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Multi-spectroscopic characterization of bitumen and its polarity-based

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 Construction and Building Materials 352 (2022) 128992

Contents lists available at ScienceDirect

Construction and Building Materials

journal homepage: www.elsevier.com/locate/conbuildmat

Multi-spectroscopic characterization of bitumen and its

polarity-based fractions
Stefan Werkovits a, Markus Bacher b, Johannes Theiner c, Thomas Rosenau b, Hinrich Grothe a, *
a
Christian Doppler Laboratory for Chemo-Mechanical Analysis of Bituminous Materials, Institute of Materials Chemistry, TU Wien, Getreidemarkt 9/E-165-01-5, 1060
Vienna, Austria
b
University of Natural Resources and Life Sciences Vienna, Division of Chemistry of Renewables, Konrad-Lorenz-Straße 24, A-3430 Tulln, Vienna, Austria
c
University of Vienna, Microanalysis Services, Austria

A R T I C L E I N F O A B S T R A C T

Keywords: Bitumen contains a complex mixture of molecules, ranging from simple linear alkanes to complex polyaromatic
Bitumen systems. Used in road construction, those structures are permanently influenced by atmospheric processes, which
SARA fractions result in oxidation, degradation, loss of volatiles and structural rearrangements. In order to understand those
NMR
31 molecular changes, a profound description of the unaltered system is needed. In this study, structural and
P NMR
HSQC
chemical peculiarities of bitumen and its polarity-based fractions were analysed using nuclear magnetic reso­
HMBC nance (NMR) techniques in combination with infrared/fluorescence spectroscopy and elemental analysis. The
DOSY bitumen sample was separated into saturates, aromatics, resins and asphaltenes. Elemental analysis provided
Elemental analysis insights into variations of heteroatom content and hydrogen-carbon ratio among the fractions. Fluorescence
Infrared spectroscopy spectroscopy showed an increase in the aromatic condensation degree with rising polarity. The combination of
Fluorescence spectroscopy heteronuclear single quantum coherence (HSQC) and elemental analysis facilitated the assessment of length and
branching extent of aliphatic side chains, throughout the fractions. Another 2D NMR technique, heteronuclear
multiple bond coherence (HMBC) found evidence for minor contributions of carboxylic esters or amides within
the resins. Hydroxyl and amino groups were analysed by 31P NMR after derivatization, revealing minor amounts
of alcohols, amides and carboxylic acids in the most polar fractions. Additionally, diffusion ordered spectroscopy
was carried out to obtain estimations on average molecular sizes and weights. Applying in-depth NMR analysis to
bitumen and combining it with complementary analytical techniques, this study provides a unique basis for
future ageing studies and demonstrates how multi-spectroscopy can give new insights into bitumen structure and
chemistry.

1. Introduction
Throughout history, a diverse palette of construction materials has
been loyal companion to mankind. Materials such as wood, glass, steel,
concrete, and asphalt became necessities in the founding of towns and
the development of complex road networks connecting those centres of
human activities. This facilitated the exchange of material goods, cul­
tural values and knowledge, accelerating the development of modern
societies. Those advances relied on the robustness and longevity of
transport structures. However, as roads are permanently exposed to
environmental influences, such as solar radiation, oxidative gases from
the atmosphere and huge temperature differences (during production
and service life), their lifetime is shortened. In order to prevent deteri­
oration, extensive research has been implemented to assure the quality,
improve mechanical performance, and understand the fundamental
physical and chemical processes [1,2] in the road construction
materials.
Regarding the chemical complexity of these materials, asphalt ranks
near the top. The predominant reasons are the varying origins and
production processes of the bituminous binder. Bitumen is a product of
the crude-oil refining process and is obtained as the residue of the vac­
uum distillation. Only two chemical elements, hydrogen and carbon,
account for over 90 wt% of the material [2]. Further elements present in
considerable amounts are sulfur, oxygen and nitrogen, and also some

Abbreviations: NMR, nuclear magnetic resonance; HSQC, heteronuclear single quantum coherence; HMBC, heteronuclear multiple bond coherence; DOSY,
diffusion ordered spectroscopy; SARA, saturates aromatics resins asphaltenes.
* Corresponding author.

