IN THE UNITED STATES PATENT AND TRADEMARK OFFICE
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`In re Inter Partes Reexamination of U.S. Patent No. 8,030,348
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`NATURAL MARINE SOURCE PHOSPHOLIPIDS COMPRISING
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`POLYUNSATURATED FATTY ACIDS AND THEIR APPLICATIONS
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`4 October 2011 to Sampalis
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`Control No.:
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`95/001,774
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`October 19, 2011
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`Bruce Campbell
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`First Supplemental Declaration by Bjorn Ole Haugsgierd, MSc, in
`Support of Reguest for Inter Partes Reexamination of
`U.S. Patent N0. 8,030,348
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`EFS WEB Filed
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`Mail Stop Inter Partes Reexam
`Commissioner for Patents
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`PO. Box 1450
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`Alexandria, VA 22313-1450
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`I, Bjnrn Ole Haugsgjerd, MSC, state as follows:
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`My present position is Deputy Manager at Nofima BioLab, Norway
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`At the request of Aker Biomarine ASA, I have extracted lipid fractions from Euphausia
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`superba and Euphausia pacg’/ica by the methods described in Beaudoin I (WO 00/23546),
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`Beaudoin II (Canadian Application 2,251,265). Following the extraction, I shipped the samples
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`to Vitas AS, Oslo, Norway, and Dr. Richard Van Breemen, of the University of Illinois for
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`analytical analysis. Frozen Euphausia superba and Euphausia paczfica were provided by Aker
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`Biomarine ASA.
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`000001
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`

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`I repeated the Beaudoin I (pages 5-6 and Table 19) and Beaudoin II (page 2 and Table
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`11) extractions with acetone in the first step and then either ethanol or ethyl acetate in the second
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`step. The extractions were performed as described in Beaudoin I and Beaudoin II. Separate
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`extractions were conducted for Euphausia superba and Euphausia pacific. I utilized the
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`following protocol:
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`0 Grind frozen krill at 4C to reduce particle size to less than 5mm.
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`0 Extract with acetone at 4C at a samplezacetone ratio of 1:6 (w/v) for 2 hours with 20 minutes
`of swirling.
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`Filter on organic solvent resistant filter paper under reduced pressure at 4C.
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`0 Wash solid material on filter with a sarnplezacetone ratio of l :2 (w/v) with pure and cold
`acetone.
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`0 Combine filtrates and evaporate solvent under reduced pressure.
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`0 Allow water residue obtained after evaporation to separate from oil phase (Fraction I) at 4C.
`Store Fraction I at 4C.
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`0 Divide solid material on filter into two aliquots, aliquot l and aliquot 2.
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`0 Extract aliquot l with pure ethanol at samplezethanol ratio of 1:2 (w/v) for 30 minutes at 4C.
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`Filter on organic solvent resistant filter paper under reduced pressure at 4C.
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`0 Evaporate solvent under reduced pressure to provide Fraction Ila.
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`0 Extract aliquot 2 with pure ethyl acetate at sample:ethyl acetate ratio of l :2 (w/v) for 30
`minutes at 4C.
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`Filter on organic solvent resistant filter paper under reduced pressure at 4C.
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`0 Evaporate solvent under reduced pressure to provide Fraction IIb.
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`

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`0 Divide Fraction I into three aliquots. In an oil bath, heat one aliquot to 60C for 5 minutes and
`another aliquot to 125C for 15 minutes under an inert atmosphere, label as E. pacifica (or
`superba) Fraction I heat treated 60C or 125C. Store at —20C until further analysis. Label the
`third aliquot as E. pacifica (or superba) Fraction I not heated. Store at -20C until further
`analysis.
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`0 Divide Fraction Ila into three aliquots. In an oil bath, heat one aliquot to 70C for 5 minutes
`and another aliquot to 125C for 15 minutes under an inert atmosphere, label as E. pacifica (or
`superba) Fraction Ila/ethanol heat treated 70C or 125C. Store at -20C until further analysis.
`Label the third aliquot as E. pacifica (or superba) Fraction Ila/ethanol not heated. Store at -
`20C until further analysis.
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`0 Divide Fraction IIb into three aliquots. In an oil bath, heat one aliquot to 70C for 5 minutes
`and another aliquot to 125C for 15 minutes under an inert atmosphere, label as E. pacifica (or
`superba) Fraction IIb/ethyl acetate heat treated 70C or 125C. Store at -20C until further
`analysis. Label the third aliquot as E. pacifica (or superba) Fraction IIb/ethyl acetate not
`heated. Store at -20C until further analysis.
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`0 The samples were marked as follows:
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`Fraction number
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`Temperature
`treatment (°C)
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`Time (min)
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`Marking of sample
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`E. superba I — E. superba 1/ Acetone Not heated
`E. superbal
`E. superba I / Acetone 60C
`E. superbal
`E. superba I / Acetone 125C
`E. superba Ila — E. superba Ha / Etanol Not heated
`E. superba Ha
`E. superba IIa/ Etanol 70C
`E.superbana
`E. superba IIa/ Etanol l25C
`Not heated _ E. superba Ilb / Ethyl acetate Not heated
`E. superba IIb
`70
`E. superba Ilb — E. superba IIb / Ethyl acetate 70C
`125
`E. superba Hb —
`E. superba Hb / Ethyl acetate 125C
`apacmcai
`—
`E. pacifica I
`
`E. pacifica I / Acetone Not heated
`E. pacifica I / Acetone 60C
`E. pacifica I/ Acetone 125C
`125
`Not heated — E. pacifica Ila / Etanol Not heated
`70
`E. pacifica Ila / Etanol 70C
`125
`E. pacifica Ila / Etanol 125C
`Not heated — E. pacifica llb / Ethyl acetate Not heated
`E. pacifica IIb / Ethyl acetate 70C
`E. pacifica Hb / Ethyl acetate 125C
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`E. pacifica I
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`E. pacifica Ila
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`E. pacifica Ila
`E. pacifica Ila
`E. pacifica Ilb
`E. pacifica IIb
`E. pacifica Ilb
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`It is my understanding that Neptune’s experts have criticized the heating procedure used
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`in the first set of repeats.
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`I understand that Dr. Gundersen heated the samples in a heat block that
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`had been placed in an oven, which is a reasonable heating method that one of skill in the art
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`would use to heat samples prior to analysis as described in Beaudoin.
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`I disagree with these
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`criticisms, but used an oil bath in this set of extractions to remove that criticism.
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`I further declare that all statement made herein of my own knowledge are true and that all
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`statements made on information and belief are believed to be true; and further that these
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`statements were made with the knowledge that willful false statements and the like so made are
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`punishable by fine or imprisonment, or both, under section 1001 of title 18 of the United States
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`Code, and that such willful false statements mayjeopardize the validity of the application or any
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`patent issued thereon.
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`Respectfully submitted,
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`Bjorn Ole Haugsgjerd, MSc
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`Date
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