Sulfhydryl Compounds
Tryptophan / analogs & derivatives,  analysis*
Grant Support
Reg. No./Substance:
0/Amino Acids; 0/Proteins; 0/Sulfhydryl Compounds; 73-22-3/Tryptophan; 7536-97-2/2,3-dihydrotryptophan

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  An enzymatic assay for activity of lipoprotein lipase.
Next Document:  Triple-column ion-exchange physiological amino acid analysis with fluorescent detection: baseline ch...