A simplified method for HPLC determination of creatinine in mouse serum
Authors Peter Yuen, Stephen Dunn, Takehiko Miyaji, Kumar Sharma, Robert Star
Submitted By Robert Star on 8/1/2003
Status Published
Journal Journal of the American Society of Nephrology : JASN
Year 2004
Date Published 6/1/2004
Volume : Pages 286 : F1116 - F1119
PubMed Reference 14970000
Abstract Mouse models are frequently used to study renal function; however, mouse serum
contains chromagens that interfere with standard picric acid-based assays for
serum creatinine. Several alternative methods exist for serum creatinine
measurements, including assay by high performance liquid chromatography (HPLC),
but only one has been adapted to mouse serum. Creatinine was measured in serum
by acetonitrile deproteinization, followed by isocratic, cation exchange HPLC.
The HPLC method was compared to a standard alkaline picrate colorimetric assay,
using serum from animals with low to moderate renal injury. Acidification of
acetonitrile with HCl in the deproteinization step produced an extra peak that
interfered with integration of the creatinine peak. Deproteinizing with
acetonitrile alone resulted in a more accurate measurement of serum creatinine,
as validated by a series of known additions of creatinine standard. The HPLC
assay was reproducible with coefficients of variations from 1.6 to 5.1%. The
picric acid assay overestimated serum creatinine, when directly compared with
the HPLC assay. The extent of overestimation, up to 6-fold, was greatest at
normal (0.1 to 0.2 mg/dl) to moderately elevated (0.5 mg/dl) serum creatinine
levels. Mouse serum contains substances that interfere with standard picric acid
assays for creatinine. Our new HPLC assay can accurately detect creatinine from
5 microliters of mouse serum. These results support the widespread adoption of
HPLC to accurately measure serum creatinine in mouse models of renal injury


Investigators with authorship
NameInstitution
Kumar SharmaUniversity of California San Diego

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