Determination of benzyladenine content of in vitro propagated dwarf J-TE-Fapple rootstock (Malus sp.) by high performance liquid chromatography

Shoots of J-TE-F dwarf apple rootstock were cultured under in vitro conditi ons on MURASHIGE-SKOOG (1962) basal medium, containing 0.25, 0.50 and 1.0 p pm BA. The (B)benzyladenine (BA) concentration of plantlets was determined by HPLC on the 14(th) and 30(th) day of subculture. The plant hormone was e xtracted by ethanol, further cleaned on SEP-PAK cartridge. The effluent was examined by reversed phase High Performance Liquid Chromatography with dio de array detection at 270 nm. Recovery and reproducibility assessment indic ates good accuracy and acceptable relative standard deviation (RSD) 5%). Li near responses (r(2)0.998) for calibration curve was obtained with BA stand ard in a range of 0.2-30 g.ml(-1) with a limit of quantification of 0.15 g. ml(-1). The highest BA concentration (5.49 ppm) was measured in the plants, which w ere cultured on 1.0 ppm BA containing multiplication medium on the 14(th) d ay. On the 30(th) day the BA concentration decreased to similar to1 ppm/dry weight.