`
`Petition for Inter Partes Review
`Of U.S. Patent 8,278,351
`Exhibit
`ENZYMOTEC - 1006
`
`
`
`O
`
`iiCiiS
`
`A PUBLICATION OF THE AMERICAN OIL CHEMISTS' SOCIETY
`
`EDITOR
`RalphT. Holman, The Hormel Institute, University of Minnesota, 801-16th Avenue N.E., Austin,
`MN 55912
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`Donna Patten, The Hormel Institute, Austin, MN. Phone: 507-433-8804
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`MANAGING EDITOR
`
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`
`Copyright 7984 by the American Oil Chemists’ Society (A 008).
`Lipids (ISSN:0024-4201) is published monthly by the American Oil Chemists’ Society at 508
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`iii-viii
`Editorial Information. Current instructions to authors are printed in Lipids, Vol. 19, pp.
`lJanuary 1984). Manuscripts for publication should be submitted, in quadruplicate, to the Editor,
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`000002
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`821
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`Lipid, Sterol and Fatty Acid Composition of Antarctic Krill
`
`(Euphausia superba Dana)
`
`H. FFlchE,a G. GERCKEN,a W. SCHREIBERb and J. OEHLENSCHLAGERPN‘
`aDepartment of Biochemistry, University of Hamburg, Hamburg, and b Institute for
`Biochemistry and Technology in the Federal Research Centre of Fisheries, Palmail/e
`.9, D-2000 Hamburg 50, Germany
`
`ABSTRACT
`
`The lipid classes, fatty acids of total and individual lipids and sterols of Antarctic krill (Euphausia
`superba Dana) from two areas of the Antarctic Ocean were analyzed by thin layer chromatography
`(TLC), gas liquid chromatography (GLC) and gas liquid chromatography/mass spectrometry (GLC/
`MS). Basic differences in the lipid composition of krill from the Scotia Sea (caught in Dec. 1977) and
`krill from the Gerlache Strait (caught in Mar. 1981) were not observed. The main lipid classes found
`were: phosphatidylcholine (PC)
`(3 3-36%), phosphatidylethanolamine (PE)
`(5-6%),
`triacylglycerol
`(TG) 83-40%), free fatty acids (FFA) (8—16%) and sterols (1.4-1.7%). Wax esters and sterol esters
`were present only in traces. More than 50 fatty acids could be identified using GLC/MS, the major
`ones being 14:0, 16:0, 16:1(n-7), 18:1(n-9), 18:1(n-7), 20:5(n-3) and 22:6(n-3). Phytanic acid was
`found in a concentration of 3% of total fatty acids. Short, medium-chain and hydroxy fatty acids
`(C < 10) were not detectable. The sterol fraction consisted of cholesterol, desmosterol and 22-dehy-
`drocholesterol.
`Lipids 19:821-827, 1984.
`
`INTRODUCTION
`
`Krill (Euphausz'a superbzz Dana) lives exclu-
`sively in cold Antarctic waters and is the central
`link in the Antarctic food web.
`Its general
`chemical and biochemical composition has been
`the subject of several investigations (1 ). A num-
`ber of contributions also have dealt with the
`lipid content and lipid composition of this
`pelagic euphausiid. Lipid contents between 1%
`and 6% have been published (2), and remark-
`ably differing data have been reported for lipid
`composition (3-12). The main lipid classes
`found by almost all investigators were phospho-
`glycerolipids,
`triacylglycerols (TG), free fatty
`acids (FFA) and free sterols. The dominating
`fatty acids reported were 1620 among saturated
`fatty acids and 18:1, 20:5 and 22:6 among un-
`saturated and polyunsaturated fatty acids. This
`investigation has been carried out
`to give
`thorough and complete analyses of lipid classes,
`fatty acids and sterols, supported by mass
`spectrometry (M S).
`
`MATERIALS AND METHODS
`
`Sample Collection and Preparation
`Antarctic krill were collected from the
`Scotia Sea on December 16, 1977 at 57° 47’ S;
`420 43' W (l 3) and from the Gerlache Strait on
`March 12, 1981 at 64° 33.7’ S;62° 32’W(14)
`during the second (1977/78) and third (1980/
`
`81) Antarctic expeditions of the Federal Re-
`*To whom correspondence should be addressed.
`
`public of Germany with FMS “Julius Fock”
`and FRV “Walther Herwig,” respectively, using
`a 1219 mesh pelagic Krill net.
