Advances in Clinical and Experimental Medicine
2016, vol. 25, nr 6, November-December, p. 1227–1239
Publication type: original article
Determination of Selected Amino Acids in Serum of Patients with Liver Disease
1 Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Czech Republic
Background. The determination of amino acids can be a reliable approach for extended diagnosis of liver diseases. This is because liver disease can be a cause of impaired amino acid metabolism. Therefore, a method for the determination of serum amino acids, applicable for clinical purposes, is necessary.
Objectives. The aim of this study was to find differences in the levels of selected amino acids between patients with liver disease and a control group.
Material and Methods. Samples of peripheral venous blood were obtained from a group of patients with liver disease (n = 131, 59 women at an average age of 60 years and 72 men at an average age of 52 years) and a control group (n = 105, 47 women at an average age of 62 years and 58 men at an average age of 58 years). Before the separation, the amino acids were derivatized with naphthalene-2,3-dicarboxaldehyde. For the separation, reverse phase column was used. The effluent was monitored with a fluorescence detector.
Results. There were significant differences in the concentrations of some amino acids between the patients and the control group, but also between women and men. Correlations between some amino acids and markers of liver blood tests and lipid metabolism were observed.
Conclusion. A simple, relatively rapid and selective HPLC method with fluorescence detection for the determination of selected amino acids in serum has been developed.
amino acids, HPLC with fluorescence detection, patients with liver disease
- Lake AD, Novak P, Shipkova P, Aranibar N, Robertson DG, Reily MD, Lehman-McKeeman LD, Vaillancourt RR, Cherrington NJ: Branched chain amino acid metabolism profiles in progressive human nonalcoholic fatty liver disease. Amino Acids 2015, 47, 603–615.
- Hagiwara A, Nishiyama M, Ishizaki S: Branched-chain amino acids prevent insulin induced hepatic tumor cell proliferation by inducing apoptosis through mTORC1 and mTORC2-dependent mechanisms. J Cell Physiol 2012, 227, 2097–2105.
- Tietge UJ, Bahr MJ, Manns MP, Böker KHW: Hepatic amino-acid metabolism in liver cirrhosis and in the longterm course after liver transplantation. Trans Int 2003, 16, 1–8.
- Borg PC, Fekkes D, Vrolijk JM, van Buuren HR: The relation between plasma tyrosine concentration and fatigue in primary biliary cirrhosis and primary sclerosing cholangitis. BMC Gastroenterol 2005, 5, 1–7.
- Prystupa A, Szpetnar M, Boguszewska-Czubara A, Grzybowski A, Sak J, Załuska W: Activity of MMP1 and MMP13 and amino acid metabolism in patients with alcoholic liver cirrhosis. Med Sci Monit 2015, 21, 1008–1014.
- Yang J, He J, Cao H, Zhao X, Fu S, Lu H, Chen Y, Pan X, Li L: Correlation between plasma amino acid profiles and the various stages of hepatitis B infection. Eur J Clin Microbiol Infect Dis 2012, 31, 2045–2052.
- Sato K, Fukushima D, Doi H, Satomi S: Higher serum methionine levels as a predictive factor in patients with irreversible fulminant hepatic failure. Transplant Proc 2013, 45, 1904–1906.
- Perucho J, Gonzalo-Gobernado R, Bazan E, Casarejos MJ, Jiménez-Escrig A, Asensio MJ, Herranz AS: Optimal excitation and emission wavelengths to analyse amino acids and optimize neurotransmitters quantification using precolumn OPA-derivatization by HPLC. Amino Acids 2015, 47, 963–973.
- Domingues DS, Crevelin EJ, de Moraes LA, Cecilio Hallak JE, de Souza Crippa JA, Costa Queiroz ME: Simultaneous determination of amino acids and neurotransmitters in plasma samples from schizophrenic patients by hydrophilic interaction liquid chromatography with tandem mass spectrometry. J Sep Sci 2015, 38, 780–787.
