Org. Geochem. Vol. 8, No. 4, pp. 293-297, 1985 0146-6380/85 $3.00 + 0.

00
Printed in Great Britain. All rights reserved Copyright 1985 Pergamon Press Ltd
Petroleum isoprenoid hydrocarbons derived from catagenetic
degradation of Archaebacterial lipids
J. ALBAIGI~S 1, J. BORBON l and W. WALKER II 2
llnstitute of Bio-Organic Chemistry (CSIC), Jorge Girona Salgado, 18-26, Barcelona-34, Spain.
2Bodega Marine Laboratory, University of California, Berkeley, CA 94720, U.S.A.
( Recei ved 24 Apr i l 1984; accept ed 5 Mar ch 1985)
Abstract--The three GC coeluting C40 bis-phytanes with head-to-tail, tail-to-tail and head-to-head
linkages considered to be archaebacterial markers, have been recognized in petroleum by enhanced
mass-fragrnentography. Supporting evidence has been obtained by the simultaneous occurrence of two
C2~-C24 series of isoprenoid and quasi-isoprenoid hydrocarbons that are supposedly formed during the
catagenetic degradation of the isomeric C40 isoprenoids. The C2~-C24 quasi-isoprenoid hydrocarbons have
been identified conclusively for the first time by comparison with authentic standards.
Key words: archaebacterial lipids, acyclic isoprenoids, synthesis of isoprenoids, mass spectra, petroleum
oils
I N T R O D U C T I O N
Acycl i c i sopr enoi ds const i t ut e a r at her compl ex
fami l y of bi ol ogi cal i ndi cat or s in pet rol eum. Even
t hough t here is good evidence t hat t he pl ankt oni c
chl or ophyl l a is the pr edomi nant source for the
r egul ar i sopr enoi ds bel ow C20 ( Di dyk e t al . , 1978), t he
recent di scovery in sedi ment s and pet r ol eums of
di phyt anyl gl ycerol ethers ( Chappe e t al . , 1982),
which are t he mai n component s of the membr ane
lipids in Ar chaebact er i a, suggests t hat a subst ant i al
cont r i but i on from these or gani sms can also be ex-
pect ed. Thi s possi bi l i t y has been st rengt hened by the
concur r ent i dent i fi cat i on of ot her lipids uni que to
Ar chaebact er i a such as di gl ycerol t et raet hers, con-
t ai ni ng two C40 head- t o- head i sopr enoi d chains (Fi g.
1, I and IV; R ~ OX, where X = hydr ophi l i c gr oup)
( Tor nabene and Langwor t hy, 1979; De Rosa e t al . ,
1977).
A series of st ruct ural l y rel at ed l ong chai n iso-
prenoi ds ( I - I V; R = H) have also been i sol at ed from
geol ogi cal sources ( Ki mbl e e t al . , 1974; Albaig6s e t
al . , 1978; Dast i l l ung and Corbet , 1978; Mol dowan
and Seifert, 1979; Albaig+s, 1980; Chappe e t al . , 1980;
Brassell e t al . , 1981; McEvoy e t al . , 1981). However,
the precise origin of some of these hydr ocar bons
( I I - I I I ) and t hei r fate in t he geosphere are still poor l y
under st ood.
We r epor t in this paper the i dent i fi cat i on of C2~ C24
quasi - i sopr enoi d al kanes ( 1 ' ~/ ) in pet rol eum. The
v C z 3 ( 5 ' )
I ~ C z l ( 1 ' )
I I
R R
C z 3 ( 3 I - 7 [
, C z l ( 1 ) m
I I I I
I I I I
I t ~
I " - C 2 2 ( 2 ' )
L Cz 4 ( 4 , ) I I
C 2 4 ( 4 ) / I l l
IV
Fig. 1. Structures of C,~ isoprenoid alkanes (R-~-H) identified in Amposta crude oil (Miocene, Spain).
Dashed lines indicate lower homologs conclusively identified in this paper.
