Separation Science and Technology

ISSN: 0149-6395 (Print) 1520-5754 (Online) Journal homepage: https://www.tandfonline.com/loi/lsst20

On-line coupling of solid-phase extraction of

phenols on porous graphitic carbon and LC
separation on C18 silica gel column via subcritical
water desorption

Dina Rashidovna Borisova, Mikhail Alexandrovich Statkus, Grigory Il’yich

Tsysin & Yury Alexandrovich Zolotov

To cite this article: Dina Rashidovna Borisova, Mikhail Alexandrovich Statkus, Grigory
Il’yich Tsysin & Yury Alexandrovich Zolotov (2016) On-line coupling of solid-phase extraction
of phenols on porous graphitic carbon and LC separation on C18 silica gel column via
subcritical water desorption, Separation Science and Technology, 51:12, 1979-1985, DOI:
10.1080/01496395.2016.1199571

To link to this article: https://doi.org/10.1080/01496395.2016.1199571

View supplementary material Accepted author version posted online: 17

Jun 2016.
Published online: 17 Jun 2016.

Submit your article to this journal Article views: 83

View Crossmark data Citing articles: 3 View citing articles

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=lsst20
SEPARATION SCIENCE AND TECHNOLOGY
2016, VOL. 51, NO. 12, 1979–1985
http://dx.doi.org/10.1080/01496395.2016.1199571

On-line coupling of solid-phase extraction of phenols on porous graphitic

carbon and LC separation on C18 silica gel column via subcritical water
desorption
Dina Rashidovna Borisova, Mikhail Alexandrovich Statkus, Grigory Il’yich Tsysin, and Yury Alexandrovich Zolotov
Moscow State University, Department of Chemistry, Division of Analytical Chemistry, Moscow, Russia

ABSTRACT ARTICLE HISTORY

An approach to the on-line coupling of solid-phase extraction on porous graphitic carbon (PGC) Received 23 October 2015
column and high performance liquid chromatography (HPLC) separation on octadecyl silica Accepted 6 June 2016
column is proposed. The approach includes extraction of phenols from 10 ml of water sample KEYWORDS
on PGC column; desorption with subcritical water at 175°C, injection of the eluate to the HPLC Liquid chromatography;
column and separation at ambient temperature with acetonitrile–water eluent. Limits of detection porous graphitic carbon;
for UV detection of phenols were 1–5 μg l−1, and analyte peaks were 20–50% narrower than for solid-phase extraction;
direct injection of 20 μl of sample. subcritical water

Introduction which can hamper effective desorption resulting in

excessive peak broadening.[4,5]
Solid-phase extraction (SPE) is widely used to enhance
For example, this approach results in peak widths 10
the capabilities of reversed-phase high performance
times greater for on-line SPE–HPLC determination of
liquid chromatography (HPLC) analysis. On-line cou-
phenols than for direct injection of 10 µl of the sample.[6]
pling of SPE and HPLC requires no manual sample
Peak broadening is most evident when SPE packing
handling between enrichment and separation steps,
differs significantly from HPLC stationary phase[2] –
thus decreasing risks of sample contamination and ana-
for example, when PGC is used for SPE, and ODS for
lyte losses.[1] On-line SPE columns are usually smaller
HPLC separation.[3] This well-known problem was
than off-line ones, thus analyte recovery could be
circumvented[7] by using only organic component of
increased only by selecting more retentive sorbents.[2]
the eluent (methanol) for desorption of analytes; the
Porous graphitic carbon (PGC) is often used for SPE
eluent was subsequently delivered to the mixing cham-
when increased retention is required compared to octa-
ber of the gradient pump and diluted to 33% of metha-
decyl silica (ODS).[3] Increased analyte retention is a
nol. This approach inevitably leads to additional peak
benefit when extraction step is carried out; however,
dispersion – first of all due to dilution, and second due to
on-line elution of the analytes from a highly retentive
mixing chamber dead volume.
sorbent could pose a problem.
Similar problems arise when coupling of solvent
Careful selection of desorption conditions of ana-
extraction from solid samples (“leaching” of soil,
lytes from SPE column is crucial for effective coupling
vegetable samples, etc.) where HPLC analysis is
of SPE and HPLC.[2] The usual approach is to use the
required.[8] Generally applied extraction procedures
same eluent for both desorption and reversed-phase
yield analytes dissolved in a large volume of organic
HPLC separation. However, effective desorption
solvent – typically from 10 to 1000 ml; and only a
requires the use of the eluent with high organic solvent
tiny fraction (about 10 µl) can be injected for HPLC
content to minimize band broadening; reversed-phase
analysis, resulting in a considerable decrease in sen-
HPLC separation is usually carried out with low or
sitivity. An original approach to overcome this pro-
medium organic solvent content in the eluent. This
blem is to use subcritical water (SW) for extraction,
contradiction is usually resolved in a compromising
desorption and HPLC separation.[9–11]
manner – by choosing a medium strength eluent,

