Org. Geochem. Vol. 18, No. 5, pp. 641--645, 1992 0146-6380/92 $5.00 + 0.

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Printed in Great Britain. All rights reserved Copyright 1992 Pergamon Press Ltd
High diahopane and neohopane abundances in a biodegraded crude
oil from the Barrow Sub-basin of Western Australia
CARIM ARMANIOS, ROBERT ALEXANDER and ROBERT I. KAGI
Centre for Petroleum and Environmental Organic Geochemistry, Curtin University of Technology,
G.P.O. Box U1987, Perth 6001, Western Australia
Abstract--Three crude oils from the Barrow Sub-basin of Western Australia have been analysed for
hopanoid biomarkers using GC-MS techniques. Enhanced abundances of nordiahopane and diahopane
and norneohopane relative to hopanes have been observed in the most severely biodegraded sample. These
differences have been attributed to the greater resistance of the diahopanes and norneohopane to microbial
degradation.
Key words- - hopanes, diahopane, norneohopanes, microbial degradation, Barrow Sub-basin, petroleum
biomarkers
INTRODUCTION
Biodegradation of crude oil involves the progressive
utilization of specific classes of hydrocarbons by
bacteria (of. Connan e t al . , 1980). Where the tempera-
ture of the reservoir is less t han 75C bacteria can
survive and function (Philippi, 1977). The reservoir
must also be in cont act with meteoric waters since this
is believed t o be the maj or mechanism by which
bacteria gain access t o the reservoir (Bailey e t al . ,
1973).
Biodegradation of a crude oil leads t o a sequential
removal of n-alkanes, isoprenoid and other branched
alkanes, and eventually some cyclic alkanes and
aromat i cs as conditions for microbial oxidation be-
come increasingly favourable (Reed, 1977; Alexander
e t al . , 1983; Vol kman e t al . , 1983a, 1984; Peters and
Mol dowan, 1991). Clearly, an understanding of the
conditions under which the common biomarkers are
altered is essential for reliable application of bio-
markers t o problems involving the correlation of
biodegraded crude oils (cf. Vol kman e t a / . , 1983a;
Peters and Mol dowan, 1991).
The hopanoi ds (Fig. 1) are pentacyclic alkanes
which are f ound widely in sedimentary rocks and
petroleum. This class of biomarker, al t hough quite
resistant t o bacterial attack, is eventually degraded
under optimal reservoir conditions (Volkman e t al . ,
1983a). It has been suggested t hat the thermally stable
17~t(H),21fl(H)-hopanes (Fig. l, I - VI I I ) in certain
circumstances can undergo a bacterially mediated
demethylation at C-10 t o form the 17~t(H),21fl(H)-
25-norhopane series (Fig. l, I X- XVI ) (Volkman
e t al . , 1983a; Peters and Mol dowan, 1991). Al t hough
this process has not been replicated so far under
l aborat ory conditions, the isolation of a hopanoi d
from recent sediments containing a carboxyl group
at C-10 (Albrecht, 1990) indicates t hat oxidation at
this position can occur under natural conditions.
Such processes, along with subsequent decarboxyla-
tion, could therefore provide a viable route t o the
17~, (H)-25-norhopanes.
Recently, a new series of C 2 9 - - - C 3 3 hopanoi ds named
diahopanes (Fig. 1, XI X- XXI I I ) has been identified
in crude oil (Mol dowan e t al . , 1991). The di ahopane
carbon skeleton differs from the hopane skeleton in
t hat it has a methyl group on C-15 instead of C-14.
This paper is concerned with the effect of biodegrada-
tion on the diahopanes and the recently identified
norneohopane (Mol dowan e t al . , 1991) (Fig. 1,
XVI I I ) in three crude oils from the Barrow Sub-
basin, Western Australia.
EXPERIMENTAL
Cr ude oi l s
Geological dat a for each of the Barrow Sub-basin
oils are given in Table 1. Detailed accounts of the
petroleum geology of the Barrow Sub-basin can be
found in papers by Crank (1973), Thomas and Smith
(1974) and Thomas (1978). Detailed studies on the
geochemistry of the crude oils have also been re-
port ed (Volkman e t al . , 1983a). In brief, the Barrow
sample is a paratfinic light crude oil which shows no
evidence of biodegradation. The Flinders Shoal and
Mardie samples are naphthenic crude oils with com-
positions consistent with their having been subjected
t o severe biodegradation and water washing. The
Di ngo Claystone of Jurassic age is the putative source
of these crude oils (Volkman e t al . , 1983b).
I s ol at i on o f br anc he d and cycl i c a l k a n e s f r o m crude oi l
The branched and cyclic alkanes were isolated
from crude oil samples using previously reported
methods (Volkman e t al . , 1983a). Briefly, the alkanes
641
642
CARIM ARMANIOS e t a l .
