Document Detail

Evaluation of amino sugar, low molecular peptide and amino acid impurities of biotechnologically produced amino acids by means of CE.
MedLine Citation:
PMID:  12008126     Owner:  NLM     Status:  MEDLINE    
As the second part of our studies on the impurity profiles of amino acids produced by biotechnological processes, a micellar electrokinetic chromatography (MEKC) was applied to determine amino sugars, low molecular peptides and amino acids as potential expected impurities at a level 0.1% w/w. 3-(4-Carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) was found to be suitable as a labeling reagent for laser induced fluorescence (LIF) detection. The labeling reaction was optimized to achieve maximum reaction yields and reproducibility for all groups of substances. The 'level 0.1%' was chosen as quantities/concentrations in such a way that the major reagent peak becomes comparable with the impurities at this level, and the minor reagent peaks (<0.01%) are not able to cover other peaks at the level of interest. The reaction optimization and the validation studies were performed with model mixtures of representative amino sugars, low molecular peptides and amino acids. A linearity range of labeling of three orders of magnitude was achieved. The total precision of the method, including the labeling reproducibility was studied and for all test substances relative standard deviation less than 5% were obtained. The accuracy was evaluated by performing recovery experiments at three concentrations covered a 50-200% interval of the level 0.1%. Confidence intervals below +/-2.5% (lambda=0.05, n=9) of the target were found sufficient for purity tests.
Nikolai Novatchev; Ulrike Holzgrabe
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of pharmaceutical and biomedical analysis     Volume:  28     ISSN:  0731-7085     ISO Abbreviation:  J Pharm Biomed Anal     Publication Date:  2002 May 
Date Detail:
Created Date:  2002-05-14     Completed Date:  2002-07-22     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8309336     Medline TA:  J Pharm Biomed Anal     Country:  England    
Other Details:
Languages:  eng     Pagination:  475-86     Citation Subset:  IM    
Institute of Pharmacy and Food Chemistry, University of Würzburg, Am Hubland, 97074, Würzburg, Germany.
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MeSH Terms
Amino Acids / analysis*,  isolation & purification*
Amino Sugars / analysis*
Electrophoresis, Capillary
Fluorescent Dyes
Hydrogen-Ion Concentration
Indicators and Reagents
Peptides / analysis*
Reproducibility of Results
Reg. No./Substance:
0/Amino Acids; 0/Amino Sugars; 0/Benzoates; 0/Buffers; 0/Fluorescent Dyes; 0/Indicators and Reagents; 0/Peptides; 0/Phosphates; 0/Quinolines; 131124-59-9/3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde

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