`Krill samples of 5 kg were quick-frozen and
`stored at -35 C until analyzed. Subsamples pre—
`pared from the core of the 5 kg samples were
`homogenized in a mortar under liquid nitrogen,
`and lipid extraction was performed according
`to Folch et al. (15). Lipids were dissolved in
`dichloromethane: methanol
`1:1
`(v/v)
`and
`stored under a nitrogen atmosphere at -23 C.
`
`Thin Layer Chromatography
`and Gas Liquid Chromatography
`
`Crude lipids were separated into classes by
`TLC on HPTLC-plates (E. Merck, Darmstadt)
`developed with n-hexane:diethyletherzglacial
`acetic acid 60:40:l (v/v) for neutral lipids, and
`with dichloromethane:methanol:glacial acetic
`acid 60:30:10 (v/v) or dichloromethanezmetha-
`nol:aqueous ammonia 6022015 (v/v) for polar
`lipids. Lipid classes were visualized by exposure
`to iodine vapor or by charting with 50% sul—
`phuric acid. After 2 dimensional TLC using the
`above mentioned solvents,
`identification was
`achieved by comparison with standard mixtures
`and lipid class specific stainings (16). After the
`silica gel was scraped off, the eluted acylglycer-
`ols were quantified by an enzymatic test for
`esterified glycerol (E. Merck, Darmstadt), and
`phosphoglycerides by phosphorus determina-
`tion (17), FFA and sterols were determined by
`GLC using heptadecanoic acid and stigmasterol,
`respectively, as internal standards.
`
`LIPIDS, VOL. 19, NO. 11 (1984)
`
`000003
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`
`
`
`822
`
`H. FRIC KE, G. GERCKEN, W. SCHREIBER AND J. OEHLENSCHLAGER
`TABLE 1
`
`Fatty acid methyl ester (FAME) of total
`lipids and individual lipid classes were prepared
`with 14% boron trifluoride in methanol (18),
`and fatty acid benzyl esters (FABE) according
`to Klemm et a1. (19). Trimethylsilylation of
`sterols was carried out as described by Ballan-
`tine et a1. (20). FAME and FABE were purified
`by TLC prior to GLC analysis. Separations and
`identifications were carried out on a polar wall
`coated (WCOT) open-tubular glass column
`(25 m) coated with SILAR 10 C (Packard in-
`struments),
`temperature
`programmed from
`110 C to 210 C (3 C/min) and on a 50 m fused
`silica column (WCOT) coated with CP SIL 5,
`temperature programmed from 100 C to 320 C
`(3 C/min) using a Packard 428 gas chromato-
`graph equipped with a FID and a HP 3371
`integrator. Helium was used as carrier gas at a
`flow of 1 ml/min with a split ratio of 100:1.
`The presence of plasmalogens and alkylglycer-
`ols was tested subsequent to hydrolysis using
`the procedure of Pugh et a1. (21).
`GLC/MS analysis of FAME and trimethyl-
`silyl
`(TMS) sterols was performed on a HP
`5985A quadrupole mass spectrometer, ioniza-
`tion energy 70 eV,
`ion source temperature
`200 C, column: 25 m WCOT coated with CP
`SIL 5 (Chrompak),
`temperature programmed
`from 140 C to 280 C (4 C/min).
`Individual FAME, FABE and TMS sterol
`peaks were identified by co-chromatography
`with standards, by comparison with calculated
`equivalent chain length (ECL) values (22) and
`by mass spectra. To ensure identification of
`unusual fatty acids, samples were hydrogenated
`and rechromatographed. For positional analy-
`sis, cleavage of PC and PE was performed with
`phospholipase A2
`from Crotalus
`durissus
`terrificus (Boehringer, Mannheim). After 24 hr
`incubation in diethylether and 0.1 M tris-buffer,
`the reaction mixture was separated by TLC into
`lysophospholipids and FFA.
`
`RESULTS AND DISCUSSION
`
`Lipid Content and Lipid Composition
`
`The total lipid content and the lipid compo-
`sition data of the 2 krill samples are given in
`Table 1. Although different lipid compositions
`have been published, there is general agreement
`as to the main lipid classes present in Euphausz‘a
`superba (3-12). The krill caught in December
`1977 has a lower fat content than the krill
`caught in March 1981. This increase in fat con—
`tent during the catching season, which co—
`incides with the sexual maturity (2) of krill, has
`been shown previously (14). Beginning with a
`low fat content of approx. 1% on a wet weight
`basis in November/December, the fat content
`
`LIPIDS, VOL. 19, NO. 11 (1984)
`
`
`
`Lipid Composition of Antarctic Krill
`(Euphausia superba Dana)
`
`Sample
`
`Total lipid content
`(% wet weight)
`
`12/1977
`
`3/1981
`
`2.7 i 0.2
`
`6.2 :r 0.3
`
`Phospholipids
`Phosphatidylcholine
`Phosphatidylethanolamine
`LySOphosphatidylcholine
`Phosphatidylinositol
`Cardrolipin.