CONTACT Statkus Mikhail Alexandrovich mstatkus@gmail.com Moscow State University, Department of Chemistry, Division of Analytical Chemistry,
119991 Moscow, Russia.
Supplemental data for this article can be accessed on the publisher’s website.
© 2016 Taylor & Francis
1980 D. R. BORISOVA ET AL.
In the recent years, there is a considerable interest in
replacing organic eluents in HPLC separations with
SW, see for example several reviews.[12,13] It was
shown[14] that pressurized water at 180°C is a
reversed-phase eluent with moderate elution strength.
Higher temperatures result in further increase of elu-
ent strength; for example, water at 250°C was used[15] for
extraction of non-polar analytes (such as polycyclic aro-
matic hydrocarbons) from soil samples.
One of the attractive features of SW is the simplicity
of required equipment and relative convenience of the
experiments when compared to the application of
supercritical fluids. Experimental set-up can be
assembled from ordinary HPLC modules with the addi-
tion of an oven from a gas chromatograph. Coupling of
SW extraction and HPLC via sorbent trapping and SW
desorption has been achieved by application of sorbent
traps packed with polystyrene-coated zirconia,[9] poly-
styrene-divinylbenzene[10] or unbonded hybrid silica
(Waters XTerra).[11] It is worth mentioning that PGC
was initially considered as a packing for the sorbent
trap,[11] but very high retention of analytes required
desorption temperatures over 200°C resulting in partial
decomposition of herbicides. Thus, unbonded silica was
chosen as a less-retentive sorbent trap packing.
Besides the preliminary data reported in Tajuddin
and Smith,[11] there is no published information on SW
desorption of analytes from SPE columns packed with
PGC. Also no information is available considering on-
line coupling of PGC-packed SPE column and HPLC
separation via SW desorption.
This present paper is devoted to further develop-
ment of the on-line coupling of SPE and HPLC via
subcritical water desorption. We have used PGC col-
umn for SPE of phenols from water samples and ODS
column for HPLC separation. HPLC separation was
carried out at ambient temperature with water–aceto-
nitrile eluent. Since SW HPLC separation of phenols
was reported previously[14,16–18] – and these substances
were relatively stable during high temperature separa-
tion – we have chosen several EPA phenols as model
substances to test the proposed coupling of PGC and
ODS columns via SW desorption.
Experimental
Chemicals
Acetonitrile (ACN) was of “Super Gradient” grade
(LabScan, Ireland). Isopropanol was of HPLC grade,
and phosphoric acid solution was of ultrapure grade
(ComponentReactive LLC, Russia). Millipore
Simplicity water purification system (Millipore, USA)
was used to produce deionized water. Stock solutions
of phenol (Ph), 4-nitrophenol (4NPh), 2-chlorophenol
(2ClPh), 2,4-dinitrophenol (24NPh), 2,4-dichlorophenol
(24ClPh), 2,4-dimethylphenol (24MPh), 2-nitrophenol
(2NPh), 4-chloro-3-methylphenol (4Cl3MPh), and
2-methyl-4,6-dinitrophenol (2M46NPh) (all reagents
were at least 98% purity from Sigma-Aldrich,
Germany) were prepared in ACN at a concentration of
2 mg L−1. Stock solutions were kept at +4°C. To prevent
possible oxidation of analytes by dissolved oxygen, water
used for high temperature desorption was purged with
nitrogen for 20 min. Tap, bottled mineral and river water
samples were filtered through a membrane filter
(0.45 µm pore size, nylon) before performing SPE. Tap
water was collected at Chemistry Department of
Moscow State University, Moscow Russia. Bottled
mineral water “Essentuki 4” was obtained at a local
shop. River water sample was collected from Moskva
River, Moscow, Russia.
Instrumentation
On-line SPE–HPLC system consisted of several mod-
ules from Aquilon JSC (Russia): two HPLC pumps
Stayer-2, automatic injection module Stayer (which
included two actuated Rheodyne 9010 six-port injec-
tion valves) and variable wavelength UV detector
Stayer UVV-104. MultiChrom 2.3 software
(Ampersend JSC, Russia) was used for data acquisi-
tion. Stainless steel tubing was used for high-tem-
perature parts of the system, and other connections
were made with PEEK tubing. Coils for preheating
and cooling of water were made from 1.5 m ×
0.25 mm i.d. stainless steel tubing.
Forced convection oven was salvaged from gas chro-
matograph LKhM-80 (Chromatograph JSC, Russia).
The oven was fitted with 1800 W heating element.
Heating was controlled by electronic thermostat
TRM-10 (Oven JSC, Russia) equipped with a thermo-
couple DTPK.
Pressure restrictors P-789 and P-455 (Upchurch
Scientific, USA) were used to keep water in liquid
state. P-789 is rated at 34 bar for 1 ml min−1 water
flow at +20°C, and P-455 at 69 bar.
Analytical procedure
Ambient temperature HPLC separation
HPLC separation was carried out at ambient tempera-
ture on a Luna C18 5 µm column (150 mm × 4.6 mm i.
d., Phenomenex, USA). The HPLC eluent was acetoni-
trile:water mixture (40:60 v/v); the mobile phase was
acidified with 0.1% (v/v) H3PO4 to prevent the

