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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 AcyI A5 Desaturation
`Intestinal Metabolism of Plasma FFA in Diabetic Rats
`
`Quantitative Analysis of Triglyceride Species
`Analysis of Sterol Esters by Capillary Gas Chromatography—Electron Impact and Chemical
`Ionization-Mass Spectrometry
`
`COMMUNICATIONS
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`902-905
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`Protein Depletion, EFA and Apoproteins of VLDL from Perfused Liver
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`ISSN:0024-4201
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`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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`000002
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`
`821
`
`Lipid, Sterol and Fatty Acid Composition of Antarctic Krill
`
`(Euphausia superba Dana)
`
`H. FFlICKE,a G. GERCKENF W. SCHREIBERb and J. OEHLENSCHLAGERPI"
`aDepartment of Biochemistry, University of Hamburg, Hamburg, and blnstitu te for
`Biochemistry and Technology in the Federal Research Centre of Fisheries, Pa/mai/le
`.9, D-2000 Hamburg 50, Germany
`
`ABSTRACT
`
`The lipid classes, fatty acids of total and individual Lipids and sterols of Antarctic krill (Euphuusiu
`superbu 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%), phosphatidylethanolam'me (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), 18:1(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 (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 (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 a1. (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 dichloromethanezmethanol:glacial 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 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
`
`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-
`01s 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 1- 0.3
`
`Phospholipids
`Phosphatidylcholine
`Phosphatidylethanolamine
`Lysophosphatidylcholine
`Phosphatidylinositol
`Cardiolipin.
`.
`Phosphatldlc a01d
`
`Neutral lipids
`Triacylglycerols
`Free fatty acids21
`Diacylglycerols
`Sterols
`Monoacylglycerols
`
`omafl
`
`Total
`
`35.6 t 0.1
`6.1 t 0.4
`1.5 :r 0.2
`0.9 i 0.1
`1.0 i 0.4
`0.6 t 0.4
`
`33.3 i 0.5
`5.2 :r 0.5
`2.8 i 0.4
`1.1 t 0.4
`1.6 i 0.2
`
`33.3 :r 0.5
`16.1 t 1.3
`1.3 t 0.1
`1.7 i- 0.1
`0.4 :r 0.2
`
`40.4 t 0.1
`8.5 i 1.0
`3.6 t 0.1
`1.4 i 0.1
`0.9 i 0.1
`
`09:01
`
`05:01
`
`98.9
`
`99.3
`
`lipids and
`Data are expressed as wt % of total
`represent means ’5 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% 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 lipolylilc
`activity against phospholipids exists even at
`temperatures of -30 C and below. Samples 0f
`
`000004
`
`000004
`
`
`
`ANTARCTIC KRlLL 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 Euphausz‘a crystal-
`lorophias 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 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
`
`1W
`ECL
`
`
`13:0i
`1420i
`15:0i
`15:0 ai
`1620i
`17:0i
`17:0 bra
`17:1 hr
`17:1 hr
`1820i
`Phytanicb
`
`326 17.7 2.82 i 0.41 1.2acid t 0.43
`
`n.d.
`tr.
`12.6
`n.d.
`0.05 10.01
`13.6
`14.6 0.19:0.00 0.31:0.15
`14.7
`0.21 $0.01
`0.24 10.07
`15.6 0.09:0.03 0.10: 0.06
`16.6
`0.54 t 0.05
`0.20 t 0.02
`16.4
`tr.
`0.09 i 0.02
`16.5
`0.05 i 0.03
`0.11 t 0.08
`16.2
`tr.
`0.10:0.05
`17.6
`tr.
`0.10:0.01
`
`
`
`
`
`
`
`228
`242
`256
`256
`270
`284
`284
`282
`282
`298
`
`
`
`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.
`aPresumably 7-methy1hexadecanoic acid.
`b3,7,11,15-tetramethylhexadecanoic acid.
`
`TABLE 2
`
`Fatty Acid Composition of Total Lipids of Euphausia superba Dana
`
`Sample
`
`12/1977
`
`3/1981
`
`Sample
`
`12/1977
`
`3/1981
`
`M:a ECLb
`Nra
`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 i 0.06
`12.0
`214
`12:0
`1922
`0.07 i 0.04
`0.04 i 0.01
`13.0
`228
`13:0
`11.33 t 1.48 15.23 i- 2.31 20:0
`14.0
`242
`14:0
`tr.