`.
`Phosphatidic acrd
`
`Neutral lipids
`Triacylglycerols
`Free fatty acids21
`Diacylglycerols
`Sterols
`Monoacylglycerols
`
`Othersb
`
`Total
`
`35.6 t 0.1
`6.1 t 0.4
`1.5 t 0.2
`0.9 1' 0.1
`1.0 t 0.4
`0.6 t 0.4
`
`33.3 t 0.5
`5.2 i 0.5
`2.8 t 0.4
`1.1 t 0.4
`1.6 r 0.2
`
`33.3 i 0.5
`16.1 t 1.3
`1.3 t 0.1
`1.7 t 0.1
`0.4 t 0.2
`
`40.4 t 0.1
`8.5 i 1.0
`3.6 t 0.1
`1.4 t 0.1
`0.9 t 0.1
`
`09:01
`
`05:01
`
`98.9
`
`99.3
`
`lipids and
`Data are expressed as wt % of total
`represent means 1 standard deviation of 3 separate
`experiments.
`aProbably mostly artifacts.
`bTraces of 1ysophosphatidylethanolamine, phos-
`phatidylserine, sphingomyelin, glycolipids, sterol es-
`ters, waxes and carotenoids were detected.
`
`increases to approx. 6% in March/April.
`Euphausz‘a superba is extremely rich in phos-
`pholipids (240% of total lipids) and TG (33
`and 40% respectively of total lipids). While the
`relative content of phospholipids is similar in
`the 1977 and 1981 samples, the percentages of
`TG differ somewhat. This is in accordance with
`the previous results of our laboratories (23),
`which show that the relative phospholipid con-
`centration will not change with varying total
`lipid contents.
`In other marine organisms an
`increase of total lipid content usually is caused
`by an increase of TG (24),
`The sterol contents of 1.4% and 1.7% fe‘
`spectively of total lipids are in the range which
`has been reported (2,25) for Krill. These are
`very low values compared with those of Clark6
`(3), who found up to 16.9% sterols of tow1
`lipids in krill from South Georgia. This differ-
`ence may be due to the methods. Clarke used
`densitometry (3) and our laboratory GLC.
`is
`In the 1977 sample the FFA content
`about twice that of the 1981 sample. The high
`value could be caused by the longer storage
`time of the 1977 sample. A residual lipolyl1C
`activity against phospholipids exists even at
`temperatures of -30 C and below. Samples of
`
`000004
`(cid:19)(cid:19)(cid:19)(cid:19)(cid:19)(cid:23)
`
`
`
`ANT A RCTIC KRlLL LIPIDS
`
`823
`
`TABLE 3
`
`Branched Chain Fatty Acid Composition
`of Total Lipids of Euphausia superba Dana
`
`Sample
`
`12/1977
`
`3/1981
`
`M"
`ECL
`
`
`228
`242
`256
`256
`270
`284
`284
`282
`282
`298
`
`n.d.
`tr.
`12.6
`n.d.
`0.05 10.01
`13.6
`14.6 0.1910.00 0.3110.15
`14.7 0.211 0.01
`0.24 1 0.07
`15.6 0.091003 0.101 0.06
`16.6 0.541005 0.201002
`16.4
`tr.
`0.09 1 0.02
`16.5
`0.05 1 0.03 0.111 0.08
`16.2
`tr.
`0.1010.05
`17.6
`tr.
`0.1010.01
`
`the same haul which were cooked on board
`immediately after hauling and stored under the
`same conditions showed a FFA content which
`was much lower, ranging from 1% to 3% of
`total
`lipids. This low FFA content of freshly
`caught krill also was confirmed by Ellingsen
`(11).
`lysophosphatidylcholine) lyso-
`In addition,
`phosphatidylethanolamine,
`phosphatidylinosi-
`tol phosphatidic acid, cardiolipin and mono-
`and diacylglycerols were detected, whereas
`phosphatidylserine, sphingomyelin, glycolipids.
