Org. Geochem. Vol. 9, No. 2, pp.

83 99, 1986
Printed in Great Britain. All rights reserved

0146-6380/86 $3.00 + 0.00 Copyright ;C, 1986 Pergamon Press Ltd

A review of sterol markers for marine and terrigenous organic matter
JOHN K. VOLKMAN CSIRO Division of Oceanography, G.P.O. Box 1538, Hobart, Tasmania, Australia 7001

(Received 23 May 1985; accepted 18 October 1985)
Abstract--A review of literature on the occurrence of 4-desmethyl sterols in unicellular algae indicates that 'few sterols are sufficiently restricted in distribution to be considered unambiguous markers for specific algal groups. Almost all of the 4-desmethyl sterols found in higher plants occur in marine algae, sometimes as major constituents, so sterol distributions do not always allow one to distinguish between marine or terrigenous organic matter. Some of the problems associated with the use of sterols as markers for specific sources are highlighted by comparison of the sterol distributions in selected marine sediments and seawater samples. Although each sediment represents a very different depositional environment the sterol distributions are surprisingly similar. The sterol distributions in a saline Antarctic lake and samples of particulate matter from oligotrophic waters off the east Australian coast show that marine phytoplankton biosynthesize a wide range of sterols, including large amounts of 24-ethylcholest-5-en-3fl-ol which is often used as a marker for terrigenous organic matter. Similar distributions occur in marine sediments from the upwelling area off Peru and in temperate intertidal sediments despite large differences in algal productivity between the two areas. In deeper sediments, most of the sterols are not derived from phytoplankton but from higher plants. These data indicate that inferences drawn from sterol distributions regarding sources of organic matter must be made with caution and should be supported using other lipid data. It further follows that in ancient sediments and crude oils a high proportion of C29 steranes need not indicate that most of the organic matter was derived from vascular plants.

Key words: sterols, phytoplankton, biological markers, Recent sediments, seawater particles, early diagenesis.

INTRODUCTION

Many studies have highlighted the advantages of sterols as biological markers and much is now known about the distributions of sterols in sediments from many different types of environments (e.g. Brassell and Eglinton, 1983a,b, Mackenzie et al., 1982). Sterols are comparatively stable and hence they have a long geological record. They also possess structural features, such as positions of double bonds and patterns of side-chain alkylation, which are restricted to a few groups of organisms. However, some sterols are widely distributed in biological systems which makes them of limited value for assigning sources of organic matter in sediments. Sterols are also excellent biomarkers for tracing diagenetic transformations in both Recent and immature ancient sediments (Mackenzie et al., 1982). In mature ancient sediments and crude oils, distributions of 5~(H)-steranes are found which can be related to precursor sterols by reasonably well documented diagenetic pathways. Sterane mixtures are often complex, as are the distributions of sterols in Recent sediments, but there has been little progress in using these distributions as source indicators. In most cases, the only parameter determined is the ratio of C27 to C2~ steranes which is thought to reflect the
OG 92 l)

relative amounts of autochthonous and allochthonous (higher plant) inputs (e.g. Huang and Meinschein, 1976; 1979). A high proportion of C29 steranes is thought to indicate a large input of terrigenous organic matter, since precursor C29 sterols such as 24-ethylcholest-5-en-3fl-ol are abundant in vascular plants. The results presented in this paper were chosen to highlight some of the strengths and ambiguities in the use of sterols as biomarkers for elucidating sources of organic matter. Marine organisms were expected to be major lipid sources in each of the samples studied, and yet C29 sterols predominate in each case. A detailed examination of the sterol distributions, combined with data for other lipid classes, indicates that marine organisms can be major sources of 24-ethylcholest-5-en-3fl-ol and other sterols presently used as biomarkers for terrigenous organic matter. The literature on sterols in unicellular algae is also reviewed and this suggests that a reassessment of many sterol biomarkers is needed.
EXPERIMENTAl,

Data for 3 sediments and 2 seawater samples are presented in this paper. Details of the sampling locations are given in Table 1. 83

(c) Antarctic lake sediments. The solvent extracts were then combined.1 0 c m ) sediments from Cranwell and V o l k m a n (1981). saline.d. Several sediment cores were obtained in June 1978 by H. Samples (a) Seawater. meromictic lake is situated in the Vestfold Hills of Princess Elizabeth Land. Burton from the middle of Ace Lake. Data from a core section 50-55 cm deep (Ace 31/50-55) are given here. Farrington and R. Seawater samples were obtained using a 81 Niskin bottles from depths of 0 m (SP11/83/23/0) and 60m (SP11/83/23/60) during cruise SP11/83 of R / V Sprightly in June. 146°20'E). 1981a). Victoria./41-50) are reported in this paper. (d) Peru sediment.. (1983) and Gagosian et al.ug/g h 1170~g/g 9. Full details of the collection and extraction of this sample are given by Volkman et al. 6 times) using ultrasonication.9'W -32 '00'S 155 0 0 ' W 32 '00'S 155' 00"W Sterols 1. Sterols from the Antarctic lake sediments and seawater samples were analyzed using a Shimadzu GC-9A gas chromatograph fitted with an SGE OCI-3 cool on-column injector and Hewlett Packard high performance grade bonded-phase fused silica column (25 m x 0. i d~ uS: sediment. Previous work with chloroform-methanol extraction of Peru sediments had shown that high concentrations of fatty acid methyl esters can be formed unless such precautions are taken.32 mm i. hConcentration o f free sterols per g o f dry sediment. sediment core was obtained in August.3 .2/1 g/I . ' D a t a for Loch Clair ( 0 . washed with saline solution and evaporated to dryness. Neutral lipids were extracted using hexanediethyl ether mixtures. Details of the distributions of sterols and other lipids in surface sediments from this site are given by Volkman (1980c. 20ml. b). . 10ml) using ultrasonication. Data from a core section 41-50 cm deep (C.4 # g / g ~ 80/~g/g 0.6 . Eastern Antarctica. This sample was collected by J. Corner Inlet. H20:CH3OH 2:1. (1983a. B./0-10 c SPI 1/83/23/0 SPI 1/83/23/60 Type ~ S S S S SW SW Depth 41-50 cm 50-55cm 16-19 cm 0-10 cm 0m 60 m W a t e r column 0m 23m 85m 32 m > 2000 m > 2000 m Location 38"49'S 6828. A dual ramp temperature program was employed with the oven heated from 40 to 150C at 30°C/min and then from 150 to 32OC at 4 C/rain.I. Information on the phytoplankton and bacteria in the lake and their lipid and pigment compositions is given by Volkman et al. After elution of fractions containing less polar lipids. This small (1. pyrex WCOT capillary column as described by Volkman et al.) coated with methyl silicone gum (0. The lake is ice-covered from March to November which prevents wind-driven mixing of the lake waters giving rise to permanent stratification. Hydrogen was used as the carrier gas at a linear velocity of 34cm/sec. Material greater than 200 #m in size was excluded using a mesh prefilter. (1986).d. 1983 from a station about 220 km off the East Australian coast north-east of Sydney. The filters were frozen for later analysis on-shore. Initial gas chromatographic analysis of the Peru sample was carried out using a Carlo Erba FTV 2150 gas chromatograph equipped with a 20 m × 0. pH 12) for 6hr. 1976 from the intertidal area of Old Yankie Beach.2'S 146"20'E 78"11.C. 1983b). A second portion was saponified in KOH solution (10 ml. (b) Intertidal sediment. SW: seawater. The initial extraction with propan-2-ol was included to inhibit any enzyme activity which might promote transesterification of lipids containing fatty acids.d.1'E 75'~30. Analyses o f sterols Aliquots of the TSE lipids from each of the sediment samples were separated into major lipid classes by column chromatography on silicic acid (7g) deactivated with 5% distilled water (Gagosian et al. a fraction containing 4-desmethyl sterols was obtained using 20% ethyl acetate in hexane. The Peru and Antarctic sediment samples were reanalyzed using a Hewlett Packard HP5890A capillary gas chromatograph with comparable results. A 2 m x 5 cm o. Gagosian using a MkIIl box corer deployed at a site (BC7) about 7 km from the Peru coast. for GC and G C MS analysis.ug/I 0. The combined extracts were washed with saline solution and a portion was then analyzed for total lipid classes using Iatroscan thin layer chromatography-flame ionisation detection (TLC-FID).11 #m fii~ thickness). Core 31 from the centre of the lake at a water depth of 23 m was sectioned into 5 cm sections at the collection site and immediately frozen (ambient air temperature <0°C).4'S 15"02. Australia (38°49'S.3 x l0 s m2). Total solvent extractable (TSE) lipids were obtained by extracting the filters 3 times with CHCI3:CH3OH (2:1. VOLKMAN Table 1. W./41-50 Ace 31/50-55 PBC7/16-19 L. Portions of each core section were thawed in December 1983 and extracted with propan-2-ol (20 ml) and then CHCI3:CH3OH (2: 1.. Sediment and seawater samples Sample code C. (1983).2 mm i. FID detection was used with a detector temperature of 350C. Each sample was filtered onboard through precleaned G F / F filters to obtain the particulate matter.I.84 JOHN K. These sterols were then converted to trimethylsilyl ether (TMS-ether) derivatives using bis(trimethylsilyl)trifluoroacetamide (BSTFA).