https://doi.org/10.1016/j.conbuildmat.2022.128992
Received 29 May 2022; Received in revised form 8 August 2022; Accepted 25 August 2022
Available online 5 September 2022
0950-0618/© 2022 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
S. Werkovits et al.
transition metals (V, Ni, Fe) are found in traces (ppm)[2]. On a molec­
ular level, several functional classes have been found, comprising
aliphatic and (poly-)aromatic structures, oxygen-containing (carbonyl,
alcohol), sulfur-containing (sulfide, (benzo)thiophene) and nitrogen-
containing (pyridine, pyrrole, quinoline and porphyrin) moieties
[3–5]. These structural entities are not isolated but rather appear in
different combinations. The fascinating question that arises is how all
those functional groups are (inter)connected and how differences in
these connections affect the material behaviour. Especially the interre­
lation between rheological and chemical behaviour and how both
change during standardized ageing procedures was emphasized in
thorough studies in the recent literature, building a robust framework to
link both worlds and showing the extraordinary importance of chemis­
try for the analysis and performance of the material [6–9]. The chemical
evaluation is, however, challenging, due to the almost unlimited number
of possible molecular structures. The task of determining a single com­
pound is both meaningless and (almost) impossible, and structural
characterization has to address the complex compound mixture by
suitable sum parameters. In this regard, structural characterization of
bitumen is similar to that of other overly complex natural products, such
as lignins, tannins or lignocellulosic chromophores, which are also
characterized as combination of characteristic structural elements and
functional groups in the bulk [10–12].
To lower the complexity of the overall bitumen system, it is
commonly separated into its polarity-based fractions, the so-called
SARA (saturates, aromatics, resins and asphaltenes) fractions [13].
This facilitates the qualitative analysis of the material and provides in­
formation on polarity distribution and ageing state of the bitumen
samples. For example, overlapping spectroscopic responses in various
spectroscopic measurements can be avoided by a preceding separation
step [4,5,8,9,14–16]. The fractionation provides, on the one hand, more
detailed information on bitumen chemistry (e.g. presence and concen­
tration of carbonyls, alcohols, aromatic structures, sulfur and nitrogen
containing groups) and, on the other hand, the possibility to monitor
ageing processes with improved accuracy.
However, the complex structure and chemistry of bitumen is far from
being fully understood and a single separation or analysis technique
alone cannot provide all the necessary information. Therefore, research
groups attempt to combine multiple spectroscopic techniques to validate
and increase the amount of obtainable information [17–19]. In this re­
gard, NMR spectroscopy stands out by the depth of molecular structural
information attainable. It provides the possibility to detect certain nuclei
(e.g. 1H, 13C, 31P) and resolves their chemical environment at a molec­
ular scale. This leads to information on directly bonded elements,
functional groups, structural motifs, molecular classes and their distri­
bution, in an ideal case providing the chemical structure of the whole
molecule. Other techniques address physicochemical molecular pa­
rameters, such as bond rotation, movements of molecular sections, size
of molecules and how fast the molecular structures are able to move in
the respective medium. It can also be used to analyse how structural and
chemical features change during the ageing process, which is particu­
larly valuable in bitumen research. In thorough studies, Pipintakos
[17,20], Lu [20], Herrington [21], Michon [22,23] and Siddiqui [24]
showed how 1H and 13C NMR techniques can be used to assess structural
changes during simulated ageing procedures in the laboratory, resulting
in proposed ageing mechanisms. The main problem in analysing
bitumen is that there is not one structure present but a rather innu­
merable quantity of different molecules, each producing a specific nu­
clear response by themselves, resulting in highly overlapping signals. In
recent years, researchers tried to solve this problem in two ways: 1)
combining separation methods (see above) with NMR analysis [5,18,25]
and 2) using 2D NMR techniques to better resolve overlapping signals
[26–28]. Silva et al. provided a comprehensive review on the usage of
NMR techniques in the analysis of heavy crude oil mixtures [29]. An
interesting example is given in the work of Shikhov [26], which
employed HSQC as an advanced characterization method to track which
2
Construction and Building Materials 352 (2022) 128992
compounds of oil mixtures are adsorbed on sandstones throughout an
ageing process. The adsorbed substances were compared to HSQC
spectra of the SARA fractions of bitumen (one part of the oil mixtures).
There is less literature on application of HMBC spectra in bitumen
research available, one example is the study of Elbaz [28]. Another
approach was implemented by Durand [30], who focused on DOSY NMR
to assess diffusion coefficients, molecular sizes and weights of hydro­
carbon mixtures and asphaltenes. Based on DOSY NMR, Vukovic et al.
[31] focused extensively on different types of asphaltenes and analysed
their aggregation behaviour in solution.
All these examples illustrate how the wide landscape of NMR tech­
niques can be utilized to analyse different structural features of the
highly complex hydrocarbon mixtures in bitumen. However, attempts to
combine all this information in one coherent picture are still largely
missing, and the present study attempts to advance in this direction,
providing not only a more detailed description of the mixture of interest,
but also a promising and robust basis of future bitumen ageing studies.
2. Materials and methods
2.1. Materials
A non-modified bitumen, as commonly used in road engineering,
classified as a 70/100 penetration graded bitumen, was employed.
Detailed specification is given in Table 1.
2.2. SARA fractionation
SARA fractions of the bitumen sample were obtained using a stan­
dardized chromatographic technique [32]. For each SARA separation
400 ± 40 mg of the binder (bitumen) was weighed in a 250 ml jar. 40 ml
of HPLC-grade n-heptane was added and the mixture magnetically
stirred for 24 h ± 2 h at 200 rpm. The separation of the asphaltenes (n-
heptane insoluble) was achieved using syringe filters (Thermo Scienti­
fic™ Titan3™ PTFE (Hydrophobic) Syringe Filters, 25 mm diameter,
0.2 µm pore size). The maltenes were collected as filtrate in four vials, of
which two were evaporated to dryness on a heating plate at a temper­
ature of 120 ◦ C, while being flushed with nitrogen. The maltene content
of the samples was calculated according to Eq. (1).
4*mmaltenes,dried
mmaltenes % = *100 (1)
mbitumen
mmaltenes%… percentual share of maltenes (%).
mmaltenes,dried … mean mass of the dried maltene fractions (g).
mbitumen… mass of the bitumen sample (g).
Subsequently, the remaining maltenes were diluted to a concentra­
tion of 3.33 mg/ml. For the solid phase extraction (SPE) of the maltenes,
SPE cartridges (Thermo Scientific™ HyperSep™ Silica Cartridges, 25 ml
volume, 40–60 μm particle size) were mounted onto a vacuum manifold
and 15 ml of the maltene solution was added to the SPE cartridge. After