`wax esters and sterol esters were present only in
`trace amounts. Wax esters were found by
`Bottino (8) in the euphausiid Euphausz'a crystal-
`Zorophias but not
`in Euphausz'a superba. The
`composition of carotenoids was not
`investi-
`gated but had been analyzed by others (26-28).
`Fatty Acid Composition of Total Lipids
`
`The composition of the fatty acids of total
`lipids of Euphausz‘a superba is similar to that of
`other marine crustaceans and some marine
`fishes (29) (Tables 2 and 3). The main fatty
`
`13:01
`14:01
`1520i
`15:0 ai
`16:01
`17:0i
`1710 hr8
`17:1br
`17:1br
`1810i
`Phytanicb
`
`17.7 2.82 1 0.41 1.2326acid 1 0.43
`Data are expressed as wt % of total fatty acids and
`represent means 1 standard deviation of 3 separate
`experiments.
`tr. = trace; n.d. = not detected; hr. = branched;
`i = iso; ai = anteiso.
`aPresumably 7-methy1hexadecanoic acid.
`b3,7,11,1 S-tetramethylhexadecanoic acid.
`
`
`
`
`
`
`
`
`
`TABLE 2
`
`Fatty Acid Composition of Total Lipids of Euphausz’a superba Dana
`
`
`Sample
`
`12/1977
`
`3/1981
`
`Sample
`
`12/1977
`
`3/1981
`
`M:a ECLb
`mra
`ECLb
`
`
`18:4(n-3)
`tr.
`tr.
`10.0
`186
`10:0
`19:0
`tr.
`tr.
`11.0
`200
`11:0
`19:1
`0.22 1 0.06
`0.23 1 0.06
`12.0
`214
`12:0
`0.07 1 0.04 1922
`0.04 1 0.01
`13.0
`228
`13:0
`15.23 1 2.31 20:0
`11.33 1 1.48
`14.0
`242
`14:0
`0.19 1 0.01 20:1(n-7)
`tr.
`13.8
`240
`14:1
`0.27 1 0.05 20:1(n-9)
`0.34 1 0.01
`15.0
`256
`15:0
`0.04 1 0.03 20:2
`tr.
`14.8
`254
`15:1
`25.91 1 2.33 31.79 1 1.73 20:4(n-3)
`16.0
`270
`16:0
`7.26 1 0.35
`7.37 1 0.34 20:5(n-3)
`15.7
`268
`16:1(n-7)
`0.09 1 0.13
`0.30 1 0.01 21:0
`15.8
`268
`16:1(n-?)
`0.82 1 0.01
`0.12 1 0.06 21:5(n-3)
`15.6
`266
`16:2(n-6)
`tr.
`0.29 1 0.01 22:0
`15.5
`264
`16:3
`0.74 1 0.06
`0.48 1 0.14 22:1(n-7)
`15.4
`262
`16:4(n-3)
`0.06 1 0.02
`0.17 1 0.15 22:1(n-9)
`17.0
`284
`17:0
`tr.
`0.41 1 0.05 22:5(n-3)
`16.7
`282
`17:1
`tr.
`0.12 1 0.06 22:5
`16.8
`282
`17:1
`1.21 1 0.18
`2.14 1 0.23 22:6(n73)
`18.0
`298
`18:0
`8.32 + 0.54
`7.49 1 0.79 23:1
`17.8
`296
`18:1(n-7)
`10.13 1 2.20
`10.52 1 0.90 24:0
`17.7
`296
`18:1(n-9)
`tr.
`0.09 1 0.05 24:1
`17.9
`296
`18:1(n-?)
`1.58 1 0.09
`0.74 1 0.38 25:0
`17.6
`294
`18:2(n-6)
`0.47 1 0.02
`0.33 1 0.07
`17.6
`292
`18:3(n-3)
`
`
`
`
`17.3 0.21 1 0.06 0.57 1 0.35 OthersC A — 3.9529218:3(n-6) 2.45
`
`Data are expressed as wt % of total fatty acids and represent means 1 standard deviation of 3 separate experi-
`ments.
`tr. 2 trace.
`aM’: molecular weight of fatty acid methyl ester as determined by GLC/MS.
`bECL: equivalent chain length, calculated by plotting chain length (as carbon number) versus retention time
`On CP SIL 5.
`“Predominantly branched chain fatty acids as given in Table 3 in detail.
