BIOMONITORING

IN PETROLEUM INDUSTRY

Zdravko Spiric
INA-Naftaplin, Croatia
Milena Horvat
IJS, Slovenia

ABSTRACT

In order to control and improve the efficiency of an industrial facility for

mercury removal from natural gas at the INA-Naftaplin gas treatment plant, Molve,
Croatia, a comprehensive environmental monitoring program, including mercury
measurement in air, was established. Generally, total mercury concentrations in air are
very low (e.g. below 10 ng.m -3) and therefore potentially subject to analytical errors
(e.g. contamination).

In the present study, an alternative methodology for monitoring mercury

concentration in air by using lichens as biomonitors was employed. The obtained
results confirm that during the study period no significant elevation of Hg in air
occurred at Molve, confirming efficient removal of Hg from natural gas.

This study also confirms that lichens can successfully be used as biomonitors
of Hg air pollution in petroleum industry. Biomonitoring, as a new field sampling and
analytical approach offer the potential for considerable time and cost savings
compared to conventional monitoring and measurement procedures, helping process
engineers and decision makers to identify and prevent potential mercury generated
hazards linked to specific industrial processes.
 MERCURY IN THE NATURAL GAS
Natural gas from gas fields around the globe often contains mercury, along
with a large number of other harmful substances (CO 2, H2S, RSH, COS, etc). Mercury
is a naturally occurring component of geological hydrocarbon deposits. It is evident
that geological/geochemical characteristics of every hydrocarbon reservoir directly
contribute and define the possible influences (level and intensity) of oil and gas
production activities on eco-systems 1. Also, it is quite certain that mobilization and/or
release of geologically bound mercury by human activities, i.e. fossil fuel combustion
(together with waste incineration and industrial processes) and especially metal ore
and hydrocarbon refining and processing are the most important sources of mercury
emission into the atmosphere on a world-wide basis 2,3.

High levels of mercury in produced gas may have harmful effects on the
environment and health not only during gas exploration, processing and transport, but
also in natural gas consumption. Pollutants (like mercury) released from
hydrocarbons into the air, depending on: process location, configuration and
performances; landscape; weather conditions and the chemical and physical
properties of the pollutants, are carried by wind patterns away from their place of
origin and could persist in the environment for many years. Mercury has long been
known to be a toxic, persistent, bio-accumulative pollutant with a wide range of
ecosystem and human health effects 4. Mercurys potentially harmful effect on
humans and on the ecological system as a whole as well as the technological risk
regarding very sophisticated and expensive petroleum process equipment and
catalysts is make its removal imperative. There is a need for the highest efficiency for
mercury removal and for permanent improvement and enhancement of technological
and control procedures, including analytical instruments and measurements 5.

At present, it is generally believed that sustainable development is the main

limiting factor, but also strong and important driving force in the same time, for
further development of the world economy and society in general. In petroleum
industry, ecological sustainability limits of hydrocarbon production can be determined
through the multidisciplinary study on relationships between geology and ecology
and evaluation of ecological risks. This knowledge is critical to our understanding,
control and optimization of existing processes, but also to planning strategies and
decision making for further operations in the oil and gas industry. Could
biomonitoring be suitable tool for this challenge?
 OUR MERCURY CONTROL EXPEERIENCE
The natural gas recovered and produced from deep wells at Podravina gas
fields contains a relatively high level of mercury along with other harmful substances.
These undesired contaminants must be removed by appropriate methods in order to
obtain safe and reliable operations of process plants, to meet the requirements related
to product specifications, and to protect the human working and living environment 6.

In order to solve potential economic, ecological and technical problems

related to the presence of mercury in processed natural gas, INA-Naftaplin
constructed appropriate process units at Gas Treatment Plant Molve, Croatia (Figure
1), that ensure complete removal of mercury from natural gas. In line with recent
international practice in petroleum industry, mercury removal from natural gas has
been accomplished by the use of sulphur-impregnated activated carbon 7. The
pollution hazards have been brought to a minimal level since a two-bed closed
mercury removal system has been constructed. The efficiency of mercury adsorption
from gas flow is 98-99 % 8.