4 1.4 TR a TR 2.9 11.0 TR TR 7.6 .1 ND 1.0 59. I 1. Data for trace constituents and some unidentified sterols have not been included but in each case 90-95% of the total 4-desmethyl sterols were identified.3 0.6 39.8 65.0 2.2 mm i. Each sterol fraction was analyzed by capillary gas chromatography mass-spectrometry (GC-MS) using a VG70/70 magnetic sector mass spectrometer coupled to a Pye 204 gas chromatograph fitted with an SGE OCI-1 on-column injector.1 3.0 1.3 0.16 .9 25. Sterols were identified from their electron impact mass spectra.C. The column was inserted directly into the ion source of the MS through an interface heated to 280°C.22E-dien-3B-ol 24-methyl-5ct -cholest-22E-en-3fl-ol 5ct-cholest-7-en-3fl-ol 24-methyl-5e-cholesta-7.2 28..2 12.12 0.7 TR 0.3 . reflecting very dissimilar amounts of organic matter contributed to each sediment. bonded phase fused silica BP-I (methyl silicone) column was used with chromatographic conditions similar to the above.8 27.0 1.5~t-cholestanol/ 24-ethylcholesterol % % % % % TR 15.0 1.8 ND ND 8.7 1.4 25.8 39.22E-alien-3fl-ol 5ct-cholest-22E-dien-3fl-ol cholest-5-en-3fl-ol 5ct-cholestan-3fl-ol 24-methylcholesta-5.8 6.07 0.5 30.3 TR 29. TR: trace ( < 0.6 0.9 62. 0-10 -11.3 ND ND 9. 1980a.6 3. 1983a.0 6. Percentage composition of 4-desmethyl sterols in sediments and seawater samples Sample" Peak" number 1 85 Structure ^ la lla lb lib Ic IIc Id lid lllc llld le lie If Ilf Ig llg lh llh IIlf Ii f lli Ij Ik f llj Ilk 11 1II Illj Identification 24-nor-cholesta-5.5ct-cholest-24(28)-en-3fl-ol 24-methylcholest-5-en-3fl-ol 24-methyl-5ct-cholestan-3fl -ol 23.21 0.3 0. ..5 0.0 39.9 3.49 1.3 TR 2.8 4. De Leeuw et al.8 ND 0. b).42 L.35 0. However.9 PBC7 16-19 0.1 4.3 55.24-dimethyl-5~t-cholestan-3fl-ol 24-ethylcholest-5-en-3fl-ol 24-ethylcholesta-5.8 1. 1. The concentrations of total sterols are very different in the three environments (Table 1). Sterol structures are given in the Appendix.24(28)E-dien-3fl -ol 24-ethyl-5ct-cholestan-3fl-ol 24-ethyl-5ct-cholest-24(28)E-en-3fl-ol 24-ethylcholesta-5.5 3.4 2.1 TR 1.0 TR 4.4 4.5 4./60 ND ND 4.4 5.- 31.7 TR 4.- 33. retention index data and coinjection with standards (Table 2). 1983.2 5.0 TR 0. L. 1984.24-dimethyl-5~-cholest-22E-en-3fl-ol 24-ethylcholesta-5.22E-dien-3fl-ol 24-nor.5%).38 Ace 31 50-55 1.1%).24-dimethylcholest-5-en-3fl-ol 23.7 3.3 ND ND ND TR Ace 31 5(~55 0.27 0..5 8.5 7. b.6 12.4 9. (~10 SPI 1/83 23/0 TR TR 5.- 5.8 0.C.6 1.5 23.2 ND ND ND 17.7 TR a 2.4 TR TR 2.22E-dien-3fl-ol 24-ethyl-5ct-cholest-22E-en-3B-ol 24-met hyl-50t-cholest-7-en-3fl-ol 23.3 0.I.I. 1.2 7.0 12..6 ND ND 35.: not reported.5 15. 41-50 C26 sterols C27 sterols C28 sterols Cz9 sterols 24*ethylcholesterol plus 24-ethyl-5ct-cholestanol Total 5~t(H)-stanols/Atstenols 5ct-cholestanol/cholesterol 24-et hyl.C.- 8.3 13.6 TR ND 6. I 0.6 TR 5.8 ./83 23/60 TR 35.- . differences in Table 3.5ct-cholest-7-en-3fl-ol C.6 0. Selected compositional data expressed in terms of structural type and carbon number are shown in Table 3.86 1.95 SPI 1/83 23/0 TR 37.4 3.4 0. RESULTS The percentage compositions of 4-desmethyl sterols in the three sediments and two seawater samples are presented in Table 2. 1981a.6 1. Capillary gas chromatograms of the sterols in two of the sediment samples are shown in Fig.1 TR 29.2 . Gagosian et al. 41 50 TR TR 2. Compositions of 4-desmethyl sterol fractions according to carbon number and structure C.12 PBC7 1619 TR 18. bStructures are given in the Appendix.2 0.9 2.6 1.2 12. The MS was scanned from 600 to 20 amu with a recycle time of 2.2 sec.3 ND 11. I TR 7./0-10: Loch Clair (~10 cm free sterols--data from Cranwell and Volkman (1981).Sterol m a r k e r s Table 2.24(28)-dien-3fl-ol 24-methyl.3 3.3 5.. I 1.- .34 SPI I.24-dimethylcholesta-5.- 18 19 20 °Peak numbers refer to Fig.6 1. CSample designations are given in Table 1.7 3.d. An SGE 25 m × 0.9 t.22-dien-3fl-ol 24-methylcholesta-5. aND: not detected ( < 0.1 .5~t-cholest-22E-en-3fl-ol cholesta-5.3 ND TR 0.5 1.44 0.- 5.5 0.9 40.- .24(28)Z-dien-3fl-ol 24-ethyl-5ct-cholest-24(28)Z-en-3fl-ol 24-ethyl.6 32.0 TR 1.8 1.15 0.0 76.0 9.5 TR 10.2 ND ND ND 27.- .20 0.7 0.05 0.2 1.22E-dien-31~-ol 23.9 3.0 TR 11.8 28.7 2.9 ND TR ND ND SP11/83 23.0 32. as described in earlier studies (Volkman et al.3 ND ND ND ND 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 .4 ND 1..

