Methanol extracts from roots and n-butanol extracts from the culture medium were hydrolyzed by 11% HCl in 70% methanol for 2h on a heating mantle under reflux as described earlier [25 (link)]. Subsequently, the hydrolysates were diluted with distilled water, methanol was evaporated in a rotary evaporator, and the obtained aqueous remaining were extracted 3 times with 40 mL portions of diethyl ether in a separation funnel. The obtained extract was washed with distilled water 3 times and evaporated to dryness.
The dried extracts were fractionated by preparative TLC on 20 cm × 20 cm glass plates, manually coated with silica gel 60H (Merck, Darmstadt, Germany). A solvent system with a chloroform:methanol (95/5,v/v) mixture was used for developing the plates. The obtained OA was methylated with diazomethane [13 (link),14 (link),28 (link)].
The dried extracts were fractionated by preparative TLC on 20 cm × 20 cm glass plates, manually coated with silica gel 60H (Merck, Darmstadt, Germany). A solvent system with a chloroform:methanol (95/5,v/v) mixture was used for developing the plates. The obtained OA was methylated with diazomethane [13 (link),14 (link),28 (link)].
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