INA-Naftaplin scientists have also created an equally important mercury-

monitoring program to obtain accurate and reliable mercury test results, which
directly indicate the effectiveness of the mercury removal process, process safety and
environmental protection against mercury contamination. The key elements of an
ecological exposure assessment include the identification of potential exposure
pathways and the estimation of exposure point concentrations. For this purpose, as a
consequence of constantly decreasing allowable levels of toxic material (along with
mercury) present in our surroundings, the sophisticated method of sampling has been
adopted. It comprises sampling of process gas streams and ambient air on Amasil
traps, which are made from gold impregnated onto silica, followed by analysis of
collected samples by atomic fluorescence spectrometry (AFS), using the PS
Analytical's Sir Galahad instrument 9. Contrary to some other analytical procedures,
this process is very accurate, but costly and time-consuming. High monitoring (field-
sampling and analytical) and site characterization cost continued to threaten
operational budgets and company management was concerned; however, if an
inadequate characterization is performed, more costly or inadequate measures may be
chosen, thereby increasing not only the risk but also the overall costs.

As a result we started to explore other possibilities. Extraordinary results and

experiences have been gathered in co-operation with Institute Joseph Stefan from
Slovenia with implementation of the bio-monitoring program involving the utilization
of lichen 10. Namely, it is well known that lichens are extremely sensitive to the
impacts of air pollution. They are useful as bioaccumulators and can reveal a lot of
information about which pollutants are present in their environment. Lichens rapidly
absorb substances from the atmosphere and permanently accumulate/concentrate
them in their tissues. Therefore, concentrations depend on ambient levels of these
pollutants, weather conditions, age of lichens and their physiological conditions.

Epiphytic lichens can be successfully used as bioindicators of Hg

concentration in air, as was first demonstrated in the Hg mining area in Idrija,
Slovenia. Concentrations of mercury in lichens represent the average long-term Hg
status in air, while Hg measurements in air represent only an average value during the
sampling period (e.g. from several minutes to the maximum daily average
concentrations).

Regarding to this, field test results for total mercury in lichens collected in the
monitored area were also obtained and taken into consideration. Measurements of Hg
in lichens are of particular importance for spatial monitoring of Hg in air and
therefore they are included in Molve Environmental Impact Assessment study.

Sampling and transplantation of lichens

In-situ lichen samples of different species were collected at several control
locations, i.e. natural gas producing sites in Croatia, Molve. Three different species of
lichens, namely Hypogymnia physodes, Parmelia caperata and Evernia sp. were
collected in nearby forest. Lichens were carefully removed from trunks of dominant
tree species, 1.5 to 2 m above the ground, stored in paper bags and transported to a
laboratory at the Joef Stefan Institute for further preparation.

At the same locations and at the same time the experiment with transplanted
lichens was also initiated using polyethylene nets filled with lichens from non-
contaminated area. Approximately one week before transplantation a lichen specie H.
physodes was collected from orchard trees at pristine area in Tuhinjska dolina
(Slovenia). In the laboratory lichens were carefully cleaned from branches and filled
in polyethylene nets. Two nets were then fastened to branches or on artificial rod
about 1.5-2 m above the ground at locations in Croatia, as well as at two reference
locations, one in Ljubljana at the Reactor site and one in Idrija. Lichens were then
exposed for a period of 6 and 12 months. Some lichen material was kept in the
laboratory to determine the initial concentration of Hg and other trace elements. One
polyethylene net from each station, including reference stations was sampled.