~
" , , U l
- XVI
17a(H),21p(H)-hopanes
I RI=H R2=CH3
II RI-C2H5 R2-CH3
Ill RI-C3H8 R2=CH3
IV RI=C4HI0 R2=CH3
V RI-C5H12 R2-CH3
VI RI-C6HI4 R2=CH3
VII RI-CTH16 R2-CH3
VIII RI-CSH18 R2-CH3
17a(H),21~(H)-norhopanes
IX RI=H R2-H
X RI=C2H5 R2=H
XI RI-C3H8 R2-H
XII RI-C4HI0 R2-H
XIII RI=C5HI2 R2-H
XIV RI=C6HI4 R2-H
XV RI=C7HI6 R2=H
XVI RI-C8HI8 R2-H
18a(H),21~(a)-neohopanes
XVII R-H
XVIII R-C2H5
I I I l l l l l l
R
I - l ~ I I I
17~( H ) - d l a h o p a n e s
. I ~ r " , , , , x x . - ~ 3 . 8
XXI R'C4HI0
XXII R'C5H12
- XXIII R=C6HI4
X - XXIII
Fig. 1. Hopanoi ds in petroleum.
w e r e s e p a r a t e d u s i n g c o l u m n c h r o m a t o g r a p h y w i t h
s i l i c i c a c i d . T r e a t m e n t w i t h m o l e c u l a r s i e v e s r e m o v e d
t h e n - a l k a n e s , l e a v i n g a f r a c t i o n e n r i c h e d i n b r a n c h e d
a n d c y c l i c a l k a n e s w h i c h w a s f u r t h e r a n a l y s e d u s i n g
G C - M S t e c h n i q u e s .
GC analysis of alkanes
The analysis of t he t ot al al kanes by gas chr omat og-
r aphy was carri ed out using a Hewl et t Packar d 5880A
gas chr omat ogr aph equi pped with a 50 m x 0.22 mm
Tabl e 1. Geochemi cal dat a pert ai ni ng t o the crude oils from the Barrow
Sub-basin (Vol kman e t a l . , 1983b)
Reservoir Reservoi r
Petroleum Type Reservoir dept h (at) age
Barrow Paraflinic Muderong 2010 Lat e
oil Shale Jurassic
Fl i nders Napht heni c Bi rdrong 700 Apt i an
Shoal oil Sands
Mardi e Napht heni c Mardi e 77 Apt i an
oil Greensand
Hopanoid biomarkers in Barrow Sub-basin crude oil 643
Table 2. Hopanoid nomenclature and abbreviations (for structures
refer to Fig. 1)
Abbreviation Structure Name
C29DIA XX 14e (H),I 7a (H),21 ~(H)-nordiahopane
C29D XI 17~ (H),21 ~(H)-25-norhopane
C~ II 17" (H),21 ~ (H)-norhopane
C29Ts XVIII 18,, (H),21,g(H)-norneohopane
C~DIA XX 14~ (H), 17,,, (H),21 ,g (H)-diahopane
C30 III 17~ (H),21fl(H)-hopane
i.d. fused silica col umn coated with BP-5 stationary
phase (SGE Australia) and an OCI-3 on-col umn
injector (SGE Australia). Hydr ogen was used as the
carrier gas at a linear flow velocity of 30 cm/s. The
oven was programmed from 70 to 300C at 4C/rain.
G C - M S analysis o f branched and cyclic alkanes
The analysis of branched and cyclic alkanes by gas
chr omat ogr aphy- mass spectrometry was carried out
using a Hewlctt Packard 5970 MSD equipped with
the RTE/ A dat a system. The GCMS was fitted with
a 60 m x 0.2 mm i.d. DB-5 (J&W) column. The oven
temperature was programmed from 50 t o 274C at
8C/rain, then t o 300C at 1 C/rain. The samples were
injected on-col umn using a HP7673A autosampler
(HP) and analysed in SIM mode.
RESULTS AND DISCUSSION
Gas chromat ograms of the total alkanes from
the three related crude oils are shown in Fig. 2, and T
are similar t o those previously published (Volkman
et al., 1983a). The mass chr omat ogr am of the unde-
graded Barrow sample shows a predominance of the
/
t t ~
n-alkanes, while those of the biodegraded crude oils [
lack n-alkanes and comprise unresolved complex / !
mixtures of branched and cyclic alkanes.
f
Fig. 3 shows partial m/ z 191 mass chromat ograms
of the crude oils. The diahopanes and C29Ts were
assigned on the basis of their published retention
times relative t o the 17~(H)-hopanes (Mol dowan
et al., 1991). These assignments were support ed by
the mass spectra in so far as all compounds assigned
as diahopanes displayed promi nent m/ z 191 and
parent ion responses. The extended diahopanes
were present in concentrations t oo low t o enable
measurements t o be made.
The partial m/ z 191 mass chr omat ogr am of the
Flinders Shoal sample, shown in Fig. 3, is similar t o
t hat of the Barrow sample, except for a small relative
increase in the peak labelled C29D. The presence of a
low concent rat i on of this compound in Flinders Shoal
crude oil has previously been interpreted as resulting
from a mixing of two crudes, one a heavily degraded
sample with some 25-norhopanes and the other a
partly degraded oil (Alexander et al., 1983; Vol kman
et al., 1983a). The remaining peaks in the Flinders
Shoal sample indicate t hat the 17~(H)-hopanes (C~,
C30), the norneohopane (C29Ts) and the two dia-
hopanes (C29DIA and C30DIA) occur in similar pro-
port i ons to those f ound in the Barrow crude oil.
