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`NOVEMBER 1984 0 815-906 0 VOLUME 19, NO. 11
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`Fatty Acids and Neutral Lipids in Developing Oyster Larvae
`Lipid, Sterol and Fatty Acid Composition of Antarctic Krill
`Comparative Study of Lipogenic Enzymes in Vertebrates
`Urinary Malondialdehyde as an Indicator of Lipid Peroxidation in the Diet and in the Tissues
`Effect of Protein and Sugar on Rabbit Lipids
`Inhibition of Fatty Acid Synthesis by TOFA in Adipocytes
`Lung Surfactant Phospholipids in Different Animal Species
`Fatty Acid Peroxidation by Peroxidase
`Effects of trans Fatty Acids on Fatty Acyl A5 Desaturation
`Intestinal Metabolism of Plasma FFA in Diabetic Rats
`
`Quantitative Analysis of Triglyceride Species
`Analysis of Sterol Esters by Capillary Gas Chromatography—E|ectron Impact and Chemical
`lonization—Mass Spectrometry
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` A PUBLICATION OF THE AMERICAN OIL CHEMISTS’SOCIETY
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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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`
`821
`
`Lipid, Sterol and Fatty Acid Composition of Antarctic Krill
`
`(Euphausia superba Dana)
`
`H. FRIcKE,3 G. GERCKEN,3 W. SCHRElBERb and J. OEHLENSCH LAG ER,br*
`3Department of Biochemistry, University of Hamburg, Hamburg, and blnstitu te for
`Biochemistry and Technology in the Federal Research Centre of Fisheries, Pa/mai//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) (33-40%), 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:l(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 19:82]-827, 1984.
`
`INTRODUCTION
`
`Krill (Euphausia 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;
`42° 43' W (13) 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 (l980/
`
`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:diethylether:glacial
`acetic acid 60:40:l (v/v) for neutral lipids, and
`with dichloromethanezmethanolzglacial acetic
`acid 60:30:10 (v/v) or dichloromethane:metha-
`nol:aqueous ammonia 60:20:5 (v/v) for polar
`lipids. Lipid classes were visualized by exposure
`to iodine vapor or by charring 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 acy1glycer-
`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
`
`000003
`
`

`
`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 al. (19). Trimethylsilylation of
`sterols was carried out as described by Ba1lan-
`tine et al. (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 al. (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 Euphausia
`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 1 0.2
`
`6.2 1 0.3
`
`Phospholipids
`Phosphatidylcholine
`Phosphatidylethanolamine
`Lysophosphatidylcholine
`Phosphatidylinositol
`Cardiolipin.
`.
`Phosphatidic acid
`
`Neutral lipids
`Triacylglycerols
`Free fatty acidsa
`Diacylglycerols
`Sterols
`Monoacylglycerols
`
`Othersb
`
`Total
`
`35.6 1 0.1
`6.1 1 0.4
`1.5 1 0.2
`0.9 1 0.1
`1.0 1 0.4
`0.6 1 0.4
`
`33.3 1 0.5
`5.2 1 0.5
`2.8 1 0.4
`1.1 1 0.4
`1.6 if 02
`
`33.3 1 0.5
`16.1 1 1.3
`1.3 1 0.1
`1.7 1 0.1
`0.4 1 0.2
`
`40.4 1 0.1
`8.5 1 1.0
`3.6 1 0.1
`1.4 1 0.1
`0.9 1 0.1
`
`09101
`
`05:01
`
`98.9
`
`99.3
`
`lipids and
`Data are expressed as wt % of total
`represent means 1 standard deviation of 3 separate
`experiments.
`3Probab1y mostly artifacts.
`bTraces of 1ysophosphatidylethanolamine, phos-
`phatidylserine, sphingomyelin, glycolipids, sterol es-
`ters, waxes and carotenoids were detected.
`
`increases to approx. 6% in March/April.
`Euphausia superba is extremely rich in phos-
`pholipids (>40% 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% re‘
`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 Clarke
`(3), who found up to 16.9% sterols of total
`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 lipolytlc
`activity against phospholipids exists even at
`temperatures of -30 C and below. Samples Of
`
`000004
`
`000004
`
`

`
`ANTARCTIC KRILL LIPIDS
`
`823
`
`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).
