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Validation Study of Two Rapeseed Oil Body Extraction Methods. Stephanie BLASER Marc ANTON, Eisabeth DAVID-BRIAND, Thibault LOISELEUX INRA 18/07/2014. Introduction. Rapeseed Brassica napus Common uses Bulk oil - canola oil High protein a nimal feed Biodiesel. Introduction.

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Validation Study of Two Rapeseed Oil Body Extraction Methods

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Validation study of two rapeseed oil body extraction methods

Validation Study of TwoRapeseedOil Body Extraction Methods

Stephanie BLASER

Marc ANTON, Eisabeth DAVID-BRIAND, Thibault LOISELEUX

INRA

18/07/2014


Introduction

Introduction

  • Rapeseed

    • Brassicanapus

    • Common uses

      • Bulkoil - canola oil

      • High proteinanimal feed

      • Biodiesel


Introduction1

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


Introduction2

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


Materials and methods

Materials and Methods

  • Extraction Method 3


Materials and methods1

Materials and Methods

  • Extraction Method 5


Materials and methods2

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


Results

Results


Cream weights

CreamWeights


Light microscope

Light Microscope

Method 3

Method 5

63x

63x


Confocal microscope

Confocal Microscope

Method 3


Granulometer

Granulometer


Nanosizer method 3

Nanosizer – Method 3

Regular

Freeze-dried


Nanosizer method 5

Nanosizer – Method 5

Regular

Freeze-dried


Lipid quantification

Lipid Quantification


Protein quantification

Protein Quantification


Dry matter

Dry Matter


Discussion

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


Discussion1

Discussion

  • Lipid Quantification

    • Slightlyhigher collection from Method 3

  • Protein Quantification

    • Slighlylower concentration from Method 3

  • Dry Matter

    • Slightlyhigher Percent Dry Matterfrom Method 3


Discussion2

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


Sustainability

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


Conclusion

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


Thank you

Thank You

Stephanie Jung, PhD

Marc Anton, PhD

Elisabeth David-Briand

Thibault Loiseleux


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