. . (cid:9)
`
`• Sc_afl*Rti• (cid:9)
`
`*OW' •
`
`SURVEY'
`
`°
`
`. (cid:9)
`
`- .
`
`SOUTHERN OCEAN
`ilization Of- Kril
`
`RIMFROST EXHIBIT 1032 page 0001
`
`OLYMSEA877ITC00163935
`
`

`

`RIMFROST EXHIBIT 1032 page 0002
`
`OLYMSEA877ITC00163936
`
`

`

`CREGOfl
`JUN 0·1 1981
`
`~ l1.ffl
`3
`
`TEE UTILIZATION OF KRILL
`
`by
`
`G.J. Grantham
`Consultant to the UNDP/FAO
`Southern Ocean Fisheries Survey Programme
`
`FOOD AND AGRICULTURE ORGANIZATION OF THE UNITED NATIONS
`
`UNITED NATIONS DEVELOPMENT PROGRAMME
`
`Rome, 1917
`
`RIMFROST EXHIBIT 1032 page 0003
`
`

`

`The designations employed and the presentation
`of material in this publication do not imply the
`expression of any opinion whatsoever on the
`part of the Food and Agriculture Organization
`of the United Nations concerning the legal
`status of any country, territory, city or area or
`of its authorities, or concerning the delimitation
`of its frontiers or boundaries.
`
`M-43
`ISBN 9 2-5-100416-1
`The copyright in this book is vested in the Food and Agriculture Orga-
`nization of the United Nations. The book may not be reproduced, in whole
`or in part, by any method or process, without written permission from
`the copyright holder. Applications for such permission, with a statement
`of the purpose and extent of the reproduction desired, should be addressed
`to the Director, Publications Division, Food and Agriculture Organization
`of the United Nations, Via delle Terme di Caracalla, 00100 Rome, Italy.
`
`©FAO 19 77
`
`RIMFROST EXHIBIT 1032 page 0004
`
`OLYMSEA877ITC00163938
`
`

`

`THE SOUTHERN OCEAN FISHERIES SURVEY PROGRAMME
`
`In view of the potential importance of the liVing resources of the Southern Ocean, the
`United Nations Development Programme (UNDP) Governing Council approved in January 1976 a
`preparatory phase of a Southern Ocean Fisheries Survey ProgramMe, to be executed by the
`Food and Agriculture. Organization of the United Nations (FAO). The long-term objective of
`the Programme is to improve the knowledge of the nature, magnitude and distribution of the
`living resources of the Southern Ocean (South of latitude 45°S), with a view to assisting in
`their eventual rational utilization.' The immediate activities of the preparatory phase (to
`continue until the end of 1977) include a review of the present information on the distribu-
`tion and magnitUde of the living resources of the Southern Ocean; a review and evaluation of
`the present state of exploitation and utilization of these resources; and the establishment
`of a system for regular compilation of statistical and other information about them.
`
`This report is one of a series prepared by FAO under the preparatory phase of the
`Programme. It gathers together current knowledge on the biochemistry, processing and
`marketing of Antarctic krill. Other reports in this series assemble current knowledge on
`the magnitUde and distribution of the living resources of the Southern Ocean, and on the
`technology of krill harvesting.
`
`E.S. Holliman
`Programme Co-ordinator
`
`RIMFROST EXHIBIT 1032 page 0005
`
`OLYMSEA877ITC00163939
`
`

`

`1/ (i) It will be observed that in several of the compositional tables, percentage
`proportions do not always total 100. This can be for several reasons:
`
`- experimental error in the derivation of the original data
`- the use of averages of several results
`- the "rounding" of results to the appropriate decimal significance
`— the inclusion of sub—totals in the data columns
`- the inclusion of one item within another. (For example, in Table 1,
`separate values are given for crude protein and chitin contents.
`However, the chemical technique used for the determination of crude
`protein will also identify chitin as protein. Similarly, in Table 11,
`the value for salt content will be included in the total ash valUe.)
`- the experimental rather than arithmetical determination of totals
`- the arithmetical correction of data not expressed on a percentage
`basis .in the original literature
`- the absence of values for particular parameters
`- the conversion of data from a dry weight to a wet weight badis,
`or vice—versa
`- the conversion of units of measurement (for example, the conversion
`of % nitrogen to % protein or of International Unite (I.U.1 for
`vitamins to microgramme % by the use of appropriate factors .
`
`As a consequence, totals are only given where it is valid to do so. They are
`expressed as true values, or as the values given in the original source. Where
`a total is not the theoretiCally expected 100, the difference gives an indica—
`tion of the likely degree of error in the data.
`
`(ii) For similar reasons, data expressed on a non—percentage basis do not always give
`the theoretical or stated total.
`
`(iii) Where data are not available for particular parameters, the entry is left blank.
`The use of the symbol " — " signifies a null result or that a component was tot
`detected.
`
`Parentheses " ( ) " are used when a value is not incorporated in a total or an
`average. This can be when:
`
`- the original figure would seem to be erroneous (in which case, a
`question mark " ? " is also used)
`- one.-omponent is included in another
`- the figure is a sub—total
`
`(v) Averages are either arithmetic means or weighted means, as appropriate
`
`(vi) Values are not generally incorporated in overall averages when they are
`expressed as ranges in the original source
`
`(vii) Specific comments are given in footnotes to the respective tables.
`
`RIMFROST EXHIBIT 1032 page 0006
`
`OLYMSEA877ITC00163940
`
`

