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Article
Advanced Aspects of Crude Oils correlating data
of Classical Biomarkers and MS Petroleomics
Jandyson Machado Santos, Flavia Micaella Lemos Santos,
Marcos Nogueira Eberlin, and Alberto Wisniewski Jr
Energy Fuels, Just Accepted Manuscript • DOI: 10.1021/acs.energyfuels.6b02362 • Publication Date (Web): 03 Jan 2017
Downloaded from http://pubs.acs.org on January 6, 2017

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1
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5 1 Advanced Aspects of Crude Oils correlating data
6
7
8 2 of Classical Biomarkers and MS Petroleomics
9
10
11 3 Jandyson Machado Santos†,‡, Flávia Micaella Lemos Santos†, Marcos Nogueira Eberlin‡,
12
13 4 Alberto Wisniewski Jr. †*
14
15 †
5 Federal University of Sergipe, Department of Chemistry, São Cristóvão, Sergipe, 49100-
16
17 6 000, Brazil
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19 7 ‡
ThoMSon Mass Spectrometry Laboratory, Institute of Chemistry, University of Campinas,
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21 8 UNICAMP, Campinas, São Paulo, 13083-970, Brazil
22
23
24 9
25
26 10 ABSTRACT
27
28
29 11 The complex geochemical characteristics of crude oils can provide important information on
30
12 oil generation, such as the input of organic matter and their depositional environments, also
31
32 13 supporting exploration, extraction and production. This work reports the first organic
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34 14 geochemical evaluation of oils from the first commercial Brazilian onshore field
35
15 (Carmópolis, Sergipe, Brazil) via both classical biomarkers as well as with petroleomics data
36
37 16 collected via ultra-high resolution and accuracy Fourier transform ion cyclotron resonance
38
39 17 mass spectrometry (FT-ICR MS). GC/MS was used to characterize the oil composition in
40
18 terms of n-alkanes, isoprenoids, terpanes and steranes biomarkers, whereas either positive or
41
42 19 negative electrospray ionization (ESI) and FT-ICR was used to profile the polar constituents
43
44 20 of the oils. GC/MS revealed geochemical characteristics that classify the oil blends at
45
21 different levels of thermal evolution and biodegradation. FT-ICR MS attributed molecular
46
47 22 formulas to more than 3,000 polar oil constituents, allowing geochemical parameters to be
48
49 23 inferred mainly from the Cn and DBE trends along for the N and O2 classes. The
50
24 Dia27(S+R)/αααC27(S+R), %C28 and %C29 parameters, S/(S+R) αααC29, αββ (S+R)/αββ +
51
52 25 ααα (S+R) C29 and %C27 biomarkers obtained from GC/MS were responsible to indicate
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54 26 similarity between samples whereas N and O2 classes provided the most distinction among
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27 the crude oil blends in terms of level of biodegradation and thermal maturity.
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58 28 INTRODUCTION
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1
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3 1 Crude oils are highly complex mixtures with molecules that range from small, volatile
4
5 2 to large, non-volatile hydrocarbons but also include a substantial fraction of polar
6 3 heteroatom-containing compounds.1 Characterization of the chemical composition and
7
8 4 correlation of the composition to predicted physical chemical properties of crude oils is
9
10 5 challenging but is key to obtain information that could guide extraction, production,
11 6 transportation, refining and distribution of products.2-4
12
13
14 7 Characterization of less polar biomarkers (mostly of the n-alkanes, isoprenoids,
15
16 8 hopanes and steranes classes) have been used extensively in geochemistry studies to infer
17 9 crude oil characteristics such as oil-oil and oil-reservoir correlations, origin, age, maturity,
18
19 10 depositional environments, and degree of biodegradation.5-11 Gas chromatography (GC/MS)12
20
21 11 as well as comprehensive two-dimensional gas chromatography (GC x GC)13 coupled to mass
22 12 spectrometry (MS) have been the major techniques used for biomarker screening in crude
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24 13 oils. Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) coupled
25
26 14 with electrospray ionization (ESI) has been used to characterize the most acid and basic polar
27 15 constituents in crude oils. The polar components, while minor, also serve as key biomarkers
28
29 16 for several major oil characteristics.14, 15
Such approach known collectively as MS
30
31 17 petroleomics has, therefore, expanded the chemical information provided by GC/MS and
32 18 GCxGC/MS.16, 17
33
34
35 19 ESI FT-ICR MS petroleomics offers the unique opportunity to sort, by class, the
36
37 20 thousands of polar compounds present in crude oils. From the assigned CnHmNxOxSx
38 21 formulas, the main classes can be identified and quantitatively compared. For example, the
39
40 22 relative abundances of N, NO, NS and O2 classes and their trends in terms of molecular
41
42 23 weight (MW), DBE (double bond equivalent) and carbon numbers can be compared and
43 24 associated with chemical properties.18-20 For instance, Vaz and co-workers17 and Liao and co-
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45 25 workers21 showed that the O and O2 classes can be used as indicators of the biodegradation