Sample preparation
In the laboratory the lichen samples were moistened with distilled water and
carefully separated from the bark substratum using nylon tweezers and lyophilized.
The samples were not washed, as our aim was to measure the elements that were
physically trapped on the surface of the thallus as well as chemically bound to the cell
wall. The samples were made brittle by immersion in liquid nitrogen and were then
crushed and ground in a zirconium mortar with Zr ball in a Fritch vibration micro-
pulverizer.
Analytical methods
Determination of total mercury by CV AAS:
About 100-200 mg of dry lichen powder was weighted directly in the Pyrex
digestion vessel and after addition of 2ml of HNO 3, conc. the vessel was sealed and
the mixture was left to react at room temperature overnight. Digestion was finished
by heating the Al block at 700C for 12 hours on a hot plate. Tubes were then cooled in
liquid nitrogen and opened. The digest was transferred to 50 ml volumetric flasks and
diluted with double distilled water to the mark. An aliquot of the digest was added to
the reduction vessel and after reduction mercury was swept from the solution by
aeration and concentrated on gold traps. Mercury was then released from the gold trap
by heating and measured by cold vapor atomic absorption spectrophotometer (CV
AAS). Detailed description of the methods used is described in two publications 11,12.
Determination of other elements (Instrumental neutron activation analysis INAA):
Tablets of ground and homogenized samples of lichens (150 200 mg) were
irradiated for 18 hours in the carousel facility of the TRIGA Mark II Reactor,
Ljubljana (thermal neutron fluence rate of 1.2 10 12 cm-2.s-1, =0-0.03). An Al - 0.1%
Au alloy wire of 1.0 mm diameter and 125 m Zr foil were co-irradiated together
with the samples as comparator and fluence rate monitor. The k o-based method 13 was
used and 53 elements were sought for. The irradiated samples were subsequently
transferred to a clean polyethylene vial and counted on a calibrated coaxial HP Ge
detector connected to a Canberra S-100 multi-channel analyzer. Samples were
measured twice, after 2 days and 8 days cooling time. For evaluation of gamma
spectrum, the Hypermet-PC, V5.0 program was used 14, for detection efficiency
calculation the SOLCOI program 15, and for elemental concentration calculations the
KEYZERO program 16.

Analytical Quality Control

The accuracy of the results was checked by the use of certified
reference material, IAEA-336, Trace and Minor Elements in Lichens.

RESULTS
In the present study, together with mercury analyser based on fluorescence
detection technique (Sir Galahad instrument, manufactured by PS Analytical), an
alternative methodology for monitoring Hg contamination in air by using lichens as
biomonitors was employed. Total mercury was measured in in-situ and transplanted
lichens.

Concentrations of Hg in air and lichens at INA-Naftaplin, Molve, were

relatively low. Highest Hg concentrations were observed at GTP station, while
concentrations at other stations were only slightly elevated. Mercury levels in air were
typically below 10 ng/m3, rarely exceeding 30 ng/m 3 (Table 1), while Hg
concentrations in lichens were in the range from 0.1 to 2.8 g/g (Table 2), which is
typical for uncontaminated sites with Hg. In Idrija, values for total Hg in lichens were
much higher (up to 5 ppm) which correlate well with much higher Hg concentrations
in air (from 10 to 300 ng/m3) 10. A very good correlation between Hg concentrations
in air and in lichens at all sampling stations around the INA-Naftaplin Gas Treatment
Plant, at Molve was observed for all lichen species, Figure 2a, 2b and 3.

It can be concluded that biomonitoring can be cost-effective and useful as

lichens increase accuracy in predictions of source releases and attenuation of
contaminants and can successfully be used as biomonitors of Hg air pollution.

Lichens may also provide data for other elements (such as selected
hydrocarbons, other trace elements and other compounds), emitted during petroleum
activities. In-situ lichen samples (such as Parmelia sulcata, Xanthoria parietina, and
Hypogymnia) were collected at various locations of the studied areas for the
determination of Hg and other 37 elements. The values for total Hg were correlated
with Hg measurements in air, while for other elements values were compared with
average concentrations found in contaminated and non-contaminated areas in Europe.
 It was shown that Ba and Br are systematically higher at Molve stations
compared to initial values and transplanted lichens. These two elements are mostly
related to the natural gas production operations. However, there are several
contributing factors of toxicity in petroleum industry. Generally, the technology of
mud mixing and treatment is recognized as a source of pollutants such as barium
(from barite). Down-hole production operations, which include all recovery methods;
but also well work-overs and well stimulations activities that uses treating chemicals:
biocides, corrosion and scale inhibitors, etc. may be sources of detected elevated
concentration of Br.

The research results, i.e. data obtained under this combined program of
biomonitoring and continuous metering of mercury in ambient air surrounding the
production and process units at GTP Molve, Croatia, have been confirmed by several
independent mercury testing authorities and institutes involved with research on
mercury present in eco-systems, with priority given to ambient air Figure 4 and 5.
During the years the GTP Molve hosted the experts participating in several field inter-
comparison exercises (Environmental Research Institute IVL Stockholm + GKSS-
Galab; Calgon Carbon Corporation, USA; St. Petersburg State University, Russia and
others). In the scope of the research programs on mercury they used various sampling
methods as well as diverse analytical and measuring techniques (Tekran Instrument
AFS, Jerome Gold Film Detector, Zemman AAS, RNAA, etc.).

