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Identification of unsaturated fatty acid from discarded Goya seeds.Okinawa National College of TechnologyBioresources engineering

*Momoko Mashiyama, Yuka Shingaki, Eisuke Kuraya and Kei Hirayama

1Our research addressed for added extra value to the unused Goya seeds that containing high amount of polyunsaturated fatty acid.Purpose add value

Functionally of Goya seeds2

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 groundHowever the seeds are discarded all the time.Is it really useless ?

Goya is summer vegetable.Very bitter3

Back groundHarvest of Goya are about 25,000 ton in Japan.[1][1] Ministry of Agriculture, Forestry and Fisheries, Research for production of local specialty vegetables. Annual statistic reports on 2008.

Large quantity of Goya seeds about 500 ton have been discarded.

This picture is Goya Seeds.4Methods

Thin layer chromatographyFlow controllerCarrier gasColumnSample injectorColumn ovenMass spectrometryGC-MSanalysisVisualized by IodineExperimental flow Soxhlet extraction Methyl esterification

To find out plants hold rich polyunsaturated fatty acid.

To prepare crude seeds oil

Analyzed by GC-MS.5TLC plate : silica gel (whatman)Solvent : hexane: diethyl ether: acetic acid (80:20:1, V/V)Development for fatty acid:visualized by Iodine.

Method Conditions for thin layer chromatographyFluid ? Wet?

6Preparation of crude Goya seeds oil.

seeds were separated from Goya fruit.

dried at 50 for 24 hours.

homogenized by blender for 30 sec.Method

Vertical angle of Goya seed

Goya seedFluid ? Wet?

7 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.Method

50248

Column DB-23Agilent Technologies(50 cyanopropyl)-methyl polysiloxane ID 0.25mm30m Film thickness 0.25m

The analytical conditions analyzing fatty acid methyl ester

JET240 DET240 Split10Total flow15.0 ml/minLinear velocity40.0 cm /s 50,1min 175,25/min 230,4/min , 5.25min (HOLD) MethodGC-17AShimadzu CorporationGC-MS analytical conditionsIDinside diameter0ml9

free fatty acid1 2 3 41. Black rice2. Goya seed3. Oleic acid4. Linoleic acidThin layer chromatography of plant oil triglycerideWe 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.Therefore, we focused on analyzing unsaturated fatty acid from Goya seeds.Results10Five 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.

Goya seeds oil analyzed by GC-MS Results11ResultsStearic acid Total ion chromatogram of fatty acid from Goya seeds oil by GC-MS.Two saturated and three unsaturated fatty acids were detected. Unknown polyunsaturated fatty acid No.5intensity

Unknown polyunsaturated fatty acid No.50.51.51.02.02.53.03.54.04.55.0010712

No.5 fatty acid51.5%Linoleic acid 9.6% Oleic acid6.8%Stearic acid28.1%Palmitic acid4.0%Ratio of fatty acid contained in the Goya seeds.Results67.9%Unsaturated32.1%Saturated

No.5 fatty acid was polyunsaturated fatty acid and which account for over half of all fatty acids.13

- linolenate methyl ester standard

No.5 polyunsaturated fatty acidResultsMass spectrum

No.5 polyunsaturated fatty acid has similar mass spectrum patern with - linolenate except peak height for 292.14Data from total ion chromatogram, no.5 polyunsaturated fatty acid has 20 carbon.

Results

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Predicted structure for no.5 polyunsaturated fatty acidThe position of double bond in this polyunsaturated fatty acid was determined by fragment ion of m/z 292.Results

16 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.Conclusionmakeacontribution to environmental conservation.

17Polyunsaturated fatty acidhas various functionalities.

For ExampleReduce risk against for cardiovascular disease and for blood clotting in the brain.Conclusion

18Discarded resource of seeds from unripe green Goya might be able to use for functional food or supplements.

Reduce 500 ton of waste every year. Contribute to environmental conservation.

Conclusion

191 Identifying peak no.5 of unknown polyunsaturated fatty acid.

2 Evaluate its bio-functionality.Future workFind out for the potential of bio-functional value to the useless Goya seeds!20Making the new bio-functional products from Goya seeds.

activate economy in Okinawa.contribute to environmental conservation.

Final goal in our research

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