dissociation of analytes. Sample loop volume was 20 µl.
Mobile phase flow rate was 1 ml min−1. UV detection
was carried out at 275 nm.[19,20]
SPE procedure
High-temperature version of Hypercarb 5 µm PGC
HPLC column (30 mm × 2.1 mm i.d., Thermo
Scientific, USA) was used for SPE of analytes. SPE
column dead volume was estimated by the injection
of a non-retained solute (NaNO3) and was found to
be 0.1 ml. SPE column was precleaned with 5 ml of
isopropanol:ACN mixture (75:25, v/v), and then pre-
conditioned with 5 ml of ACN. Water samples (acid-
ified with 0.1% (v/v) H3PO4) with volumes ranging
from 10 to 50 ml were passed through the SPE column.
The flow rate was 1 ml min−1 in all these steps. The
analytes were desorbed by either ACN or SW; see the
“Results and discussion” section for details.
For selection of maximum sample volume that could
be percolated through SPE column, deionized water
samples (10–50 ml) were spiked with 4 µg of each
analyte. The desorption was carried out in an off-line
mode with 5 ml of ACN; the eluate was collected in a
vial and then analysed by ambient temperature HPLC.
Subcritical water desorption
The practice of high-temperature HPLC revealed that
mobile phase temperature could be different from the
oven temperature.[13,19] This difference is related to the
limited rate of heat transfer from the air in the oven to
the column hardware and, finally, to the mobile phase.
During our preliminary research, we have found out
that for the oven and the column hardware used, the
optimum eluent flow rate was 0.5 ml min−1, and a static
preheating period of 10 min was required when the
oven was heated up to 150–200°C from the ambient
temperature (see supplementary materials, Fig. S1).
During the preheating period, the eluent flow was
stopped. We used the same optimum flow rate and
preheating duration in the current work.
Results and discussion
The aim of this paper was to develop an approach to
on-line coupling of SPE on PGC and HPLC separation
on ODS. The development of an effective on-line cou-
pling of SPE and HPLC requires several important
factors to be studied, e.g. maximum sample volume
that could be safely passed through SPE column at the
required recovery level; the minimum volume of the
eluent that is required to desorb the analytes, etc. Some
of these factors can be studied in off-line mode for the
sake of experiment simplicity and clear data evaluation.
SEPARATION SCIENCE AND TECHNOLOGY 1981
First of all, maximum sample volume should be estab-
lished, which can be percolated through SPE column
without significant losses of the analyte. Then, deso-
rption conditions should be optimized. Next step is the
optimization of the on-line coupling of SPE and HPLC.
Finally, the overall performance of the proposed tech-
nique should be evaluated and compared to earlier
published results.
Sample volume selection
Recovery values were obtained for off-line SPE proce-
dure to determine the highest sample volume that
could be percolated through SPE column in order to
decrease the limits of detection (LODs) of the overall
procedure. The recovery vs. sample volume plots for
phenol, 4-nitrophenol and 2-chlorophenol are pre-
sented in Fig. 1; similar plots for other analytes are
presented in Fig. S2. Recovery of phenol dropped
from 70% (10 ml sample) to 14% (50 ml sample).
Recovery values for dinitrophenols also dropped from
70% to 20%; one of the reasons for poor recovery and
reproducibility of these phenols is that their desorption
with 5 ml of ACN was not quantitative – see the section
“Desorption conditions”. For the rest of analytes, the
recoveries were 80–95% for the whole volume range
studied. Sample volume was fixed at 10 ml for the