46
47 26 level. Crude oils from some Brazilian basins have also been investigated via petroleomics
48 27 approach using ESI FT-ICR MS.22, 23
49
50 28 The field of Carmópolis in the state of Sergipe ranks as the second largest onshore
51
52 29 production site in Brazil.24 But despite its high historical, economic and social importance, of
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54 30 the crude oils from the Sergipe-Alagoas sedimentary basin have the geochemistry not been
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57 31 thoroughly studied.25-28 Rodrigues and co-workers25 used GC/MS to classify marine
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1
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3 1 evaporitic oils from Sergipe-Alagoas basin that are constituted of immature organic matter.
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5 2 They suggested that the formation of the detected alkyl-steranes and monoaromatic alkyl-
6
7
3 steroids was due to decarboxylation of carboxylic acids during diagenesis. Rodrigues and co-
8
9
10 4 workers26 also found via GC/MS a series of cyclohexyl-undecanoic acids in marine evaporitic
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12 5 oil samples from the same basin. More recently, Sousa Junior and co-workers28 focused their
13
14 6 study on the class of isoprenoid biomarkers, and found that the low GC/MS pristane/phytane
15
16 7 ratios in crude oils from the Sergipe-Alagoas Basin point to a depositional anoxic and saline
17
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19 8 environment. Santos and co-workers29 has previously studied crude oil blends from Sergipe-
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21 9 Alagoas basin by the view of the application of classic techniques such as thermogravimetric,
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23 10 infrared and ultraviolet spectroscopy and energy dispersive X-ray fluorescence spectrometry.
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25 11 It is also known that crude oils blends from onshore wells in Sergipe-Alagoas basin presents
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12 distinct physicochemical characteristics (higher concentration of water and gas from the
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29
30 13 upstream process and viscosity), and the injection of greater amounts of demulsifiers is
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32 14 required to treat some blends.29
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34 15 In view of the importance that the Carmópolis field for the Brazilian production of
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36 16 crude oil, and the limited knowledge of the chemical composition of its crude oils, we
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38
39 17 investigated the geochemistry of four crude oil blends from this field using both the classical
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41 18 geochemical GC/MS biomarker analysis, and advanced FT-ICR MS petroleomics approach
42
43 19 to profile the most polar constituents.
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46 20
47
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49 21 EXPERIMENTAL SECTION
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22 Sergipe-Alagoas basin. The four onshore crude oils blends (CEOL1, CEOL2,
52 23 CEOL3 and CEOL4) were provided by Petrobras UO-SEAL and were collected from the
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54 24 Oiteirinhos II station, in the Carmópolis Field on Sergipe-Alagoas basin, Sergipe state,
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25 Brazil. The blends are a combination of oils from many wells arriving at the primary
57 26 treatment station through four pipelines and were collected in the same time. The blends were
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3 1 identified as CEOL1, CEOL2, CEOL3 and CEOL4, respectively, for lines 1, 2, 3 and 4 of the
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5 2 station. These samples have been studied and have been described in detail in a previous
6 3 work.29
7
8
9 4 Gas chromatography/mass spectrometry (GC/MS). The GC/MS analysis were
10
11 5 performed in a Shimadzu, GCMS-QP5050A instrument operated with a capillary column
12 6 [DB-5MS 5% diphenyl, 95% dimethyl polysiloxane - 30 m; 0.25 mm ID; 0.25 µm]. The
13
14 7 oven temperature program was 60 °C (3.0 min); 7 °C/min at 310 °C (14.0 min), and the
15
16 8 injector temperature was 250 °C. He (99.999%) as used as the carrier gas (flow of 1.20
17 9 mL/min – 71.8 kPa) with an interface temperature of 300 °C and a total analysis time of
18
19 10 52.71 min. Analyses were performed in the linear scan mode (SCAN) in the range of m/z 40-
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21 11 550. Selective ion monitoring (SIM) was also performed for the ions of m/z 191 and 217
22 12 (biomarker ions for terpanes and steranes, respectively). Samples were prepared with
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24 13 solutions at a concentration of 10 mg/mL in n-heptane and using an injection volume of 1 µL
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26 14 in the splitless mode (1.0 min).
