スライド 1

Identification of unsaturated fatty acid
from discarded Goya seeds.
Okinawa National College of Technology
Bioresources engineering
*Momoko Mashiyama,
Yuka Shingaki, Eisuke Kuraya and Kei Hirayama
Purpose
Our research addressed for
added extra value to the unused
Goya seeds that containing high
amount of polyunsaturated fatty
acid.
Back ground
Unripe green Goya
・largely cultivated in Okinawa.
・can harvest from spring to
autumn in Okinawa.
・containing full of vitamin C.
・well consumed by people in Okinawa.
Back ground
Harvest of Goya are
about 25,000 ton in Japan.[1]
Large quantity of Goya seeds
about 500 ton have been discarded.
[1] Ministry of Agriculture, Forestry and Fisheries, Research for production
of local specialty vegetables. Annual statistic reports on 2008.
Methods
Experimental flow
①Thin layer
chromatography
② ・Soxhlet extraction
・Methyl esterification
Visualized by Iodine
Sample
injector
①To find out plants hold rich
polyunsaturated fatty acid.
③GC-MS analysis
Flow
controller
②To prepare crude seeds oil
③Analyzed by GC-MS.
Carrier
gas
Column
Column
oven
Mass
spectrometry
Method①
Conditions for thin layer chromatography
・TLC plate : silica gel (whatman)
・Solvent :
hexane: diethyl ether: acetic acid
(80:20:1, V/V)
・Development for fatty acid:
visualized by Iodine.
Method②
Preparation of crude Goya seeds oil.
・ seeds were separated from Goya fruit.
・dried at 50 ℃ for 24 hours.
・homogenized by blender for 30 sec.
Goya seed
Vertical angle of
Goya seed
Method②
・ Soxhlet extraction
crude oil from unripe green Goya seeds.
・ Methyl esterification
n-Heptadecanoic acid was using as a internal standard.
Fatty acid methyl ester was analyzed by GC-MS.
Method③
GC-MS analytical conditions
 Column
DB-23（Agilent Technologies）
(50％‐ cyanopropyl)-methyl polysiloxane
ID 0.25mm×30m Film thickness 0.25µm
 The analytical conditions
GC-17A
（Shimadzu Corporation）
analyzing fatty acid methyl ester
JET
240 ℃
DET
240 ℃
Split
Total flow
10
15.0 ml/min
Linear velocity 40.0 cm /s
① 50℃,1min
② 175℃,25℃/min
③ 230℃,4℃/min ,
5.25min (HOLD)
Results
Thin layer chromatography of plant oil
1. Black rice
triglyceride
2. Goya seed
Therefore, we focused on analyzing
free fatty acid
3. Oleic acid
unsaturated fatty acid from Goya seeds.
4. Linoleic acid
1
2
3 4
We analyzed about 30 kinds of plants inhabited in Okinawa Island.
Picture Showing triglyceride from black rice and Goya seed.
Unripe Goya hold high amount of polyunsaturated fatty acid.
Results
Goya seeds oil analyzed by GC-MS
Five fatty acids were detected from unripe
green Goya seeds.
These fatty acids were identified by mass
spectrogram for using typical unsaturated
and saturated fatty acid standards.
Results
Total ion chromatogram of fatty acid
from Goya seeds oil by GC-MS.
intensity
×107
5.0
4.5
4.0
Stearic
acid
Unknownpolyunsaturated
polyunsaturated
Unknown
fattyacid
acid（No.5）
（No.5）
fatty
3.5
3.0
2.5
2.0
1.5
1.0
0.5
0
Two saturated and three unsaturated fatty acids were detected.
Results
Ratio of fatty acid contained
in the Goya seeds.
No.5 fatty acid
51.5%
Linoleic acid
9.6%
Oleic acid
6.8%
Stearic acid
Palmitic acid
28.1%
4.0%
67.9%
Unsaturated
32.1%
Saturated
No.5 fatty acid was polyunsaturated fatty acid and
which account for over half of all fatty acids.
Results
Mass spectrum
α- linolenate methyl ester standard
No.5 polyunsaturated fatty acid
No.5 polyunsaturated fatty acid has similar mass spectrum
patern with α- linolenate except peak height for 292.
Results
Data from total ion chromatogram,
no.5 polyunsaturated fatty acid has 20
carbon.
Results
Predicted structure for
no.5 polyunsaturated fatty acid
The position of double bond in this
polyunsaturated fatty acid was determined
by fragment ion of m/z 292.
Conclusion
Five fatty acid were detected.
→Goya seeds hold three unsaturated fatty
acids.
・
・Peak
no.5 polyunsaturated fatty acid has
account for over half of all fatty acids.
Conclusion
Polyunsaturated fatty acid
…has various functionalities.
For Example
Reduce
…risk against for cardiovascular disease
and for blood clotting in the brain.
Conclusion
1.Discarded resource of seeds from
unripe green Goya might be able to
use for functional food or supplements.
2. Reduce 500 ton of waste every year.
Contribute to environmental conservation.
？
Future work
1． Identifying peak no.5 of unknown
polyunsaturated fatty acid.
2． Evaluate its bio-functionality.
Find out for the potential of
bio-functional value to the
useless Goya seeds!
Final goal in our research
Making the new bio-functional
products from Goya seeds.
・activate economy in Okinawa.
・contribute to environmental conservation.

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