`

`- v -
`
`TABLE OF'CONTENTS
`
`ABSTRACT
`
`1. BACKGROUND
`2.
`BIOCHEMICAL COMPOSITION
`
`2.1 Introduction
`2.2 Proximate Composition
`2.3 Component Composition
`2.4 Protein
`2.5 Fat
`2.6 Vitamins
`2.7 Minerals
`2.8 Chitin
`2.9 Calorific Values
`2.10 Contaminants
`
`3. PROCESSING
`
`3.1 Introduction
`3.2 Raw Material Properties
`3.3 Handling and Pre-processing
`
`3.3.1 Conveying
`3.3.2 Raw Material Storage
`3.3.3 Sorting
`3.3.4 Grading
`
`3.4 Processing Options
`3.4.1 Gut Removal
`3.4.2 Intact Flesh Removal
`3.4.3 Minced Flesh Removal
`3.4.4 Structuring
`3.4.5 Concentrates, Isolates and Meal
`3.4.6 StabiiIzation
`3.4.7 Preservation
`3.4.8 By-product Recovery
`
`4.
`
`PRODUCTS AND MARKETS
`
`4.1 Introduction
`4.2 Whole Krill
`4.3 Whole Tail Meats
`4.4 Coagulated Paste
`4.5 Krill Mince
`4.6 Concentrates, Extracts and Ferments
`4.7 Miscellaneous Food Products
`4.8 Human Nutrition
`4.9 Legislation
`4.10 Non-food and By-products
`
`4.10.1 Krill Meal
`4.10.2 Chitin
`4.10.3 Other By-products and Residues
`
`Page
`
`1
`2
`
`3
`
`3
`3
`5
`5
`11
`11
`16
`16
`16
`16
`
`18
`
`18
`18
`19
`19
`20
`21
`21
`
`21
`
`21
`22
`23
`24
`27
`28
`30
`31
`
`33
`
`33
`34
`37
`38
`41
`41
`43
`43
`44
`44
`
`44
`45
`49
`
`RIMFROST EXHIBIT 1032 page 0007
`OLYMSEA877ITC00163941
`
`

`

`- vi
`
`5.
`CONCLUSIONS
`
`6.
`REFERENCES
`
`li
`
`LIST OF TABLES-/
`
`1. The composition of krill
`
`Variation of water, fat, protein and ash content of E. superba
`2.
`3:. The components of krill
`4. The protein composition of krill
`5. The amino acids of krill
`4. The lipids of krill (cid:9)
`composition of crude lipid
`7,. The lipids of krill - fatty acid composition
`8'. The vitamins of krill
`9,. The minerals of krill
`10. Flow sheet and estimated materials balance for krill meal production
`11. The composition of boiled, frozen krill
`12. The proximate, mineral and amino acid analyses of Russian krill paste
`13. Russian patents for products manufactured from krill paste
`14. The composition of krill mince
`15. The composition of krill meal
`16. The amino aoid composition of krill meal
`
`page
`
`50
`
`52
`
`4
`6
`7
`8
`9
`12
`13
`15
`17
`29
`36
`39
`40
`42
`46
`47
`
`RIMFROST EXHIBIT 1032 page 0008
`
`OLYMSEA877ITC00163942
`
`

`

`THE VIIILIZILTION OP BILL
`
`The report reviews available information on the technology
`f ijmolt:ar8tic jkrill. ona1 racoon , ' given-
`
`of
`to the bioOhmeicalianOechitelogioal 'properties of krill,
`to the reported processiig teohniques, and to product appr_
`tleeshiSis,and markets. While process and product development
`is proceeding rapidly, no one obvious product application
`has emerged. Likely product spectra will vary from country
`to country. Process technology is advanced, products are
`generally sophisticated and costly. Assistance will be
`necessary to develop alternative technologies and products
`suited to the developing countries.
`
`RIMFROST EXHIBIT 1032 page 0009
`
`OLYMSEA877ITC00163943
`
`

`

`1. BACKGROUND
`
`This report was. prepared during a ten week consultancy to the preparatory phase of the
`UNDP/FAO Southern Ocean Fisheries Survey Programme, from May to July, 1977. Its aim is to
`review the information available on the technology of the utilization of the Antarctic krill,
`and on product and market opportunities. It does not attempt any assessment of feasibility,
`but it provides the information, indicates sources, and identifies areas where further
`knowledge is needed for such an assessment to be made in any particular situation.
`
`Chapter 2 of the report considers the biochemical composition of krill, as related to
`its technological properties in process, product and nutritional performance. Chapter 3
`enumerates the various processing techniques that have been applied to krill to date, and
`suggests areas worthy of further research or development. Chapter 4 discusses product
`applications reported for krill, and, where possible, indicates likely markets, values,
`and tonnages. Finally, Chapter 5 assesses the current state of the art and likely commercial
`developments. It also identifies the aspects of krill utilisation that might require further
`investigation at international, national, or industrial levels prior to successful exp-
`loitation.
`
`The findings of the report should be considered in the light of the prevailing'
`situation in krill exploitation. The immense size of the resource has been confirmed, at
`least to the extent that the sustainable yield will exceed by orders of magnitUde the
`likely demand for many years to come. It has also been confirmed that krill can be located
`and harvested on a commercial basis, given economically viable outlets for the catch. With
`a few exceptions, hoiever, this latter reqUirement has yet"to be met. In other words, it
`can be said that successful exploitation of the resource is now dependent on the development
`of suitable process and product technology and on the adoption of appropriate marketing or
`disposal strategies.
`
`It is recognised that some of the tentative conclusions drawn here may be out of date
`or erroneous. This is because complete information could not always be obtained (often an
`indication in itself of the imminence of commercial activity). It must also be noted that,
`while factual details are recorded from published literature and other sources, the opinions,
`conclusions and recommendations are those of the writer. The specific mention of manufacturers'
`equipment, plant or processee does not necessarily imply approbation, nor does it preclude
`the value of un-named. alternatives.
`
`Acknowledgement is due to Unilever. Research Laboratories through whese resources much
`of the information incorporated in this report was collected.
`
`RIMFROST EXHIBIT 1032 page 0010
`
`OLYMSEA877ITC00163944
`
`