`lyso-
`lysophosphatidylcholine,
`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 Euphausia crystal-
`Zorophias but not
`in Euphausia 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 Euphausia superba is similar to that of
`other marine crustaceans and some marine
`fishes (29) (Tables 2 and 3). The main fatty
`
`TABLE 3
`
`Branched Chain Fatty Acid Composition
`of Total Lipids of Euphausia superba Dana
`
`Sample
`
`12/1977
`
`3/1981
`
`1\/I“
`
`ECL
`
`13:01
`14:0i
`1S:0i
`15:0 ai
`1620i
`17:0i
`17:0 bra
`17:1 hr
`17:1 hr
`18:0i
`Phytanicb
`acid
`
`228
`242
`256
`256
`270
`284
`284
`282
`282
`298
`
`326
`
`n.d.
`tr.
`12.6
`n.d.
`0.05 10.01
`13.6
`14.6 0.1910.00 0.3110.1S
`14.7 0.2110.01 0.24 10.07
`15.6 0.0910.03 0.101 0.06
`16.6
`0.54 1 0.05
`0.20 1 0.02
`16.4
`tr.
`0.09 1 0.02
`16.5
`0.05 1 0.03 0.111 0.08
`16.2
`tr
`0.1010.0S
`17.6
`tr
`0.1010.01
`
`17.7
`
`2.82 1 0.41
`
`1.2
`
`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; br. = branched;
`i = iso; ai = anteiso.
`3Presumab1y 7—methylhexadecanoic acid.
`b3,7,11,1S—tetramethylhexadecanoic acid.
`
`TABLE 2
`
`Fatty Acid Composition of Total Lipids of Euphausia superba Dana
`
`Sample
`
`12/1977
`
`3/1981
`
`Sample
`
`12/1977
`
`3/1981
`
`Mm ECLb
`
`Nra EcLb
`
`10:0
`11:0
`12:0
`13:0
`14:0
`14:1
`15:0
`15:1
`16:0
`16:1(n-7)
`16:1(n—‘.7)
`16:2(n—6)
`16:3
`16:4(n—3)
`17:0
`17:1
`17:1
`18:0
`18:1(n-7)
`18:1(n—9)
`18:l(n-7)
`18:2(n—6)
`18:3(n—3)
`18:3(n—6)
`
`186
`200
`214
`228
`242
`240
`256
`254
`270
`268
`268
`266
`264
`262
`284
`282
`282
`298
`296
`296
`296
`294
`292
`292
`
`10.0
`11.0
`12.0
`13.0
`14.0
`13.8
`15.0
`14.8
`16.0
`15.7
`15.8
`15.6
`15.5
`15.4
`17.0
`16.7
`16.8
`18.0
`17.8
`17.7
`17.9
`17.6
`17.6
`17.3
`
`18:4(n—3)
`tr.
`tr.
`19:0
`tr.
`tr.
`19:1
`0.22 1 0.06
`0.23 1 0.06
`1922
`0.07 1 0.04
`0.04 1 0.01
`11.33 1 1.48 15.23 1 2.31 20:0
`tr.
`0.19 1 0.01
`20:1(n—7)
`0.34 1 0.01
`0.27 1 0.05 20:1(n—9)
`tr.
`0.04 1 0.03 20:2
`25.91 1 2.33 31.79 1 1.73 20:4(n—3)
`7.26 1 0.35
`7.37 1 0.34 20:5(n—3)
`0.09 1 0.13
`0.30 1 0.01
`21:0
`0 82 1 0 01
`0.12 1 0.06 2l:5(n—3)
`tr.
`0.29 1 0.01 22:0
`0.74 1 0.06
`0.48 1 0.14 22:l(n—7)
`0.06 1 0.02
`0.17 1 0.15 22:1(n—9)
`tr.
`0.41 1 0.05 22:5(n—3)
`tr.
`0.12 1 0.06 22:5
`1.21 1 0.18
`2.14 1 0.23 22:6(n—3)
`8.32 1 0.54
`7.49 1 0.79 23:1
`10.13 1 2.20
`10.52 1 0.90 24:0
`tr.
`0.09 1 0.05 24:1
`1.58 1 0.09
`0.74 1 0.38 25:0
`0.47 1 0.02
`0.33 1 0.07
`0.21 1 0.06
`0.57 1 0.35 Others‘:
`
`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 11.57
`tr.
`0.42 1 0.03
`0.14 1 0.03
`0.29 1 0.17
`0.51 1 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 11.27
`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
`all 1 0.07
`tr.
`0.15 1 0.11
`tr.
`
`—
`
`3.95
`
`2.45
`
`Data are expressed as wt % of total fatty acids and represent means 1 standard deviation of 3 separate experi-
`ments.
`tr. =trace.
`3M: 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 S.
`“Predominantly branched chain fatty acids as given in Table 3 in detail.