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28 15 Petroleomics analysis by FT-ICR MS. Solutions for the four crude oils were
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30 16 prepared using 1 mg of the crude oil sample in toluene/methanol mixture (1:1 v:v) with a
31
32 17 final concentration of 1 mg mL-1. The FT-ICR MS analysis was performed using a 7.2T LTQ
33 18 FT Ultra mass spectrometer (Thermo Scientific, Bremen, Germany) equipped with a direct
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35 19 infusion electrospray ionization source (ESI) in both the positive and negative ion modes.
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37 20 The follow conditions were used: capillary voltage +3.5 and -3.1 kV, tube lens + and - 160 V
38 21 and temperature 300 °C. N2 was used as the nebulization gas. Mass resolving power was
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40 22 400,000@m/z 400 and the acquisition in the ICR cell was done with 100 µscan in each run.30
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42 23 Data acquisition was performed along the range of m/z 100-2000 by the Xcalibur 2.0
43 24 software. Identification of the ions was done by comparing its m/z values with a library of
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45 25 compounds present in the database of the PetroMS18 software based on literature search and
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47 26 standards. We considered a match between the experimental and the theoretical m/z values
48 27 from our library when the mass error was < 1 ppm and then the molecular formulas were
49
50 28 assigned for each m/z.
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52
53 29 Multivariate analysis. For the multivariate approach, we used the percentage of
54 30 classes assigned for the mass spectrum of crude oils from the software PetroMS. The data
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56 31 were aligned and preprocessed by autoscaling (data mean-centering followed by variance
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58 32 scaling), and were exploited via multivariate statistical analysis using principal component
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3 1 analysis (PCA) and hierarchical cluster analysis (HCA). The software Statistica for Windows
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5 2 v. 7.0 (StatSoft Inc., USA) was used for processing the data.
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7 3
8
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10 4 RESULTS AND DISCUSSION
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12
13 5 Distribution of alkanes. Figure 1 displays the distribution of n-alkanes and branched
14 6 alkanes (isoprenoid) in the four samples studied as obtained by GC/MS using the full SCAN
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16 7 mode. All the four crude oil blends showed bimodal chromatographic profiles which are
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18 8 characteristic of low MW paraffinic oils. Note that the alkanes were found to vary from n-C10
19 9 to n-C35, with n-C15 and n-C17 as the major n-alkanes. For the isoprenoid class, 2,6-
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21 10 dimethylundecane; 2,6,10-trimethylundecane; 2,6,10-trimethyldodecane; 2,6,10-
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23 11 trimethyltridecane; 2,6,10-trimethylpentadecane; pristane and phytane were clearly identified.
24 12 While the TICC profiles of Figure 1 are similar to each other, they differ from previous
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26 13 reports. Rocha and co-workers27 analyzed crude oil samples from the same SE-AL basin, and
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28 14 abundant low MW n-alkanes in the n-C13 to n-C19 range were detected only in two of the
29 15 eight analyzed oils. Sousa Júnior and co-workers28 observed even more contrasting GC/MS
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31 16 results and an unresolved complex mixture (UCM) for crude oils from the same basin. They
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33 17 attributed these findings to the mixing of biodegraded and non-biodegraded oils. Such UCM
34 18 behavior was not observed for the oils studied herein, and the clear bimodal GC/MS
35
36 19 distribution observed from our data provides no indication of extensive biodegradation of the
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38 20 blends. Peters and Fowler3 showed that bimodal n-alkane distributions, particularly those
39 21 skewed toward the n-C23-n-C31 range, are usually associated with terrigenous higher-plant
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41 22 waxes, which indicates that the oil has reached the early to middle oil window based on
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43 23 thermal maturation.
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45 24 Carbon preference index (CPI) and isoprenoids ratio. From the relative areas in
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47 25 TICC for the n-alkanes and isoprenoid identified in Figure 1, it was possible to calculate the
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49 26 CPI31, Pr/Ph, Pr/n-C17 and Ph/n-C18 ratios for each crude oil blend. These ratios, which are
50 27 summarized in Table 1, can be used to predict organic geochemistry properties for crude oils.