The partial m/ z 191 mass chr omat ogr am from the
Mardie sample shows a domi nant peak for 25-
norhopane (C29D) and the peaks for nor hopane (C29)
and hopane (C3o). Interestingly, the diahopanes
(C29DIA, C~JDIA) and the norneohopane (C29Ts)
have a similar distribution t o those in the less de-
graded samples. This indicates t hat the diahopanes
and norneohopanes are more resistant to biodegrada-
tion t han the 17~(H)-hopanes.
Evidence t hat significant depletion of diahopanes
and norneohopane also occurs was provided by
compari ng the relative abundances of hopane and
norneohopane in the undegraded Barrow sample
with those of nor hopane and norneohopane in the
Mardie sample. I f hopane had been converted t o the
demethylated 25-norhopane without alteration of
norneohopane, then the peak labelled C29D in the
mass chr omat ogr am from Mardie should be only
about one hal f t hat labelled C30 in the Barrow mass
Barrow UJ
2O
i t
I. Fllnders Shoal
L
RETENTION TIME
Fig. 2. Capillary gas chromatograms of the alkanes isolated
from the crude oils. C2o n-alkane is indicated by (20), the
isoprenoid alkanes pristane and phytane by (#), the Ci 4~6
bicyclanes by (*), and the 25-norhopane series by (^).
644 CARIM AglvL~baOS et al.
chromatogram, since the 191 fragment is derived only
from the D/ E ring moiety of C29D while the peak
labelled C30 has a 191 contribution from both the A/B
and D/E ring moieties. Clearly the response from
C29D, shown in Fig. 3, is much stronger (compared
with C29Ts) than predicted, suggesting that nor-
neohopane and the diahopanes are degraded but at
a slower rate than the 17n(H)-hopanes.
An attempt was made to identify degradation
products from C29Ts and the diahopanes assuming
that they had undergone similar degradation pro-
cesses to the 17~(H)-hopanes. However, a detailed
analysis of the Mardie crude oil provided no evidence
of 25-nordiahopanes nor of a 25,30-bisnorneoho-
pane. It is possible that these rearranged hopanes,
like the steranes, are degraded to non-hydrocarbon
products.
Although the evidence strongly supports bio-
degradation effects as the most likely cause of the
elevated relative abundances of 25-norhopanes, nor-
neohopane and diahopanes in the Mardie sample,
an alternative explanation, based on differences in
thermal stability of hopanoids can be proposed.
Moldowan et al. (1991) reported that the rearranged
hopanes are more stable than the l%t(H)-hopanes
due to relief of steric strain when the methyl group at
Ci4 is relocated to Cl5 (diahopanes), or when the
methyl group Ct8 is relocated to C~7 (neohopanes).
Using similar reasoning it is also probable that
removal of the methyl group at C,0 would enhance
the stability of the 25-norhopanes in comparison with
the 17~(H)-hopanes. Hence, a crude oil subjected to
extreme thermal conditions might eventually become
relatively enriched in 25-norhopanes and rearranged
hopanoids.
Maturity assessment of severely degraded crude
oils such as the Mardie sample is difficult because
many of the compounds used as maturity indicators
are either absent or severely affected by biodegrada-
tion. Alkylnaphthalenes and alkylphenanthrenes in
Mardie are either of very low abundance or have been
significantly altered by biodegradation rendering
them unreliable for maturity ratios. Volkman et al.
(1984) however, reported that the relative proport-
ions of short chain (C2~, C22) and long chain (C29)
monoaromatic steranes were similar in all three
crude oils and typical of mature source rocks. These
results suggest to us that the Mardie crude oil has
l
m / Z 191
C30
C29
~ c 2 9 r , f ' l
c29 hI^ [~A %o hI^ ~IL
C29D.
1
L
I
I
I
Fl/nders S ~ a l
Mardie
w
R E T E N T I O N TIME
Fig. 3. Partial m/ z 191 mass chromatograms showing comparison of relative distributions of diahopanes
to hopanes in selected crude oils from the Carnarvon Basin, (refer to Table 2 for hopanoid nomenclature).
Hopanoid biomarkers in Barrow Sub-basin crude oil 645
not been subj ect ed t o unus ua l l y severe t her mal con-
di t i ons sufficient to cause t he obser ved hopa noi d
di st r i but i on.
CONCLUSION
The rel at i ve a bunda nc e s of di ahopanes a nd 30-
nor ne ohopa ne rel at i ve t o t he 17g( H) - hopanes have
been s hown to be enhanced i n a severely bi odegr aded
cr ude oil f r om t he Bar r ow Sub- basi n. The effect has
been pr e domi na nt l y at t r i but ed to t he sl ower rat e of
mi cr obi al degr adat i on of t he nor ne ohopa ne a nd
di ahopanes compar ed wi t h t he 17~t(H)-hopanes.
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