/41 50 it is less abundant than 24-ethylcholest-5-en-3fl-ol (9. Matsumoto et al.I.8 /~g/g." . Volkman et al. C29 sterols collectively comprise ~ 56% of the total sterols whereas C27 sterols represent only 15. 1981a)..T 36 38 i 40 42 I 44 I 46 I 48 I 50 Time ( rain ) Fig. Antarctic lake sediments The concentration of4-desmethyl sterols in the Ace Lake sediment is very high ( l l 7 0 ~ g / g . 1978).22E-dien-3fl-ol (6. 29. and even in organic rich sediments deposited in highly productive upwelling areas off Namibia and Peru and sterol concentrations fall typically in the range 80-1201~g/g (Wardroper et al. At the surface.86 Ace Loke 5 0 .6/~g/g which is only slightly less than in surface sediments from this area (1.2% cf. Gagosian et al. VOLKMAN 17 13 5 114 '°11 11'511 12 18 20 3 ~L Peru Box Core 7 16 . Table 2) reflecting high algal productivity.. 24-methylcholest-5-en-3flol (7.9%. GC conditions are given in the experimental section.. more organicrich.. Total concentrations show little change with depth. 1981a). A previous study of Antarctic lake sediments from the dry valleys of Victoria Land found much lower concentrations of sterols (range: 0.. Corner Inlet intertidal sediment Sterols are present in very low concentrations in this sediment compared with other. In the 41 50 cm section. The total concentration of 4-desmethyl sterols is only 1.3%. I..19 cm 13 17 18 24 3 ~ ' . Table 2). 1982). but the proportions of individual sterols vary considerably (unpublished data). 1983a).. good preservation and the absence of benthic fauna. environments. seagrass detritus and vascular plants with minor contributions from other algae. Volkman et al. In surface sediments diatoms are major contributors (Volkman et al. bacteria. but in deeper sediments vascular plants and seagrass detritus are clearly more important contributors.4%). Sterol identifications are listed in Table 2.7%). Major sources of lipids in the intertidal zone include diatoms. with somewhat . Other major sterols include 24-ethyl-5~cholestan-3fl-ol (11.7/~g/g. stanol/stenol ratios (Table 3) are surprisingly small when one considers the large variation in depositional conditions and redox potentials.13-16. and 24-ethylcholesta-5. 1981a) but in C..8%... the n-alkane and fatty acid distributions are dominated by C20-C30 components typical of higher plants (Johns et al.' . the major 4-desmethyl sterol is 24-methylcholesta-5. 1978.3%). 1982. Smith et al.22E-dien-3fl-ol (24.55 cm JOHN K.. Partial capillary gas chromatograms of free sterols (as TMS-ethers) in (a) Ace Lake 50 55 cm sediment and (b) Peru coastal sediment 16-19 cm.

1983). although complex sterol distributions are found in a few freshwater lacustrine environments (e. This is surprising since the two environments are very different.6%) and 24-ethyl-5~-cholest-7-en-3fl-ol (4.1% of the 4-desmethyl sterols and yet there are few diatom cells in the lake. ~sJtypical of marine ecosystems. but these organisms are not present in Ace Lake so the A 7- 87 sterols must be derived from algal sources. but the source of these sterols in Ace Lake is uncertain at this stage. (1982) only 9 sterols were reported... nor are they abundant in diatomaceous oozes from Walvis Bay and Peru (Wardroper et al.. Surface sediments from the same general area contain much higher concentrations of sterols and the distributions are considerably more complex with C28 sterols such as 24-methylcholesta-5. 1982). Sterols with As. . 1974. 1983b).g. this sediment section contains high concentrations of n-alkanes and fatty acids whose distributions clearly indicate substantial inputs of terrigenous organic matter (Volkman et al. Marine invertebrates such as sponges and asteroids are common sources of A7-sterols in the marine environment (Faulkner and Anderson.. 1983a. 1980a. The distribution of sterols in this sediment section resembles that found in lacustrine sediments such as those from Loch Clair (Cranwell and Volkman.22E-dien-3fl-ol is also abundant (10. 1983a. but more recent studies of diatoms have not identified these sterols as significant components (Ballantine et al. Cholest-5-en-3/~-ol is the only other sterol to exceed 10% in relative abundance (Table 2). 1983a.22E-dien3fl-ol represents 11. The high concentrations of cholest-5-en-3fl-ol in the sediments are probably mainly derived from the only animal in the lake. Burton. b). These 4 algae must contribute most of the sterols in the sediment. have been identified in a few marine sediments (lkan et al. This diversity of structures.5% of the total sterols. 1975).. 1981.. The identifications of 24-methyl-5~-cholesta-7. but data from sediment trap studies clearly show that 24-ethylcholest-5-en-3/~-ol and other C29 sterols are minor constituents of phytoplankton in this region (Gagosian et al..9%) were confirmed by coinjection with authentic standards. gelidicola (unpublished data) so this green alga is probably not the source of AT-sterols in Ace Lake. 1983b). There are also smaller populations of a dinoflagellate. Small amounts of 5c~-cholest-7-en-3fl-ol and other A7-sterols. consisting of over 25 identified constituents and several sterols of unknown structure (Table 2). Another common plant sterol. 1983a.. 24-ethylcholesta-5.. other minor A7-sterols may be present but standards were not available for confirmation.0%) despite the fact that vascular plants are not present in the Vestfold Hills. Cyanobacteria are a potential source of A7-sterols in some environment (Table 4)... Ballantine et al. Scales of the green alga P y r a m i m o n a s gelidicola are abundant throughout the core (H. AT-Sterols were not detected in P. Robinson et al. but the distribution of sterols is similar in many respects (Table 3). 1978. Cyanobacterial mats fringe the lake but since cells have not been found in the water column it seems unlikely that they are contributors of lipids in sediments deposited in the centre of the lake. 1981 ) where much of the organic matter is derived from vascular plants (Table 2). 1981a). R. 1986). 1983). In the Antarctic lake sediments studied by Matsumoto et al. Smith et al. Gagosian et al. the copepod Paralabidocera antarctica. A~-Sterols have been reported in several species of diatoms (Orcutt and Patterson.. since in common with most copepods it contains cholest5-en-3fl-ol as the major sterol (unpublished data). On the basis of inferences drawn from the Antarctic sediment data. There are very few reports of A7-sterols in sediments. 1982. a Cryptophyte and a small biflagellate alga which may be Prymnesiophyte (Volkman et al. 1982. The major sterol is 24-ethylcholest-5-en-3fl-ol (35... personal communication) so one would expect this alga to be a major source of lipids. b.22E-dien3/~-ol.4%). Previously studies have suggested that phytoplankton could contribute some of the 24ethylcholest-5-en-3/~-ol in surface sediments from this area (Gagosian et al. 24-methyl-5~-cholest-7-en-3fl-ol 6. 1981a)... b. 1983a.. The 4-desmethyl sterol distributions in the Ace Lake sediments are very complex. Smith et al. Also. many studies have established that the organic matter deposited in the Peru coastal area results mainly from high phytoplankton productivity (Gagosian et al.and AS~14)-unsaturation have similar mass spectra but these are readily identified from retention index data (Itoh et al.24(28)-dien-3/~-ol as major components (Gagosian et al. AT-Sterols are found in several species of green algae (Table 4) and thus these sterols have been proposed as markers for lipids from this algal class (Volkman et al. Peru sediment The concentration of sterols in the Peru sediment is 125 times less than in the Antarctic core section (Table 1). which comprise 49. Gillan et al. Volkman et al. 24-ethyl5~-cholestan-3/~-oi and 24-ethylcholesta-5.. 1975.9%). 24-Methylcholesta-5.Sterol markers higher concentrations occurring at the sedimentwater interface (Smith et al. and these were not detected in the sediment... An unusual feature of the sterol distribution is the presence of sterols with AT-unsaturation (Table 2)... 1983b) including surface sediments from Corner Inlet (Volkman et al. 1979. one might argue that most of the Cz9 sterols in the Peru sediments are also derived from marine algae but this is highly unlikely. Smith et al. 1983a).22E-dien-3/3-ol and 24methyicholesta-5.22Edien-3fl-ol (2. 1982).. Smith et al. Lin et al. The major constituents are the Cz9 sterols 24-ethylcholest-5-en-3/~-ol. 1984).. Smith et al. although the use of packed GC columns in that study would have made it difficult to recognize any minor sterols which might have been present. 1979).