further optimization of desorption conditions.
Desorption conditions
Desorption of analytes should be quantitative and car-
ried out with minimum eluent volume. The main fac-
Figure 1. The recovery vs. sample volume plots for SPE of
phenols. Deionized water samples (10–50 ml) were spiked
with 4 µg of phenol, 4-nitrophenol and 2-chlorophenol. The
desorption was carried out in an off-line mode with 5 ml of
ACN at +20°C. n = 4, P = 0.95. Other details of the experiment
are presented in the section “Sample volume selection”.
1982 D. R. BORISOVA ET AL.
tors affecting desorption with SW are: temperature,
flow rate and the duration of static preheating before
the actual desorption begins. According to the pub-
lished data,[13] the most important of these factors is
temperature. Phenols were extracted with SW from soil
samples and other objects[9,15] at the temperature range
of 100–200°C. The PGC column is rated for tempera-
tures up to 200°C, so we decided to study the deso-
rption of phenols at temperatures from 150°C to 200°C.
Desorption was studied in off-line mode by collect-
ing fractions of the eluate and analysing them sepa-
rately. SPE procedure was performed as described in
the section “SPE procedure”; 10 ml samples of deio-
nized water were spiked with 4 µg of each analyte. The
experimental set-up used for the off-line study of des-
orption is shown in Fig. S3. Desorption recovery was
calculated as the ratio of the desorbed amount of ana-
lyte to the total extracted amount of the analyte. Off-
line SW desorption graphs were plotted by analysing
0.5 ml fractions of the eluate (see Fig. 2 and Fig S4).
Similar graphs were plotted for desorption carried out
with ACN.
Desorption efficiency is usually associated with
minimum eluent volume required for quantitative des-
orption. This volume depends on both retention of the
analyte during desorption and broadening of the ana-
lyte peak due to axial dispersion. SW at 150°C was
more effective than ACN at +20°C for desorption of
Figure 2. Off-line desorption graphs for phenol (A), 4-nitrophenol (B), 2-chlorophenol (C) and 4-chloro-3-methylphenol (D).
Desorption was carried out with acetonitrile (ACN) at +20°C and subcritical water (SW) at 150–200°C. Other details of the experiment
are presented in the section “Desorption conditions”.
phenol and 2-chlorophenol – both in terms of broad-
ening and retention. A temperature of 175°C was
necessary for SW desorption of 2-nitrophenol and
2,4-dimethylphenol. More hydrophobic 4-chloro-3-
methylphenol, 4-nitrophenol and 2,4-dichlorophenol
required the desorption temperature to be raised to
200°C. Desorption efficiency in this case was similar
or better than with ACN at room temperature.
However, desorption of dinitrophenols (2,4-dinitro-
phenol and 2-methyl-4,6-dinitrophenol) was less effi-
cient with both ACN and SW at different
temperatures. We were unable to elute these dinitro-
phenols quantitatively with 5 ml SW even at 200°C;
desorption with ACN produced a wide peak for
2,4-dinitrophenol and no peak of adequate height for
2-methyl-4,6-dinitrophenol.
It is yet unclear which properties of analytes make
it difficult to perform an effective desorption from
Hypercarb with either SW or ACN. The hydrophobic
properties of the molecular forms of 2,4-dinitrophenol
and 2-methyl-4,6-dinitrophenol in terms of log Ko/w
are 1.55 and 2.06, respectively, which is less than that
of, e.g., 2-chlorophenol (2.27) which is effectively des-
orbed. We hope to shed some light on this question
by the application of a so-called solvation parameters
model[20] to link the physico-chemical properties
(dipolarity, polarizability, etc.) of the analytes and
their retention and desorption.
 SEPARATION SCIENCE AND TECHNOLOGY 1983

