Plant Genetic Conservation Project under Royal Initiation by Her
Royal Highness Princess Maha Chakri Sirindthorn at Kok Phutaka
area, Wiang Kao District, Khon Kaen Province, and was identified
by W. Saksirirat as N. nambi. The voucher specimens (PW1 and
PW2) were deposited at the Department of Plant Science and
Agricultural Resources, Faculty of Agriculture, Khon Kaen University,
Khon Kaen, Thailand. The mushroom culture was incubated in
a dark room without shaking with 2 h of light per day at 25 C for
30 days.
3.3. Extraction and isolation
3.3.1. N. nambi (PW1). Extraction of culture liquid of PW1 (2.025 L)
with EtOAc (2.5 L) yielded 4.6 g of crude extract, from which 2.1 g
of yellow crystals of Aurisin A (6) were obtained after crystallization
from EtOAc. The filtrate was evaporated to dryness and the
residue (2.5 g) was then separated over silica gel flash column
chromatography (FCC), eluted with a gradient system of hexane:
EtOAc and EtOAc:MeOH to give 5 fractions, EF1-EF5. Fraction
EF1 was washed with EtOAc to give an additional amount of
compound 6 (2.04 g). Fraction EF4 was separated on silica gel
column chromatography (CC), eluted with an isocratic system of
hexane:EtOAc:MeOH (50:45:5) to give 3 subfractions, EF4.1eEF4.3.
Subfraction EF4.2 was purified by preparative TLC, using EtOAc:
hexane:MeOH (45:50:5) as eluent to give a colorless oil of
Fig. 4. ORTEP plot of aurisin A.
Fig. 5.
Plant Genetic Conservation Project under Royal Initiation by Her
Royal Highness Princess Maha Chakri Sirindthorn at Kok Phutaka
area, Wiang Kao District, Khon Kaen Province, and was identified
by W. Saksirirat as N. nambi. The voucher specimens (PW1 and
PW2) were deposited at the Department of Plant Science and
Agricultural Resources, Faculty of Agriculture, Khon Kaen University,
Khon Kaen, Thailand. The mushroom culture was incubated in
a dark room without shaking with 2 h of light per day at 25 C for
30 days.
3.3. Extraction and isolation
3.3.1. N. nambi (PW1). Extraction of culture liquid of PW1 (2.025 L)
with EtOAc (2.5 L) yielded 4.6 g of crude extract, from which 2.1 g
of yellow crystals of Aurisin A (6) were obtained after crystallization
from EtOAc. The filtrate was evaporated to dryness and the
residue (2.5 g) was then separated over silica gel flash column
chromatography (FCC), eluted with a gradient system of hexane:
EtOAc and EtOAc:MeOH to give 5 fractions, EF1-EF5. Fraction
EF1 was washed with EtOAc to give an additional amount of
compound 6 (2.04 g). Fraction EF4 was separated on silica gel
column chromatography (CC), eluted with an isocratic system of
hexane:EtOAc:MeOH (50:45:5) to give 3 subfractions, EF4.1eEF4.3.
Subfraction EF4.2 was purified by preparative TLC, using EtOAc:
hexane:MeOH (45:50:5) as eluent to give a colorless oil of
Fig. 4. ORTEP plot of aurisin A.
Fig. 5.
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