1979.1/~g/1. (14) Paoletti et al. Saliot et al. (11) Seckbach and lkan (1972). (1984).g. Saliot and Barbier. unpublished data (1984). (6) Prahl et al. 1974).5/~g/l.04#g/l. (3) Lenton et al. 1983b). unpublished data (1985) (10) Reitz and Hamilton (1968). Seawater samples Sterols were present in low concentrations in seawater particles from the East Australian coast (Table 1). Calothrix sp. The factors determining the relative rates of degradation of marine vs. G--24-methylcholesta-5. such as western North Atlantic (0. These concentration values (0. 1982. . (5) Goad et al. 1983).. 1983b). Nostoc commune 36 + B 2 48 31 28 30 23 23 C 8 33 3 16 7 1 13 D E F 26 19 G 7 H Others 21" Reference 1 2 3 1 64 4 1 56 60 72 64 20 3 4 80" 4 4 4 5 6 + 5 1 34 99 48 2 58 t 1 7 51 95 5 23 7 55 6 90 3 2 20 1 8 9 10 10 II 12 13 14 14 14 14 29 57 + 55 8 6 15 10 10 6 6 2 1 4 7 3 48 36 39 20 51 33 48 4 13 7 8 10 4 22 8 1 3 Notes: ~A---cholest-5-en-3/%ol.. 1983. 1976) and Arabian Sea (0. The two C28 sterols are common in seawater where they are derived mainly from diatoms (e. (1979). References cited: (1) Ballantine et al. B--24-methylcholest-5-en-3~-ol.~. (12) Nishimura and Koyama (1977). but there has been uncertainty as to its origin. C--24-ethylcholest-5-en-3//-ol.22E-dien-3/~-ol.g. (13) Teshima and Kanazawa (1972). and their concentrations would be expected to vary in response to fluctuations in phytoplankton abundances and species distributions. 1979). (8) L i n e t aL (1982). so the enhanced abundance of lipids from terrigenous sources probably results from selective diagenetic removal of marine lipids (Volkman et al. . For reviews of sterols in green algae see Patterson (1971) and Holden and Patterson (1982). D--cholesta-5.g. hstationary phase.24(28)-dien-3/~-ol. Quantitative data for major constituents are shown in Table 2. In surface waters. which prevented positive identification of some minor sterols by GC-MS. Gagosian and Nigrelli. VOLKMAN Table 4. Gagosian and Nigrelli. 1983 and refs therein).3/~g/1. The relative importance of these two sources will depend on the type of environment. F--24-ethylcholesta5.2 and 0.24(28)-dien3/3-ol and 24-methylcholesta-5... 24-methylcholesta-5.. but others have suggested the possibility of a phytoplankton origin (e. (4) Patterson (1974)..24(28)E or Z-dien-3/~-ol. terrigenous organic matter are still poorly understood but the association of plant lipids with resistant biopolymers may enhance their survival compared with more readily decomposed algal cells. Gagosian et al. (1973). composition differs in exponential phase.22E-dien-3/~-ol. 1982).450.5-Unsaturated 4-desmethyl sterols in unicellular green algae (Chlorophyceae and Prasinophyceae) and cyanobacteria Sterol" A Chlorophyceae Dunaliella minuta ~ Coccomyxa elongata Trebouxia decolorans Chlorella ellipsoidea Chlorella saccharophila Chlorella pringsheimii Trebouxia sp. emainly AT-sterols.. (9) Volkman. Dunaliella primolecta Prasinophyceae Tetraselmis tetrathele Tetraselmis chui Tetraselmis sueica c Pyramimonas gelidicola Cyanobacteria Anacystis nidulans a Fremyella diplosiphon d Cvanidium caldarium Microcystis aeruginosa Anabaena cylindrica Spirulina platensis Spirulina sp. (7) Volkman. Cholest-5-en3/3-ol is usually the major sterol in seawater. 1979). Major sterols include cholest-5-en-3/3-ol. Gagosian and Heinzer.1-1. Gagosian et al. most of the sterols are associated with particulate organic matter (e. Cas Platymonas sueica. Black Sea (0.g. H--24-ethylcholesta-5. (1972).88 JOHN K. (1976). Meyers et al. (2) Patterson (1974). with minor contributions from phytoplankton (e. Gagosian. and it is thought to originate mainly from zooplankton lipids. for cyanobacteria see also De Souza and Nes (1968) and Martiniz Nadal (1971).22E-dien-3/~-ol.g. Saliot et al.5-2. Some authors have used it as a marker for terrigenous organic matter (e. Volkman et al. Sargasso Sea (0. 24-Ethylcholest-5-en-3/3-ol is often present in significant concentrations in seawater. (10). 24-ethylcholest-5-en-3/3-ol. 3 in Ref. Gagosian.22E-dien-3/3-ol (Table 2). aapproximate values calculated from Fig. There is no reason to suspect that this core section received significantly less marine organic matter than other sections in the core. 1984).3/~g/l) are comparable to values obtained for other seawater samples.. the proximity of the site to sources of land-derived organic matter and the abundance and type of phytoplankton present. 1976). E--24-methylcholesta-5.

1983. 20:5o~3 and 22:6~3. whereas others contain a high proportion of 18:1~ 9 (Piorreck et al. 1977). including 24-ethylcholest-5-en-3fl-ol. Other lipid classes such as fatty acids. Palmitoleic acid (16:leJ7) is a relatively minor component of the fatty acid distribution (Fig. 2) gives an indication as to likely sources of the organic matter and hence sterols. The phytoplankton in these oligotrophic waters are mainly small flagellates (Prasinophyceae and Prymnesiophyceae) and cyanobacteria (Hallegraeff. but 4-methyl sterols typical of these algae such as dinosterol are very minor constituents of the sterols in these seawater samples.. 155°W off the east coast of Australia. Partial capillary gas chromatogram of free fatty acids (methyl esters) in particulate matter from surface waters at 32°S. One complication in any comparison with the fatty acid compositions of the different phytoplankton groups is that some of the saturated fatty acids in the particulate matter.9 i 15:0 o 15:0 16:1 I I I I I I '12 16 20 Time 25 ( rain ) 30 35 40 Fig. the distribution is most similar to those found in Prymnesiophycean algae (Volkman et al.2 mm i. perhaps as much as 50% in this case. GC conditions: 20 m × 0. but it is usually the major fatty acid in diatoms indicating that these algae cannot be the major source of the fatty acids. are probably derived from phytoplankton since the sampling site is far from land. 1981.. It is of interest that fatty acids typical of bacteria such as vaccenic acid (18:1~ 7) and branched fatty acids comprise less than 3% of the total fatty acids. In the two samples studied here most of the sterols. whereas in areas of upwelling and high productivity diatoms are usually abundant. alcohols or hydrocarbons did not contain significant quantities of biomarkers indicative of higher plant lipids (unpublished data) precluding inputs from airborne terrigenous sources (cf. 2. With this exception. 100 to 320°C at 4°C/min. 1975). The nomenclature used is number of carbon atoms: number of double bonds. Similar distributions are also found in some dinoflagellates (Joseph. Unicellular green algae typically synthesize a high proportion of 18:1e~9. are associated with detritus and not living cells. 2). 18:2(o6 and 18:3co3 fatty acids but they usually also contain significant amounts of 16:4co3 which is barely detectable in the seawater samples. The distribution of free fatty acids in the surface waters (Fig. 18:5~o3.Sterol markers SP 11/83 surface total ( methyl fatty Station particulate acids ) esters 23 matter 89 16:0 18:0 14: ( 18:1 _ --/. 1981b). fused silica methyl silicone capillary column.d. particularly with respect to the presence of 18:4~3. H 2 carrier gas. It should be noted that in phytoplankton the fatty acid 18:5~3 has only been found in Prymnesiophycean algae and dinoflagellates. 1984). 1984 and refs therein). Cyanobacteria produce different types of fatty acids: some species contain 16:0 and 16:1o~7 as major components. Simoneit. The fatty acid data do not exclude a significant contribution from these algae. .