`

`-3-
`
`2. BIOCHEMICAL COMPOSITION
`
`2.1 (cid:9)
`
`Introduction
`
`The biochemical composition of krill will determine its technological and nutritional
`properties and thus directly influence the selection of processing and product options.
`Commercial catches of krill would seem to consist predominantly of Euphausia superba.
`Therefore the biochemical composition of the catch will be characterized by the euphausiid,
`although the other minor components of the krill crop cannot always be ignored, as will be
`discussed later.
`
`Only the composition of whole krill is considered in this chapter•, the analyses of the
`various components are enumerated in the chapters on processing and products (3 and 4), as
`well as the nutritional implications.
`
`SoMe of the biochemical properties of krill have been extensively spalymed, others less
`so. This chapter attempts to correlate and assess the results available and to identify
`areas where further information is needed. It will be seen at once that the chemical com—
`position and food qualities of gisuperba are very similar to those of known related species—
`shrimps, crabs,' lobsters, etc. It will also be noted that precise agreement between the
`various authors is rare. This is almost certainly due to the complex interactions of age,
`season, location, sex, physiological condition and diet, in the various samples that have,
`been studied and analysed. Correlation of these factors is difficUlt and in fact probably not
`necessary to successful exploitation.
`
`2.2 Proximate Composition
`
`The results of some twenty authors are shown in Table 1 and summarized below:
`
`THE PROXIMATE COMPOSITION OF WHOLE E.SUPERBA: SUMMARY OF LITERATURE VALUES
`
`Moisture
`
`% dry weight
`Crude
`Crude 1/ (cid:9)
`Protein (cid:9)
`Fat (cid:9)
`
`Ash
`
`1, wet weight
`Crude
`Crude 1/ (cid:9)
`Protein (cid:9)
`Fat
`
`Average
`
`80.1
`
`Mean Maximum :'83.1
`
`Mean minimum 77.9
`
`65.1 (cid:9)
`
`77.5 (cid:9)
`
`59.7 (cid:9)
`
`14.2 (cid:9)
`
`13.9
`
`13.0 (cid:9)
`
`26.0 (cid:9)
`
`16.7
`
`15.4 (cid:9)
`
`2.8
`
`5.1
`
`6.7 (cid:9)
`
`11.7
`
`11.9 (cid:9)
`
`1.3
`
`1/ Total nitrogen x 6.25, includes non—protein material
`
`It can be seen, then, that the composition of krill is well balanced in terms of a
`potential foodstUff. As a pelagic crustacean, it has a higher moisture and fat content
`than bottom living (benthic) species — and proportionally less crude protein.
`
`The composition is variable. Juvenile krill possibly have a higher water and fat
`content than adults with a proportionally lower protein content, but available results are
`contradictory and insufficient. The fat content would also seam to vary with the season,
`increasing from an average of 5 percent up to 30 percent dry weight as the summer progresses
`(Il'ichey1967, Littlepage 1964),. This seasonal increase is associated with a decrease in
`water and ash contents, and probably serves to build up energy reserves for the winter
`period of food scarcity.
`
`RIMFROST EXHIBIT 1032 page 0011
`
`OLYMSEA877ITC00163945
`
`

`

`- 4 -
`
`Table 1
`The Composition of Krill
`Reported literature values for the proximate oomposition of Whole Euphausia superba
`
`Souroe
`
`% K2O
`
`Crude Protein
`0 x 6.25)
`
`Crade-
`Fa*
`
`Carbohydrate
`
`Ash
`
`Chitin
`
`% Dry Weight
`
`Arai 1976
`Burkholder 1967
`Bykov 1975
`Egorova 1970
`Gilberg 1971
`Il'iohev 1967
`Krytiohkava 1969 a
`Kryaohkava 1969 b
`Kryaohkava 1970
`KAMM 1975 a and 1976
`Lagunov 1974
`Wonaka 1964
`Pieroe 1969
`Raymaat 1971 Range
`Average
`Rosohke 1976 a'
`Sidhu 1970
`Srinivasagam 1971
`Vinogradays, 1960
`Watanabe 1976
`Tanase 197i
`Tanase 1974 a
`
`Range
`Average
`Wean Minimum
`Mean Nexieqm
`
`a/ Insufficient data
`
`76.9
`.63.9
`51 .4-80.5
`61.1-77.9
`53.1
`73.0-74.5
`57.0-78.0
`56.0..80.0
`65.0-80.5
`65.2
`80.4
`
`61.8
`50.0
`52.7
`60.1
`49.'0
`44.0
`55.6-60.8
`66.5-86.9
`56.0
`67.3-68.6
`
`44.0-86.9
`65.1
`59.7
`77.5
`
`.
`
`73.7...82.3
`
`75.6-81.6
`
`73.1.782.3
`79.0-82.6
`79.6
`79.0
`
`82.0
`78.4
`- (cid:9) 71.4
`84.2
`85.3-85.8
`79.8-85.2
`81.6
`78.7-81.7
`
`71.4,-85.2
`80.1
`77.9
`83.1
`
`7.1
`17.8
`5.0..26.4
`
`21.9
`6.513.9
`
`5.0-26.0
`5.7-16.7
`12.3
`5.7
`19.4
`18.7
`6.0-33.0
`13.3
`14.1
`24.6
`
`19.0?
`
`4.2
`
`5.0
`4.7
`
`2.5-26.0
`7.3-19.9
`18.4
`15.2-19.9
`.
`
`2.5-33.0
`14.2
`6.7'
`26.0
`
`6.028.47
`
`10.9
`3.6-5.4
`-
`3.6-10.9
`5.7
`2/
`
`15.4
`13.1
`10.5-18.2
`
`10.4
`13.1...19.0,
`
`10.0-15.0
`11.0-16.1
`13.7
`10.9
`
`18.9
`
`17.0
`10.4
`9.8
`
`6.7... 8.7
`14.7-21.3
`14.7
`16.1-18.9
`
`6.7-21.3
`13.9
`11.7
`16.7
`
`6.1
`12.3?
`
`2.3
`
`4.1
`
`2.5
`
`2.3-6.1
`' (cid:9) 3.8
`A/
`
`RIMFROST EXHIBIT 1032 page 0012
`
`OLYMSEA877ITC00163946
`
`

