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Rapid Determination of Creatinine in Human Urine by Microchip Electrophoresis with LED Induced Fluorescence Detection

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Abstract

In clinical medicine, urine creatinine concentration is an important marker in the evaluation of renal function and muscular dysfunctions. Herein, we reported a novel method for rapid determination of creatinine in urine by microchip electrophoresis with light-emitting diode induced fluorescence detection. Creatinine was derivatized by fluorescein isothiocyanate, and then quantitatively detected by the developed microchip LED induced fluorescence detection system. The excitation and emission wavelengths were 490 and 523 nm, respectively. The urine samples were analyzed after centrifuge and filtration. A baseline separation was obtained in <30 s using 10 mM borate buffer (pH 9.0, containing 45 mM sodium dodecylsulfate), with separation voltage of 1.5 kV. Good linearity was obtained (r 2 = 0.9978) in the concentration range of 10.0–2.00 × 103 μM, and the limit of detection was 2.87 μM (S/N = 3). The recovery was 96.0–107 %, and the interday precision was <4.5 % (n = 6). To validate assay results, we compared the present method with the Jaffe’s colorimetric assay by measuring real urine samples. The method was reliable, sensitive, high-speed, low-cost and suitable for the routine analysis of creatinine in biofluids.

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Acknowledgments

Financial supports from the National Natural Science Foundation of China (Nos. 20727006, 21075139), the Fundamental Research Funds for the Central Universities, and Guangdong Provincial Science and Technology Project (No. 2008A030102009) are gratefully acknowledged.

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Authors and Affiliations

  1. School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan East Road of Higher Education Mega Centre, Guangzhou, 510006, People’s Republic of China

    Shuping Wang, Xinchun Li, Jianping Yang, Xiujuan Yang, Fenghua Hou & Zuanguang Chen

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  1. Shuping Wang
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  2. Xinchun Li
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  3. Jianping Yang
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  4. Xiujuan Yang
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  5. Fenghua Hou
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Correspondence to Zuanguang Chen.

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Wang, S., Li, X., Yang, J. et al. Rapid Determination of Creatinine in Human Urine by Microchip Electrophoresis with LED Induced Fluorescence Detection. Chromatographia 75, 1287–1293 (2012). https://doi.org/10.1007/s10337-012-2324-3

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  • DOI: https://doi.org/10.1007/s10337-012-2324-3

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