yeasts. benthic fauna and bacteria.90 DISCUSSION JOHN K. there is still a surprising diversity of sterol structures found in the sediments of the lake. fungi. the major sterol is 24-ethylcholest-5-en-3fl-ol which is usually thought of as a marker for terrigenous organic matter. and 3 minor ones. Although there is only one major phytoplankter in Ace Lake. to the extent that the distribution of lipids may bear little resemblance to that produced in the overlying water column. As new data are obtained it is apparent that many sterols are more widely distributed than had been thought and some earlier sterol identifications are now known to be in error. phytoplankton. For reasons of space. as seems to be the case in the deeper Peru and Corner Inlet sediments. These include for example. principally diatoms. (1979). (3) Baitantine et al. These examples highlight the need to integrate data from a variety of lipid classes with other chemical and biological information such as physical characteristics of the sediment. Even in this comparatively simple ecosystem. d _ Cricosphaera carterae: " = Co'stallolithus hyalinus. ~ as Monochrysis lutheri: ~as Pseudoisocho'sis paradoxa. lower plants.22E-dien-3[~-ol. . Although marine organisms would be expected to be the most important sources of organic carbon.' lsocho'sis sp. presence of specific phytoplankton species and carbon isotope data. These results highlight a major difficulty which may occur when attempts are made to reconstruct palaeoenvironments from lipid data (Didyk et al. so clearly there are important algal sources for this sterol. I have summarized much of the recent literature concerning the distribution of sterols in unicellular algae.. (4) Volkman et al. Each of the sediment samples studied in this paper represents a different depositional environment and yet the sterol distributions are remarkably similar. Furthermore. AS-Unsaturated 4-desmethyI sterols in Prymnesiophycean algae Sterol" B C26 6 1 18 23 73 D E F G H Others Reference 1 2 I I 3 2 4 4 2 2 5 1 6 1 4 4 Chrysochromulina po(vlepis Prvmnesium patellifi. In Tables 4-8. (1982). and some undoubtedly have several sources. (1984). If these lipids do not reflect the bulk of the organic matter deposited in the sediment. VOLKMAN the fatty acid. macroalgae. 1978). (1981b). Indeed.24-dimethylcholesta-5. (2) Ein et al. The similarity of the sterol distributions is even more surprising when one considers the great differences in species compositions in the different environments. vascular plants. it is highly unlikely that any geochemist given Table 5. which could make it difficult to assign origins to even some of the major sterols. protozoa. then any reconstruction of the palaeoenvironment is unlikely to be accurate. the tables In any sedimentary environment there are many potential sources of organic matter. Vascular plants do not contribute to the lipids in the Ace Lake sediments or the seawater samples. hydrocarbon and sterol distributions in the Peru sediment would have predicted that this sample came from an area of upwelling and high productivity. References cited: (I) Marlowe et al. I _ mainly 23. ra Ochrosphaera neapolitana Ochrosphaera verrucosa Pavlova lutheri h Pavlova lutheri Emiliania huxleyi Isochrvsis galbana lsocho'sis galbana lsochrysis sp. zooplankton. Cho'sotila lamellosa Cho'sotila lamellosa Cho'sotila stipitata l-lymenomomas carterae '~ Coccolithus pelagicus" 74 94 59 36 27 + 2 4 1 2 I 4 3 18 16 12 8 1 98 96 97 98 97 86 49 50 54 88 12 35 31 10 + 141 3t + 2 + 6 ~ + I0 44 47 33 12 13 t Notes: "sterols are given in Table 4. In the Corner Inlet and Peru sediments the major source of 24-ethylcholest-5-en-3fl-ol is vascular plants despite the fact that most of the organic matter in surface sediments from these areas is derived from marine sources. It is axiomatic that the lipids found in ancient sediments are those which best resist chemical and biological degradation. An assessment of sterol biomarkers for unicellular algae A major difficulty facing organic geochemists has been the small proportion of algal species which have been analysed for sterols using modern analytical techniques. This material can be produced in situ or brought in from distant areas by currents or from aeolian inputs. The composition of the organic matter is further affected by chemical and biological modifications. the taxonomy of many algal groups is in a state of flux and it is not uncommon to see some species transferred to completely different classes and species names changed several times. it is not possible to assign unequivocal origins for many of these sterols. (5) Berenberg and Patterson (1981): (6) Raederslorff and Rohmer (1984). In most oceanic and lacustrine environments the number of potential sources of sterols are likely to be orders of magnitude greater.

(1 I) Kates et al. (1980).. (12) Morris and Carre (1984). In this group.b). (1978). References cited: (1) Orcutt and Patterson (1975). and a cyanobacterium (De Souza and Nez. (1982). (6) Volkman et al. (3) Withers et al. '~mainly A 7 or A~191unsaturated sterols. although some dinoflagellates do not contain this sterol (Teshima et al. In a few cases the name of a species or its taxonomic classification has been amended according to lists published by Parke and Dixon (1976). (1980). (1978). (2) Alam et al. (1979a). Asterionella glacialis Biddulphia sinensis Cvclotella nana h Novicula pelliculosa Nitzschia closterium Phaeodactylum tricornutum Phaeodactylum tricornutum Stauroneis amphioxys Nitzschia ovalis Biddulphia aurita Chaetoceros sp.Thalossiosira pseudonana. (10) Berenberg and Patterson (1981). 89 67 60 42 40 25 25 I 2 2 + B 5 21 3 4 4 17 6 39 1 C 5 2 I 17 II 95 82 7 100 98 98 3 1 D E 6 4 54 F I 2 12 43 40 58 1 +' 6 G H 6 23 Others Re~rence I 2 3 1 4 3 5 1 6 7 8 1 I 91 I 59d 5 4 2 1 21 I1 2 4 2 + 17 + 33 4 1 33 91 91 79 54 21 12 + + 3 9a 2 18" 2 6 83 80 99 + 100 26 13 57 J 4 3 + 12 3 1 9 I I 5 I 10 1 10 I1 12 18 38 36 9 41 d 40 4 4 I 2 9 4 Notes: %terols are given in Table 4. tmainly 23. A 5'7 and A 7'22 sterols (Holden and Patterson.Sterol markers Table 6. (1981).fi'ustulum Chaetoceros simph'x Skeletonema costatum Thalassiosira [luviutilis Fragilaria sp. 1980. Withers (1983). (1981). A possible exception is dinosterol which appears to be a reliable biomarker for dinoflagellates. Data from some early studies have not been included because the identifications are suspect or have proved to be erroneous. 1982). . are restricted to the 8 most commonly encountered AS-unsaturated sterols found in sediments. Jones et al.[oliaeeum Gonyaulax diegensis' Pyroeystis lunula Gonyaulax polygramma Gymnodinium wih'zeki Glenodinium hallii Noetiluca milaris a Gymnodinium simplex Proroeentrum cordatum 100 1O0 80 39 76 36 26 8 B C D E F G H Others Reference I 2 3 4 5 6 7 8 9 10 I1 20 29" 6 1 39 50 1 2 9 35 I 5 I 7 42 73 53 5 6 63 14 47 25t 32 15 t 47 t 7 Notes: %terols are given in Table 4. References cited: (I) Alam et al. (1984). (7) Volkman et al. or even to define a typical sterol distribution for each group of algae. (9) Gillan et al. (1984). (1979. (5) Lin et aL (1982). (10) G o a d and Withers (1982). 24-methylcholest-5-en-3fl-ol and 24-ethylcholesta- Table 7. (I I) Robinson et al. (2) Nishimura and K o y a m a (1977). (4) Boutry and Barbier (1974). (1978.. but a few contain mainly A 5unsaturated sterols (Table 4). Thalassiosira pseudonana Thalassiosira pseudonana Amphora exigua Amphora sp. (5) Kokke et al.24-dimethyl substituted sterols. among the better studied algal classes some trends can be discerned: (i) many species of green algae (Chlorophyta) contain A 7. Wengrovitz et al. "isofucosterol. (4) Alam et al. Withers et al. (1979).. For other analyses of sterols see Swenson et al. 1982). 'Tucosterol. Species which do not contain significant amounts of AS-sterols such as many green algae (Holden and Patterson. (8) K a n a z a w a et al. ' + = < I%. (6) Volkman and Gagosian (1982)--unpublished results. has Gh. (9) Tesbima et al. 1981a). 'as Gonyaulax diagenesis. (1981). 1981a. Nonetheless. Goad and Withers. bCyclotella nana . (1971). (1980a). nodinium[bliaceum. ALUnsaturated 4-desmethyl sterols in diatoms (Bacillariophyceae) Sterol" A Nitzschia hmgissima Melosira granulata Thalassionema nitzschoides Nitzschia . ALUnsaturated 4-desmethyl sterots in dinoflagellates (Dinophyceae) Sterol ° A Gonyaulax spp. 1981a. There are still too few published sterol analyses to state that a particular sterol is characteristic of a given algal class. (1984). (1979). (7) Nichols et al. das Noctiluca milialis. Kokke et al. 1982). (1979b). 1968) have also been excluded. Peridinium foliaceum h Peridinium . Nitzschia alba Rhizoselenium spp. (8) Alam et al. 1982). (3) Ballantine et al. Kokke et al. some dinoflagellates (Kokke et al. (1983).