the addition of 10 ml of n-heptane, the cartridges were moved to the next
position on the manifold. In this position the mobile phase was changed
and 25 ml of a mixture of toluene and n–heptane (80:20, v/v) was added.
The cartridges were moved to a third position, where 40 ml of a mixture
of dichloromethane and methanol (90:10, v/v) was added. The obtained
SAR-fractions were evaporated on a heating plate under N2 flushing
Table 1
Rheological parameters of the bitumen sample.
Rheologic parameter Obtained value
Penetration at 25 ◦ C 91 mm
Performance grade 58–22
Softening point 47 ◦ C
S. Werkovits et al.
until constant masses were achieved. The remaining asphaltene frac­
tions were dissolved in toluene, and the solution transferred into glass
vials and evaporated in a similar way.
2.3. Elemental analysis of bitumen and its fractions
Carbon, hydrogen, nitrogen and sulfur were analysed in a single run
on an EA 3000 CHNS-O analyser (Eurovector) with flash combustion at
1000 ◦ C. The combustion products N2, CO2, H2O and SO2 were analysed
by on line-gas chromatography with thermal conductivity detection. Data
were recorded by the Callidus software. The analysis of oxygen is based
on high temperature pyrolysis of material in a HT-1500 high temperature
oven (Hekatech) using granulated carbon as the reducing agent. Diges­
tion at 1480 ◦ C creates only carbon monoxide from all kinds of oxygen in
the samples. Data evaluation was performed with in-house software tools
optimised for low-level contents of N, S and O. The LOQ (limit of quan­
tification) was 0.05 wt% (500 ppm) for C, H, N and O and 0.02 wt% (200
ppm) for S, according to long term laboratory evaluations. The uncer­
tainty of element quantification is smaller than 0.3 wt%. Quantitative
manipulation of small amounts of high viscous and sticky bitumen ma­
terial might be unreliable and was therefore replaced by a two-step
sample preparation: The sample was dissolved in cyclohexane and
transferred into tin crucibles used for analysis by a micro-pipette. The
solvent was removed by evaporation at 105 ◦ C over 30 min. Attenuated
total reflection infrared spectroscopy (ATR-IR) spectroscopy was used to
verify the complete removal of the solvent. The constant sample mass
after drying was recorded on an ultra-micro-balance (Cubis MCA2,
Sartorius, ±0.1 µg) and used as 100 % reference. Reference measure­
ments directly with the native material verified the validity of this sample
preparation. Triplicate analysis was performed. A sum of 100 ± 0.6 wt%
confirmed complete coverage of elemental constituents.
2.4. Infrared spectroscopy
IR spectra of the fractions were obtained on a Vertex 80v FTIR
spectrometer (Bruker, Karlsruhe) in attenuated total reflection mode
with a diamond crystal. The spectral range was 680–4000 cm− 1, a
spectral resolution of 1 cm− 1 was chosen and 48 scans were accumulated
for each spectrum. The spectra were normalized at 2920 cm− 1. To
measure the saturates, aromatics and resins, a small sample size of each
fraction was applied directly on the crystal with a spatula. The asphal­
tenes were applied in solution (in dichloromethane) and IR spectra were
recorded after complete evaporation of the solvent (fading signal in the
IR spectrum). For each fraction, four samples were applied on the crystal
and each sample was measured four times.
2.5. Fluorescence spectroscopy
Photoluminescence spectroscopy was carried out with a fluorescence
spectrometer (FLSP920, from Edinburgh Instruments) equipped with a
XE900 xenon arc lamp, double Czerny-Turner monochromators (type
TMS300) and a S900 single photon photomultiplier (type R928) as the
detector. The measurements in solution were done in dichloromethane at
concentrations of 0.5–1 mg/l in a quartz glass cuvette (500 µl, Hellma™
Suprasil™ quartz, Germany). A non-fluorescent metal substrate was used
for front-face measurements of non-dissolved samples. Fluorescence
Table 2
Parameters of the fluorescence measurements.
Method Excitation wavelength Emission wavelength Emission filter
(nm) (nm) (nm)
Emission 270 340–600/850a 340
scan
EEM 230–500/650a 340–600/440-850a 340/440a
a
solution/front-face set up.
3
Construction and Building Materials 352 (2022) 128992
excitation-emission mapping (FEEM) and emission scans were performed
with wavelength increments of 5 nm (1 nm for emission scans) and dwell
times of 0.25 s. Further experimental parameters are given in Table 2.
2.6. NMR spectroscopy
All NMR spectra were recorded on a Bruker Avance II 400 (resonance
frequencies 400.13 MHz for 1H and 100.63 MHz for 13C) equipped with
a 5 mm N2-cooled cryo probe head (Prodigy) with z–gradients at room
temperature with standard Bruker pulse programs. The sample was
dissolved in 0.6 ml of CDCl3 (99.8 % D). Chemical shifts are given in
ppm, referenced to residual solvent signals (7.26 ppm for 1H, 77.0 ppm
for 13C). 1H NMR data were collected with 32 k complex data points. All
two-dimensional experiments (HSQC, HMBC) were performed with 1 k
× 256 data points, while the number of transients and the sweep widths
were optimized individually. HSQC experiments were acquired in edited
mode using an adiabatic pulse for inversion of 13C and GARP-sequence
for broadband 13C-decoupling, optimized for 1J(CH) = 145 Hz. For the
HMBC spectra a long-range coupling constant of JCH = 8 Hz was used.
2.6.1. DOSY NMR spectroscopy
For measurement of the diffusion coefficients with diffusion-ordered
NMR spectroscopy (DOSY) a pulse sequence with bipolar gradient pulses
for diffusion and eddy current delays was used. The samples were dis­
solved in 0.6 ml CDCl3 and for relative calibration of the spectra one
drop of CH2Cl2 was added. CDCl3 itself was not suited as calibration
reference because of overlap with aromatic signals leading to not clearly
defined peaks in the 2D DOSY spectra. The temperature during mea­
surement was set to 303 K and the diffusion specific parameters were
determined being Δ = 0.15 s and δ = 600 μs. Data were acquired with 16
k data points and a recycle delay of 2 s whereas the gradient strength
was incremented in 16 linear steps from 2 % to 98 % attenuation. Es­
timations on the hydrodynamic radius RH and the average molecular
weight were made according to Eqs. (2) and (3). The dynamic viscosity η
was approximated with the viscosity of the solvent (CDCl3) at 303 K. The
diffusion coefficient D is strongly dependent on the analyte concentra­
tion [30]. To guarantee only minor deviations, the weight percentage
was kept low, within a range of 0.5–1.5 wt% (manalyte/msolvent) for all
samples. Finally, to obtain estimations of molecular weights throughout
the fractions the shape factor α (empirical exponent in equation (2)) was
set to 0.6[30,33].
kB *T
RH = (2)
6*π*η*Di
Di… Measured diffusion coefficient of the sample i (m2/s).
kB… Boltzmann constant (J/K).
T… Temperature (K).
η… Dynamic viscosity of the sample at 303 K (Pa*s).
RH… Hydrodynamic radius (Å).
( )1
Mi = Mref /
Di α
(3)
Dref
Mi… Molecular weight of the sample i (g/mol).
Mref… Molecular weight of dichloromethane (g/mol).
Di… Measured diffusion coefficient of the sample i (m2/s).
Dref… Diffusion coefficient of dichloromethane (m2/s).
α… Average shape factor (-).
2.6.2. 31P NMR spectroscopy
The measurement used a derivatization approach, adapted from a
standard method in lignin chemistry, in which acidic groups (OH, NH,
COOH) are chemoselectively converted into phosphite esters [34]. A
mixture of 15–20 mg sample and 3–4 mg of internal standard
S. Werkovits et al. Construction and Building Materials 352 (2022) 128992

