Abstract
The activity of glutamic pyruvic transaminase (GPT) is an important clinical evidence for some acute diseases such as acute hepatopathy and myocardial infarction. Thus, there is a demand for rapid determination of GPT in small formats at point-of-need. Herein, we describe a novel method of electrochemical determination of GPT with microfluidic technique. GPT activity was indirectly determined via the electrochemical (EC) detection of nicotinamide adenine dinucleotide (NADH) produced from the GPT transdeamination reaction. A type of microfluidic chip was developed, in which a passive mixer comprising 100 sub-ribs and a three-electrode strip for EC were integrated. To verify the response to NADH, a series of NADH concentrations varying from 19 µM to 5 mM were calibrated with cyclic voltammetry within the microfluidic chip. And a linear relationship with R 2 0.9982 between the peak current and the concentration of NADH was obtained. Then, the GPT activity was determined using the chips containing and not containing a ribs-type mixer. And a linear relationship which contained two sections between the GPT activity and the peak current was obtained. The chip with a ribs-type mixer exhibited the sensitivity of 0.0341 μA U−1 L in the range of 10–50 U L−1 and 0.0236 μA U−1 L in the range of 50–250 U L−1. And the detection limit of the chip with a ribs-type mixer was 9.25 U L−1. The complete detection process of GPT activity within the microfluidic chip was realized, and the time-consuming problem was remarkably improved too.
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Alam S, Ahmad N, Mustafa G, Shrestha A, Alam A, Khan M (2011) Evaluation of normal or minimally elevated alanine transaminase, age and DNA level in predicting liver histological changes in chronic hepatitis B. Liver Int 31(6):824–830
Bard AJ, Faulkner LR (2001) Electrochemical methods: principles and applications, 2nd edn. Wiley, New York
Gholizadeh A, Shahrokhian S, Irajizad A, Mohajerzadeh S, Vosoughi M, Darbari S, Koohsorkhi J, Mehran M (2012) Fabrication of sensitive glutamate biosensor based on vertically aligned CNT nanoelectrode array and investigating the effect of CNTs density on the electrode performance. Anal Chem 84(14):5932–5938
He YN, Chen HY (1997) The kinetics-based electrochemical determination of serum glutamate pyruvate transaminase activity with a gold microelectrode. Anal Chim Acta 353:319–323
Huang XJ, Choi YK, Im HS, Yarimaga O, Yoon E, Kim HS (2006) Aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) detection techniques. Sensors 6(7):756–782
Jing-Li Z, Ping-Ping N, Zheng H-T, Zhang J-M (2009) Progress of electrochemical biosensors based on nicotinamide adenine dinucleotide (phosphate)-dependent dehydrogenases. Chin J Anal Chem 37(4):617–623
Kim W, Flamm SL, Di Bisceglie AM, Bodenheimer HC (2008) Serum activity of alanine aminotransferase (ALT) as an indicator of health and disease. Hepatology 47(4):1363–1370
Moolla Z, Anderson F, Thomson SR (2013) Use of amylase and alanine transaminase to predict acute gallstone pancreatitis in a population with high HIV prevalence. World J Surg 37(1):156–161
Morimoto K, Upadhyay S, Higashiyama T, Ohgami N, Kusakabe H, Fukuda J, Suzuki H (2007) Electrochemical microsystem with porous matrix packed-beds for enzyme analysis. Sens Actuators B: Chem 124(2):477–485
Ohgami N, Upadhyay S, Kabata A, Morimoto K, Kusakabe H, Suzuki H (2007) Determination of the activities of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase in a microfluidic system. Biosens Bioelectron 22(7):1330–1336
Raj CR, Behera S (2005) Mediatorless voltammetric oxidation of NADH and sensing of ethanol. Biosens Bioelectron 21(6):949–956
Ren H-X, Huang X-J, Kim J-H, Choi Y-K, Gu N (2009) Pt/Au bimetallic hierarchical structure with micro/nano-array via photolithography and electrochemical synthesis: from design to GOT and GPT biosensors. Talanta 78(4–5):1371–1377
Sampson EJ, Whitner VS, Burtis CA, McKneally SS, Fast DM, Bayse DD (1980) An interlaboratory evaluation of the IFCC method for aspartate aminotransferase with use of purified enzyme materials. Clin Chem 26(8):1156–1164
Schindhelm RK, Diamant M, Dekker JM, Tushuizen ME, Teerlink T, Heine RJ (2006) Alanine aminotransferase as a marker of non-alcoholic fatty liver disease in relation to type 2 diabetes mellitus and cardiovascular disease. Diabetes/Metab Res Rev 22(6):437–443
Wang J (2002) Electrochemical detection for microscale analytical systems: a review. Talanta 56(2):223–231
Wooten M, Gorski W (2010) Facilitation of NADH electro-oxidation at treated carbon nanotubes. Anal Chem 82(4):1299–1304
Wu S, Zhou Z, Xu L, Su B, Fang Q (2014) Integrating bipolar electrochemistry and electrochemiluminescence imaging with microdroplets for chemical analysis. Biosens Bioelectron 53:148–153
Xu J-J, Wang A-J, Chen H-Y (2007) Electrochemical detection modes for microchip capillary electrophoresis. TrAC Trends Anal Chem 26(2):125–132
Xu Z, S-l Hu, W-f Tian, D-z Wang, J-s Liu, Liu C, L-d Wang (2015) Cyclic voltammetric determination of glutamic–pyruvic transaminase activity based on transdeamination. Anal Methods 7(22):9421–9425
Acknowledgements
The authors are grateful for the financial support from National Natural Science Foundation of China (No. 51475080, 51321004) and Funds of Key Laboratory of Liaoning Education Department (No. LZ2014005).
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Wang, J., Xu, Z., Zou, Hq. et al. Electrochemical determination of glutamic pyruvic transaminase using a microfluidic chip. Microfluid Nanofluid 21, 27 (2017). https://doi.org/10.1007/s10404-017-1869-8
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DOI: https://doi.org/10.1007/s10404-017-1869-8