22E-dien-3fl-ol are found in several species. ALUnsaturated 4-desmethyl sterols in other unicellular algae Sterol" A Chrysophyceae Ochromonas danica Ochromonas malhamensis Ochromonas sociabilis Unidentified sp. (1983). Seckbach and Ikan. VOLKMAN Table 8. amainly ergosterol. Despite this. In most species. 1983 and refs therein). Moderate amounts of 24-ethylcholest-5en-3fl-ol and 24-ethylcholesta-5. 1975). (ii) Cyanobacteria (blue-green algae) have not been regarded as a major source of sterols in seawater or sediments but this view may need to be revised. 1972). (8) Beastall et al.22E-dien-3fl-ol predominates (Table 6).g.22E-dien-3fl-ol predominates (Table 5). (11) Raederstorff and Rohmer (1984). (9) Beastall et al. ~see also Cargile et al. emainly C~0 sterols. albeit in low concentration compared with many unicellular algae. (7) Brothers and Dickson (1980). sterol distributions in sediments . Some species of Gonyaulax contain large amounts of cholest-5-en-3fl-ol but there does not appear to be one distribution of 4-desmethyl sterols which characterizes these algae.24dimethyl substitution.9 9 10 I1 12 + 63 4 12 + + 6 25 + 62 2 Notes: °sterols are given in Table 4. 4-Methyl sterols have not been detected in either Prymnesiophycean algae or diatoms. although in more than half of those analyzed a single sterol represents over 80% of the total. (1971). Nematochrvsopsis roscoffensis Raphidophyceae Fibrocapsa japonica c B 5 C 9 D E 13 I 20 10 F 58 98 90 10 4 G H Others 15a 1 Reference I 1 2 3a 3b 4 4 4 2 1 17 10 22 13 1 I 6 28" 5Y 14 33 31 12 27 24 3 45 86 67 69 16 45 44 + 13 95 2 2 3 6 5 + 69 t 14 27 34d 38a 5 53: 38 3 5 5 6 7 8. 5. (iv) Dinoflagellates (Dinophyceae) are generally considered to be the major source of 4-methyl sterols in sediments (e. Three species of Tetraselmis contain 24-methylcholest-5en-fl-ol and 24-methylcholesta-5. and many species contain unusual sterols having A7 or A8 double bonds and unusual patterns of side-chain alkylation such as 23..22E-dien-3fl-ol usually predominate with moderate amounts of 24-ethylcholest-5-en-3fl-ol (Table 4).. but they are also potential contributors of 4-desmethyl sterols. (4) Mercer et al. 1968. (1974).24(28)-dien-3fl-ol but in Amphora species 24-ethylcholesta-5. (1980). Although earlier research had suggested that sterols are not present in cyanobacteria. 4-methyl sterols predominate but exceptions are known (e. (5) Brandt et al. References cited: (1) Gershengorn et al. In most species the major sterol is either 24-methylcholesta-5. 1980). (3b) Kokke et al.92 JOHN K .22E-dien- 3fl-ol. (1974). Complex mixtures of sterols are usually found (Table 7). Most research has concentrated on the Chlorophyceae and not other classes which is unfortunate since the Prasinophyceae can be important constituents of the phytoplankton in oceanic waters. (iii) Prymnesiophytes usually contain from 1 to 5 major sterols. /mainly A7sterols. Some species have been reported to contain large amounts of A7-unsaturated sterols (Orcutt and Patterson. (1970). Diatoms (Bacillariophyceae) contain a wider variety of sterols (Table 6). (6) Rohmer and Brandt (19733. but this has not been substantiated by more recent studies. but A7 and AS'7-sterols including ergosterol are also found (De Souza and Nez. De Leeuw et al. Cas FCRG51--chemotaxonomic data suggest a biochemical link to dinoflagellates. cholest-5-en-3fl-ol or 24-methylcholesta-5. and commonly cholesterol or 24-methylcholesta-5.g.24(28)-dien-3fl-ol as major sterols (Table 4). Generally. From the preceding discussion it is apparent that many sterols are widely distributed and few can be considered as characteristic of a particular algal class. (1975). one would not expect dinoflagellates to be a major source of 4-desmethyl sterols in a sediment where 4-methyl sterols are not abundant. (1968). Teshima et al. Xanthophyceae Botrydium granulatum Monodus subterraneus Tribonema aequale Euglenophyceae Euglena gracilis Astasia Ionga Bangiophyceae Goniotrichum elegans Porphyridium cruentum Porphyridium aeurigeurn h Cryptophyceae Crvptomonas sp. Most species seem to contain simple mixtures in which cholesterol and 24-ethylcholest-5-en-3fl-ol predominate (Table 4). (12) Nichols et aL (1983). (10) Goad et al. more recent studies have confirmed their presence. (3a) Rohmer et al. (1984). (2) Avivi et aL (1967).