`
`LIPIDS, VOL. 19, NO. 11 (1984)
`
`000005
`
`000005
`
`

`
`824
`
`H. FRICKE, G. GERCKEN, W. SCHREIBER AND J. OEHLENSCHLAGER
`
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`826
`
`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:l(n-7)
`(8%), 20:5(n-3) (8-13%) and 22:6(n-3) (3-5%).
`Odd-numbered fatty acids with chain lengths
`ranging from C-11 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 iso- or
`anteiso series. Among the multi-branched-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 1420 and
`1620 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-11) series (36).
`The data found suggest that a (n-7) monoene
`series is present carrying from 16: l(n-7)through
`l8:l(n-7) and 20:l(n-7) to 22:l(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 (19).
`
`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 TG are mostly satu-
`rated or monounsaturated with 14:0, 16:0,
`l6:l(n-7), 18:1(n-7) and l8:1(n-9)as dominat-
`ing species. Polyunsaturated fatty acids were
`
`LIPIDS, VOL. 19, NO. 11 (1984)
`
`found only in small amounts.
`In phospholipids phytanic acid was detected
`only in traces, but it represented 5.6% 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 Euphausia superba Dana (1977 Sample)
`
`Phospholipid
`sn-position
`
`PC
`
`PE
`
`sn—1
`
`srz—2
`
`sn-1
`
`14:0
`16:0
`l6:1(n—7)
`18:0
`18:1(n-7)
`18:1(n-9)
`18:2(n—6)
`20:5(n-3)
`22:6(n-3)
`Others
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`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
`Desmosterolb
`22—Dehydrocholesterol‘3
`Others
`
`70.0 : 5.9
`18.2 : 1.4
`11.5 i 4.8
`0.8 : 0.5
`
`75.5 : 3.7
`17.7 : 1.1
`6.0 i 3-5
`1.0 : 0.7
`
`Total
`
`100.5
`
`100.2
`
`3Cholesta—S—en—3{3—ol
`bCholesta-S,24—dien—3{i—ol
`C22—cis/trans—ch0lesta—5,22-dien—3{3—ol
`
`000008
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`

`
`ANTARCTIC KRILL LIPIDS
`
`827
`
`North—Holland Publishing Company, Amsterdam.
`Broekhuyse, R.M.
`(1968) Biochim. Biophys.
`Acta 152, 307-315.
`Morrison, W.R., and Smith, L.M. (1964) J. Lipid
`Res. 5, 600-608.
`Klemm, H.P., Hintze, U., and Gercken, G. (1973)
`J. Chromatogr. 75, 19-27.
`Ballantine, J.A., Roberts, J.C., and Morris, R.J.
`(1980) J. Exp. Mar. Biol. Ecol. 47, 25-33.
`Pugh, E.L., Kates, M., and Hanahan, D.J. (1977)
`J. Lipid Res. 18, 710-716.
`Heckers, H., Dittmar, K., Melcher, F.W., and
`Kalinowskj, H.D.
`(1977)
`J. Chromatogr. 135,
`93-107.
`Fricke, H., and Schreiber, W. (1983) Naturwiss.
`70, 308-309.
`in Proceedings of the
`(1982)
`Ackman, R.G.
`Second international Conference on Aquaculture
`Nutrition: Biochemical and Physiological Ap-
`proaches to Shellfish Nutrition (Pruder, G.D.,
`Langdon, C.J., and Conklin, D.E., ed.) Baton
`Rouge, Louisiana, pp. 358-376.
`Kubota, K. (1980) J. Jap. Soc. Food Nutr. 33,
`191-193.
`Czeczuga, B., and K,lyszejko, B. (1978) Pol. Arch.
`Hydrobiol. 25, 657-662.
`Czerpak, R., Jackowska, H., and Mical, A. (1980)
`Pol. Polar Res. 1,139-145.
`Yamaguchi, 1(., Miki, W., Toriu, N., Kondo, Y.,
`Murakami, M., Konosu, S., Satake, M., and
`Fujita, T.
`(1983) Bull. Jap. Soc. Sci. Fish. 49,
`1411-1415.
`(1980) in Advances in Fish Sci-
`Ackman, R.G.
`ence and Technology (Connell, J.J., ed.) pp. 86-
`103, Fishing News Books Ltd., Farnham, Surrey,
`U.K.
`Chanmugam, P., Donovan, J., Wheeler, D.J., and
`Hwang, D.H. (1983) J. Food Sci. 48,1440-1441
`and 1462.
`Bell, M.V., Simpson, C.M.F., and Sargent, J.R.