51
52 28 The CPI > 1 for all crude oils show the predominance of odd n-paraffins, suggesting that
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54 29 these samples display medium thermal maturity levels and display characteristics of
55 30 lacustrine oils, associated to the deposition of initial organic matter.32
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3 1 The Pr/Ph < 1 ratios for all samples points to crude oils derived from anoxic
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5 2 depositional environments.31 According to Didyk and co-workers,33 such crude oils can also
6 3 be classified as coming from marine carbonate and hypersaline environments, due the
7
8 4 predominance of phytane. The Pr/n-C17 < 1 ratios suggest also an environment of deposition
9
10 5 related to swampy conditions and water washing, whereas the combination of the Pr/n-C17 <
11 6 1 and Ph/n-C18 > 1 ratios indicates that the samples studied are indeed a mixture of
12
13 7 biodegraded and non-biodegraded oils.
14
15
16 8 In previously studies, Asif and co-workers10 correlated Pr/n-C17 versus Ph/n-C18 and
17 9 Pr/n-C17 versus API gravity (or °API) as indicative of biodegradation process in crude oils.
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19 10 The value of °API is associated with the density of the crude oil, where oils with lower °API
20
21 11 are classified as heavy crude oils. The same correlation in Figure 2 (dotted arrow) shows that
22 12 the Pr/n-C17 and Ph/n-C18 ratio increases in oils with higher levels of biodegradation. Such
23
24 13 ratios for the four crude oils studied herein indicates partial biodegradation with a mixture of
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26 14 mature and immature oils. The same correlation can be seen in Figure 2 (filled arrow) with a
27 15 smaller °API and the Pr/n-C17 ratio tending to one, pointing to crude oils with intermediate
28
29 16 biodegradation levels.
30
31
32 17 Terpanes and steranes biomarkers. The n-alkanes, CPI index, °API, and Pr/Ph,
33 18 Pr/n-C17, Ph/n-C18 ratios previously discussed herein have pointed to oils with quite similar
34
35 19 geochemical properties. To further test such similarity, geochemical ratios obtained from
36
37 20 terpanes and steranes biomarkers were calculated. The terpanes and steranes biomarkers were
38 21 identified from the GC/MS data in SIM mode using theoretical references7, 8, 10-12, 34, 35 and
39
40 22 retention indexes described previously by Hwang36 and Oudot and co-workers.37 Figures 3
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42 23 and 4 show the chromatograms used to selectively detect biomarkers for the class of terpanes
43 24 (SIM mode for the ion of m/z 191) and steranes (SIM mode for the ion of m/z 217),
44
45 25 respectively.
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47
48 26 From the identification of terpanes and steranes biomarkers in Figures 3 and 4, it was
49 27 therefore possible to estimate the levels of thermal evolution and depositional environment of
50
51 28 such crude oils (Table 2). The Ts/Ts+Tm ratios were lower than 1 suggesting high thermal
52
53 29 evolution levels for the all four crude oils. Asif and co-workers10 showed that values for
54 30 Ts/Ts+Tm ratio in the range of 0.31-0.73 indicate oils with medium to high thermal evolution
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56 31 levels. In a previously study of crude oils from the Sergipe-Alagoas Basin, Lima and co-
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3 1 workers34 obtained Ts/Ts+Tm on the range of 0.30-0.83 also indicating medium to high
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5 2 thermal evolution levels.
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7 3 The biomarker C29Ts/(H29+C29Ts) ratio also provided an indication of thermal
8
9 4 evolution because it is known that C29Ts is more thermally stable than H29.7 This ratio was
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11 5 quite similar for all blends (0.27-0.28, Table 2) indicating that all four crude oils exhibited
12 6 medium levels of thermal evolution. Sousa Júnior and co-workers28 have, however, reported
13
14 7 data pointing to low thermal maturity for crude oils of the Carmópolis field at Sergipe-
15
16 8 Alagoas basin. Therefore, crude oils from the same field and same basin may present
17 9 different geochemical characteristics, and the exact information is essential to plan the proper
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19 10 upstream and midstream process in oil industry.
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21
22 11 It has been reported that S/(S+R)αααC29 ratios (Table 2) with values > 0.55 indicate
23 12 partial degrees of biodegradation.7 The CEOL2 sample showed a S/(S+R)αααC29 ratio <
24
25 13 0.55, indicating that steranes were not affected by the biodegradation process.
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27 14 S/(S+R)αααC29 ratios > 0.55 for other three samples, however, indicate considerable
28 15 steranes biodegradation. According to Peters and co-workers,31 such biodegradation is caused
29
30 16 by the selective removal of organic epimer αααC29R by bacteria.
31
32
33 17 Frequently, the percentage of cholestanes, ergostanes and stigmastanes classes of