1980a). 1979a. Cargile et al.. high concentrations of cholest-5-en-3/~-ol are generally attributed to zooplankton or other marine fauna since this is the major sterol of most marine animals. and in many species it is the major sterol present. It is usually absent from the Prymnesiophyceae and Xanthophyceae and a very minor constituent of a few cyanobacteria. (1981b) and Marlowe et al. and in almost half of the species examined to date it represents over 80% of the total sterols (Table 5). and a cyanobacterium Anabaena eylindrica (Table 4). 24-Methylcholesta-5. Prahl et al. It is not adequate to simply check for the presence or absence of a particular sterol. It is the major sterol (82%) in the common marine diatom Biddulphia sinens& (Volkman et al. the demonstration that copepods excrete significant amounts of cholest-5-en-3/t-ol (up to 4 ng/pellet. 1980b. unicellular red algae and dinoflagellates. Xanthophyceae and Euglenophyceae also contain large amounts (Table 8) and it appears to be a major sterol in most cyanobacteria although reliable quantitative data are lacking. In sediments. (1975) regarding the use of cholest-5-en-3//-ol as a "zoogenous" marker seems to be well founded. Studies by Volkman et al. a wide diversity of phytoplankton also contain cholest-5-en3[:~-ol (Tables 4 8). In marine environments. In C. Very few dinoflagellates do not contain cholest-5-en-3/~-ol and in many species it is the major 4-desmethyl sterol present (Table 7). It is the major sterol in 2 dinoflagellates Gymnodinium simplex and Noctiluca milialis. This is probably valid in areas of high productivity where diatoms predominate but this sterol is also found in many other algal groups. the latter associated with blooms known as red tides. The cholest-5-en-3/~-ol found must therefore be associated with the smaller size fractions which are mainly phytoplankton and cyanobacteria. The dominant flagellates off the East Australian coast are Prymnesiophytes and many of these are species of Chrysochromulina (Hallegraeff. (1984) established that 24-methylcholesta-5. Thus. 1983). 24-methyl- . Also.. In oligotrophic waters such as those studied in this paper. Limited data are available for the Cryptophyceae but in those species examined. in some environments they could be significant sources of cholest-5-en-3//-ol if those species rich in cholest-5-en-3//-ol happen to predominate.. 1983b). Small amounts of cholesta-5. Phytoplankton must be considered as potentially major sources of the cholest-5-en-3/~-ol present in the smaller size fractions ( < 100/xm) of marine particu- 93 late matter. polylepis. but these algae are not abundant in seawater. 1982). Volkman et al. significant amounts could be derived from zooplankton faeces and moults (Gagosian et al. In oligotrophic waters a significant amount of cholest-5-en-3/~-ol may be derived from these algae. Small amounts are also found in some Chrysophycean algae.. However.22Edien-3/3-ol than diatoms. 1984) indicates that zooplankton faecal pellets are a major source of cholest-5-en-3/t-ol in marine sediments.22E-dien-3/~-ol are found in most marine sediments and in seawater.22E-dien-3//-ol is the major sterol in unicellular red algae of the genus Porphyridium (Beastall et al. it samples containing high concentrations of dinoflagellate sterols it is possible that much of the cholest-5-en-3/~-ol is also derived from these algae.. Cholesta-5.Sterol markers and seawater can still be useful indicators of possible sources of organic matter provided that the distribution as a whole is studied. and they also contain significant amounts of ergosterol which has not been identified in marine sediments or seawater.. and it would be valuable to establish whether high concentrations of cholest-5-en-3/~-ol are typical of this genus. Diatoms therefore appear to be the most likely primary source for this sterol in phytoplankton samples. Notwithstanding this.22E-dien-3//-olhas often been used as a biomarker for diatom lipids since it is present in most species and often represents over 90% of the total sterols (Table 6). Nonetheless.. and yet this sterol is not common in unicellular alga.22E-dien-3/~-ol is also common in Prymnesiophycean algae. Cholest-5-en-3/~-ol is not common in the Chrysophyceae or Chlorophyceae. A few species from the Bangiophyceae. cholest-5-en-3/~-ol represents 74% of the total sterols (Table 5). Probable sources for selected AS-unsaturated 4-desmethyl sterols arc discussed below. b). 24-Methylcholest-5-en-3/t-ol is much less widely distributed than cholesterol and only rarely is it the major sterol in algae. and the caution expressed by Cargile et al. 1975). Significant amounts are found in a few diatoms and dinoflagellates and it is common in those green algae which do not biosynthesize predominantly AT-unsaturated sterols. In the seawater samples examined here most zooplankton were excluded using a prefilter although some juvenile stages may have been included. 1974.. Although high concentrations of cholest-5-en-3/~-ol are not typical of this group it is nonetheless the major sterol in several important marine species.. It is also a minor constituent of a few dinoflagellates (Table 7). A source of cholest-5-en-3/3-ol which has largely been ignored is Prymnesiophycean algae. It is also abundant in cultured zooxanthellae (dinoflagellates) from marine invertebrates (Withers et al. zooplankton are undoubtedly a major repository of cholest-5-en-3/~-ol in the marine environment and they convert much of the sterols produced by algae into cholest-5-en-3/~-ol. Prymnesiophytes are probably a more likely source of 24-methylcholesta-5. Cholest-5-en-3/~-ol is also present in many diatoms but most do not contain large amounts (Table 6). rather one should determine the most reasonable source of each sterol and examine whether this is consistenl with all information known about the sample and other lipid data. and it is present in at least 3 other diatoms (Table 6). High concentrations are found in Gonyaulax species which occasionally form massive blooms known as red tides (Alam et al.

but it seldom occurs in dinoflagellates.24(28)-dien-3/3-ol is the major sterol in only a few diatoms (Table 6) and it cannot be said to be a typical constituent of these algae.. A few Prymnesiophycean algae and some Chlorophycean algae contain moderate amounts of 24-ethylcholest-5-en-3fl-ol (Tables 4 and 5) and thus are potential sources of this sterol in cholesta-5. 24-Ethylcholesta-5.g.. unpublished data). (1981a) suggested that diatoms might be important whereas in Antarctic lake sediments green algae and cyanobacteria are thought to be major sources (this paper and Matsumoto et al. Green algae from the Prasinophyceae can be important constituents of the phytoplankton in oligotrophic waters (e.22E-dien-3/3-ol is the major sterol (Goad et al. 1972). 1983).. its place being taken by sterols with 23. 1983. and it is found in many green algae from the Chlorophyceae but not from other genera of the Chlorophyta. in a diatomaceous ooze from Walvis Bay. and a few plants contain (24/3)-24-methylcholesta.94 JOHN K. b).. For example.24(28)-dien-3/3-ol ( ~ 9 0 % .for plant inputs The m a j o r sterols found in higher plants (Goad and Goodwin.g. C29 sterols are not common in these algae (Table 6).22E-dien3/3-ol (stigmasterol) and (24~)-24-methylcholest-5en-3/3-ol (campesterol). Table 8)..3/3-ol (brassicasterol). Many Prymnesiophycean algae however do contain moderate amounts and this probably account for some of the 24-ethylcholesta-5.. inferences about sterol origins derived from studies of more typical algal sterol distributions are of limited value. . A5'24~28~-and A-~'25unsaturation also occur as minor components in some plants (Goad and Goodwin. However.24(28)-dien-3/3-ol is abundant in some sediments.22E-dien-3/3-ol is also common in marine sediments although only rarely does it represent more than 10% of the 4-desmethyl sterols. but too little is known about the sterols of these algae to generalize at this stage. Other researchers have speculated that unicellular algae could be significant sources of 24-ethylcholest5-en-3/3-ol in sediments (e. which accounts for the predominance of this sterol in seawater and sediment samples from this region. Diatoms are major components of the phytoplankton of this region and Thalassiosira eccentrica is often abundant. the presence of 24-ethylcholestane in ancient sediments and crude oils is not unambiguous evidence for organic matter derived from land plants. Lee et al..24(28)E isomer (fucosterol) is the major sterol of nearly all macroscopic brown algae (Phaeophyceae). Hallegraeff. 24-ethylcholest-5-en-3/3-ol was only 4% of the total 4-desmethyl sterols (Wardroper et al.24(28)-dien-3fl-ol is not common in other phytoplankton groups and thus in most samples it is probably derived from diatoms. Walvis Bay (Wardroper et al. However. VOLKMAN An assessment o f sterol biomarkers . Volkman et al. Small amounts of cholesterol are also found in many species.5. The C29 sterols 24-methylcholesta-5. 1981a. The major sterol of this species is 24-methylcholesta5. Hallegraeff. However 24-methytcholesta5. 1980.. 1983a). Volkman. With the possible exceptions of differences in C-24 stereochemistry. Small amounts of both isomers have been found in several diatoms and a moderate amount of isofucosterol occurs in the dinoflagellate Gonyaulax diegensis ( = diagenesis) (Alam et al. high concentrations have been found in two dinoflagellates and 2 species of Tetraselmis (Prasinophyceae). 1972) are (24ct)-24-ethylcholest-5-en3fl-ol (sitosterol). green algae (mainly Prasinophyceae) and cyanobacteria usually dominate the phytoplankton (e. The type of environment will have a major influence on the species of phytoplankton present. 1982. 1983a. but it has not been clear which phytoplankton species might be potential contributors. Significant amounts have also been found in a species of Cryptophyceae (Table 8). all of the common higher plant sterols are major constituents of the Ace Lake sediments and seawater samples and hence cannot be considered as unambiguous biomarkers for terrigenous organic matter. but more studies are required to establish whether this sterol is common in these species.22E-dien-3/3-ol found in oligotrophic waters. 24-Methylcholesta-5. Diatoms often dominate the phytoplankton in upwelling areas and thus they are potential contributors of 24-ethylcholest-5-en-3fl-ol in such environments. K. so we would expect to find proportionally more C27 and C2~ sterols. these species are rarely abundant in seawater. 1982). However. Gagosian et al. The 5. Note that this sterol is only a minor constituent of diatomaceous oozes from another area of upwelling. 24-Methylcholesta-5. In oligotrophic environments.g. 1978). All of these sterols are found in unicellular algae. 1981). particularly those from the Peru upwelling area (Gagosian et al. Sterols with Av-. 1978).22E-dien. and hence will determine the distribution of sterols incorporated into the sediment. 1972). This sterol also predominates in species of Ochromonas (Chrysophyceae. Prymnesiophycean algae. (24cQ-24-ethylcholesta-5. 1978). Smith et al.. whereas the 24(28)Z isomer (isofucosterol) is the major constituent of some macroscopic green algae (Charophyceae. Most C> sterols have the 24ct configuration but recent studies have demonstrated that the C2s sterols can occur as both 24~ and 24/3 epimers. In such cases.. 1983). Matsumoto et al.. Patterson.24-dimethyl side-chain substitution. J. It is a major sterol in a few diatoms. By extension. Lee et al. (1980) and Volkman et al. usually on the basis that terrigenous sources seemed unlikely.24(28)-dien3/3-ol can exist as either the 24(28)E or 24(28)Z isomers but neither is common in unicellular algae. and the usefulness of 24-ethylcholest-5-en-3/3-ol as a biomarker for vascular plants is seriously open to question particularly in samples where most of the organic matter is derived from marine organisms.

