1

USE OF BIOMARKERS AS A PROXY FOR SOURCE ROCK-OIL

CORRELATION AND PALEOCLIMATIC CHANGES IN THE
KUNDAL OIL FIELD, MIANWALI PAKISTAN
SULEMAN ORAKZAI
SUPERVISED BY: DR SAMINA SIDDIQUE
 2
Presentation format

 Introduction
 Study Area
 Aims & Objectives
 Significance
 Methodology
 Results
 Discussion & Conclusion
 3
Introduction

 Crude oil and source rock include biomarkers that are produced by the breakdown of lipids,
nitrogenous chemicals, algal C3/C4 type of vegetation, and marine creatures. Leaf-wax lipids
biomarkers accumulated in the thick marine/deltaic or both or terrestrial sediments of a petroleum
system act as an archive of depositional environment, kerogen type, and thermal maturity of
organic matter.
 Under the current scenario of climate change, warming and disturbance is accelerating at an
alarming scale regionally and globally. Paleo-vegetation type (C3/C4) biomarkers are used as a
proxy to reconstruct the paleoclimate changes occurred under geological timescale

 C3/C4 vegetation types are long chain hydrocarbons (nC25-nC35).

 4
Study Area

 Potwar Plateau oil seeps provide an interesting information about the

underground petroleum system for exploration.
 Kundal was one of the first well drilled in the area.
 It was drilled by British Petroleum in 1866 but due to less oil potential crude oil
extraction was not possible
 The seepages are still present in the area.
 Kundal Oil Seeps are located next to Kaala Pani Naala. Kundal oil seeps touches
the northeast of Khisor Range and is the western part of Kundal village.
5
6
 7

 Starting from the start of the Kundal Village up to 6+ kilometers only river deposits
(conglomerates) can be found on the left side while on the right side its through out
variegated clays (red and yellow) color.
 The water channel running in between these two formations is reddish which indicates the
presence of iron in it.
 The Tredian Formation belongs to Musa Khel Group (Triassic Period)which has 3
members.
1. Mianwali Formation
2. Tredian Formation
3. Kingriali Formation
 8

 Mianwali Formation is eroded and can not be seen at the

sampling site, whereas the Tredian formation has a
confirmable contact with the upper Kingriali formation..
 Only a small portion of Tredian and Kingriali Formations
are exposed in the area.
 Tredian formation has sand stones with few interbedded
beds of shale with total exposed area from top to bottom is
200m while the Kingriali shales are very less and only 6m
bed can be found there.
 9
AIMS & OBJECTIVES

The present study aims to reconstruct the paleo-climate changes to predict future climate change
using vegetation type biomarkers as a proxy in oil seep, source rock and soil of Kundal oil field.

Objectives are given as follows.

1. To evaluate the geochemical characteristics of soil and rock samples.

2. To identify diverse group of biomarkers in crude oil/source rock/soil samples and establish
correlation between oil and source rock.

3. To reconstruct the paleo-climatic changes using vegetation type biomarkers such as diterpenoids
and triterpenoids as a proxy.
 10
SIGNIFICANCE

 Biomarkers are used as an indicator prior to decide new exploration site within a
petroleum system.
 The presence of biomarkers is commonly used to establish correlation oil to
source rock within a petroleum system.
 New trends in biomarkers study revealed the contention that biomarker may be
used to reconstruct paleoclimate changes of petroleum system occurred over
geological time scale.
 11
METHODOLOGY

Soil and rock

geophysical
characteristics

EDS (SEM)

Field Lab TOC and Rock Eval

Work work
Pyrolysis

Sampling
Vitrinite Reflectance

Biomarkers analysis
 12
FIELD WORK

 Kundal oil Seeps are present between Essa Khel and Kundal village.
 The area is mostly conglomerates on the left side whereas on the right-side
extrusive deposition of clays (reddish-yellowish) can be seen
 Kundal Oil seeps are almost 6Km away from the main Kundal Village road
 Oil seeps, water, sludge, soil and source rock samples were collected from
Kundal oil Field area.
 The samples from the oil seeps were carefully collected and stored in plastic
bottles, while the source rock samples were placed in secure zipper bags and
brought to the laboratory.
 13

Field Pics
 Conglomerate beds can be
seen in left picture whereas
on right side reddish and
yellow clays can be seen
 14
Oil Sample Collection

Oil Spill from the Formation

 15
Lab work:- Soil and Rock Geophysical
Characteristics

 A bulk soil sample was spread over the polyethylene sheet and
were air dried for 24 hours.
 These subsamples were then subjected to sieving using a mesh size
of 0.2 millimeters.
 The powdered rock and soil samples underwent detailed analysis
to assess their physicochemical characteristics.
 Hydrometer method of (Gavlak;Horneck and RO, 2005) was used
to determine the texture of soil and powdered rock samples.
 16
EDS by SEM

