TY - JOUR
T1 - Simultaneous determination of guanidinosuccinic acid and guanidinoacetic acid in urine using high performance liquid chromatography/tandem mass spectrometry
AU - Saigusa, Daisuke
AU - Suzuki, Naoto
AU - Takahashi, Mai
AU - Shiba, Kanako
AU - Tanaka, Satoshi
AU - Abe, Takaaki
AU - Hishinuma, Takanori
AU - Tomioka, Yoshihisa
PY - 2010/9
Y1 - 2010/9
N2 - 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 0.1mM and 20mM 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-13C6 hydrochloride and creatinine-d3 (methyl-d3) were used. The calibration ranges were 0.50-25.0μgmL-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 25.0μgmL-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.
AB - 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 0.1mM and 20mM 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-13C6 hydrochloride and creatinine-d3 (methyl-d3) were used. The calibration ranges were 0.50-25.0μgmL-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 25.0μgmL-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.
KW - Arginine
KW - Creatinine
KW - Diabetes mellitus
KW - Guanidinoacetic acid
KW - Guanidinosuccinic acid
KW - Liquid chromatography/tandem mass spectrometry
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U2 - 10.1016/j.aca.2010.08.005
DO - 10.1016/j.aca.2010.08.005
M3 - Article
C2 - 20837184
AN - SCOPUS:77956614754
VL - 677
SP - 169
EP - 175
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
IS - 2
ER -