`(1983) Lipids 18, 720-726.
`Hansen, R.P. (1969) Aust. J. Sci. 32, 160-161.
`Hansen, R.P., and Meiklen, S.M. (1970) J. Sci.
`Fd. Agric. 21, 203-206.
`Ackman, R.G. (1968) Comp. Biochem. Physiol.
`24, 549-565.
`Ackman, R.G. (1970) J. Fish. Res. Bd. Canada
`27, 513533.
`Golovnya, R.V., Kuzmenko, T.E., Samusenko,
`A. L., and Grigoreva, D.N. (1981) Appl. Biochem.
`Microbiol. 17, 47-53.
`Ratnayake, W.N., and Ackman,
`Lipids 14, 795-803.
`Ratnayake, W.N.,
`Lipids 14, 804-810.
`Lilly, M.L., and Bottino, N.R. (1981) Lipids 16,
`871-875.
`Dembitskii, V.M. (1979) Sov. J. Mar. Biol. 5(5),
`86-91.
`Brockerhoff, H., Yurkowski, M., Hoyle, R.J.,
`and Ackman, R.G.
`(1964)
`J. Fish. Res. Ed.
`Canada 21, 1379-1384.
`(1977) Oceanogr.
`Morris, R.J., and Culkjn, F.
`Mar. Biol. Ann. Rev. 15, 73-102.
`Gordon, T.
`(1982) J. Am. Oil Chem. Soc. 59,
`536-545.
`Sargent, J.R., and Falk-Petersen, S. (1981) Mar.
`Biol. 62, 131-137.
`
`and Ackman, R.G.
`
`(1979)
`
`R.G.
`
`(1979)
`
`Sterols
`
`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, desmosterol 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 norvegica,
`whereas
`the herbivorous
`copepod Calanus
`fmnmarchicus contained
`l4.l% 22-dehydro-
`cholesterol
`and
`27.7% desmosterol besides
`cholesterol as main sterol (44).
`
`ACKNOWLEDGMENTS
`
`I. Wasum and Dr. W. Konig 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, l-1.17., thanks the latter for financial support.
`
`REFERENCES
`
`9.
`
`(1980) J. Exp. Mar. Biol. Ecol. 43,
`
`1. Grantham, G.J. (1977) Southern Ocean Fisheries
`Survey Progr. GLO/SO/77/2, FAO, Rome.
`2. Shibata, N. (1983) Bull. Jap. Soc. Sci. Fish. 49,
`259-264.
`3. Clarke, A.
`221-236.
`4. Rzavskaja, F.M., Sakaeva, E.A., and Dubrovskaja,
`T.A.
`(1979) Rybnoe
`chozjajstvo
`1979(10),
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`S. Bottino, N.R. (1973) Fed. Proc. Fed. Am. Soc.
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`6. Bottino, N.R. (1974) Mar. Biol. 27, 197-204.
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`8. Pierce, R.W., van der Veen, J., and Olcott, H.S.
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`van der Veen, J., Medwadowskj, B., and Olcott,
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`Soc. Sci. Fish. 30, 630-634.
`11. Ellingsen, T.E. (1982) Biokjemiske studier over
`antarktisk krill,
`Ph.D. Thesis, University of
`Trondheim, 239-316.
`12. Mori, M., and Hikichi, S. (1976) The Report of
`the Central Res. Lab. of Nippon Suisan Co., Ltd.
`No. 11,11-17.
`13. Hempel, G., Sahrhage, D., Schreiber, W., and
`Steinberg, R. (1979) Arch. FischWiss. 30 (Beih.
`1), 1-119.
`14. Christians, O., Birnbaum, A., Leinemann, M.,
`Manthey, M.,
`and Oehlenschlager,
`J.
`(1982)
`Arch. FischWiss. 33 (Beih. 1), 143-170.
`15. Folch, J., Lees, M., and Sloane-Stanley, G.H.
`(1957) J. Biol. Chem. 226, 497-509.
`16. Kates, M.
`(1972) Techniques of Lipidology,
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`18.
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`19.
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`20.
`
`21.
`
`22.
`
`23.
`
`24.
`
`25.
`
`26.
`
`27.
`
`28.
`
`29.
`
`30.
`
`31.
`
`32.
`33.
`
`34.
`
`35.
`
`36.
`
`37.
`
`38.
`
`39.
`
`40.
`
`41.
`
`42.
`
`43.
`
`44.
`
`[Received April 30, 1984]
`
`LIPIDS, VOL. 19, NO. 11 (1984)
`
`000009
`
`000009