293
294 J. ALBAIG[S et al.
simultaneous occurrence of these compounds with
the C2~-C24 (1~1) isoprenoid series provides evidence
of the catagenetic degradat i on of three C40 isomeric
isoprenoid hydrocarbons ( I - I I I ) considered to be
bacterial or Archaebacterial markers. These markers
which coelute in gas chr omat ogr aphy were also
identified in the same petroleum samples by enhanced
mass-fragmentography.
EXPERI MENTAL
S a mp l e p r e p a r a t i o n a n d anal y s i s
The crude oils belong t o the off-shore fields Am-
post a and Tarraco, in the Western Mediterranean,
and are reservoired respectively in Lower Cretaceous
and Upper Jurassic limestones. They have been
recognized to be dissimilar (Albaigrs and Torradas,
1977) al t hough the actual source of the Ampost a oil
remains unpublished. However, the possible source
rocks in the area (Middle Miocene and Mesozoic)
consist in carbonat es generated in a restricted marine
environment, which provide favourable conditions
for the preservation of biological markers.
Branched-alkane fractions of these crudes were
obt ai ned in the usual way: silica-gel col umn chro-
mat ography, urea and thiourea adduct i on of the
saturated fractions and alumina col umn chro-
mat ogr aphy of the latter adduct (Albaigrs, 1980).
Gas chromat ographi c analysis of extracts and ref-
erence compounds was carried out in a Carlo Erba
FW 4160 GC instrument equipped with glass capil-
lary columns (25 m x 0.25 mm. i.d.) coated with SE-
52 (Bayona and Albaigrs, 1982).
The COM- GC- MS system comprised a GC
Fi nni gan 9610, equipped with a 30 m x 0.3 mm i.d.
DB-5 fused silica col umn (J&W Scientific), linked to
a Finnigan 4023 MS with an I NCOS dat a system.
Operating conditions are given elsewhere (Bayona e t
al . , 1983; Albaigrs e t al . , 1983).
Pr e p a r a t i o n o f r e f e r e nc e c ompounds
The regular C24 isoprenoid (4) was produced by
hydrogenat i on (PtO2) of the allylic alcohol obtained
in a Gri gnard reaction between phyt al dehyde and
butyl bromide, in a manner similar to t hat described
previously for other homol ogs in the series (Albaigrs
e t al . , 1978) (yield 53%; RIAp i .... L 2145; MS in
Fig. 4).
The quasi-isoprenoid hydrocarbons C2~ (1'), C22
(_2') and C24 (4') were synthesized following the
scheme outlined-in Fig. 3, basically a coupling (via
Wittig reactions) of the adequate syntons, a series of
methyl ketones, using the trimethylene-bis-
triphenylphosphonium-ylide (Friedich and Henning,
1959) (m.p. 334-336C). 6-Methyloct-5-en-2-one
(E + Z) was produced in 78% yield by a Carroll
sequence (Mori e t al . , 1969) from
3-methylpenten-3-ol; homol ogat i on of this ketone by
the ylide of (4,4-ethylenedioxy)-pentyltriphenyl-
phosphoni um iodide (m.p. 217-218C) (Cavill e t aL,
1969) rendered 6,10-dimethyldodeca-5,9-dien-2-one
(51% yield). Pentan-2-one and heptan-2-one were
commercially available (Fluka, A.G.). Product s from
the Wittig reactions, namely the alkene C21H36 and
two alkene mixtures, in the case of C22 and C24
hydrocarbons, were hydrogenat ed with PtO2 as the
catalyst. After purification by silica-gel chro-
mat ogr aphy (eluent: n-pentane), the C2~ isoprenoid
(_1') was obtained in pure form (23% yield with respect
t o the correspondi ng ketone; RIAp ~ .... L 1982),
whereas the C22 (_2') and C24 (4') isoprenoids were
obtained in mixtures of hydrocarbons with abun-
dance of 50 and 54% respectively, when the high and
low molecular weight ketones were reacted in a 2:1
ratio (absolute yield 15-18% and RIAp i .... L 1978 and
2156, respectively).
