A Method for Analysis of Threonine and Accompanying Amino Acids by Quantitative High Performance Thin-Layer Chromatography in Cultural Broths

Автор: O.E. Mel'kina, B.V. Tyaglov, I.V. Manukhov, S.V. Antonova, E.D. Barsukov, I.I. Malakhova, V.D. Krasikov, A.S. Yanenko, and S.P. Sineokii

Страница: 89-96

A Method for analysis of threonine and Accompanying Amino Acids by quantitative high performance thin-layer chromatography in cultural broths

Biotekhnologiya, 2012, N 4, P. 89-96

UDC 547.466. 543. 544

Section:  “Metrology, Standartization, and Control”

O.E. Mel'kina ¹,  B.V. Tyaglov ^{1, *},  I.V. Manukhov ¹,  S.V. Antonova ¹,  E.D. Barsukov ¹,  I.I. Malakhova ²,  V.D. Krasikov ²,  A.S. Yanenko ¹,  and  S.P. Sineokii ¹

¹  The Research Institute for Genetics and Selection of Industrial Microorganisms,  117545, Moscow Russia

²  The research and Technical Center “Lenchrom”,  199004, St.-Peterburg Russia

e-mail:  b_tyaglov@genetika.ru

Quantitative high performance thin-layer chromatography has been applied to the L-threonine and concomitant amino acids analysis in cultural broths of a threonine-producing strain. Using a “PRIZMA” model, a mobile phase (containing acetone, propanol-2, water and 25% aqueous ammonia solution) was designed that makes it possible to separate threonine from other amino acids occurring in the cultural broth; the analysis takes 25—30 min. Detection limit for threonine is 0,1 μg in spot. The conditions for the application of the ninhydrin reagent to the chromatograms, performance of color reaction and treatment of the chromatograms on scanning densitometers were determined. It was established that the errors (n=3, P 0,95) did not exceed 5%. The validation of the developed method was carried out.

Key words:  alanine,  amino acids,  glycine,  glutamic acid,  homoserine,  lysine,  ninhydrin reagent,  quantitative high performance thin-layer chromatography,  scanning densitometry,  threonine,  validation.

The full English version of the article was published in “Biotechnology in Russia”, 2012, Issue 4, pp. 89-96 as O.E. Mel'kina, B.V. Tyaglov, I.V. Manukhov, S.V. Antonova, E.D. Barsukov, I.I. Malakhova, V.D. Krasikov, A.S. Yanenko, S.P. Sineokii “A Method for analysis of threonine and Accompanying Amino Acids by quantitative high-performance thin-layer chromatography in cultural broth”.

It is contained at the Russian Scientific Electron Library website:  http://elibrary.ru/item.asp?id=21532469

08.05.2015, 1362 просмотра.