`

`-5—
`
`However, the absence of waxes — discussed later— implies that the krill do not fast over the
`winter. The carbohydrate content remains fairly constant at around 6 percent dry weight; this
`is slightly higher than most zooplankton, and probably reflects the herbivorous habit of _
`krill (Raymont, Srinivasagam and Raymont 1971, Ferguson and Raymont 1974). The variations of
`water, protein and fat content with age and sex have been quantified by Russian workers.
`These are given in Table 2 which also gives Clarke's (1976) correlation between fresh weight,
`dry weight and ash weight.
`
`It can only be concluded, therefore, that the proximate compobition of krill is
`essentially variable, and that allowancvfor this should be made in the development of any
`process or compositional specification:
`2.3 (cid:9)
`
`Component Composition
`
`In gross terms, whole raw krill can be said to comprise of approximately 28% tail meat,
`34% cephalothorax (head section) and 26% carapace (tail shell); the,balance of 12% being lost
`on separation. The proximate composition of these components is given in Table 3 but again
`data are limited.
`
`2.4 Protein
`
`It has been shown that the content of the nitrogenous eUbstances in krill varies
`around 13% wet weight, expressed as Nx6.25. Information regarding the quantity, species and
`molecular structure of the proteins present is somewhat confused and contradictory: Bykov
`(1975) and Srinivasagam et al (1971) report that some 80% of the total nitrogen is contained
`in true protein, while Yanase (1971) states that about 55% of the crude protein is hot water
`extractable (i.e. suggesting low molecular weight peptides, free amino acids, stroma proteins
`and possibly tropomyosin). Similarly Gilberg .(1971) records a notably high proportion of free
`amino acids—over 50% as compared with 30% in crayfish and 10-15% in round fish. Some of
`these discrepancies (and some of the soluble nitrogen) can probably be accounted for by the
`rapid autolysis of the krill after catching and, to a lesser extent, during frozen storage
`of the samples. Table 4 attempts to correlate these results and concludes that the 13% wet
`weight of total nitrogen, expressed on a crude protein basis, is made up of 8% true protein
`and 2.5% free amino acids; the balance of the nitrogen will be volatile bases,ohitin, and nucleic
`acids. However, further work is needed before these figures can be accepted with confidence.
`
`Qualitative disc electrophoresis has identified actin, myosin and actomyosin in the
`muscle protein, of forms reportedly different from shrimp (Maekawa et al 1975a, Matsumoto
`et al 1976a). Thermal denaturation of the myosin occurs between 40 and 65°C; the albuminous
`material in the flesh denatures at 60-68°C; the blood proteins denature at 70-74°C (Niohiro
`Gyogyo ICaisha Ltd. 1976).
`
`quamtative determinations of content and molecular weights would be desirable, together
`with analyses of reactivity and identification of other protein species (myogen, tropomyosin,
`stroma proteins).
`
`The amino acid composition of krill protein has been well studied, and is characterized
`by its relatively high content (0%) of the essential amino acids and by the presence of most of
`the unessentials; these are tabulated against some reference standards in Table 5. It will
`be seen that glutamine, aspartic acid, lysine, leucine and proline are predominant (about
`half the total), while cysteine and possibly trytophaneare limiting. The relatively high
`free glycine possibly serves as protection from the sub—zero sea temperatures (Srinivasagam
`et al 1971). Japanese workers have also found significant quantities of anserine and betaine.
`
`1/ The situation is simplified, however, by the reportedly low genetic variability of the
`composition of the krill stocks (Ayala, Valentine and Zumwalt 1975, Valentine and Ayala
`1976)
`
`RIMFROST EXHIBIT 1032 page 0013
`
`OLYMSEA8771TC00163947
`
`

`

`Table 2
`
`Variation of water, fat, protein and ash contents of
`E. superba with size, age and sex, at the end of the summer
`
`% Fat
`
`Bykov 1975
`Lagunov 1974W
`Bykov 1975
`Lagunov 1974 2/
`
`EN x 6.25
`
`' Bykov-1975
`
`Ash
`
`.Bykov 1975
`
`Adolescents
`<35 mm
`
`80.0-82.0
`73.7
`
`2.5- 4.1
`7.2
`
`12.8-15.0
`
`2.67 1.1
`
`Male
`
`79.3
`
`1.8
`
`Adults 35-55 mm
`Female
`
`75.0
`
`4.3
`
`77.6-80.5
`
`4.1- 5.8
`
`14.5-16.3
`
`2.7- 3.1
`
`2/ After Kryuchkova 1969 b (cid:9)
`
`See also Vinogradava 1960
`
`Clarke (1976) has found a positive correlation between whole fresh weight and
`moisture content (dry wt. (mg) = 0.21 x wet wt. (mg) - 2.27) and between ash weight
`and dry weight (ash wt. (mg) = 0.12 x dry wt. (mg) + 0.94) for adolescent krill
`
`RIMFROST EXHIBIT 1032 page 0014
`
`OLYMSEA877ITC00163948
`
`