34 18 steranes (%C27, %C28 and %C29 αββ) have been used to indicate conditions of deposition
35
36 19 from organic matter to crude oils.38 From Table 2, the distribution of such compounds
37
38 20 showed the greatest value for %C29αββ (approximately 50%), indicating for all crude oils a
39 21 terrigenous characteristic for the deposition of organic matter.10, 12, 39
These geochemical
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41 22 aspects were consistent with the origin for the crude oil blends under study, because they all
42
43 23 came from the process of onshore exploration.
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45 24 Multivariate analysis applied to biomarkers ratio. The biomarker data so far points
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47 25 to subtle differences in geochemical properties for all samples. Multivariate data analysis
48
49 26 tools were then applied to better assess such similarities using parameters indicative of
50 27 thermal evolution, biodegradation and depositional environments summarized in Table 2. The
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52 28 standardization method of data was performed using autoscaling (data mean-centering
53
54 29 followed by variance scaling). Note that the geochemical parameters Ts/(Ts+Tm),
55 30 C29Ts/(H29+C29Ts) and Ts/H30 failed to contribute to the multivariate data analysis and
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57 31 thus were excluded.
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3 1 Figure 5(A) shows the PCA plot for from the biomarker parameters presented in
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5 2 Table 2 for the crude oils blends, with a variance of 57.14% to PC1 and 39.91% to PC2, and
6 3 a total cumulative variance of 97.05%. Note that to obtain the correlation, parameters with
7
8 4 correlation values above ±0.6 were considered in Figure 5(B). From Figures 5(A-B), PC1
9
10 5 was found to most significantly contribute to differentiation with a positive weight for the
11 6 following parameters: Dia 27(S+R)/αααC27 (S+R) in correlation with the crude oil CEOL2,
12
13 7 and %C28 and %C29 in correlation with the crude oil CEOL3. However, a more significant
14
15 8 discrimination was attained in PC2, where the parameters influenced the formation of two
16 9 distinct groups. The first group was formed by CEOL3 and CEOL2 samples with a positive
17
18 10 correlation with the Dia27(S+R)/αααC27(S+R), %C28 and %C29 parameters. The second
19
20 11 group was formed by the CEOL1 and CEOL4 samples with a positive correlation with the
21 12 S/(S+R) αααC29, αββ (S+R)/αββ + ααα (S+R) C29 and %C27 parameters. As described in
22
23 13 the last section, these parameters are attributed to conditions of organic matter deposition
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25 14 during formation of crude oils.
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27 15 We also constructed a HCA dendrogram (Figure 5 (C)) to better visualize contrasting
28
29 16 features among the four crude oils blends using the Euclidean distance for Ward's method.
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17 The dendogram reinforces the trends pointed by the PCA analysis in Figure 5 (A-B). In
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32 18 Figure 5(C) the CEOL4 and CEOL1 samples were shown to be approximately 95% similar in
33
34 19 terms of thermal maturity and biodegradation, whereas the CEOL3 samples not presented
35
36
20 significant similarity with the another samples. In all, we can conclude from the GC/MS data
37 21 that the four crude oils blends studied here are formed by a mixture of different crude oils
38
39 22 with distinct geochemical characteristics, despite being from the same field of production and
40
41
23 the same basin.
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43 24 Petroleomics approach. Figure 6(A) shows the ESI(+) FT-ICR MS for the crude oils
44
45 25 blends with high abundant ions centered around m/z 550. From these spectra, the PetroMS
46
26 software was able to assigned 1610, 1477, 1622 and 2296 different molecular formulas to the
47
48 27 CEOL1, CEOL2, CEOL3 and CEOL4 samples, respectively, with an m/z error < 1 ppm. The
49
50 28 chemical formulas were considered to correspond to CcHhNxOxSx with c=1-100; h=1-1000
51
29 and x-1-4. Figure 6B summarizes the abundance of each class. Note that for all crude oils the
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53 30 N-class was far the most abundant, which correspond to mainly pyridines and their
54
55 31 derivatives. Minor abundant NO, N2 and N2S2 classes were also detected for all four samples.
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32 The NOS and O4S classes were only detected for CEOL4, indicating the presence of higher
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3 1 MW sulfur compounds. This finding corroborates with a previously study using energy
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5 2 dispersive X-ray fluorescence spectrometry technique.29
6
7 3 Figure 6(C) shows the DBE relative abundance distribution plot for the N-class,
8
9 4 which are used to indicate the thermal evolution and aromaticity levels in crude oils.40, 41 A
10
11 5 higher thermal evolution level over geologic time tends to form aromatic rings, and
12 6 consequently increases the DBE levels of crude oils. Figure 6(C) shows that CEOL2 and
