biologia plantarum

Biologia plantarum 4:226-231, 1962 | DOI: 10.1007/BF02933101

New Methods of Chromatographic Separation of Gibberellins A₁ and A₃

Milan Kutáček^1,2, Jan Rosmus^1,3, Zdeněk Deyl^1,3

¹ Research Institute of Plant Production, Czechoslovak Academy of Agricultural Siences, and Central Research Institute of Food Industry, Praha

² Výzkumný ústav rostlinné výroby ČSAZV, Praha-Ruzyně

³ Ústřední výzkumný ústav potravinářského průmyslu, Praha-Smý chov

New Chromatographic methods, chromatography in centrifugal field and thin-layer chromatography on alumina, were used for separating physiologically active gibberellins A₁ and A₃. Paper electrophoresis was found unsuitable for the separation of the two gibberellins. For centrifugal chromatography the system benzene-acetic acid-water (4:1:2) was found convenient. In both the centrifugal and thin layer chromatography the overflowing technique was applied with great success. In the case of centrifugal separation bromophenol blue was used to indicate the position of the solvent front. Development was stopped when the spot of the indicator reached 3/4 of the paper length. For separating on a thin layer of alumina the solvent system benzene-acetic acid (100:23) was used. Here again the solvent was left to overflow; for dimensions of the thin layer of 10 X 23 cm 60 ml of the solvent mixture were used. The spots were detected in both cases with 0.5% potassium permanganate. The detection of the spot of gibberellin A₃ using 70% sulphuric acid after thin layer chromatography was found unsufisfactory. Sulphuric acid may be replaced by 70% perchloric acid. Both analyses take about 6 hrs. which reduces the time necessary for their completion to one working day.

Received: November 8, 1961; Published: September 1, 1962  Show citation