HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 286/6/F1116    most recent 00366.2003v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (27) Citing Articles via Google Scholar Google Scholar Articles by Yuen, P. S. T. Articles by Star, R. A. Search for Related Content PubMed PubMed Citation Articles by Yuen, P. S. T. Articles by Star, R. A.

A simplified method for HPLC determination of creatinine in mouse serum

¹Renal Diagnostics and Therapeutics Unit, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-1268; and ²Dorrance Hamilton Research Laboratories, Division of Nephrology,Medicine, Thomas Jefferson University, Philadelphia, Pennsylvania 19107

Submitted 17 October 2003 ; accepted in final form 13 February 2004

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 with 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 variable results, including an extra peak that interfered with integration of the creatinine peak or loss of the creatinine peak. Deproteinizing with acetonitrile alone resulted in a more reliable measurement of serum creatinine, which was validated by a series of known additions of creatinine standard. The HPLC assay was reproducible with coefficients of variation 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 sixfold, 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 µl of mouse serum. These results support the widespread adoption of HPLC to accurately measure serum creatinine in mouse models of renal injury.

Jaffé assay; mouse models; renal failure

This article has been cited by other articles:

				K. Doi, P. S.T. Yuen, C. Eisner, X. Hu, A. Leelahavanichkul, J. Schnermann, and R. A. Star Reduced Production of Creatinine Limits Its Use as Marker of Kidney Injury in Sepsis J. Am. Soc. Nephrol., June 1, 2009; 20(6): 1217 - 1221. [Abstract] [Full Text] [PDF]

				H. Geng, R. Lan, G. Wang, A. R. Siddiqi, M. C. Naski, A. I. Brooks, J. L. Barnes, P. Saikumar, J. M. Weinberg, and M. A. Venkatachalam Inhibition of Autoregulated TGF{beta} Signaling Simultaneously Enhances Proliferation and Differentiation of Kidney Epithelium and Promotes Repair Following Renal Ischemia Am. J. Pathol., April 1, 2009; 174(4): 1291 - 1308. [Abstract] [Full Text] [PDF]

				A. Leelahavanichkul, H. Yasuda, K. Doi, X. Hu, H. Zhou, P. S. T. Yuen, and R. A. Star Methyl-2-acetamidoacrylate, an ethyl pyruvate analog, decreases sepsis-induced acute kidney injury in mice Am J Physiol Renal Physiol, December 1, 2008; 295(6): F1825 - F1835. [Abstract] [Full Text] [PDF]

				M. D. Breyer Stacking the Deck for Drug Discovery in Diabetic Nephropathy: In Search of an Animal Model J. Am. Soc. Nephrol., September 1, 2008; 19(9): 1623 - 1624. [Full Text] [PDF]

				H. Yasuda, A. Leelahavanichkul, S. Tsunoda, J. W. Dear, Y. Takahashi, S. Ito, X. Hu, H. Zhou, K. Doi, R. Childs, et al. Chloroquine and inhibition of Toll-like receptor 9 protect from sepsis-induced acute kidney injury Am J Physiol Renal Physiol, May 1, 2008; 294(5): F1050 - F1058. [Abstract] [Full Text] [PDF]

				J. Roberts, B. Chen, L. M. Curtis, A. Agarwal, P. W. Sanders, and K. R. Zinn Detection of early changes in renal function using 99mTc-MAG3 imaging in a murine model of ischemia-reperfusion injury Am J Physiol Renal Physiol, October 1, 2007; 293(4): F1408 - F1412. [Abstract] [Full Text] [PDF]

				S. M. Malakauskas, H. Quan, T. A. Fields, S. J. McCall, M.-J. Yu, W. M. Kourany, C. W. Frey, and T. H. Le Aminoaciduria and altered renal expression of luminal amino acid transporters in mice lacking novel gene collectrin Am J Physiol Renal Physiol, February 1, 2007; 292(2): F533 - F544. [Abstract] [Full Text] [PDF]

				H. J. Zhao, S. Wang, H. Cheng, M.-z. Zhang, T. Takahashi, A. B. Fogo, M. D. Breyer, and R. C. Harris Endothelial Nitric Oxide Synthase Deficiency Produces Accelerated Nephropathy in Diabetic Mice J. Am. Soc. Nephrol., October 1, 2006; 17(10): 2664 - 2669. [Abstract] [Full Text] [PDF]

				M. D. Breyer, E. Bottinger, F. C. Brosius III, T. M. Coffman, R. C. Harris, C. W. Heilig, K. Sharma, and for the AMDCC Mouse Models of Diabetic Nephropathy J. Am. Soc. Nephrol., January 1, 2005; 16(1): 27 - 45. [Abstract] [Full Text] [PDF]