`
`290
`312
`310
`308
`326
`324
`324
`322
`318
`316
`340
`330
`354
`352
`352
`344
`344
`342
`366
`382
`380
`396
`
`
`
`17.4
`19.0
`18.8
`18.7
`20.0
`19.8
`19.7
`19.6
`19.5
`19.3
`21.0
`20.2
`22.0
`21.6
`21.5
`21.2
`21.4
`21.1
`22.5
`24.0
`23.6
`25.0
`
`0.67 1 0.07
`tr.
`0.12 1 0.04
`tr.
`0.04 1 0.00
`0.40 1 0.01
`0.77 1 0.04
`tr.
`0.46 1 0.10
`12.71 1 1.57
`tr.
`0.42 1 0.03
`0.14 1 0.03
`0.29 1 0.17
`0.51 + 0.06
`0.54 1 0.09
`tr.
`5.41 1 0.51
`tr.
`tr.
`tr.
`tr.
`
`
`
`
`
`0.62 1 0.49
`0.11 1 0.16
`0.20 1 0.09
`0.07 1 0.05
`0.19 1 0.14
`0.50 1 0.09
`1.35 1 0.23
`0.08 1 0.06
`0.22 1 0.06
`7.83 1 1.27
`tr.
`0.30 1 0.18
`tr.
`0.41 1 0.16
`1.22 1 0.33
`0.24 1 0.11
`0.04 1 0.03
`2.60 1 0,79
`0.11 1 0.07
`tr.
`0.15 1 0.11
`tr.
`
`LIPIDS, VOL. 19, NO. 11 (1984)
`
`000005
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`LIPIDS, VOL 19, NO. 11 (1984)
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`LIPIDS, VOL. 19, NO. 11 (1984)
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`H. FRICKE, G. GERCKEN, W. SCHREIBER AND J. OEHLENSCHLAGER
`
`(26-32%).
`16:0
`(ll-15%),
`14:0
`are
`acids
`(10%), 18:1(n-7)
`(7%), 18:1(n-9)
`16:1(n—7)
`(8%), 20:5(n-3) (8—13%) and 22:6(n-3) (36%).
`Odd—numbered fatty acids with chain lengths
`ranging from C-ll
`to C—25 also were found in
`trace amounts and verified by GLC/MS. In the
`unsaturated fatty acids the species of the (n-3)
`series are dominant, while (n-6) fatty acids are
`found only to a limited extent. This also has
`been reported for marine shrimps and fish (29—
`31). Several branched-chain fatty acids ranging
`from C-13 to C-18 (straight-chain length) were
`found, most of them belonging to the 150- or
`anteiso series. Among the multi-branchcd-chain
`fatty acids phytanic acid (32-34), which was
`the main component, amounted up to 3% of
`total fatty acid content. The samples from the
`early season 1977 contain more unsaturated
`fatty acids, especially 20:5(n—3) and 22:6(n—3),
`and less saturated fatty acids such as 14:0 and
`16:0 than the sample from March 1981. This
`difference in the fatty acid compositions seems
`to be a seasonal phenomenon which also was
`reported by Shibata (2).
`In most of the investigations of krill lipids
`the fatty acids were determined only by their
`retention behavior (3,35). In this study it was
`possible to determine the mass, and hence the
`chain length and number of double bonds, for
`all fatty acids by the combination of GLC/MS.
`The number of 57 analyzed fatty acids exceeds
`that reported by Golovnya et al, (36), who used
`the same technique. According to their ECL
`values 20:1 and 22:1 belong to the (n-7) and
`(n-9) series and not to the (n-ll) series (36).
`The data found suggest that a (n—7) monoene
`series is present carrying from 16: 1(n—7)through
`18:1(n-7) and 20:1(n-7) to 22:1(n-7) (37,38),
`Arachidonic acid which was found by Clarke
`(3), Golovnya (36) and Bottino (5) in krill, and
`by Bottino in a shrimp (39) as a minor compo-
`nent, was not found. Dembitskii (40) showed
`that marine crustacea contained high levels of
`lipids with alkenyl side chains. In the samples
`investigated neither free aldehydes nor dimeth~
`ylacetals after derivatization could be detected.
`Short chain, medium chain and hydroxy fatty
`acids (<C—12) were not detectable even after
`transesterification to the corresponding benzyl
`esters (l9).
