Quantitative Determination of AICAR, a New Wide-Therapephtic-Use Substance, by Chromatographic Techniques in Cultural Broths

Автор: S.V. Antonova, B.V. Tyaglov, E.D. Barsukov, N.G. Demina, N.F. Rumiantseva, L. Arrayse-Lopes, N.V. Korol'kova, K.V. Lobanov, R.S. Shakulov, and A.S. Mironov

Страница: 87-92

Quantitative Determination of AICAR, a new wide-Therapephtic-Use substance, by chromatographic techniques in cultural broths

Biotekhnologiya, 2012, N 2, P. 87-92

UDC 577.123;577.18

Section:  “Metrology, Standartization, and Control”

S.V. Antonova,  B.V. Tyaglov ^*,  E.D. Barsukov,  N.G. Demina,  N.F. Rumiantseva,  L. Arrayse-Lopes,  N.V. Korol'kova,  K.V. Lobanov,  R.S. Shakulov,  and  A.S. Mironov

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

e-mail:  b_tyaglov@genetika.ru

A method of quantitative high-performance thin-layer chromatography has been used for the first time for the AICAR analysis in cultural broths. Using a “PRIZMA” model, a mobile phase (containing  chloroform, methanol and 25% aqueous ammonia solution) that makes it possible to separate AICAR, inosine and hypoxanthine was designed; the time of separation made up 25 min, and the detection limit or AICAR was 0,1 g per spot. The quantitative analysis of the chromatograms was performed using scanning densitometers of Shimadzu CS-920 and Camag TLC scanner 3 at 265 nm. High performance liquid chromatography was used as a reference method. A validation of the suggested method was carried out; it was established that the errors (n = 3, and P = 0,95) did not exceed 5%. A good correlation between the results obtained by both techniques (r = 0, 99350) was observed.

Key words:  AICAR,  HPLC,  hypoxanthine,  inosine,  quantitative thin-layer chromatography,  scanning densitometry,  validation.

The full English version of the article was published in “Biotechnology in Russia”, 2012, Issue 2, pp. 87-92 as S.V. Antonova, B.V. Tyaglov, E.D. Barsukov, N.G. Demina, N.F. Rumiantseva, L. Arrayse-Lopez, N.V. Korol’kova, K.V. Lobanov, R.S. Shakulov, A.S. Mironov “Quantitative Determination of AICAR, a new wide-Therapephtic-Use substance, by chromatographic techniques in cultural broths”.

It is contained at the Russian Scientific Electron Library website:  http://elibrary.ru/item.asp?id=21092716

15.06.2015, 1322 просмотра.