Analysis of creatinine in mouse and rat serum by ion exchange high performance liquid chromatography for in vivo studies of renal function.
An ion exchange high performance liquid chromatography method was developed for determining creatinine levels in both mouse and rat serum samples. Separation of creatinine from other serum components was achieved in 10 min using a 100 x 4.1-mm, 10 microm strong cation exchange column following acetonitrile precipitation of serum proteins. Incorporation of a guard cartridge placed in-line prior to the analytical column was employed to prevent interference from compounds used in renal disease animal trials. Creatinine levels in normal and diseased animals were accurately determined in the 0.01-10 mg/dL range, and average recovery of the method was approximately 85% for both mouse and rat serum. Addition of 0.5-1.0% acetic acid to the acetonitrile used for protein precipitation significantly improved creatinine recovery to above 97% in mouse serum. The method was used for routine preclinical diagnosis of rat and mouse model renal function, and for the evaluation of renal disease treatment efficacy.
Fountain
Kenneth J
KJ
Analytical Research and Development, Drug and Biomaterial Research and Development, Genzyme Corporation, 153 2nd Avenue, Waltham, MA 02451, USA. ken.fountain@genzyme.com
Kloss
Alla
A
Garibyan
Ilya
I
Blitshteyn
Bella
B
Brezzani
Alexander
A
Kyostio-Moore
Sirkka
S
Zuk
Anna
A
Sacchiero
Robert
R
Cohen
Aharon S
AS
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003404:Creatinine / Q000097:blood*; D007677:Kidney Function Tests; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D051381:Rats; D015203:Reproducibility of Results
D003404:Creatinine