RESULTS AND DISCUSSION
The C40 b i s - p h y t a n e (I) and C27-C39 isoprenoid
hydrocarbons resulting from its catagenetic de-
gradat i on have been identified in several petroleums
(Mol dowan and Seifert, 1979; Albaigrs, 1980). How-
ever, in examining the branched-alkane fractions
previously isolated from the Spanish Ampost a and
Tarraco crude oils (Albaigrs, 1980) we realized t hat
the chromat ographi c peak correspondi ng t o b i s -
phyt ane (I) was composed of more t han one com-
pound (Albaigrs e t al . , 1981). Using high resolution
gas chr omat ogr aphy- mass spectrometry and passing
the recorded signal t hrough smoot hi ng and peak
enhancement, we proved the presence of three mai n
component s (Fig. 2A). These were identified as the
head-to-head b i s - p h y t a n e (I, R= H) , the regular
C40-isoprenoid (II) and the tail-to-tail C40-isoprenoid
(lycopane, III) by reference to the retention times and
mass spectra of st andard compounds (Chappe e t al . ,
1980; Albaigrs e t al . , 1978; Kimble e t al . , 1974). In
addition, at slightly higher retention times we ob-
served two other peaks (IV and V; Fig. 2B), with very
similar spectra containing fragment ions at m / z 165,
166, 167, 194 and 195, which are characteristic of C40
saturated isoprenoids with one pentacyclic ring
(Chappe e t al . , 1980). The mass spectrum of the first
of these compounds appeared to be identical to t hat
reported for the hydr ocar bon IV ( R=H) , produced
during the degradat i on of kerogen (Chappe e t al . ,
1980). The structural relationship of this hydro-
carbon (IV) to the isoprenoid moieties of glycerol
tetraethers found in Archaebact eri a (De Rosa e t al . ,
1977) provides, as in the case of b i s - p h y t a n e (I;
R= H) , additional evidence of the cont ri but i on of
these micro-organisms to the format i on of petroleum.
The biological sources of the regular (240 isoprenoid
(II) and of lycopane (III) are unclear. They could be
formed by diagenetic reduction of polyprenols and
carotenoids (e.g. lycopene or spirilloxanthin) re-
spectively, which is compatible with the processes
occurring during the format i on of petroleum. But,
as their structure resembles the widely distributed
Petroleum isoprenoid hydrocarbons from archaebacterial lipids 295
II
_ _ _ 9 1
~ V ~
4 2 0 0
7 0 : 0 0
IV
V
i
~ . ~ - - , , , . . . - - , , . ( B )
- - , , , . - ' i - A , , ",'~',
RIC
. . . . I . . . . I ' ' ' ' i
4 2 5 0 4 3 0 0 4 3 5 0 4 4 0 0 4 4 5 0
7 0 : 5 0 7 1 : 4 0 7 2 : 3 0 7 3 : 2 0 7 4 : 1 0
Fig. 2. Enhanced (A) and partially reconstructed (B) (~m/z 166, 167, 194, 195) gas chromatograms of
an isoprenoid fraction of Amposta crude oil. (Column: fused silica DB-5, 30 m (J&W); temperature:
60-300C at 2C/min; Finnigan Incos data system). Peaks are identified in Fig. 1.
( A )
4 5 0 o 4 5 5 0 Sc a n
7 5 : 0 0 7 5 : 5 0 T i m e
0 0
+ OE t =
OH
I . - - ( ~ 3 P C H 2 C H 2
0 2 . - - H 2 / P t O 2
1 . - I - [ ~ L - ~ 0 / D M S O -
+ p o 3 O Y
2 . - - p - - T s O H
5- 5-
i . ( ~ 3 P C H 2 C H 2 C H 2 P ~ 3 ) 2 B r - / D M S O -
4,- C21H44(1' )
1. ( ~ 3 P C H z C H 2 C H z P O a ) 2 B r - / D M S O - , , 2. H z / P I " O 2
l
C 2 2 H 4 6 ( 2 ' ) C 2 4 H 5 0 ( 4 ' )
Fig. 3. Synthetic pathways for isoprenoids C21H44 (1'), C22H46 (2') and C 2 4 H 5 0 (4').