`
`Fatty Acid Composition of Lipid Classes
`
`The analysis of fatty acids of individual lipid
`classes indicates different fatty acid composi-
`tions
`for phospholipids
`(Table 4) and TG
`(Table 5). Fatty acids in TO are mostly satu-
`rated or monounsaturated with 14:0, 16:0,
`16:1(n-7), 18:1(n-7) and 18:1(n-9)as dominat-
`ing species. Polyunsaturated fatty acids were
`
`LIPIDS, VOL. 19. NO. 11 (I984)
`
`found only in small amounts.
`In phospholipids phytanic acid was detected
`only in traces, but it represented 56% of TG
`fatty acids. The phospholipids and FFA have
`16:0, 20:5(n-3) and 22:6(n-3) as principal fatty
`acids. In the individual lipid classes a difference
`can be seen between the December samples
`(1977) and the March samples (1981). The
`lipid classes of the December samples contain
`more saturated fatty acids and less unsaturated
`fatty acids than the March 1981 samples. The
`discrepancy in the seasonal changes of the fatty
`acid composition of total lipids as mentioned
`above and that of the individual lipid classes is
`caused by the different lipid class composition
`with varying relative amounts of TG.
`The positional analysis of the fatty acids in
`the main phospholipids PC and PE (Table 6)
`shows that saturated fatty acids are commonly
`linked to the sn-1 position and that the sn-2
`position is preferred by unsaturated fatty acids.
`In this respect krill has the same fatty acid dis-
`tribution as other marine animals (41 ).
`TABLE 6
`
`Fatty Acid Positional Analysis in Phosphatidylcholine
`(PC) and Phosphatidylethanolamine (PE)
`of Euphausz’a superba Dana (1977 Sample)
`
`Phospholipid
`PC
`PE
`sn-position
`sn-l
`srz»2
`sn-l
`sn»2
`
`
`0.8
`0.7
`2.3
`3.5
`14:0
`3.3
`45.4
`4.7
`60.8
`16:0
`05
`0.7
`5.4
`1.5
`16:1(ne7)
`1.5
`5.6
`0.8
`1.9
`18:0
`0.9
`24.0
`tr.
`11.1
`18:1(n-7)
`2.9
`4.8
`22.0
`3.5
`18:1(ne9)
`0.6
`0.6
`4.8
`0.6
`18:2(n-6)
`31.3
`5.7
`27.7
`5.6
`20:5(n-3)
`41.3
`3.5
`11.1
`2.1
`22:6(nA3)
`
`
`
`
`9.4 21.2 9.0Others 16.9
`Data are expressed as wt % of fatty acids in one
`position from one experiment.
`
`TABLE 7
`
`Composition of the Free Sterol Fraction
`in Euphausia superba Dana
`
`
`
`
`
` Sample 12/1977 3/1981
`
`Cholesterola
`Desmosteroib
`22-Dehydrocholesterol‘5
`Others
`
`70.0 t 5.9
`18.2 +. 1.4
`11.5 t 4.8
`0.3 i 0.5
`
`75.5 t 3.7
`17.7 11.1
`6.0 i 3-5
`1.0 i 0-7
`
`100.2
`100.5
`Total
`
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`
`ANTARC TIC KRILL LIPIDS
`
`827
`
`Sierols
`
`samples contained 3 sterols as
`The krill
`major components identified by GLC/MS and
`traces of other sterols and sterol esters with un-
`known structure. The proportions of choles-
`terol, desmostero] and 22-dehydrocholesterol
`are given in Table 7. Cholesterol, which cannot
`be synthesized de novo in marine crustaceans
`(42),
`is the main sterol. Desmosterol and 22—
`dehydrocholesterol levels are very high. These
`sterols are assumed to be intermediates in the
`conversion of dietary sterols to cholesterol (42,
`43). A small amount of 22-dehydrocholesterol,
`but no desmosterol, also was detected in the
`Arctic euphausiid Meganyctiphanes norvegz’ca,
`whereas
`the herbivorous
`copepod Calanus
`finnmzzrchz‘cus
`contained
`14.1% 22-dehydro-
`cholesterol
`and
`27.7% desmosterol besides
`cholesterol as main sterol (44).
`
`ACKNOW LEDGMENTS
`
`I. Wasum and Dr. W. Ko'nig did the GLC/MS
`Mrs.
`analyses. This work was supported by the Federal
`Ministries of Food, Agriculture and Forestry and the
`Federal Ministries of Science and Technology. One of
`us, H. F., thanks the latter for financial support.
`
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`
`LIPIDS, VOL. 19. NO. 11 (1984)
`
`000009
`(cid:19)(cid:19)(cid:19)(cid:19)(cid:19)(cid:28)
`
`