Document Detail


In-capillary determination of creatinine with electrophoretically mediated microanalysis: characterization of the effects of reagent zone and buffer conditions.
MedLine Citation:
PMID:  19046749     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Previous work has demonstrated proof-of-concept for carrying out the clinically useful Jaffe reaction between creatinine and picrate within a capillary tube using electrophoretically mediated microanalysis (EMMA). Here, it is shown that careful control of reagent plug length as well as concentration and pH of the background electrolyte (BGE) can result in a marked improvement in the sensitivity of this assay. Increasing the length of the picrate reagent zone is shown to give rise to as much as a 3-4-fold enhancement, and increasing the concentration and/or pH of the borate buffer also results in an additional, albeit modest, improvement in sensitivity. Interestingly, borate BGE concentrations approaching 100mM give rise to an unexplained drop in reaction efficiency, an effect which can be avoided by utilizing lower borate concentration with higher pH. The improvements appear to primarily minimize electrodispersion of the picrate reagent, allowing higher picrate concentration in the reaction zone. The same conditions also appear to minimize the electrodispersion of the in-line product as well. With optimized EMMA parameters, the sensitivity of the in-line Jaffe chemistry can be enhanced to an extent that there is no need for the two capillary "high sensitivity" detection system required in previous work. Using optimized conditions, three different human serum samples spanning the expected clinical range of creatinine concentrations were successfully analyzed. Overall, this work illustrates the importance of systematically characterizing the conditions under which EMMA analyses are carried out.
Authors:
Philip E Mason; Derek C Schildt; Timothy G Strein
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2008-11-19
Journal Detail:
Title:  Journal of chromatography. A     Volume:  1216     ISSN:  0021-9673     ISO Abbreviation:  J Chromatogr A     Publication Date:  2009 Jan 
Date Detail:
Created Date:  2008-12-22     Completed Date:  2009-03-20     Revised Date:  2013-06-04    
Medline Journal Info:
Nlm Unique ID:  9318488     Medline TA:  J Chromatogr A     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  154-8     Citation Subset:  IM    
Affiliation:
Department of Chemistry, Bucknell University, Lewisburg, PA 17837, USA.
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MeSH Terms
Descriptor/Qualifier:
Borates / chemistry
Buffers
Creatinine / blood*
Electrophoresis, Capillary / methods*
Humans
Hydrogen-Ion Concentration
Indicators and Reagents / chemistry
Microchemistry / methods*
Picrates / chemistry
Sensitivity and Specificity
Time Factors
Grant Support
ID/Acronym/Agency:
R15 EB003854-01/EB/NIBIB NIH HHS; R15 EB003854-02/EB/NIBIB NIH HHS; R15-EB003854-02/EB/NIBIB NIH HHS
Chemical
Reg. No./Substance:
0/Borates; 0/Buffers; 0/Indicators and Reagents; 0/Picrates; 60-27-5/Creatinine
Comments/Corrections

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