31
Fig. 1. P NMR spectrum of derivatized asphaltenes with characteristic chemical shift regions indicated [35].

Table 3 3. Results and discussion

Yields of the SARA fractions.
Fraction Unaged (wt%) 3.1. SARA fractionation
Saturates 21.8 ± 0.6
Aromatics 45.6 ± 0.4 The SARA fractionation was repeated four times. The average weight
Resins 17.5 ± 0.2 percentages are shown in Table 3. The most abundant fraction was the
Asphaltenes 15.1 ± 0.2 aromatic one followed by saturates, resins and asphaltenes. The content
of low-polar fractions (saturates + aromatics) was approximately 30 wt
% higher than that of the more polar ones (resins and asphaltenes).
(N–hydroxy-5-norbornen-2,3-dicarboxylic acid imide, 97 %) were dis­
solved in 800 μl of a 1:1.6 (v/v) mixture of CDCl3 and pyridine (dried
over molecular sieve, 3 Å). After thorough mixing, 100 μl of phosphi­ 3.2. Elemental analysis of bitumen and its fractions
tylation reagent (2-chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane,
TMDP) and 100 μl of a solution of relaxation reagent (4 mg Cr(III)(acac)3 The elemental composition of the investigated bitumen sample and
/ml CDCl3) were added through a septum into the vial to avoid any its SARA fractions is shown in Table 4. Carbon and hydrogen were the
contact between reagents and moisture. After the addition of TMDP, the most abundant elements in the samples with more than 91 wt% in all
sample was shaken for 1 h at room temperature and subsequently fractions. The H/C-ratio, reflecting the degree of saturation, had the
transferred into a moisture-free NMR tube. For quantitative measure­ highest value in the saturates. The ratio decreased for the fractions with
ments, inverse gated 1H decoupled 31P NMR spectra were acquired with higher polarity. However, contrary to previous publications [4,18], the
64 k data points, a relaxation delay of 14 s and 160 scans. The charac­ order found in this study was: Saturates > Resins > Aromatics >
teristic 31P NMR spectrum of an asphaltene sample in the range between Asphaltenes. This divergence of order compared to other studies in
120 and 180 ppm is shown in Fig. 1. The educt (TMDP), the internal literature is probably correlated to the type of SARA fractionation and to
standard and characteristic chemical shifts [35] of specific OH- the origin of the analysed bitumen. To understand this observation on a
containing groups are highlighted. molecular basis, one has to emphasize at first that the amount of aro­
matic structures and their degree of condensation is increasing from
saturates to asphaltenes (according to literature [2]), resulting in a
saturation order of Saturates > Aromatics > Resins > Asphaltenes.
Assuming that this is also true for our binder, the higher H/C-ratio in
resins compared to aromatics points to the amount and length of

Table 4
Elemental content of the SARA fractions and bitumen.
Fraction C (wt%) H (wt%) N (wt%) O (wt%) S (wt%) H/C-Ratio