`

`Table 3
`
`The Components of Krill
`
`Reported literature values for the proportional and proximate composition of krill body components, As % wet weight
`
`% Proportion
`
`(1)
`
`(2)
`
`(4)
`
`26.2
`
`34.4
`26.4
`
`29.0
`
`27.6
`
`61.5
`
`34.4
`26.4
`
`(1)
`76.3
`80.5
`60.4
`
`% H20
`
`(3)
`77.3
`
`% Total 1
`
`% Crude Pat
`
`% Ash
`
`(3)
`2.9
`
`(4)
`76.8
`80.5
`60.4
`
`(1)
`3.1
`2.0
`1.5
`
`(4)
`3.0
`2.0
`1.5
`
`(1)
`0.2
`0.7
`0.6
`
`(3)
`2.0
`
`(4)
`
`1.1
`0.7
`0.6
`
`(1)
`
`(3)
`
`(4)
`
`1.8
`
`4.2
`6.0
`29.8
`
`3.0
`6.0
`29.8
`
`Tail neat
`
`Cephalothorax
`
`Carapace
`
`Separating Losses
`
`12.8
`
`9.5
`
`11.2
`
`Total
`
`99.8
`
`100.0
`
`99.6
`
`Columna: (1) Illichow 1967 (2) Lagunov 1974 (3) Kryuchkova 1969 b (4) Combined results
`
`RIMFROST EXHIBIT 1032 page 0015
`
`6176E9 1, 0001ILL8V3SVINIO
`
`

`

`Table 4
`
`The Protein Composition of Krill
`
`Reported literature values for the nitrogenous constituents of E. superba, As % N wet weight x 6.25
`
`Seki 1975
`
`Yanase 1974 a
`
`Yanase 1971
`
`Gilberg 1971
`
`Pierce 1969
`
`Average
`
`True Protein
`
`Free Amino Acids
`
`Total Volatile Bases
`
`8.5
`
`) )4.5
`
`8.3
`
`1.6
`
`8.9
`1.5
`
`0.075
`
`0.125
`
`7.5
`1.4
`0.063
`
`)
`)5.7
`
`>5.3
`
`) )12.3
`)
`
`
`8.3
`2.5
`0.088
`
`Total
`
`13.0
`
`10.0
`
`10.4
`
`9.0
`
`14.0
`
`13.6
`
`12.4
`
`11.8
`
`10.9
`
`RIMFROST EXHIBIT 1032 page 0016
`
`096E9 1, 0001ILL8V3SVINIO
`
`