- Mudiam MK, Ratnasekhar C: Ultra sound assisted one step rapid derivatization and dispersive liquid-liquid microextraction followed by gas chromatography-mass spectrometric determination of amino acids in complex matrices. J Chromatogr A 2013, 1291, 10–18.
- Srazin C, Delaunay N, Costanza C, Eudes V, Mallet JM, Gareil P: New avenue for mid-UV-range detection of underivatized carbohydrates and amino acids in capillary electrophoresis. Anal Chem 2011, 83, 7381–7387.
- Holub M, Tuschl K, Ratschmann R, Strnadová KA, Mühl A, Heinze G, Sperl W, Bodamer OA: Influence of haematocrit and localisation of punch in dried blood spots on levels of amino acids an acylcarnitines measured by tandem mass spectrometry. Clin Chim Acta 2006, 373, 27–31.
- Wilcken B, Wiley V, Hammond J, Carpenter K: Screening newborns for inborn errors of metabolism by tandem mass spectrometry. N Engl J Med 2013, 348, 2304–2312.
- Müller C, Fonseca JR, Rock TM, Krauss-Etschmann S, Schmitt-Kopplin P: Enantioseparation and selective detection of D-amino acids by ultra-high-performance liquid chromatography/mass spectrometry in analysis of complex biological samples. J Chromatogr A 2014, 1324, 109–114.
- Drábková P, Šanderová J, Kovařík J, Kanďár R: An assay of selected serum amino acids in patients with type 2 diabetes mellitus. Adv Clin Exp Med 2015, 24, 447–451.
- Kang X, Xiao J, Huang X, Gu Z: Optimization of dansyl derivatization and chromatographic conditions in the determination of neuroactive amino acids of biological samples. Clin Chim Acta 2006, 366, 352–356.
- Buha SM, Panchal A, Panchal H, Chambhare R, Kumar S, Jain M, Patel PR: HPLC-FLD for the simultaneous determination of primary and secondary amino acids from complex biological sample by pre-column derivatization. J Chromatogr Sci 2011, 49, 118–123.
- Ziegler J, Abel S: Analysis of amino acids by HPLC/electrospray negative ion tandem mass spectrometry using 9-fluorenylcarbonyl chloride (Fmoc-Cl) derivatization. Amino Acids 2014, 46, 2799–2808.
- Sharma G, Attri SV, Behra B, Bhisikar S, Kumar P, Tageja M, Sharda S, Singhi P, Singhi S: Analysis of 26 amino acids in human plasma by HPLC using AQS as derivatizing agent and its application in metabolic laboratory. Amino Acids 2014, 46, 1253–1263.
- Kuo YH, Defoort B, Getahun H, Tekle Haimanot R, Lambein F: Comparison of urinary amino acids and trace elements (cooper, zinc and manganese) of recent neurolathyrism patients and healthy controls from Ethiopia. Clin Biochem 2007, 40, 397–402.
- Fischer JE, Yoshimura N, Aguirre A, James JH, Cummings MG, Abel RM, Deindoerfer F: Plasma amino acids in patients with hepatic encephalopathy. Effects of amino acid infusions. Am J Surg 1974, 127, 40–47.
- Dodd KM, Tee AR: Leucine and mTORC1: a complex relationship. Am J Physiol Endocrinol Metab 2012, 302, 1329–1342.
- Holeček M: Ammonia and amino acid profiles in liver cirrhosis: Effects of variables leading to hepatic encephalopathy. Nutrition 2015, 31, 14–20.
- Tietge UJ, Bahr MJ, Manns MP, Böker KHW: Plasma amino acids in cirrhosis and after liver transplantation: Influence of liver function, hepatic hemodynamics and circulating hormones. Clin Transplant 2002, 16, 9–17.
- Morgan MY, Marshall AW, Milsom JP, Sherlock S: Plasma amino-acid patterns in liver disease. Gut 1982, 23, 362–370.