 Powdered rock and soil sample were analyzed using energy

dispersive X-ray spectroscopy (EDS) and scanning electron
microscopy (SEM). This analysis is performed in SEM laboratory
of NCEG.
 17
TOC And Rock Eval Pyrolysis

 To assess a rock's potential as a source rock for hydrocarbons, it is

essential to determine the total amount of organic carbon (TOC) that is
present in the sample. The Oil and Gas Development Company Limited
(OGDCL) uses an ELTRA HELIOS CS-580A analyzer for this
procedure.
 Then, a high frequency induction furnace is used to burn one gram of
the powdered sample up to 1200°C.
 The common requirement is that a rock contains more than 1% (wt%)
TOC in order to be regarded as a good source rock for hydrocarbons
 18
Vitrinite Reflectance

 The vitrinite reflectance determines the maturity and maximum

paleotemperature and its duration in geological time scale.
Therefore, vitrinite reflectance is referred to paleo geothermometer.
 Epoxy is used to make whole-rock or kerogen concentration
pellets, which are then allowed to cure overnight. Pellets are
polished and put overnight in a desiccator to eliminate moisture.
The fraction of light reflected off the vitrinite maceral at 500X
magnification in oil immersion is measured as vitrinite reflectance
(%Ro).
 19

Biomarkers
 20
Biomarkers extraction from Water samples

 Water samples, approximately one liter in volume, were collected for analysis.
The water samples were subjected to extraction three times using 30 mL of
dichloromethane (DCM) in each extraction. This extraction process helps
separate the hydrocarbon compounds from the water.
 To eliminate any remaining moisture from the hydrocarbon extract, the extracted
solution was passed through anhydrous sodium sulfate. Sodium sulfate is a
desiccant that effectively absorbs water.
 The dichloromethane solvent containing the extracted hydrocarbons was then
evaporated. This was achieved by gently heating the solution while
simultaneously purging it with a stream of nitrogen gas.
 21
Biomarkers extraction from Sludge, Oil and
soil samples

 The extraction of organic matter from sludge samples was conducted. In a glass beaker, 50
milliliters of dichloromethane were added to 50 grams of the sludge sample. The sludge sample
and dichloromethane were vigorously mixed for 2 to 3 minutes to ensure thorough contact and
extraction of organic matter from the sludge. After the initial mixing, the sample was sonicated
for 15 minutes in an ultrasonic bath. Sonication uses high-frequency sound waves to aid in the
extraction process, facilitating the release of organic compounds from the sludge.
 To eliminate any remaining moisture from the hydrocarbon extract, the extracted solution was
passed through anhydrous sodium sulfate. Sodium sulfate is a desiccant that effectively
absorbs water.
 The dichloromethane solvent containing the extracted hydrocarbons was then evaporated.
This was achieved by gently heating the solution while simultaneously purging it with a
stream of nitrogen gas.
 22
Results & Discussion
Particle Size Distribution

Particle size Soil Powdered Oily

in % sample rock Sludge
sample
Sand 20 55 53
Clay 55 1 4
Silt 25 44 42
Texture Clayey Silty loam Silty loam
soil
 23

 The particle size distribution of Tredian Formation and oil seep sludge of this formation of

this study was silty loam in nature along with presence of very little clay is (Table mentioned

above). The greater content of silt is more likely because of shales in the rock sample, whereas

presence of sand is also because of sandstone the dominant particle size features of the Tredian

Formation. The fine-grained particle in the rock suggests that the rocks undergo geological

processes and can influence properties like porosity and permeability.

 24
EDS of Rock Samples

Elements Mass% Atom% Sigma Net K Ratio Line

Carbon(C.) 43.42 54.28 0.05 31121 0.0076517 K
Oxygen (O.) 40.37 37.89 0.16 28021 0.0314843 K
Magnesium (Mg) 0.20 0.12 0.02 1011 0.0004435 K
Aluminium (Al) 2.38 1.33 0.03 15340 0.0053789 K
Silica (Si) 8.86 4.73 0.06 59609 0.0201910 K
Sulphur(S) 1.17 0.55 0.02 7362 0.0029466 K
Potassium (K) 0.45 0.17 0.02 2556 0.0013262 K
Calcium (Ca) 0.67 0.25 0.02 3883 0.0021179 K
Iron (Fe) 2.47 0.66 0.04 5746 0.0066572 K
Total 100 100
 25
Elements Mass% Atom% Sigma Net K Ratio Line
Carbon (C.) 8.09 13.47 0.03 5924 0.0007472 K
Oxygen (O.) 47.57 59.47 0.10 102792 0.592491 K
Magnesium 0.25 0.22 0.02 1327 0.0003106 K
(Mg)