13
14 7 CEOL3 display lower thermal evolution levels as indicated by highest DBE at the 4-12 range,
15
16 8 whereas for the CEOL1 and CEOL4 higher DBE numbers were found at the 13-26 range.
17 9 This finding suggests higher unsaturation levels for the CEOL1 and CEOL4 oils as compared
18
19 10 to CEOL2 and CEOL3 oils. The ESI(+) FT-ICR MS data is also in agreement with the
20
21 11 GC/MS biomarker results. In general, the four samples oils displayed characteristics of crude
22 12 oils with high to medium thermal maturity levels which is slightly higher for the CEOL1 and
23
24 13 CEOL4 oils.
25
26
27 14 Figure 7(A) shows now the ESI(-) FT-ICR MS data for the crude oils blends, where
28 15 most abundant anions are centered around m/z 420. From such MS data, a total of 1935,
29
30 16 2349, 2326 and 2876 molecular formulas could be assigned for samples CEOL1, CEOL2,
31
32 17 CEOL3 and CEOL4, respectively with an m/z error < 1 ppm when considering CcHhNxOxSx
33 18 formulas with c=1-100; h=1-1000 and x-1-3. Figure 7(B) shows the main classes assigned for
34
35 19 the four samples. Note that when ESI(-) was used, O2-class was the major class, which
36
37 20 corresponds to fatty acids, naphthenic acids and/or derivatives, followed by the N-class which
38 21 corresponds to pyrroles and their derivatives. Minor components from the NO, NO2, O and
39
40 22 O3-classes could also be assigned for all four blends. According to Pan and co-workers,42 low
41
42 23 abundances for the N-class in ESI(-) suggested more biodegraded oils. Since the four crude
43 24 oils displayed quite contrasting abundances for the N-class during ESI(-), they seem to
44
45 25 possess different levels of biodegradation. Higher degree of biodegradation was attributed to
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47 26 the CEOL1 and CEOL4 samples due to lower abundance of N-class. According to Vaz and
48 27 co-workers,17 biodegraded oils also display lower abundances for the NO-class which is
49
50 28 generated from oxidation of the N-class compounds due to aerobic or anaerobic
51
52 29 biodegradation. From Figure 7(B) it is clear that this characteristic can be assigned to the
53 30 CEOL1 and CEOL4 samples. Liao and co-workers21 suggested that the selective removal of
54
55 31 N-class and/or the generation of O2-class is more evident when the biodegradation level
56
57 32 increase, as observed herein for the CEOL1 and CEOL4 samples.
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3 1 The biodegradation level can also be assessed from the oxygen content, with the most
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5 2 biodegraded oils displaying higher abundances for the O2-class and consequently lower
6 3 abundances for the O-class due the O-to-O2 oxygenation that occurs during organic matter
7
8 4 deposition.17, 42, 43 Again, the CEOL1 and CEOL4 oils were, using the O-to-O2 ratio, more
9
10 5 biodegraded than the CEOL2 and CEOL3 samples. Higher abundances of the N-class
11 6 (carbazoles and derivatives) were also observed by ESI(-) FT-ICR MS (Figure 7) for CEOL2
12
13 7 and CEOL3 oils, in accordance with higher abundances of the O2-class. The CEOL3 is the
14
15 8 most emulsified oil, as showed previously by Santos and co-workers29, with seems therefore
16 9 to indicate that higher N-class abundances results in higher tendency to form stable
17
18 10 emulsions. The trends in biodegradation and thermal maturity suggested by the ESI(±) FT-
19
20 11 ICR MS have therefore indeed complemented the geochemical predictions based on GC/MS
21 12 classical biomarkers.
22
23
24 13 Figure 7 (C) shows plots of DBE versus relative abundance distributions for the O2-
25
26 14 class. Note that for all four crude oil samples, O2-class is formed mainly by fatty acids (DBE
27 15 = 1), naphthenic acids with 1-6 rings (DBE = 2-7) and higher aromatic compounds (DBE >
28
29 16 7). The four samples also showed highest O2-class abundances in the lower DBE range, with
30
31 17 maximum abundances for DBE = 5 (a carboxylic acid with four rings) for CEOL1, CEOL3
32 18 and CEOL4 in DBE = 1 for CEOL2 (aliphatic carboxylic acid). Note also that we have found
33
34 19 the higher abundances for naphthenic acids with DBE = 2-7.
35
36
37 20 Biodegradation levels were also evaluated via van Krevelen diagrams constructed as a
38 21 function of carbon number (Cn) for the O2-class (Figure 8), where specific regions for fatty
39
40 22 acids (H/C > 2.0), naphthenic acids (H/C from 1.7 to 2.0) and aromatics (H/C < 1.7) can be
41
42 23 clearly delineated. Naphthenic acids are predominant for all oils, but such constituents are of
43 24 highest abundance for CEOL4 as demonstrated by the heat map.
44
45
46 25
47
48
49 26 CONCLUSIONS
50
51 27 A comprehensive geochemical characterization of onshore crude oils blends from
52
53 28 Sergipe-Alagoas sedimentary basin has been performed using both the classical approach of
54
55
29 GC/MS comparison of biomarker profiles or ratios such as those represented by the n-
56 30 alkanes, isoprenoids, terpanes and steranes, as well as geochemical predictions based on
57
58 31 profiles of polar compounds as revealed by ESI(±) FT-ICR MS.
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3 1 The combination of the GC/MS and FT-ICR MS data suggested that the four crude oil
4
5 2 blends investigated were comprised of paraffinic and lacustrine crude oils with intermediate
