Simultaneous determination of guanidinosuccinic acid and guanidinoacetic acid in urine using high performance liquid chromatography/tandem mass spectrometry

• Published in: Analytica chimica acta Vol. 677; no. 2; pp. 169 - 175
• Main Authors: Saigusa, Daisuke, Suzuki, Naoto, Takahashi, Mai, Shiba, Kanako, Tanaka, Satoshi, Abe, Takaaki, Hishinuma, Takanori, Tomioka, Yoshihisa
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 16.09.2010; Elsevier
• Subjects: Analytical chemistry; Animals; Arginine; Arginine - urine; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, High Pressure Liquid - methods; Chromatography, High Pressure Liquid - standards; Creatinine; Creatinine - urine; Diabetes mellitus; Diabetes Mellitus - metabolism; Diabetes Mellitus, Experimental; Disease Models, Animal; Exact sciences and technology; Glycine - analogs & derivatives; Glycine - urine; Guanidines - urine; Guanidinoacetic acid; Guanidinosuccinic acid; Homoarginine - urine; Linearity; Liquid chromatography/tandem mass spectrometry; Male; Mass spectrometry; Mathematical analysis; Mice; Mice, Inbred C57BL; Monitoring; Noise levels; Other chromatographic methods; Spectrometric and optical methods; Succinates - urine; Tandem Mass Spectrometry - methods; Tandem Mass Spectrometry - standards; Urine; Creatinine; Guanidinoacetic acid; Guanidinosuccinic acid; Liquid chromatography/tandem mass spectrometry; Arginine; Diabetes mellitus; Performance evaluation; Ammonium; Chemical analysis; Metabolite; Internal standard; HPLC chromatography; Precipitation; Accuracy; Hydrochlorides; Monitoring; Liquid chromatography/tandem mass; Trace analysis; Cation exchange; Urine; Gradient; Rodentia; Liquid chromatography; Acetate; Concentration; Calibration; Protein; Positive ion; Vertebrata; Mammalia; Mouse; Linearity; Mass spectrometry MS/MS; Simultaneous measurement; Analytical method; Succinic acid; spectrometry; Negative ion
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2010.08.005

We present a method for the simultaneous determination of guanidinosuccinic acid (GSA) and guanidinoacetic acid (GAA) from urine by protein precipitation and liquid chromatography/tandem mass spectrometry. The chromatographic separation was performed using a cation exchange column with an elution gradient of 0.1 mM and 20 mM ammonium acetate buffers. GSA was detected with the mass spectrometer in negative ion mode monitoring at m/ z 174.1, and GAA, creatinine, arginine, and homoarginine were in positive ion mode monitoring at m/ z 118.1, 114.1, 175.1, and 189.1, respectively. As an internal standard, l-arginine- 13C 6 hydrochloride and creatinine-d 3 (methyl-d 3) were used. The calibration ranges were 0.50–25.0 μg mL −1, and good linearities were obtained for all compounds ( r > 0.999). The intra- and inter-assay accuracies (expressed as recoveries) and precisions at three concentration levels (1.00, 5.00 and 25.0 μg mL −1) were better than 83.8% and 7.41%, respectively. The analytical performance of the method was evaluated by determination of the compounds in urine from male C57BL/J Iar db/ db diabetes mellitus (DM) mice. The values of GSA and GAA corrected by the ratios of the individual compounds to creatinine were significantly increased in DM mice compared with control mice. These results indicated that the newly developed method was useful for determining urinary guanidino compounds and metabolites of arginine.