`

`The Amino Acids of Krill
`Reported literature vanes for the amino acid content of E. suPerba protein, against reference proteins, in g/100 g amino acids
`
`Table 5
`
`Total Nitrogenous Substances
`
`r- ON
`
`40
`i
`..4 el
`
`3.3
`3.2
`5.4
`2.9
`6.6
`
`0.5
`1.6
`2.2
`2.5
`
`3.2
`6.6
`3.1
`9.9
`k.4
`3.2
`5.5
`tr
`
`q.
`a
`4.
`0
`
`o .....
`
`5.1
`4.9
`8.3
`4.5
`10.2
`
`0.8
`2.5
`3.4
`3.8
`
`43.5
`4.9
`10.2
`4.8
`15.2
`6.8
`4.9
`8.5
`tr
`
`In
`IA
`I..
`
`0.
`
`5.9
`5.1
`7.7
`4.7
`8.6
`1.5
`1.5
`3.0
`4.1
`6.5
`
`6.2
`12.2
`5.0
`14.6
`4.2
`4.7
`5.5
`2.3
`
`Essential a.a.s.
`Val
`Ilo
`Len
`Thr
`14ye
`Trp
`Cys
`Net
`Tyr
`Plus
`Total Essentials %
`
`Arg
`Asp
`Ser
`Glu
`Pro
`Gly
`Ala
`His
`Tau
`(NH3)
`
`'''"Ci' a
`4. 0
`
`('-•
`
`5.4
`4.0
`7.7
`4.3
`8.1
`1.7
`1.4
`3.1
`4.6
`5.4
`
`......
`1
`I;
`
`i
`a
`1
`
`q
`o
`-P 0
`
`o 4.--.
`
`pi o.
`03 4 •
`
`0
`.-....,
`
`A a 10
`r- I
`i (cid:9) ,,..:. .
`4 .r,L•
`
`o
`'0; ....
`0. e e m
`Q
`
`q
`
`i
`E
`-.
`
`o r-
`cn
`e
`k
`ri
`
`.c. r-
`
`ON
`
`.8 d
`1 d
`
`1)
`k
`to• 4
`
`I
`W
`
`5.2
`5.6
`5.0
`4.9
`7.7
`7.4
`4.2
`4.5
`8.2 10.0
`1.4
`1.4
`1.2
`1.4
`2.8
`2.9
`3.9
`6.2
`
`5.0
`
`7.5
`
`4.7
`4.6
`7.0
`3.8
`9.1
`1.3
`1.1
`2.5
`
`4.6
`
`38.7
`6.8
`
`46.4
`
`5.9
`11.7
`4.8
`13.9
`4.0
`4.5
`5.3
`2.3
`
`3.5
`
`2.8
`
`6.1
`4.5
`8.7
`4.9
`9.2
`1.9
`1.6
`3.5
`5.2
`6.1
`
`7.5
`10.4
`4.6
`15.7
`5.6
`8.6
`6.9
`2.4
`
`6.8
`7.1
`8.0
`7.6
`9.6 10.1
`4.9
`4.7
`7.7
`8.1
`
`1.1
`1.7
`4.4
`5.3
`
`1.2
`1.8
`4.6
`5.6
`
`4.5- 5.9
`4.5- 5.3
`6.7- 8.0
`3.3- 4.5
`6.1-12.6
`0.7- 1.1
`
`2.0- 2.7
`2.9- 3.9
`3.8- 6.1
`
`51.4
`45.7
`6.0
`6.6
`6.3
`3.6- 7.7
`8.7-12.3
`9.2 10.9 11.4
`2.2- 3.6
`3.7
`3.5
`4.1
`13.8 10.5 11.0 10.8-13.6
`1.9- 6.1
`4.6
`4.9
`4.4
`4.7- 6.5
`4.2
`7.6
`4.0
`6.1
`5.7- 7.4
`4.9
`5.1
`2.1
`2.2
`1.3- 1.8
`2-1
`
`5.6
`6.0- 6.4
`4.5- 7.1
`5.2
`4.0- 8.0
`5.2- 5.4
`7.8
`6.7-10.1
`7.5- 7.8
`3.3- 5.1
`4.1- 5.1
`4.3
`9.2
`6.1-12.6
`9.3-11.0
`1.1
`0.7- 1.7
`0.7- 1.3
`1.2
`1.2- 1.3
`0.8- 1.4
`2.5
`1.8- 3.1
`1.9- 2.7
`3.7
`3.0- 3.6
`2.9- 4.6
`3.8--6.2
`4.9
`4.1- 4.3
`44.2-47.5 38.7-51.4 45.6.
`7.1- 8.5
`3.6- 8.5
`6.4
`9.8-12.0
`8.7-12.3
`9.5
`3.8
`2.2- 4.8
`3.4- 4.1
`9.9-10.9 10.8-15.2 12.4
`4.0- 7.6
`5.8- 7.6
`6.7
`4.2- 9.5
`6.0- 9.5
`6.0
`5.1- 8.5
`6.5
`6.6- 7.3
`2.0
`1.3- 2.8
`1.3- 2.3
`
`1.1
`
`1.7
`
`1.4
`
`1.3
`
`2.9- 4.9
`
`1.3- 4.9
`
`2.7
`
`Total
`Unaccounted for
`
`65.0 100.5 104.7 100-c
`35.0
`-4.7
`
`113.4 100.1
`-13.4
`
`95.2 99.9
`4.8
`
`98.9
`
`RIMFROST EXHIBIT 1032 page 0017
`
`[96E9 [0001ILL8V3SVINIO
`
`

`

`Table 5 / koont'd)
`The Amino Acids of Krill
`Reported literature values for the amino acid content of E. superba protein, against reference proteins, in g/100 g amino acids
`
`Fres Amino-Acids (TCARrtracts)
`
`l',
`
`......
`
`3
`t
`41:1
`01 s-
`
`'f
`E 0
`
`.....
`
`o
`C--
`ZN
`
`e i
`g
`M
`
`True Protein (TCA Precipitate) Referanoe Proteins
`.o. y-
`
`1••
`t-a•
`
`if
`i
`.3
`,...
`t
`0
`.4
`
`0
`0
`g,
`It
`
`• f..
`.4
`
`7.4
`4tie
`
`ci
`0
`
`<4
`0
`
`
`5.6
`4.1- 6.4
`6.5
`5.5 4.1
`4.6
`4.9 '3.2- 5.7
`4.4 3.1
`6.6
`7.0 2.6
`4.1
`5.8- 9.3
`6.0
`4.5- 6.6
`4.9 3.8
`5.5
`16.8 8.4 13.2 12.9-15.5, 14.6
`2.0
`2.0
`tr- 0.8
`3.2
`.tr- 1.3 al 0.2
`0 tr
`tr
`1.0
`1.9- 3.0
`2.3
`3.3 0.5
`0.8
`2.8- 4.9
`3.5
`1.5
`2.7
`3.2
`1.9
`2.7 0.9
`3.4- 5.2
`47.3
`8.7 8.1
`1.1
`3.8
`2.2
`17.3
`7.6
`12.0
`0
`
`1.4
`40.1
`12.8 10.4-22.5
`2.9- 3.3
`1.3- 3.9
`1.9- 4.8
`'7.4-10.8
`7.6- 9.8
`7.6-10.3
`0.8- 2.3
`
`3.0
`
`48.6
`13.6
`2.4
`3.0
`3.0
`11.8
`8.3
`10.0
`1.5
`
`1.9
`
`2.0
`2.1
`3.4
`1.8
`1.7
`
`1.6
`5.5
`1.9
`6.5
`1.2
`1.8
`2.4
`.
`tr
`
`
`4.
`, 0
`4%.
`1
`
`5.4
`5.7
`9.2
`5.0
`4.6
`
`5.5
`5.8
`8.0
`4.5
`9.8
`1.9
`1.0
`2.5
`4.9
`4.6
`
`0.5
`2.7
`4.0
`5.1
`42.2
`6.8
`4.3
`14.8 11.0
`5.1
`4.7
`17.5. 12.5
`3.2
`3.2
`4.9
`4.5
`6.5
`6.2
`2.7
`0.8
`
`2.8
`
`4.2 12 6.5
`5.5
`5.8
`4.2
`4.0
`5.5
`8.6. 4.8
`7.5
`8.3
`4.8
`2.8 '3.8
`4.4
`7.2.
`4.2
`8.0
`7.4
`1.5
`1.9
`1.4
`ali
`0.6 al
`2.0
`2.6 , 2.2
`2.6
`2.5
`3.2
`5.7
`4.5
`4.9
`3.7
`4.5
`37.2 46.6
`46.4
`5.6 (4.1) 10.9
`3.7
`10.8
`12.9
`6.4
`4.0, 5.7
`4.9
`15.0
`16.3 20.3
`9.6
`3.2
`3.5
`8.8
`1.8
`4.7
`5.0
`2.9
`6.4
`2.7 (2.2) 1.7
`2.8
`0.8
`
`iv
`:!
`0
`
`5.0
`4.0
`6.4
`4.5
`5.3
`
`0
`3.0
`2.5
`2.5
`
`7.9
`1.0
`3.5
`2.0
`15.8
`6.9
`10.9
`0
`
`Essential a.a.s.
`'Val
`Ito
`Lou
`Thr
`Lyn
`Trp
`078
`Met
`Tyr
`Phe
`- Total Essentials %
`Arg
`Asp
`Ser
`Gin
`Pro
`Sly
`Ala
`His
`Tau
`(NE3)
`
`-
`
`Total
`Unaccounted for
`
`Z96£9 [0001ILL9V3SINA10
`
`1.0
`
`1.1
`
`91.2 101.1
`7.8
`
`1.1
`
`0.6
`
`1.6
`
`103.3
`
`37.1
`62.9
`
`100.1 99.0 102.5
`1.0
`
`1.1
`
`99.3 99.8
`
`RIMFROST EXHIBIT 1032 page 0018
`
`