CONCLUSION
The corrosion and environmental pollution hazards on the Croatian Podravina
natural gas fields have been brought to a minimal level since an efficient, closed
mercury removal system has been constructed. Mercury concentration measurements
in the ambient air, performed by combined atomic fluorescence spectrometry
technique and biomonitoring, confirm that the design of the system at the Molve gas
plant is adequate to meet the mercury removal objectives. During the study period no
significant elevation of Hg in air occurred at Molve, confirming efficient removal of
Hg from natural gas. Moreover, concentration levels of Hg in in-situ lichens further
confirm that during the last few years no significant Hg contamination occurred in
this area.

This study confirms that Biomonitoring is an accurate, comprehensive,

flexible, time and many saving package for environmental, health & safety
compliance and management. Biomonitoring can also act as an early warning of
harmful conditions (as result of poor efficiency or malfunction of the processes)
and/or as fingerprint of local environmental pollution. By using lichens it is
possible to identify almost all pollutants that are normally very hard to measure
because they are present in very small amounts (ppm, ppb, ppt level), or is difficult to
detect them by other methods. Finally, the biomonitoring on Molve site provided us
with more information and environmental data than traditional analytical procedures,
but what is more important, it reduced mercury-monitoring costs for more than 80 %.
 References:

1. Spiric Z. and Mashyanov N.R., Mercury measurements in ambient air near

natural gas processing facilities, special issue paper, Fresenius J Anal Chem.
366 (2000) 5, 429-432
2. Ebinghaus R, Tripathi RM, Wallschlager D, Lindberg SE (1999) Natural and
anthropogenic mercury sources and their impact on the air-surface exchange
of mercury on regional and global scales. Ibid., pp 3-50
3. Mason RP, Fitzgerald WF, Morel FMM (1994) Geochim & Cosmochim Acta
58(10): 3191-3198
4. Agency for Toxic Substances and Disease Registry (ATSDR) (1992) Case
studies in environmental medicine, Mercury toxicity, Atlanta, GA: US
Department of Health and Human Services, Public Health Service
5. Spiric Z, Dragas M, Vadunec J, Mashyanov NR and Ozerova N. (1999)
Investigation of mercury content in Podravina gas fields and environment,
6th International Petroleum Environmental Conference, November 16-19,
Houston, USA
6. Spiric Z, Vadunec J (1996) Protection of Process Plant and Environment
against Mercury during Natural Gas Production, 12-th International Congress
of Chemical and Process Engineering - CHISA' 96, Prague
7. McNamara J.D. and Wagner N. J., Removal of Mercury from Natural Gas to
Ultra Low Levels using Type HGR Activated Carbons, Paper presented at the
A.I.Ch.E. Spring National Meeting in New Orleans, LA, (February 1996)
8. Spiric Z. and Hraste M, (1998) Mercury saturation profile across the sulphur
impregnated activated carbon bed, In Ebinghaus R, Turner RR, Lacerda D,
Vasiliev O. and Salomons W. (eds), Mercury Contaminated Sites:
Characterization, Risk Assessment and Remediation, 409-417, Springer
Environmental Science, Springer Verlag Heidelberg,
9. Spiric Z. and Stockwell P.B., Ambient Air Mercury measurements during
Natural Gas Production, American Environmental Laboratory Vol. 10, No. 5.,
16-20 (1998)
10. Horvat M, Jeran Z, Spiric Z, Jacimovic R and Miklavcic V.: Mercury and
other elements in lichens at INA-Naftaplin gas treatment plant, Molve,
Croatia, Journal of Environmental Monitoring, 2000, volume 2, issue 2, 139-
144.
11. Horvat M, Zvonaric T, Stegnar P (1986) Optimization of a wet digestion
method for the determination of mercury in blood by cold vapour absorption
spectrometry (CV AAS), Vestn. Slov. Kem. Drus. 33(4), 475 486.
12. Horvat M, Lupsina V, Pihlar B (1991) Determination of total mercury in coal
fly ash by gold amalgamation cols vapour atomic absorption spectrometry.
Anal. Chim. Acta 243, 71 79.
13. Smodis B, de Corte F, de Wispelaere A: J. Radioanal. Nucl. Chem., Letters,
186 (1994) 183.
14. Hypermet-PC V5.0, users Manual, Copyright 1997, Institute of Isotopes,
Budapest, Hungary.
15. Hypermet-PC, User's Manual, 1997
16. Xilei L, de Corte F, Moens L, Simonits A, Hhoste J: J. Radioanal. Nucl.
Chem., Articles, 81 (1984) 333.
Table 1. Results obtained with PSA's Sir Galahad Atomic Fluorescence
Spectrophotometer
The 24-hour ambient air samples for mercury content in Molve area