296 J. ALBAIG~S et al.
_I
F x 2 1 '
2 6 7 2 9 6
2 '
1 9 7 211
1 2 7 141 2 6 7 281 3 1 0
4 '
1 9 7 2 3 9
I 127 1 6 9 I , 2 6 7 = ~ 9
~ ~ o u = 3 3 8
113
I / [ 1 8 a
253 323 338
1 0 0 2 0 0 3 0 0
Fig. 4. Mass spectra of isoprenoid hydrocarbons synthesized and identified in petroleum. Numbers refer
to Fig. 1.
Archaebacterial isoprenoids previously described, a
more direct biological origin is also conceivable.
Lycopane has been identified in several geological
settings (Dastillung and Corbet, 1978; McEvoy e t a L,
1981; Brassell e t al . , 1981); and in the recent sedi-
ments where it has been reported, methanogenic
activity has also been document ed (Brassell e t a L,
1981). Moreover, Ourisson e t aL (1982) have
suggested the existence of different families of
polyterpenoid compounds in the membranes of
microorganisms yet to be identified.
On the ot her hand, petroleum is formed at a depth
where thermal degradat i on of organic matter be-
comes i mport ant (Tissot and WeRe, 1978). Hence,
degradat i on product s of the parent hydrocarbons
would be expected to occur. I n light of this, we have
focused our attention on the hydrocarbons 1-4
and 1' - 4' (Fig. 1). The regular series C2~-C23 (1-3)
has been reported in crude oils (Han and Calvin,
1969; Albaigrs and Torradas, 1977) and in shale
(Spyckerelle e t al . , 1972), and has been attributed t o
the catagenetic degradat i on of regular isoprenoid
hydrocarbons (Albaigrs e t al . , 1978). However, the
format i on of the C22 member of this series by this
process in highly unlikely, requiring simultaneous
cleavage of two geminal C- C bonds. The presence of
the C22 member (2) in ancient sediments has been
considered to indicate a cont ri but i on from lycopane
(Han and Calvin, 1969). The presence of the C24
homol og (4) would support this hypothesis; we were
able to identify a constituent with mass spectra and
retention time correspondi ng to an authentic stan-
dard.
Other standards, the quasi-isoprenoid hydro-
carbons C21H44 ( l ' ) , C22H46 (2' ) and C24H50 (4'), were
synthesized according t o the scheme outlined in Fig.
3 and then analyzed by GC- MS and high resolution
gas chromat ography.
The mass spectra (see Fig. 4) and retention indices
on two different columns (Apiezon L and DB-5) were
found to be identical to those of the correspondi ng
peaks in alkane fractions isolated from the Spanish
crude oils.
The C23 member of the series (3'), recognized on the
basis of the mass spectrometric data, was found only
in mi nor quantities. This fact strongly suggests t hat
the origin of these C21-C24 quasi-isoprenoid hydro-
carbons is from catagenetic degradation of the regu-
lar isoprenoid II, al t hough their format i on from a
parent head-to-head isoprenoid (1) cannot be com-
pletely ruled out. This implies t hat there is no func-
tional group in the original molecule that may induce
a preferential cleavage at one end, as is the case of
liberation of phytol from kerogen. Thus, we favor the
idea t hat the regular isoprenoid II is of direct biolog-
ical origin, as is probabl y true with lycopane (III).
The format i on of quasi-isoprenoid aikanes not
significantly different from the normal isoprenoid
alkanes was also predicted by mathematical models
(Waples and Tornheim, 1978) which assumed t hat
Petroleum isoprenoid hydrocarbons from archaebacterial lipids 297
cr acki ng of a pr e- exi st i ng i s opr enoi d hydr oc a r bon
occur s pr ef er ent i al l y at t he mor e hi ghl y subst i t ut ed
c a r bon at oms , and t er mi nal cl eavage yi el di ng met h-
ane t akes pl ace less f r equent l y.
Acknowl edgement --Thi s work was partially supported by
the Spanish-North American Cooperative Project No.
793028.
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