`

`-11-
`
`The Japanese have investigated the enzymic activity of whole krill and homogenates.
`An unspecified protease has been identified, of molecular weight 26 000, with maximum activity
`at 40°01 OH 6.0. (Yanase (1971) reports a whole krill pH of 7.8). It displays a sharply
`defined activity curve with high activity between 20 and 45°C, falling off rapidly outside this
`range. The protease is 80% inactivated by a ttypsin inhibitor, and completely inactivitated
`by boiling the extract for two minutes or the whole krill for fifteen minutes. Tyrosinase,
`polyphenol oxidase and ATPase have also been studied. The polyphenol oxidate and tyrosinase
`are implicated in the discolouration problems of pigment darkening. The tyrosinase is found
`mainly in the blood, and is active and thermally stable at temperatures up to 70°C; it can
`be deactivated at 78-800C. Pierce, van der Veen and Olcott (1969) also suggest the presence
`of cellulase activity.
`
`Further work is probably necessary on the kinetios of protease activity at ambient holding
`temperatures (say -5 to +2500, and on lipid oxidases and hydrolases both in the:fresh and.
`frozen states.
`
`2.5 (cid:9)
`
`Fat
`
`Although the fat content of krill varies markedly, its composition would. seem to
`remain fairly constant. It is characterized by its high content of complex (phospho) lipids
`(50%, mainly lecithins (phosphatidyl-oholine) and cephalins (phosphatidyl ethanolamine)),
`about 30-40% neutral fats (glycerides), and about 8% unsaponifiable elements (see Table 6).
`Unlike other Antarctic zooplankters, it contains no waxes. Cholesterol is the only major
`sterol found, although traces of vitamin D and appreciable quantities of the pro-vitamin are
`also present.!/ The fatty acid composition is tabulated in Table 7, where it, will be seen that
`a considerable proportion (around 70%) is unsaturated- iodine value 110-1903/, Most of the
`common fatty acids are present, notably oleic, 020:5/ palmitic and myristic. The three
`'essentials'. are found, totalling 5%. The erucic acid content is low. Several unusual species
`are also evident. The occasional reports of high free fatty acid content (high acid value)
`are probably due to lipolysis during frozen storage of samples. The similarity between whole
`body, organ and remaining carcass lipid composition suggests that there is little differentiation
`of organ lipids (Bottino 1974). The greatest part of the lipid is found in the thoracic region.
`
`2.6 (cid:9)
`
`Vitamins
`
`Krill is potentially a significant source of vitamins Al D, and the B group complex.
`Of most interest is vitamin A ansi,,it&precursors. Astaxanthin is present in _appreciable
`quantities(up,to 3 mg °%), Mainly in the exoskeleton and the eyes (170 mg %)..1/ It is this
`pigment that gives the krill its characteristic orange/red colouration, audit may have some
`provitamin A activity. Provitamin A ( 0-carotene) is present to a lesser extent (20 pg%),
`although some research has suggested that vitamin A is formed from the. non-carotenoid fraction
`of the oil (Gilberg 1971); vitamin A is stored in the eyes (20 mg %), with a net Concentration
`in the whole krill of 140 pee
`
`Vitamin B12 is present, at levels higher _than usually associated with crustacea
`(Hirano, Kikuchi and Okada 1964).
`
`The complete vitamin composition is given in Table 8.
`
`iirEtIoDhiet (cid:9)
`(1976) Chokii et:al(1976 Kuwano et 21 (1975b & 1976) Matsumoto 21 al
`(1975) Nagaku, Nishimuro and Mori (1976 Seki, Ozawa and Arai (1975) Nichiro Gyogyo
`Kaisha Ltd. (1976)
`Bottino (197311974 and 1975), Gilberg (1971), Bykov (1975)
`.3/ /Tanaka and Koizumi (1964), Watanabe et g (1976), Rosohke (1976a), Saiki and Mori (1953)
`AL/Mauchline and Fisher (1969), Fisher, FCon and Thompson (1954, 1955), Fisher et al (1955)
`Arai. Watanabe and Kinisnaki (1976)
`
`RIMFROST EXHIBIT 1032 page 0019
`
`OLYMSEA877ITC00163953
`
`(cid:9)
`(cid:9)
`

