Validation Study of Two Rapeseed Oil Body Extraction Methods

1. Validation Study of TwoRapeseedOil Body Extraction Methods Stephanie BLASER Marc ANTON, Eisabeth DAVID-BRIAND, Thibault LOISELEUX INRA 18/07/2014

2. Introduction • Rapeseed • Brassicanapus • Common uses • Bulkoil - canola oil • High proteinanimal feed • Biodiesel

3. Introduction • Oil bodies • Energystorage structures in plant seeds • 0.5- 2.5 µm diameter • Components • Triaclyglycerolcore • Outer phospholipidmonolayer • Charged surface proteins - oleosins • Structure givesphysical and chemicalstability

4. Introduction • Potential applications in the humanfood system • Deliver stable, preemulsifedoilintoappropriatefoodsystems • Create a poorly-digestedemulsion, through high pressure processing, for the increasing obese population • Have an oil source extractedwithoutsolvents • Objective • Find the methodthatextracts the highestquantity of oil bodies with the highestpurity

5. Materials and Methods • Extraction Method 3

6. Materials and Methods • Extraction Method 5

7. Materials and Methods • Extraction 3 and 5 treatments • Regular - freshcream in phosphate buffer (100 mg/ml) • Freeze-driedcream in phosphate buffer • Freeze-driedcream in water • Analyses • Dry Matter • Size • Microscope • Granulometer • Nanosizer • Protein quantification • BSA • Lipid quantification • Isopropanol/hexane extraction

8. Results

9. CreamWeights

10. Light Microscope Method 3 Method 5 63x 63x

11. Confocal Microscope Method 3

12. Granulometer

13. Nanosizer – Method 3 Regular Freeze-dried

14. Nanosizer – Method 5 Regular Freeze-dried

15. Lipid Quantification

16. Protein Quantification

17. Dry Matter

18. Discussion • Cream Collection Weights • No major differencebetween Method 3 and 5 • Size • Method 3 vs 5 • Bothdisplayedsimilarpresence of floculation and particle sizes- granulometer, nanosize, and light microscope • Regular vs. Freeze-dried • Light microscope • No major visualdifferences • Granulometer • More floculation present in samplesfreeze-driedwithbuffer • Nanosizer • Freeze-driedsamplesshoweda wider range of particle sizes • Differences in freeze-driedsamplespotentiallydue to destruction of oil bodies duringharshtreatment

19. Discussion • Lipid Quantification • Slightlyhigher collection from Method 3 • Protein Quantification • Slighlylower concentration from Method 3 • Dry Matter • Slightlyhigher Percent Dry Matterfrom Method 3

20. Discussion • Challenges • Cream collection • Cream can stick to cap of centrifuge tube • Cream in Method 3 is, in general, not as firm and durable as Method 5 • Size measurement instrumentation • Granulometer and nanometer have size detectionlimitsthat are on bothsides of the oil bodies upper and lowerdiameter range • New equipmentarrivingnextmonth • Freeze-drying • Low volume of final product • Time consuming • Slightlylowerprotein content • With buffer • Powdery final product - easier to collect • Contained phosphate buffer salts • Room for errors in calculations • With water • Waxy final product - more difficult to collect

21. Sustainability • Future options • Usingsustainabiliygrownseeds • 2010 Unilever Sustainable Agriculture Code • Put intoeffect in Germany under Cargill • Using the valueableprotein for a humanfood source and not just for animal meal • Hurdles: glucosinolates, phenolics, phytates, and high amount of fiber • Benefits: balanced amino acid profile, functional properties (emulsifying, foaming, and gelling), and new alternative to feed increasing population • Creatingindustrialscale extraction methodswithout the use of dangeroussolvents

22. Conclusion • Method 3 • More consistent • Less time consuming • Lessprotein contamination • Method 5 • Easier handling of cream • Long soakstep • More protein contamination • Freeze-drying • Time consuming • Few addedbenefits • Nextstep • Compare againstThibault’s data • Increase collection volume to largerbenchscale

23. Thank You Stephanie Jung, PhD Marc Anton, PhD Elisabeth David-Briand Thibault Loiseleux