`0.19 i 0.01 20:1(n-7)
`13.8
`240
`14:1
`0.34 i- 0.01
`0.27 i- 0.05 20:1(n-9)
`15.0
`256
`15:0
`tr.
`0.04 i 0.03 20:2
`14.8
`254
`15:1
`25.91 i 2.33 31.79 i 1.73 20:4(n-3)
`16.0
`270
`16:0
`7.26 :r 0.35
`7.37 x 0.34 20:5(n-3)
`15.7
`268
`16:1(n-7)
`0.09 t 0.13
`0.30 t 0.01
`21:0
`15.8
`268
`16:1(n-7)
`0.82 t 0.01
`0.12 i 0.06 21:5(n-3)
`15.6
`266
`16:2(n-6)
`tr.
`0.29 i 0.01 22:0
`15.5
`264
`16:3
`0 74 t 0.06
`0.48 1- 0.14 22:1(n-7)
`15.4
`262
`16:4(n-3)
`0.06 i 0.02
`0.17 i 0.15 22:1(n-9)
`17.0
`284
`17:0
`tr.
`0.41 i 0.05 22:5(n-3)
`16.7
`282
`17:1
`tr.
`0.12 i 0.06 22:5
`16.8
`282
`17:1
`1.21 i 0.18
`2.14 t 0.23 22:6(n-3)
`18.0
`298
`18:0
`8.32 i 0.54
`7.49 t 0.79 23:1
`17.8
`296
`18:1(n-7)
`10.13 i 2.20
`10.52 t 0.90 24:0
`17.7
`296
`18:1(n-9)
`tr.
`0.09 i- 0.05 24:1
`17.9
`296
`18:1(n-7)
`1.58 i- 0.09
`0.74 i 0.38 25:0
`17.6
`294
`18:2(n-6)
`18:3(n-3)
`292
`17.6
`0.47 i 0.02
`0.33 t 0.07
`
`
`
` 18:3(n-6) 292 17.3 0.21 i 0.06 0.57 t 0.35 Othersc —— — 3.95 2.45
`
`
`
`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 t 0.07
`tr.
`0.12 t 0.04
`tr.
`0.04 i 0.00
`0.40 t 0.01
`0.77 i 0.04
`tr.
`0.46 t 0.10
`12.71 i 1.57
`tr.
`0.42 i 0.03
`0.14 1- 0.03
`0.29 i 0.17
`0.51 i 0.06
`0.54 t 0.09
`tr.
`5.41 i- 0.51
`tr.
`tr.
`tr.
`tr.
`
`0.62 t 0.49
`0.11 1- 0.16
`0.20 t 0.09
`0.07 t 0.05
`0.19 t 0.14
`0.50 t 0.09
`1.35 t 0.23
`0.08 t 0.06
`0.22 i' 0.06
`7.83 i 1.27
`tr.
`0.30 i 0.18
`tr.
`0.41 i 0.16
`1.22 t 0.33
`0.24 t 0.11
`0.04 i 0.03
`2.60 t 0.79
`0.11 i 0.07
`tr.
`0.15 t 0.11
`tr.
`
`290
`312
`310
`308
`326
`324
`324
`322
`318
`316
`340
`330
`354
`352
`352
`344
`344
`342
`366
`382
`380
`396
`
`
`
`
`
`
`
`
`
`Data are expressed as wt % of total fatty acids and represent means i 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 5.
`CPredominantly branched chain fatty acids as given in Table 3 in detail.
`
`LIPIDS, VOL. 19, NO. 11 (1984)
`
`000005
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`H. FRICKE, G. GERCKEN, W. SCHREIBER AND J. OEHLENSCHLAGER
`
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`
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`
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`
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`
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`
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`LIPIDS, VOL. 19, NO. 11 (1984)
`
`000007
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`000007
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`
`
`826
`
`H. FRICKE, G. GERCKEN, W. SCHREIBER AND J. OEHLENSCHLAGER
`
`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 Euphausz’a superba Dana (1977 Sample)
`
`PE
`PC
`Phospholipid
`
`sn-2sn-lsn-position sn-l
`
`
`
`
`
`(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) (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-l3 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 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 a1. (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-l 1) series (36).