Mg
Al
0.26
6.86
0.21
5.08
0.02
0.04
2234
74395
0.0005027
0.0133818
K
K
EDS of Soil
Si 20.00 14.24 0.07 229123 0.0398134 K samples
K 7.23 3.70 0.04 69038 0.0183728 K
Ca 0.83 0.42 0.02 8161 0.0022833 K
Mn 0.48 0.18 0.03 2126 0.0011408 K
Fe 8.43 3.02 0.06 34293 0.0203838 K
Total 100.0 100.0
 26

 The EDX study of Tredian Formation shows that nearly

83.79% of the Formation was carbon and oxygen followed
by Silica (8.86%) and then Al and Fe (2.38 and 2.47%)
whereas magnesium, potassium and calcium were present
in traces. However, Sulphur was found at 1.127%. Whereas
soil was dominated by oxygen and silica followed by
carbon and then aluminum, iron and potassium and traces
of calcium, magnesium, manganese, and sodium
 27
Sludge Rock
Parameters
Sample 1 (R001) Sample 2 (ROO4)
TOC(%) 16.63 0.38
S1(mg/g) 96.55 0.02
S2(mg/g)
S3(mg/g)
Tmax(°C)
72.91
0.26
414
0.17
0.04
437
TOC &
Rock Eval
PI 0.57 0.13
HI 438 45
OI 2 11
PC(%) 14.25 0.02

Pyrolysis
RC(%) 2.38 0.36
MINC(%) 3.25 3.56
pyroMINC(%) 1.16 0.17
oxiMINC(%) 2.09 3.39
OICO 7 16
S3CO(mg/g) 1.2 0.06
S3'CO(mg/g) 5.7 0.2
S3'(mg/g) 38.1 6.1
S5(s) 582532 8726781
Qty(mg) 7.6 70.2
 28

 The presence of nearly 50% of carbon in Tredian

Formation impregnated with oil seeps revealed
the contention that such formation has Quality Wt. % TOC Wt. % TOC
hydrocarbon potential. This is further confirmed (Carbonates) (Shales)
from the TOC results of this study that oil seep
Excellent > 2.0 > 4.0
contains nearly 16% of organic carbon. However,
Tredian Formation without oil seep has only Very Good 1.0-2.0 2.0-4.0
0.38% of TOC which suggests that the Formation Good 0.5-1.0 1.0-2.0
has low hydrocarbon potential
Fair 0.2-0.5 0.5-1.0
 Total organic carbon (TOC) provides information
about the kerogen type and thermal maturity of Poor < 0.2 < 0.5
the organic matter (Hunt et al., 1996)
 29

 Rock Eval analysis for the rock sample yielded S1, S2, and S3 values of
96.55 mg/g, 72.91 mg/g, and 0.26 mg/g, respectively. In contrast, the
sludge sample exhibited substantially lower values of S1 (0.02 mg/g), S2
(0.17 mg/g), and S3 (0.04 mg/g).
 The mineral carbon content (MINC) in both samples was relatively similar,
with the rock sample having a MINC of 3.25% and the sludge sample
having a slightly higher MINC of 3.56%. This suggests that both samples
do not contain a appropriate proportion of carbonates. The higher organic
matter content in the sludge from the oil seep site implies that the source
rock was likely not composed of carbonates.
 30

Vitrinite
Reflectance
 31

Thermal Maturity Stages Vitrinite Rflectance (Ro

 The results of this study shows that (%) %) Tmax (oC)
VR was 0.77% which lies between 0.2-0.6 >435
Immature
0.5-1.1% of oil window and
because of fine grain texture, low Mature 0.6-1.35 435-470

organic matter content revealed the

0.6-0.65 435-445
contention that Tredian formation Early
oil seeps is not of good vitrinite 0.65-0.9 445-450
Peak
particle to measure.
0.9-1.35 450-470
Late
>1.35 >470
Post mature
 32
Pw-19811 Pw-19812 Px-19813 Pr-19814
Sr. No Parameters
(Waste Water) (Rock Sample) Sludge (Soil Sample)
1 Isoalkanes (Pr/ph) - - - -
2 Iso Alkanes (Pr/nC17) - - - 0.45
3 Isoalkanes (Ph/nC18) - - - 0.30
4 OEP29 - - - 0.95
5 Triterpanes (Ts/Ts+Tm) 0.57 0.58 0.60 0.52
6 Triterpanes (C19/C19+C23) 0.08 0.17 0.17 0.09
7 Triterpanes (C29/H30) - - - 1.82
8 Gammacerane/H30) - - - 0.72
9 Moretane (M30/H30) - - - 0.34
10

11
Oleanane (O/H30)

Homohopanes (H32S/ H32S+ H32R)