`

`Table 6
`
`The Lipids of Krill
`
`Reported literature values for the composition of crude lipid, as weight % of total lipid
`
`Ls., r-
`alr,
`a-
`
`o
`
`o m
`SI
`Tct; (cid:9)
`v- cat
`
`W I-3
`k
`0 ;4
`
`ON
`•.--
`
`0
`0
`
`[^-
`01
`
`rn
`LrN
`ON ON
`a- tr‘
`-.4 (cid:9) ....-
`
`xi
`t
`'4D
`01
`
`u) r-
`
`01-
`
`-r-I
`
`\o r-
`ar.
`a-
`
`W
`ra
`
`00
`rig g
`544
`M
`g ci
`
`r (cid:9)
`
`°di
`
`.
`
`d .
`
`g .
`
`C) S
`
`0
`
`.4
`
`Waxes
`Sterols
`Steriod Esters
`Triglycerides
`Diglycerides
`(E Complex Lipids)
`PhosPhatidyloholine
`PhosPhatidylethanolanine
`LysoPhosphatidylcholine
`Phosphatidylglycerol
`Phosphatidyl Serine
`Sphingomyelin
`PhosPhoinositide
`Free Fatty Acids
`Unsaponifiable
`
`•
`2/ Cholesterol + Traces Xanthophyll
`
`x = Reported Present
`
`20
`
`—
`—
`8-36
`4-17
`(54-58) (50)
`48
`8
`1
`1
`
`x
`x
`
`x
`x
`
`(29.9)
`
`4.7
`0.8
`50.3
`tr
`(8.9?)
`
`x
`x
`
`x
`x
`x
`x
`
`31.7
`8.6
`
`4.3--6.9
`
`27.4
`
`0.0
`1.7-7.62/ 0.0- 7.6
`4.7
`0.0— 0.8
`0.4
`28.6
`8.0-50.3
`10.5
`4.0-17.0
`(29.9-58.0) (48.0)
`48
`8
`1
`1
`x
`x
`x
`29'.6
`7.5
`
`27.4-31.7
`4.3-13.1
`
`4.5-13.1
`
`RIMFROST EXHIBIT 1032 page 0020
`
`1796E9 1, 0001ILL8V3SVINIO
`
`

`

`- 13 -
`
`Table 7
`
`The Lipids of Krill
`
`Reported literature values for fatty acid composition, as weight % of total fatty acids
`
`Total Lipids.- Unfractionated
`
`o e- cs,
`
`tfl e-
`C-
`°N
`
`c., (cid:9) 6
`
`U.I.- cr,
`
`1
`+.
`4
`
`• t.
`a:
`1: C
`
`ON
`Cr, T.
`VD
`0 o
`1.,
`.,-i 04
`
`vt I.-
`0 \
`.,-.
`
`-1
`
`m
`
`0 \ In
`
`cr„ .-
`
`.,4
`
`m
`
`SATURATED
`
`Capric (cid:9) 1/
`
`Laurio
`
`Myristic
`
`Palmitic
`Stearic
`Araohidic
`Behenio
`Total Saturated
`
`0.2
`
`9:0
`10:0
`11:0
`12:0
`013:0
`014:0 14.0
`0.4
`015:0
`016:0 22.3
`1.0
`018:0
`020:0
`22:0
`
`37.9
`
`0.3
`
`0.5
`
`tr
`
`11.9
`
`4.1
`tr
`24.4 14.4
`1.7
`1.4
`1.0
`
`14.6 12.0
`0.6
`23.0 18.0
`1.1
`0.9
`
`0.1
`39.7 30.9
`
`'0
`
`ON -
`
`0
`
`r••• cr, .-
`G .. m
`
`o I.--
`C71 _
`
`I i
`
`tr
`
`0.4
`0.1
`12.0 12.2
`0.5
`0.9
`16.1 18.2
`1.2
`1.2
`1.6
`
`•
`
`31.7 26.3
`
`43.8 29.8 34.6
`
`UNSATURATED
`Essentialsainoleic 18:2
`18:3
`Linolenio
`20:4
`Arachidanio
`(Total Essentials)
`Non - Essential
`
`4.0
`2.5
`2.4
`1.2
`1.3
`1.2
`0.8
`1.3
`0.4
`(4.0) (4.6) (6.5)
`
`1.7
`1.3
`
`2/
`a/
`• 2/
`
`2.2
`
`0.7
`
`4.2
`1.2
`1.3
`(6.7)
`
`st
`
`VD cr. 1-.
`
`\o
`C-
`0 \
`....
`m
`1
`
`g
`
`5.8
`
`11.1
`10.2?
`4.2?
`0.2
`31.8
`
`.d./
`A/
`2/
`
`10,4
`0.9
`19.1
`1.3
`
`31.7
`
`3.9
`1.1
`1.7
`(6.7)
`
`0.4
`
`0.8
`
`14.5?
`
`6.3
`1.4
`A/9.7
`)
`1/
`) (cid:9) 2.9
`A/
`I/
`2/
`!/ 22.7
`
`0.0
`
`0.8
`
`15.8
`67.1
`98.8
`
`11:1
`12:1
`13:1
`14:n
`1