Saturates 85.01 ± 0.14 12.95 ± 0.04 < 0.05 0.79 ± 0.17 1.03 ± 0.09 1.81
Aromatics 83.63 ± 0.20 9.61 ± 0.11 0.58 ± 0.02 1.45 ± 0.08 4.54 ± 0.15 1.37
Resins 81.59 ± 0.50 9.96 ± 0.06 0.99 ± 0.03 2.61 ± 0.06 4.88 ± 0.01 1.45
Asphaltenes 82.37 ± 0.21 8.74 ± 0.07 1.01 ± 0.07 1.98 ± 0.03 5.42 ± 0.18 1.26
Bitumen 84.34 ± 0.24 10.24 ± 0.07 0.56 ± 0.03 1.13 ± 0.02 4.33 ± 0.27 1.45

4
S. Werkovits et al.
saturated aliphatic side chains covalently bonded to aromatic structures.
From the heteroatom content in Table 4 it is evident that no
detectable nitrogen groups and only rather little (≤1 wt%) of oxygen
and sulfur-containing moieties were present in the saturates fraction.
The higher N, O and S content in the other fractions is another
contributor to their increased polarity. All three elements are (highly)
electronegative and thus will be bound in groups of high polarity that, in
addition, engage in electrostatic, van der Waals and/or H-bond in­
teractions. The heteroatom content reached a maximum of approx. 9 wt
% in resins and asphaltenes. The most abundant heteroatom in all
samples was sulfur followed by oxygen and nitrogen. Furthermore, the
oxygen content in resins exceeded that in asphaltenes.
3.3. Infrared spectroscopy
Infrared spectra of the fractions showed distinct chemical charac­
teristics (Fig. 2). In the saturates fraction almost all signals were
apparently related to the aliphatic constituents. Minor aromatic bands
were observable at 1600 cm− 1 and 815 cm− 1 indicating the presence of
aromatic structures in the saturates fraction. The signal intensity of the
major aromatic signal increased from saturates to asphaltenes.
Furthermore, sulfoxide signals at 1030 cm− 1 were found within the ARA
fractions, with the highest intensities in resins and asphaltenes. This is
evidently in agreement with the sulfur contents from elemental analysis.
Further indications of oxygen-containing groups came from bands at
Fig. 2. FTIR spectra of the SARA fractions. Left: Fingerprint area. Right: OH/NH region.
Fig. 3. Fluorescence emission (at 270 nm excitation) of the SARA fractions. Left: Dissolved in DCM; right: Front-face measurement of non-dissolved fractions.
5
Construction and Building Materials 352 (2022) 128992
1670 cm− 1 (resins), at 1735 cm− 1 (resins and asphaltenes) and from
broad bands above 3100 cm− 1, corresponding to quinolone [14] con­
taining-, carboxylic acid [14]/ester containing-, and OH/NH-containing
structures, respectively. A detailed analysis of COOH/OH/NH structures
was implemented using 31P NMR (see chapter 3.5.4).
3.4. Fluorescence spectroscopy
The fluorescence measurements of the highly diluted fractions in DCM
are presented in Fig. 3. The position of the emission maximum shifted to
higher wavelengths from saturates to asphaltenes (Fig. 3, left). This
characteristic red shift reflects differences of aromatic systems in the
fractions with an increasing degree of polycondensation of the aromatic
structures. The front-face measurements (Fig. 3, right) of the fractions
showed a red shift in emission for aromatics, resins and asphaltenes.
Those fractions contain high amounts of different aromatic structures
which interfere with each other by reabsorption processes (inner-filter
effects) [36,37]. The reason for the almost unaltered saturates spectrum
was the low abundancy of aromatic structures in this fraction. The high
content of saturated hydrocarbons, which acted as a non-fluorescent
matrix in which the fluorescent aromatic molecules are diluted, pre­
vents such inner-filter effects. Table 5 shows how the spectral regions of
maximum fluorescence intensity in the diluted samples are shifted to­
wards higher emission wavelength with increasing polarity.
S. Werkovits et al. Construction and Building Materials 352 (2022) 128992

Table 5 Table 6
Spectral regions with maximum fluorescence intensity and estimated aromatic Average diffusion coefficient D, hydrodynamic radius RH and molecular weight
condensation degree. Mw of the SARA fractions from 1H DOSY NMR experiments.
Fraction Emission (nm) Number of condensed aromatic rings [38] Sample D (m2/s) RH (Å) Mw (g/mol)

Saturates 350–370 ± 5 1–3 Saturates 4.77*10-10 9.15 1175 ± 38

Aromatics 370–405 ± 5 2–4 Aromatics 4.18*10-10 10.40 1452 ± 71
Resins 390–450 ± 10 2–5 Resins 3.74*10-10 11.69 1770 ± 150
Asphaltenes 445–500 ± 10 5–7 Asphaltenes 2.10*10-10 20.69 4570 ± 105
Bitumen 370–415 ± 5 – Bitumen 4.03*10-10 10.80 1545 ± 52

