Page 228 - Demo

232 PROCEEDINGS OF INTERNATIONAL SCIENTIFIC CONFERENCE ON APPLIED BIOTECHNOLOGYlabeled E2.2.1 to E2.2.4. Fraction E2.2.3 was separated by normal phase silica gel column chromatography, with a solvent system of n-hexane:EtOAc (2:1), yielding compound 1 (18 mg) in the form of yellow needle-shaped crystals.Fraction E5 was separated using normal phase silica gel column chromatography, with a solvent system of n-hexane:EtOAc (2:1), yielding 5 fractions labeled E5.1 to E5.5. Fraction E5.3 was separated using normal phase silica gel column chromatography, with a solvent system of dichloromethane:methanol (20:1), yielding 4 fractions labeled E5.3.1 to E5.3.4. Fraction E5.3.2 was separated using normal phase silica gel column chromatography, with a solvent system of chloroform:methanol (10:1), yielding 3 fractions labeled E5.3.2.1 to E5.3.2.3. Fraction E5.3.2.2 was purified by reversed-phase RP-18 column chromatography with a solvent system of methanol:water (3:1), yielding compound 2 (8,5 mg) as a pale yellow solid.The water extracts (75g) of avocado leaves was subjected to separation through Diaion HP-20 column chromatography with a gradient solvent system of water:methanol (100:0, 70:30, 50:50, 30:70, 0:100), yielding 5 corresponding fractions labeled M1 to M5.Fraction M5, after solvent removal, was separated using normal phase silica gel column chromatography, with a solvent system of EtOAc:methanol (10:1), yielding 6 fractions labeled M5.1to M5.6.Fraction M5.4 was separated using normal phase silica gel column chromatography with a solvent system of EtOAc:methanol (7:1), yielding 3 fractions labeled M5.4.1 to M5.4.3. Fraction M5.4.2 was separated using normal phase silica gel column chromatography with a solvent system of dichloromethane:methanol:water (5:1:0,05), yielding 3 fractions labeled M5.4.2.1 to M5.4.2.3. Fraction M5.4.2.2 was purified using reversed-phase RP-18 column