6 3 levels of thermal evolution and biodegradation The geochemical similarity was quite high for
7
8 4 the CEOL1 and CEOL4 samples, which are samples with higher thermal maturity levels and
9
10 5 more biodegraded than the CEOL2 and CEOL3 crude oil blend, and the data has also showed
11 6 valuable insight to the field of organic geochemistry. For GC/MS and FT-ICR MS data
12
13 7 herein presented should provide key information to understand the behavior of the crude oils
14
15 8 from Sergipe-Alagoas sedimentary basin, and may generate important information for the
16 9 upstream and midstream processes in the Brazilian oil industry.
17
18
19 10
20
21
11 AUTHOR INFORMATION
22
23
24 12 Corresponding Author
25
26 13 *E-mail: albertowj@ufs.br. Tel.: +55 79 2105 6654.
27
28
29 14 Notes
30
31 15 The authors declare no competing financial interest.
32
33
34 16
35
36
37 17 ACKNOWLEDGMENTS
38
39 18 We would like to thank the Fundação de Amparo à Pesquisa do Estado de São Paulo
40
41 19 (FAPESP) for the scholarship awarded to J.M.S. (process number 2013/19161-4),
42
20 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Conselho
43
44 21 Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (Brazilian research councils)
45
46 22 for fellowships. The authors are grateful to PETROBRAS/UOSEAL/ENGP/LABF for the
47
23 financial support through the cooperation term – 0050.0078683.12.9.
48
49
50 24
51
52
53 25
54
55 26
56
57
58 27 REFERENCES
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25 21 2973.
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29
30 25
31
32 26
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34 27
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36 28
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38 29
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40 30
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42 31
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44 32
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46 33
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53 36
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55 37 List of Tables and Figures
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57 38
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3 1 Table 1. CPI and Pr/Ph, Pr/n-C17, Ph/n-C18 ratios of the four crude oils blends obtained by relative areas in
4 2 TICC for the n-alkanes and isoprenoid using GC/MS
5
6
Sample CPI* Pr/Ph Pr/n-C17 Ph/n-C18
7
8
CEOL1 1.13 0.70 0.61 1.19
9
10 CEOL2 1.15 0.78 0.67 1.10
11
12 CEOL3 1.15 0.82 0.72 1.19
13 CEOL4 1.15 0.71 0.59 1.16
14
15 3
16
4 Table 2. Geochemical parameters (based on the peak heights from GC/MS data in SIM mode) from the class of
17
18 5 terpanes and steranes biomarkers for the crude oils blends of the Oiteirinhos II Sation, Carmópolis-SE
19
20
Sample
21 Parameters
CEOL1 CEOL2 CEOL3 CEOL4
22
23 Ts/(Ts+Tm)a 0.45 0.44 0.45 0.44
24 C29Ts/(H29+C29Ts)b 0.27 0.28 0.27 0.28
25 Ts/H30c 0.56 0.58 0.56 0.58
26 S/(S+R) αααC29d 0.70 0.40 0.69 0.85
27 αββ (S+R)/αββ + ααα (S+R) C29e 0.73 0.63 0.74 0.76
28
Dia 27(S+R)/ααα C27 (S+R)f 1.34 1.97 1.51 1.49
29
30 %C27g 29.49 29.35 26.99 29.59
31 %C28h 20.10 20.42 22.10 20.07
32 %C29i 50.41 50.23 50.91 50.34
33 6 a
Ts = C27 18α(H)-22,29,30-Trisnorneohopane; Tm = C27 17α(H)-22,29,30-Trisnorhopane
34 7 b
C29Ts = 18α(H)-30-norneohopane C29; H29 = 17α,21β(H)-30-norhopane
35 c
36 8 Ts = C27 18α(H)-22,29,30-Trisnorneohopane; H30 = 17α, 21β(H)-hopane
d
37 9 αααC29(S) = 5α,14α,17α (H) - Stigmastane 20S; αααC29(R) = 5α,14α,17α (H) - Stigmastane 20R
38 10 e
αββC29 (S+R) = 5α,14β,17α (H) - Stigmastane 20S + 5α,14β,17α - Stigmastane 20R; αααC29(S+R) = 5α,14α,17α -
39 11 Stigmastane 20S + 5α,14α,17α - Stigmastane 20R
40 f
41 12 Dia 27(S+R) = 13β,17α – Diacholestane 20S + 13β,17α – Diacholestane 20R; αααC27 (S+R) = 5α,14α,17α - Cholestane
42 13 20S + 5α,14α,17α - Cholestane 20R
43 14 g
[C27 5α,14α,17α - Cholestane 20S + C27 5α,14α,17α - Cholestane 20R / C28 5α,14β,17β - Ergostane 20R + C28
44 15 5α,14β,17β - Ergostane 20S + C29 5α,14β,17α - Stigmastane 20R + C29 5α,14β,17α - Stigmastane 20S] x 100
45 h
46 16 [C28 5α,14β,17β - Ergostane 20R + C28 5α,14β,17β - Ergostane 20S / C28 5α,14β,17β - Ergostane 20R + C28 5α,14β,17β
47 17 - Ergostane 20S + C29 5α,14β,17α - Stigmastane 20R + C29 5α,14β,17α - Stigmastane 20S] x 100
48 18 i
[C29 5α,14β,17α - Stigmastane 20R + C29 5α,14β,17α - Stigmastane 20S / C28 5α,14β,17β - Ergostane 20R + C28
49 19 5α,14β,17β - Ergostane 20S + C29 5α,14β,17α - Stigmastane 20R + C29 5α,14β,17α - Stigmastane 20S] x 100.
50
51
52 20
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45
46
47
48
49
50 1
51
52 2 Figure 1. Total ion current chromatogram (TICC) of four crude oils obtained in the SCAN mode, showing the
53 3 distribution in terms of n-alkanes (n-C10-35) and specific isoprenoids (a: 2,6-dimethylundecane; b: 2,6,10-
54
55 4 trimethylundecane; c: 2,6,10-trimethyldodecane; d: 2,6,10-trimethyltridecane; e: 2,6,10-trimethylpentadecane;
56 5 Pr: pristane and Ph: phytane).
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3 1
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7
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18
19 3
20
21 4 Figure 2. (A) Pr/n-C17 versus Ph/n-C18 and (B) Pr/n-C17 versus °API as biodegradation indicative for crude oils
22
5 blends studied.
23
24
25 6
26
27 (x100,000)
1.75 191.00 (1.00)