`The data found suggest that a (n-7) monoene
`series is present carrying from 16: 1(n-7)through
`l8:1(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 (SC-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,
`16:1(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)
`
`000008
`
`14:0
`16:0
`16:1(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-Dehyd1'ocholesterolc
`Others
`
`70.0 t 5.9
`18.2 i 1.4
`11.5 i 4.8
`0.8 i 0.5
`
`75.5 t 3.7
`17.7 t 1.1
`6.0 i 3-5
`1.0 t 0.7
`
`100.2
`100.5
`Total
`
`
`aCholt-zsta-S-en-313-ol
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`
`000008
`
`
`
`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, DJ. (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 ]nternational 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 KJyszejko, B. (1978) P0]. Arch.
`Hydrobio]. 25, 657-662.
`Czerpak, R., Jackowska, H., and Mica], A. (1980)
`P0]. Polar Res. 1,139-145.
`Yamaguchi, K., 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 TechnOIOgy (Connell, J.J., ed.) pp. 86-
`103, Fishing News Books Ltd., Farnham, Surrey,
`U.K.
`Chanmugam, P., Donovan, 1., Wheeler, DJ, and
`Hwang, DH. (1983) J. Food Sci. 48,1440-1441
`and 1462.
`Bel], M.V., Simpson, C.M.F., and Sargent, J.R.
`(1983) Lipids 18, 720-726.
`Hansen, RP. (1969) Aust. J. Sci. 32, 160-161.
`Hansen, R.P., and Meiklen, SM. (1970) J. Sci.
`Fd. Agric. 21, 203-206.
`Ackman, R.G. (1968) Comp. Biochem. Physio].
`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) App]. Biochem.
`Microbiol. 17, 47-53.
`Ratnayake, W.N., and Ackman, R.G.
`Lipids 14, 795-803.
`Ratnayake, W.N.,
`Lipids 14, 804-810.
`Lilly, M.L., and Bottino, NR. (1981) Lipids 16,
`871-875.
`Dembitskii, V.M. (1979) Sov. J. Mar. Biol. 5(5),
`86-91.
`Brockerhoff, H., Yurkowski,
`and Ackman, R.G.
`(1964)
`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.
`
`(1979)
`
`and Ackman, R.G.
`
`(1979)
`
`M., Hoyle,
`Fish. Res.
`
`R.J.,
`Bd.
`
`J.
`
`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 t0 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
`finnmarchz‘cus contained
`14.1% 22-dehydro-
`cholesterol
`and
`27.7% desmosterol besides
`cholesterol as main sterol (44).
`
`ACKNOWLEDGMENTS
`
`I. Wasum and Dr. W. Kdnig 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.
`
`REFERENCES
`
`9.
`
`(1980) J. Exp. Mar. Bio]. Ecol. 43,
`
`1. Grantham, G.J. (1977) Southern Ocean Fisheries
`Survey Progr. GLO/SO/77/2, FAO, Rome.
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`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, NR. (1973) Fed. Proc. Fed. Am. Soc.
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`6. Bottino, NR. (1974) Mar. Biol. 27, 197-204.
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`(1975) Comp. Biochem. Physio].
`50 B, 479-484.
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`(1969) J. Ag'r. Food Chem. 17, 367-369.
`van der Veen, J., Medwadowski, B., and Olcott,
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`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.
`l3. Hempe], G., Sahrhage, D., Schreiber, W., and
`Steinberg, R. (1979) Arch. FischWiss. 30 (Beih.
`1), 1-119.
`14. Christians, 0., Birnbaum, A., Leinemann, M.,
`Manthey, M.,
`and Oehlenschla'ger,
`J.
`(1982)
`Arch. FischWiss. 33 (Beih. 1), 143170.
`15. Folch, J., Lees, M., and Sloane-Stanley, G.H.
`(1957) J. Biol. Chem. 226, 497-509.
`16. Kates, M.
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`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
`
`