3.5. NMR spectroscopy
3.5.1
. 1H DOSY NMR
Fig. 4 shows a multiple display plot of the experimental 1H DOSY
spectra of the fractions. The vertical axis corresponds to the chemical
shift of protons (1H) and the horizontal axis to the logarithmic repre­
sentation of the diffusion coefficient. Peaks at δH 5.31 and 7.26 ppm
corresponded to the diffusion standard dichloromethane and the solvent
deuterochloroform, respectively. The signals in the aliphatic region
(0.5–3 ppm) were the most intense and therefore used for the evaluation
of the average diffusion coefficients. These were extracted after cali­
bration of the spectra onto CH2Cl2 using the software Dynamics Center
2.8.0.1 (Bruker). Based on Eqs. (2) and (3), the average hydrodynamic
ratio RH and the average molecular mass Mw were estimated (Table 6).
The calculated results revealed that both the hydrodynamic radius and
the molecular weight increased with higher polarity. The measurement
of the bituminous samples (Table 6) showed that both values were close
to the values of the aromatic fraction.
3.5.2. HSQC NMR
The HSQC spectra reveal information on carbon atoms directly
bound to protons within organic molecules. An exemplary HSQC spec­
trum of bitumen is shown in Fig. 5. The horizontal axis depicts the
chemical shifts of protons and the vertical axis those of carbons. The
intrinsic characteristic of the edited HSQC pulse sequence is that
methine (CH) and methyl (CH3) groups give rise to a different phase
Fig. 4. 1H DOSY NMR spectra of the SARA fractions. The horizontal axis de­
picts the chemical shift (ppm) of protons and the vertical axis the diffusion
coefficient D (m2/s).
than methylene (CH2) groups, resulting in either positive or negative
signals. This is represented in the 2D contour plot with different colours:
CH3 and CH are given in blue and CH2 in red. Intrinsically, HSQC is
unable to report carbon atoms without H being directly attached, leav­
ing functional groups, such as ketones, carboxylic acids or esters, and
quaternary carbon moieties in aliphatic chains or condensed aromatic
ring systems undetected. Table 7 gives a summary of important spectral
areas [29] within the HSQC plots. The share of characteristic CH-
structures in specified chemical environments was calculated by
comparing the integrals in specific areas to the total integrals of all
signals.
The HSQC spectra revealed that throughout all fractions aliphatic
structures exceed aromatic ones (shown in Fig. 6). The total methylene
content was greater than the methyl content and no evidence of olefinic
structures was observed throughout all samples (in agreement with
other publications [4,20]). The ratio between methylene and methyl
groups decreased from saturates to resins, aromatics and asphaltenes.
This result suggests that longer aliphatic chains are present in the
saturates and the shortest aliphatic structures are found in the
asphaltenes. The highest aromatic CH content was found in the aro­
matics and asphaltenes (~20 %) followed by the resins (~15 %) and
the saturates (~5%). In order to understand this trend, one has to
emphasize that only CHn groups are observable with HSQC. Conse­
quently, not just the number of aromatic compounds but mainly the
aromatic condensation degree and the number of substituents at the
aromatic core have an impact on the amount of detectable aromatic CH
groups. The higher the degree of condensation or substitution the lower
is the share of measurable aromatic CH in the molecule, while the
‘HSQC-invisible’ quaternary carbons increase at the same time. Those
effects might overcompensate the numerical increase of aromatic
structures in resins and asphaltenes resulting in an overall lower aro­
matic CH percentage.
HSQC also provides the opportunity to differentiate aliphatic sub­
stituents directly attached to aromatic rings (see Fig. 7). The content of
aromatic methylene groups exceeded the aromatic methyl content in all
samples other than the resins. The saturates were the fraction with the
lowest intensities of aromatic CH which is primarily due to the lack of
aromatic molecules in general. The share of aromatic CH and directly
bonded CHn groups (aromatic methyl and methylene) was the highest in
the aromatics fraction which would indicate that this fraction comprises
the most aromatic molecules.
Tendencies for aliphatic side chain lengths and branching were
derived by comparing the share of aromatic alkyls (directly attached to
aromatic rings) to the total content of the respective alkyl (Table 8). The
decreasing share of aromatic methylene groups from aromatics,
asphaltenes, resins to saturates suggests that the chain length of
aliphatic substituents increases in the same order. The share of aromatic
methyl groups in the aromatics fraction exceeded the share in resins and
asphaltenes by a wide margin, indicating that a significant portion of the
methyl groups is directly attached to aromatic rings and/or that the
branching in the high-polar fractions is higher (the higher the branching
of an aliphatic substituent the more CH3 groups are present). Addi­
tionally, the obtained values for the bituminous sample are shown in

6
S. Werkovits et al. Construction and Building Materials 352 (2022) 128992

Fig. 5. Exemplary HSQC NMR spectrum of bitumen. Left: Aliphatic region. Right: Aromatic region. Blue coloured domains depict positive signals (CH3, CH groups)
and red domains negative signals (CH2-groups). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of
this article.)

Table 7
Characteristic 1H and 13
C chemical shift regions and structural assignments.
1 13
H-shift [ppm] C-shift [ppm] Structural assignments [29]

0.5–1.5 10–25 Terminal CH3 in aliphatic chains

0.9–1.9 24–45 CH2 of alkyl chains
1.3–1.5 31–34 CH group in aliphatic chains
2.0–3.0 10–24 Aromatic CH3
2.3–3.1 26–38 Aromatic CH2
4.5–6.0 120–135 CH in olefins
6.0–9.3 115–145 CH of aromatic compounds

Fig. 7. Share of aromatic CH and directly bonded aromatic methyl and meth­
ylene groups.

