Simultaneous determination of guanidinosuccinic acid and guanidinoacetic acid in urine using high performance liquid chromatography/tandem mass spectrometry
Simultaneous determination of guanidinosuccinic acid and guanidinoacetic acid in urine using high performance liquid chromatography/tandem mass spectrometry
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DOI：
10.1016/j.aca.2010.08.005
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发表时间：
2010-09-16
影响因子：
6.2
通讯作者：
Tomioka, Yoshihisa
中科院分区：
文献类型：
--
作者：
Saigusa, Daisuke;Suzuki, Naoto;Tomioka, Yoshihisa
建立了一种同时测定尿液中胍丁二酸（GSA）和胍乙酸（GAA）的蛋白质沉淀-液相色谱/串联质谱法。色谱分离采用阳离子交换柱，洗脱梯度为0.1 mM，乙酸铵缓冲液为20 mM。GSA在m/z 174.1的负离子模式下检测，GAA、肌酐、精氨酸和同型精氨酸分别在m/z 118.1、114.1、175.1和189.1的正离子模式下检测。内标采用l-精氨酸- c -13(6)盐酸盐和肌酐-d(3)（甲基-d(3)）。标定范围为0.50 ~ 25.0 μ g mL(-1)，所有化合物均具有良好的线性关系（r < 0 0.999）。3个浓度水平（1.00、5.00和25.0 μ g mL(-1)）下的组内和组间准确度（以回收率表示）和精密度分别优于83.8%和7.41%。通过对C57BL/ jlar雄性糖尿病（DM）小鼠尿液中化合物的测定，评价了该方法的分析性能。与对照小鼠相比，DM小鼠的GSA和GAA值（由单个化合物与肌酐的比值校正）显著增加。结果表明，该方法可用于尿中胍类化合物和精氨酸代谢产物的测定。(c) 2010 Elsevier B.V.保留所有权利。
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-C-13(6) hydrochloride and creatinine-d(3) (methyl-d(3)) were used. The calibration ranges were 0.50-25.0 mu g 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 mu g 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 lar 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. (c) 2010 Elsevier B.V. All rights reserved.