-

-
-

-
-

-
0.50

0.63
Biomarkers
12 Steranes (C27/C27+C29) 0.46 0.54 0.54 0.51
13 Steranes (C29S/C29S+C29R) 0.40 0.54 0.57 0.40
14 Steranes (C29ββ/C29ββ+C29αα) 0.46 0.59 0.60 0.55
15 Bisnorhopnae (C28,30 BNH) Prominent peak Prominent peak Prominent peak Prominent peak
 33

 It is observed in this study

that all single chain
biomarkers ranged from
nC10 to nC44 are
completely absent in the oil
seeps of Kundal. This
suggests that this oil is
biodegraded as per
classification of (Wenger
and Isaksen, 2002)
 34
Single Chain Biomarkers

 This study also shows that pristane/phytane/ and other single chain carbon biomarkers are
completely absent in the oily sludge, oil water emulsion but the pristane/nC17,
phytane/nC18 ratio in soil was 0.45 and 0.30, whereas was not determined in other
samples.
 Odd Even Preference index (OEP29) was 0.95 in soil samples and cannot be calculated in
other samples more likely because the odd and even biomarkers were completely absent
in sludge, rock and wastewater samples.
 The absence of isoprenoids also represents that the oil is biodegraded. This is suggested
that Kundal oil seep is severely degraded oil
 35
Aromatic Hydrocarbons

 Triterpanes and steranes are those biomarkers which are used to provide information about
the organic matter input and depositional environments of oil.
 Their presence in rock or oil seeps or soil usually indicates the vegetative carbon source of
such biomarkers and also the climate of the environment during the time of deposition of
such vegetation nearly millions of years before.
 Triterpanes (Ts/Ts+Tm) and Triterpanes (C19/C19+C23) were present in the rock, oily sludge
and wastewater and soil samples but Triterpanes (C29/H30) were absent and were present
only in soil at 1.8 ratio.
PETROLEUM CARBON SEDIMENTATION TYPE ORGANIC MATTER SOURCE DOMINANT
COMPOUNDS CHAIN ALKANES

n-alkanes C9-C21 Marine/Lacustrine Algal (C3)-lipids and nitrogenous compounds C15,C17 and
C19

C25-C37 Terrestrial plant Plant wax (C4)-cellulose and lignin C27, C29 and
C31

C12-C24 Marine/Lacustrine Bacteria

Carbonates/evaporites Saline anoxic environment C20-C24

Acyclic C19, C20 Marine Prokaryotes/chlorophyll phytyl side-chain and/or α-

isoprenoids tocopherol (vitamin E)
Pristane,
Phytane

36
 37

In our study the Carbon preference index is missing which is essential for paleoclimatic reconstruction
and shows about the input either anthropogenic or biogenic. Based on the results of the CPI, which is missing,
input source for paleoclimatic reconstruction cannot be predicted
Similarly other factors used for paleoclimate reconstruction are Di-terpenoids and Tri-terpenoids. Di-
terpenoids are missing in all the samples whereas Tri-terpenoids ratio can be seen in all of the samples. Based
on the results of biomarker, Triterpanes (Ts/Ts+Tm) ratio is 0.57 for water samples which indicates that organic
matter is thermally mature. marine organic matter input seems dominant because marine biomarker C23 is in
high concentration as compared to terrestrial organic matter biomarkers C19 and C20. The C19/C19+C23 ratio
is 0.08 which indicates the low terrigenous organic matter input.
Triterpanes (Ts/Ts+Tm) ratio is 0.58 for rock sample, which indicates that organic matter is thermally
mature. Marine organic matter input seems dominant because marine biomarker C23 is in high concentration as
compared to terrestrial organic matter biomarkers C19 and C20. C19/C19+C23 ratio is 0.17 which indicates the
high marine organic matter input.
 38
Conclusion

 Biomarkers Consistency: Ion chromatograms for all samples, including water,

rock, soil, and sludge, consistently show the presence of the same set of
biomarkers. This suggests a strong genetic correlation among these samples,
indicating that the biomarkers characteristic of the Tredian Formation is present in
all of them. Notably, Pr-19814 (soil) stands out as it contains a complete set of
terpanes, unlike the other samples, which may be indicative of unique depositional
conditions.
 Biodegradation Patterns: The N-alkane profiles, as observed in GC-FID
chromatograms, indicate signs of biodegradation in all samples, except for soil.
The absence of biodegradation in soil suggests that this particular sample has
experienced different preservation conditions.
 39

 Thermal Maturity and Organic Matter Input: All samples exhibit signs of
thermal maturity, indicating that they have been subjected to heat and pressure
over geological time. Additionally, they display mixed/marine organic matter
input, which suggests that the source of organic material in these samples likely
includes both terrestrial and marine sources. The presence of C28,30
Bisnorhopane (BNH) biomarker in higher percentages in all samples further
supports this interpretation.
 This is concluded that oil sludge and crude oil samples are genetically same, and
their genetic origin is shale.
 40

Thank You