CEOL 1
1.1
(x100,000)
191.00 (1.00)

CEOL 3
28 1.50
1.0

0.9

29 1.25 0.8

0.7

30 1.00
0.6
10
10
31 0.75

1
5 7 9
0.5

0.4
1 3 4
5
6
7
8
9

1112
2 3 4 6 2 13 14 15
32 0.50
8 11 12 13 14 15
0.3

0.2
0.25

33 36.0 37.0 38.0 39.0 40.0 41.0 42.0 43.0 44.0 45.0 46.0 47.0
0.1

36.0 37.0 38.0 39.0 40.0 41.0 42.0 43.0 44.0 45.0 46.0 47.0

34
(x100,000) (x100,000)
191.00 (1.00) 191.00 (1.00)
1.4
1.3
CEOL 2 1.3
CEOL 4
1.2

35 1.1
1.2

1.1
1.0

36 0.9

0.8
1.0

0.9

37 0.7

0.6
10
0.8

0.7 10

38
0.6
9 5 7 9
0.5 5 7
1 3 6
0.5
1 3 6
0.4 2 4 11 2 4 8 1112
8 12 15 0.4
13 14 15
39 0.3

0.2
13 14
0.3

0.2

40 0.1

36.0 37.0 38.0 39.0 40.0 41.0 42.0 43.0 44.0 45.0 46.0 47.0
0.1

36.0 37.0 38.0 39.0 40.0 41.0 42.0 43.0 44.0 45.0 46.0 47.0

41 7
42
43 8 Figure 3. GC/MS in SIM mode for the ions of m/z 191 (terpanes biomarkers) for the crude oils blends of the
44
45 9 Oiteirinhos II Sation, Carmópolis-SE. 1: 18α(H)-22,29,30-Tris-nor-neo-hopano; 2: 17α(H)-22,29,30-Tris-nor-
46 10 hopano; 3: 17α,21β(H)-30-nor-hopano; 4: 18α(H)-30-nor-neo-hopano; 5: 17α, 21β(H)-hopano; 6: 17β, 21α (H)-
47 11 hopano; 7: 17α, 21β(H)-29-homo-hopano; 8: 17α, 21β(H)-29-homo-hopano; 9: 17α, 21β(H)-29-bis-homo-
48
49 12 hopano; 10: 17α, 21β(H)-29-bis-homo-hopano; 11: 17α, 21β(H)-29-tris-homo-hopano; 12: 17α, 21β(H)-29-tris-
50 13 homo-hopano; 13: 17α, 21β(H)-29-tetrakis-homo-hopano; 14: 17α, 21β(H)-29-tetrakis-homo-hopano; 15: 17α,
51
52 14 21β(H)-29-pentakis-homo-hopano.
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2 (x100,000) (x10,000)

3 1.4

1.3
217.00 (1.36)

CEOL 1 9.0
217.00 (1.57)

CEOL 3

4 1.2

1.1
6
8.0

1
6
1 7.0

5
1.0

0.9 6.0

0.8
2
2 7
6 0.7

0.6
12 5.0

4.0
7
12
10
7 0.5

0.4
3 4 8
9
10 11

13
14 3.0
3 4 8
9
11

13
14
5 5
8 0.3

0.2
2.0

1.0

9 0.1

34.50
(x100,000)
34.75 35.00 35.25 35.50 35.75 36.00 36.25 36.50 36.75 37.00 37.25 37.50 37.75 38.00 38.25 38.50 38.75 39.00 39.25 39.50 39.75 40.00 34.50
(x100,000)
34.75 35.00 35.25 35.50 35.75 36.00 36.25 36.50 36.75 37.00 37.25 37.50 37.75 38.00 38.25 38.50 38.75 39.00 39.25 39.50 39.75 40.00

1.1 217.00 (1.53) 217.00 (1.69)

10 1.0
CEOL 2 1.3

1.2
CEOL 4
6
11 0.9

0.8
1 1.1

1.0
1 6

12 0.7
2
7
0.9

0.8 2
0.6 7
13 0.5
10
12 0.7

0.6
10
12

11 11
14 0.4

0.3
3 4
5
8
9
13
14 0.5

0.4
3 4
5
8
9 13
14

15
0.3
0.2
0.2

0.1
0.1

16 34.75 35.00 35.25 35.50 35.75 36.00 36.25 36.50 36.75 37.00 37.25 37.50 37.75 38.00 38.25 38.50 38.75 39.00 39.25 39.50 39.75 40.00 34.50 34.75 35.00 35.25 35.50 35.75 36.00 36.25 36.50 36.75 37.00 37.25 37.50 37.75 38.00 38.25 38.50 38.75 39.00 39.25 39.50 39.75

17 1
18
19 2 Figure 4. GC/MS in SIM mode for the ion of m/z 217 (steranes biomarkers) for the crude oils blends of the
20 3 Oiteirinhos II Sation, Carmópolis-SE. 1: 13β, 17α – Diacolestano 20S; 2: 13β, 17α – Diacolestano 20R; 3: 5α,
21
22 4 14α, 17α - Colestano 20S; 4: 5α, 14β, 17β - Colestano 20R; 5: 5α, 14β, 17β - Colestano 20S; 6: 5α, 14α, 17α -
23 5 Colestano 20R; 7: 5α, 14α, 17α - Ergostano 20S; 8: 5α, 14β, 17β - Ergostano 20R; 9: 5α, 14β, 17β - Ergostano
24
6 20S; 10: 5α, 14α, 17α - Ergostano 20R; 11: 5α, 14α, 17α - Estigmastano 20S; 12: 5α, 14β, 17α - Estigmastano
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26 7 20R; 13: 5α, 14β, 17α - Estigmastano 20S; 14: 5α, 14α, 17α - Estigmastano 20R.
27
28 2.5
(A) 1.0 (B)
29 2.0
%C28
30 CEOL3 Dia 27(S+R)/aaa C27 (S+R)
1.5
31 CEOL2
0.5 %C29
32 1.0
Factor 2 : 39.91%

33
PC2: 39.91%

0.5
34
0.0 0.0
35
36 -0.5

37 -1.0 CEOL1
aßß-0.5
(S+R)/aßß + aaa (S+R) C29
38 -1.5
CEOL4 S/(S+R) aaaC29 %C27
39
-2.0
40 -1.0
41 -2.5
-4 -3 -2 -1 0 1 2 3 4 -1.0 -0.5 0.0 0.5 1.0
42
PC1: 57.14% PC1: 57.14%
43
44 (C)
45
46 CEOL1

47
48 CEOL4
49
50
51 CEOL2
52
53
CEOL3
54
55
56
0 20 40 60 80 100 120
57
(Dlink/Dmax)*100
58 8
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