L-lysine determination in animal feed using microbiological microtiter plate assay

L-lysine determination feed microbiological assay HPLC

Chromatographic methods are most commonly used for the analysis of amino acids; however, there is growing need for faster, simpler and more price-effective assays. In this paper, the applicability of a rapid microbiological assay for quantification of the total content of L-lysine in feed samples was evaluated. The assay relies on the dependency of bacterial growth of Pediococcus acidilactici on the presence of L-lysine. Microbiological microtiter plate assay method for the quantitative determination of total (added and natural) L-lysine in feed samples has been verified, and parameters such as accuracy, precision, limit of detection, and limit of determination were evaluated. Results of total L-lysine determination in different feed samples have been compared with results of validated HPLC method. The microbiological microtiter plate assay method can be employed as a qualitative and quantification method for total L-lysine determination with detection and determination limit of 0.040 % and 0.085 %, respectively. However, further research on the influence of sample matrix on the determination of low lysine levels is required.

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Bartolomeo MP, Maisano F (2006) Validation of reversed-phase HPLC method for quantitative amino acid analysis. J Biomol Tech 17:131-137.
Chalova VI, Zabala-Diaz IB, Woodward CL, Ricke SC (2008) Development of a whole cell green fluorescent sensor for lysine quantification. World J Microb Biot 24:353-359.
Cottingham LS, Smalidge RL (1988) Reverse-phase liquid chromatographic determination of lysine in complete feeds and premixes, using manual precolum derivatization. J AOAC Int 71:1012-1016.
Couch JR, Thomas M, Agric J (1976) A comparison of chemical methods for the determination of available lysine in various proteins. Food Chem 24:943-946.
Culea M, Scrob S, Suvar S, Podea P, Has I, Muste S (2015) Determination of amino acids in corn seed by gas chromatography- mass spectrometry. Anal Lett 48:37-46.
European Commission (2009) Commission Regulation (EC) No 152/2009 of 27 January 2009 laying down the methods of sampling and analysis for the official control of feed, Off J Eur Union L54:30-31.
Fiechter FG, Mayer HK (2014) A novel UHPLC method for the determination of amino acids using either fluorescence or ultraviolet detection, Current Analytical Chemistry 10: 366-374.
Fontaine J, Eudaimon M (2000) Liquid chromatographic determination of lysine, methionine, and threonine in pure amino acids (feed grade) and premixes: collaborative study. J AOAC Int 83:771-783.
Fountoulakis M, Lahm HW (1998) Hydrolysis and amino acid composition analysis of proteins. J Chromatogr A 826:109-134.
Garcia-Villar N, Saurina J, Hernandez-Cassou S (2001) Potentiometric sensor array for the determination of lysine in feed samples using multivariate calibration methods. Fresen J Anal Chem 371:1001-1008.
Hasani M, Yaghoubi L, Abdollahi H (2007) A kinetic spectrophotometric method for simultaneous determination of glycine and lysine by artificial neural networks. Anal Biochem 365:74-81.
Herman RA, Scherer PN, Shan G (2008) Evaluation of logistic and polynomial models for fitting sandwich-ELISA calibration curves. Journal of Immunological Methods, 339, 245-258.
Horn MJ, Jones DB, Blum AE (1946) Microbiological determination of methionine in proteins and foods. J Biol Chem 166:321-326.
Horn MJ, Jones DB, Blum AE (1947) Microbiological determination of lysine in proteins and foods. J Biol Chem 169:71-76.
Intitut für Produktqualität, VitaFast L-Lysin. Microbiological microtiter plate test to quantitate L-lysine, Art. No.: P1012 (2016) http://www.produktqualitaet.com/en/products-test-kits/microtiter-plate-vitafast/amino-acids.html [23 August 2016]
Jajić I, Krstović S, Glamočić D, Jakšić S, Abramović B (2013) Validation of an HPLC method for the determination of amino acids in feed. J Serb Chem Soc 78:839-850.
Khan AS, Faiz F (2008) Amino acid analysis using ion exchange resins. J Natur Sci Math 48:1-17.
Latorre RM, Saurina J, Hernandez-Cassou S (2001) Sensitivity enhancement by on-line preconcentration and in-capillary derivatization for the electrophoretic determination of amino acids. Electrophoresis 22:4355-4361.
Latorre RM, Saurina J, Hernandez-Cassou S (2002) Continuous flow derivatization system coupled to capillary electrophoresis for the determination of amino acids. J Chromatogr A 976:55-64.
Le A, Ng A, Kwan T, Cusmano-Ozog K, Cowan TM, (2014) A rapid, sensitive method for quantitative analysis of underivatized amino acids by liquid chromatography–tandem mass spectrometry (LC–MS/MS) Journal of Chromatography B 944: 166-174.
Liao SF, Wang T, Regmi N (2015) Lysine nutrition in swine and the related monogastric animals: muscle protein biosynthesis and beyond. SpringerPlus 4:1-12.
Loy HW, Wright WW (1959) Microbiological assay of amino acids, vitamins, and antibiotics. Anal Chem 31:971-974.
Reason AJ (2003) Validation of amino acid analysis methods. In: B.J. Smith (Ed.). Protein Sequencing Protocols. Methods Mol Biol (Volume 211), Humana Press, Totowa, NJ: pp 181-194.
Redshaw MS, Fickler J, Fontaine J, Heimbeck W, Hess V, Reimann I (2010) Amino Acid Composition of Feedstuffs, Evonik Industries. Saurina J, Hernandez-Cassou S, Fabregas E, Alegret S (1999) Determination of tryptophan in feed samples by cyclic voltammetry and multivariate calibration methods. Analyst 124:733-737.
Sugawara A, Matsui D, Yamada M, Asano Y, Isobe K (2015) New enzymatic methods for selective assay of L-lysine using anl-lysine specific decarboxylase/oxidase from Burkholderia sp. AIU 395. J Biosci Bioeng 119:369-374.
Thompson M, Ellison SL, Wood R (2002) Harmonized guidelines for single laboratory validation of methods of analysis. Pure Appl Chem 74:835-855.
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