1982). oleanane and ursane skeletons (e. but as yet this sterol has not been identified in lacustrine sediments.. Thompson et al. Bohlin et al.g. two Prymnesiophycean algae and two Cryptophycean algae. have been shown to produce (24~)-24-methylcholesta-5. One possibility is to look at the ratios of the three sterols 24-methylcholest-5-en-3fl-ol. The predominance of C29 sterols in cyanobacterial mats (Boon et al. Further work on this aspect would clearly be a valuable aid to assigning the sources of sterols in sediments (Maxwell et al. Conversely. Several algal species. but long analysis times are involved.24-dimethyl. mid-chain alkyl ketones. in sediments where 24-ethylcholest-5-en-3/3-ol was less abundant than the other two sterols one would be reasonably confident that algal (not necessarily marine) sources of organic matter are important. CONCLUSIONS Studies of the sterol composition of sediments from . 1983a). and to date the method has not been exploited by organic geochemists. saturated even-chain C40-C60 wax esters (e. propylidine or cyclopropyl are present. 1974. 24ethytcholesta-5.g.Sterol markers seawater and sediments. Both ~ and fl isomers have been identified in an unidentified alga believed to be a Chrysophyte (Kokke et al. so the method is fallible.. 1982. The presence of organic matter from terrestrial plants can be confirmed from such biomarkers as long-chain alkanes showing a high proportion of odd chain-lengths. Maxwell et al.g.6: 6. whereas green algae and dinoflagellates biosynthesize sterols with the 24fl configuration (Goad et al. Goad and Withers. The presence of A-nor steroids and sterols with shortened side-chains in sponges (Bohlin et al. hydroxy acids and dicarboxylic acids characteristic of cutin and suberin. C . With few exceptions. 1982 and refs therein) may provide biomarkers for oxic marine conditions since only a few sponges are found in freshwater environments. In the Peru and Loch Clair sediments the ratios are I: 1. High concentrations of gorgosterol (22. 24-Ethylcholest-5-en-3fl-ol has not been identified in the Prasinophyceae although many species common in seawate~ have not yet been studied. it is important to recognize that different algal groups produce sterols with either 24~ or 24fl stereochemistry. 1983. 1980. 1978). 1979) and it remains to be shown whether they are also common in freshwater species. and coral reef sediments (unpublished data). 1984). Goad and Withers. Cranwell and Volkman. 1980).22-dien-3fl-ol (epibrassicasterol. polar glass capillary columns (Maxwell et al. Such sterols are not unique to marine organisms but appear to be mainly found in the marine environment. 1983).. 1984).. cyanobacteria have been shown to contain 24-ethylcholest-5-en-3fl-ol in high concentration relative to other sterols (Table 4)... since the two epimers are not separable on conventional (25 m or less) apolar capillary columns. Clearly there are many environments where high concentrations of 24-ethylcholest-5-en-3fl-ol could be derived from sources other than vascular plants. 1984). 1974. 1981. Many are derived from marine dinoflagellates (e.2% of 4-desmethyl sterols compared with 3. but not conclusive since the C-24 stereochemistry of 24-ethylcholest-5-en-3fl-ol in cyanobacteria has not been determined.0% for the 24/3 epimer. but even with this development we may still be unable to assign unequivocal origins for many sterols. (24~)-24-ethylcholest-5-en-3fl-ol comprised 10. with the latter being fairly typical of sediments where most of the sterols are derived from higher plants. It seems likely from biosynthetic considerations that any C29 sterols produced by these algae will also have the 24~ configuration although work by Raederstorff and Rohmer (1984) suggests that this may not always be true.24-dimethylcholest-5-en-3/3-ol) characterize some sediments from Walvis Bay (Wardroper et al. the C29 sterols of vascular plants generally have the 24~ configuration.. Distinguishing between sterols derived from vascular plants or marine phytoplankton is a difficult task as illustrated by the samples reported in this paper. Sterols differing in C-24 stereochemistry can be separated on very long.22E-dien-3/3-ol and 24-ethylcholest5-en-3/3-ol. Rubinstein and Goad.. Goad et al. certain triterpanoids based on friedelane. Similar values are found in some of the samples studied here. However.6. 1981).4:4. Brassell and Eglinton. 1980. as shown by the sterol data for the Peru and Corner Inlet coastal samples (Table 2). The presence of (24fl)-24-ethylcholest5-en-3fl-ol in sediments would probably be good evidence for lipid contributions from green algae. 27-nor. However.0 and 1: 1. indicates that cyanobacteria can be a major source of this sterol in such environments and perhaps in lacustrine and marine environments as well.2 4 stereochemistry In discussions on the sources of sterols in sediments little attention is usually given to the stereochemistry of the alkyl group at C-24.. Another possibility is to check whether sterols with unusual side-chains such as 23. including a diatom. 1981). fl-di-ketones and estolides amongst others. High concentration of C30 4-desmethyl sterols have also been found in two species of marine Cryptophyceae (Raederstorff and Rohmer..23-methylene-23.. Green algae are common in both freshwater and saline lakes and can be major sources of 24-ethylcholest-5-en-3/3-ol and other "higher plant" sterols. 95 A better approach is to compare inferences drawn from the sterol data with information derived from other lipid classes. but rarely both. Withers et al. 24-nor.. For example. Raederstorff and Rohmer. a predominance of 24-ethylcholest-5-en-3fl-ol does not necessarily indicate a lacustrine environment.

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