Table 8
Share of aliphatic groups directly attached to aromatic rings.
Region Saturates Aromatics Resins Asphaltenes Bitumen

Aromatic 4.92 ± 25.05 ± 15.51 ± 12.77 ± 18.50 ±

CH3/total 0.70 1.90 0.83 0.76 0.39
CH3
Aromatic 1.99 ± 8.12 ± 4.55 ± 5.14 ± 0.34 7.25 ±
CH2/total 0.68 0.09 0.84 0.67
Fig. 6. Ratio of methyl, methylene and aromatic groups in the fractions. CH2
Aromatic 2.50 ± 12.25 ± 7.12 ± 7.15 ± 0.11 9.87 ±
CHn/total 0.66 0.28 0.80 0.56
CHn
Table 8. Being a mixture of the above fractions, the contents of the
molecular groups ranged (as expected) between the obtained extrema of
the fractions.
Additionally, there were several unique signals within the resin in the 13C domain (see Fig. 8). Those signals had high intensities in
spectrum. Those signals appeared at 4.11/65.39 ppm (OCH2), 5.34/ the saturates, resins and asphaltenes but were hardly observable in
138.2 ppm and 5.57/101.0 ppm. The former was interpreted as an the aromatic fraction and the base binder. The shift values indicate
alkoxy moiety in an ether or ester, whereas the chemical shifts of the steric crowding [40], possible structures being CH2 groups that bridge
latter signals would suggest the presence of alkenes (olefins) [39]. branching points as highly substituted carbons; e.g. the bridging CH2
Another interesting feature were three methylene signals (CH2) with in 2,4 dimethyl pentyl or next to quaternary carbons like in
shifts of 1.19, 1.04 and 0.86 ppm in the proton domain and 45.5 ppm neopentyl.