Mol Mol Mol Mol Mol

GTP
-9 - 10 - 11 - 12 - 14
Distance from
- 525 1000 825 1575 1475
GTPs, meter
In direction off the
- NE S W SE N
GTPs,
Number of samples
171 32 79 25 23 20
taken, total
Minimum value,
0,1 0,1 0,1 0,1 0,1 0,1
ng/m3
Maximum value,
56,2 14,7 26,9 36,1 18 6,6
ng/m3
Average value,
10,6 5,7 5,0 5,4 3,7 2,3
ng/m3

Table 2. Results for total mercury in in-situ lichens (Parmelia sulcata,

Xanthoria parietina, Hypogymnia physodes) (g/g, dry weight)
collected on GTP Molve area sampling locations

Concentration Hg
Sampling location Lichen species (g/g)
After 6 and 12 months

Parmelia sulcata 0.53 0.20

MOL 9
Xanthoria parietina - 0.29

0.15 0.16
Parmelia sulcata
MOL 10 Hypogymnia
0.20 0.20
physodes

0.34 0.23
Parmelia sulcata
MOL 11
0.20 0.21
Xanthoria parietina

0.11 0.17
MOL 12 Parmelia sulcata

0.54 1.17
Parmelia sulcata
MOL 14
2.80 2.27
Xanthoria parietina
IAEA, Lichen 336
Certified value: 0.20 g/g 0.21 0.21
 CO2 /H2S
1. Hg ADSORBER REMOVAL

BATTERY LIMITS

NATURAL aMDEA
GAS WATTER
WELLS
WASH

THREE
PHASE
SEPARATOR

2. Hg ADSORBER

C2+

BRAZED MOLECULAR
ALUMINIUM SIEVE
HEAT. DRYERS
EXCH.
2 pcs

C1
SALES CRYOGENIC PROCESS
GAS

GTP MOLVE III - NATURAL GAS PROCESSING

Figure 1. Scheme of the GTP Molve Natural gas processing plant

Mercury content in air on the GTP MOLVE

100
90
80
70
60
Hg ng/m3

50
40
30
20
10
0
19.12.1995.
05.01.1996.

16.01.1996.

09.02.1996.

01.03.1996.

07.03.1996.

21.03.1996.

27.03.1996.

20/21.05.1996.

06/07.06.1996.

27/28.06.1996.

13/14.01.1997.

25/26.02.1997.

14.01.1998.

29/30.03.1999.

20/21.05.1999.

16/17.06.1999.
8/9.05.1996

28/29.01.1997.

17/18.03.1999.

13/14.04.1999.

03/04.05.1999.

03/04.06.1999.
11/12.12.1996.
13/14.11.1996.

DATE

Figure 2a. Mercury concentration measurements in ambient air on GTP

Molve
 1,0
Initial Hg conc. MJ
Exposed from April 10 - Sept. 24
0,8
Exposed from April 10 - Dec. 8

0,6
Hg conc. (g/g)

0,4

0,2

0,0
CPS CPS, V3102 MOL 10 MOL 11 MOL 14
Sampling Location

Figure 2b. Mercury concentrations in transplanted lichens

Hypogymnia ph. at Molve stations

10000

1000

100
C o n c . H g i n a i r ( n g m- 3 )

10

0 ,1

0 ,0 1
100 1000 10000

Figure 3. Correlation between mercury concentrations in transplanted

lichens Hypogymnia physodes in Molve and Idrija after 6 months of
exposure. Horizontal lines represent the mean and concentration ranges
(min.-max-) of Hg in air; vertical lines represent mean standard deviation
(n=3) of Hg concentrations in lichens 10.