7
S. Werkovits et al.
Fig. 8. Aliphatic region of HSQC spectra (only CH2 signals). Black: bitumen.
Green: saturates. (For interpretation of the references to colour in this figure
legend, the reader is referred to the web version of this article.)
3.5.3. HMBC NMR
The use of another 2D NMR technique, HMBC, enabled us to gain
additional information not obtainable with HSQC. It detects carbon
atoms which are not directly bound to protons but are separated from
them through a number (mostly 2 – 4) of bonds. Especially quaternary
carbons in aromatic structures and carbon atoms of carbonyls (e.g. ke­
tones, amides, esters) can be detected with this experiment. The
knowledge about those functionalities is crucial to improve the under­
standing of molecular structures within bitumen but is also important in
the analysis of ageing processes. The most intense carbonyl signals were
observed in the resins at chemical shifts of 175.5 ppm, indicating the
presence of carboxylic acids, esters or amides, and 2 different H-regions:
2.24 and 2.37 ppm, indicating aliphatic structures. This is in good
agreement with the intense carbonyl band found in infrared spectros­
copy at 1670 cm− 1 (Fig. 9).
Another interesting feature is that the 13C shift in the aliphatic region
was extended towards 57 ppm with the highest intensities in saturates,
but only minor contributions in the aromatics, asphaltenes and resins
fraction. The 1H shift of this signal correlated with the frequent peak at
45.4 ppm (for all fractions, see HSQC), providing evidence of neopentyl-
Fig. 9. Evidence for amide groups in HMBC and IR spectra of the resin fraction.
8
Construction and Building Materials 352 (2022) 128992
Fig. 10. HMBC spectrum of bitumen. Highlighted areas are distinguishable
spectral regions. Multiple bond C–H correlations are shown for the aromatic
regions. Quaternary C-Atoms greater than 130 ppm.
type structure with the quaternary carbon of this structure resonating at
that frequency. There are three spectral regions for CH correlations
involving aromatic rings in HMBC: Cali–Haro, Caro–Haro and Caro–Hali (see
Fig. 10). The Caro-Hali region (bottom right in Fig. 10), with almost all
cross-peaks with 1H shifts over 2 ppm, comprises aromatic alkyl groups,
i.e. aliphatic chains directly attached to the aromatic core. However,
there were also some signals between 1 and 1.5 ppm which have 13C
shifts that exceeded 130 ppm. Those originated from quaternary aro­
matic carbons, which are correlated with the chain protons in β-position.
A qualitative comparison of the fractions showed that the aromatic re­
gions of resins and asphaltenes consisted almost exclusively of quater­
nary carbon signals (confirming the higher degree of condensation, see
above), whereas in the aromatics fraction additional correlations to ar­
omatic CH were found. Finally, also correlations in the Caro-Haro region
were found showing the correlations of aromatic protons with carbons of
the aromatic ring systems.
3.5.4. 31P NMR
Quantitative 31P NMR spectra of the derivatized fractions showed
minor contributions of accessible functional groups (OH, NH) in the
S. Werkovits et al.
Table 9
OH–/NH-containing groups in the SARA fractions.
Sample Carboxylic acid (%) Alcohols + Amides (%)
Saturates 0.00 0.00
Aromatics 0.00 ≤0.01
Resins ≤0.96 ≤0.91
Asphaltenes ≤0.65 ≤0.32
Bitumen 0.00 0.00
resins and asphaltenes, never exceeding a total of 2 % for all detectable
compounds. The regions of the phosphorous ester derivate signals (see
Fig. 1) corresponded to aliphatic OH (alcohols, carboxylic acids) and NH
(amines, amides) groups. The quantitative assessment (Table 9) showed
that only resins and asphaltenes contained trace amounts of those
functional groups. The resins contained the highest share of carboxylic
acids and alcohols/amides which is supported by the elemental
composition (highest O-content in resins) and the characteristic IR
bands (amide and carboxylic acid band in resins). No corresponding
signals were detected in the bituminous sample, due to the high share of
saturates and aromatics, which pushed the concentration of the groups
below the detection limit.
3.6. Summary
The main results of this study can be subdivided in structural and
chemical findings. The relative molecular sizes of the bitumen fractions
were estimated using DOSY experiments, revealing an increase from
saturates to asphaltenes. Furthermore, the combination of elemental
analysis with HSQC NMR and fluorescence spectroscopy showed dif­
ferences in aromatic content throughout all fractions. The red-shifting
emission wavelengths (from saturates to asphaltenes) in the fluores­
cence spectra correlate with increasing degrees of aromatic condensa­
tion. HSQC and elemental analysis revealed a change of order among
aromatics and resins (lower saturation in aromatics and higher content
of aromatic compounds in aromatics), the main reasons for these dif­
ferences being number and length of aliphatic substituents. The peak-
integration results of HSQC suggest more and longer side chains
within the resins compared to the aromatics. Furthermore, information
of branching tendencies was gathered, with indications of neopentyl-
like structures (in HSQC). The abundance of these groups throughout
the fractions and the share of aliphatic CH3 groups (as an indirect in­
dicator) give first hints that the aromatics contain the aliphatic side
chains with the least extent of branching.
Looking at the chemical side, sulfur was the main heteroatom
throughout all fractions, reaching a maximum of roughly 5.5 wt% in the
asphaltenes. Sulfur was observed in the infrared spectra of aromatics,
resins and asphaltenes as sulfoxides. The lack of sulfoxide signal in
saturates and its very low intensity within the aromatics (at elemental
analysis values comparable to resins and asphaltenes) suggest the
presence of low-polar sulfur-containing compounds, such as aliphatic
and aromatic sulfides, disulfides and heterocyclic species, such as
(benzo)thiophenes. The oxygen content increases from saturates to
resins (maximum), but interestingly decreases again from resins to
asphaltenes. According to infrared spectroscopy we found oxygen
mainly as carboxylic acids and their derivatives (amides/esters) and
some alcohols. Carboxylic acids were found in resins and asphaltenes,
whereas aromatic amides were found only in the resins. Further evi­
dence of amide and ester structures were provided by NMR (HSQC/
HMBC). The analysis of OH/NH-containing groups by 31P NMR revealed
low amounts of aliphatic alcohols and carboxylic acids within the resins
and asphaltenes. The lack of clear differentiation between NH and OH
groups with all these techniques is still a problem, and a presence of both
types of functionalities is most likely. However, looking at the strikingly
low content of nitrogen throughout all fractions (<1%), future attention
should focus rather on carboxylic acid esters.
9
Construction and Building Materials 352 (2022) 128992
4. Conclusions and outlook
The focus of this study was to widen the knowledge basis regarding
molecular structures in bitumen. By pairing polarity-based separation
with a combination of spectroscopic techniques, distinct structural and
chemical features were revealed. Such assignments would not have been
possible for the base binder (non-fractionated bitumen). The use of 2D
NMR techniques provided important molecular features and how they
change with increasing polarity. Throughout the fractions notable dif­
ferences in aromatic condensation and alkyl substituents (length, num­
ber and branching) were revealed. DOSY NMR spectroscopy provided
another aspect of the structure of bitumen and its fractions, the average
molecular sizes and weights, showing an increase from saturates to
asphaltenes. Combining the information of NMR and IR spectroscopy
helped to validate functional groups and how they are distributed within
the fractions. The use of a selective chemical reaction in combination
with 31P NMR showed great potential to classify and quantify OH– and
NH– groups in the SARA fractions. However, with regard to general­
ization of the results, caution is advised as only one binder was analysed
with the full set of analytical techniques in this study. Therefore, it is
mandatory to emphasize, that varying the crude oil source and/or
changing the manufacturing process will influence the composition of
bitumen and the distribution of fractions and molecular species in the
bitumen. Thus, while a limitation of generalized conclusions with regard
to the obtained numbers has to be acknowledged, the demonstration of
the analytical power of the combined spectroscopic approach for
bitumen characterization remains fully upright.
The sensitivity of the employed techniques to bitumina variability
and minor specific changes can also be used as an advantage, especially
in ageing investigations. The gained knowledge in the assessment of
molecular structures will be of great benefit for the elucidation of
structural changes upon ageing processes. In particular, the comparison
of naturally or artificially aged with unaltered bitumen samples will
provide new insights on a wide range of chemical parameters, going
beyond a mere evaluation of the main oxidation products. Application of
the presented extended multi–spectroscopic approach to the ageing
chemistry of bitumen will be the topic of an upcoming account that will
be reported in due course.
CRediT authorship contribution statement
Stefan Werkovits: Conceptualization, Methodology, Investigation,
Formal analysis, Validation, Data curation, Writing – original draft,
Visualization. Markus Bacher: Methodology, Investigation, Formal
analysis, Writing – review & editing. Johannes Theiner: Methodology,
Investigation, Writing – review & editing. Thomas Rosenau: Writing –
review & editing. Hinrich Grothe: Resources, Supervision, Project
administration, Funding acquisition, Writing – review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgements
The financial support by the Austrian Federal Ministry for Digital and
Economic Affairs, the National Foundation for Research, Technology
and Development and the Christian Doppler Research Association is
gratefully acknowledged. Furthermore, the authors would also like to
express their gratitude to the CD laboratories company partners BMI
S. Werkovits et al.
Group, OMV Downstream and Pittel + Brausewetter for their financial
support.
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