Incomplete desorption of dinitrophenols could
result in systematic analysis errors due to carryover:
analytes remaining after incomplete desorption from
the previous sample could be eluted during next deso-
rption. To eliminate this problem, we have introduced
a wash step with 5 ml of isopropanol:ACN (75:25, v:v)
and 5 ml of ACN. This step was carried out after
desorption, simultaneously with HPLC separation to
reduce the overall duration of the procedure.
For subsequent experiments, we choose the mini-
mum volume of SW required for complete desorption
of phenols (5 ml) and optimum SW temperature (175°
C). The most realistic approach to reduction of the
eluent volume required for desorption of analyte is to
reduce the size of the SPE column used.
On-line coupling of SPE and HPLC
During preliminary experiments, we found that 5 ml of
the aqueous sample could be injected directly into the
HPLC column without a significant peak broadening.
However, large volume direct injection of real samples
is impractical due to quick deterioration of LC column
performance because of clogging and fouling with par-
ticulate matter in the sample and the presence of highly
retained compounds. These problems are solved by
using a separate SPE column. Thus we decided to try
a simple approach to the on-line coupling of SPE and
HPLC: the whole volume of the effluent after deso-
rption is introduced into HPLC column, then enacting
the HPLC separation with ACN–water eluent. The
schematic of the used equipment is presented in Fig. S5.
SPE was carried out as described in the section “SPE
procedure”, with 10 ml of the sample being spiked with
the appropriate amount of each phenol. Analytes were
desorbed in the following manner: oven was set to
+175°C, and turned on for 10 min preheating. Then
5 ml of water was passed through the SPE column at a
flow rate of 0.5 ml min−1.
A typical on-line SPE–HPLC chromatogram
obtained via SW desorption is presented in Fig. 3C.
Similar experiments were carried out with neat ACN
and ACN:water (40:60, v:v) as the eluents for deso-
rption. After SW desorption, the eluate is cooled
down to an ambient temperature; thus, injecting 5 ml
of water eluate results in focusing of the analytes on the
frontal part of the HPLC column. Desorption with
ACN does not provide the possibility to focus the
analytes, thus no peaks could be detected on the chro-
matogram, see Fig. 3D. Desorption with ACN:water
(40:60, v:v) resulted in poor baseline and wider peaks
(Fig. 3B) than desorption with SW.
A comparison was made between peak widths
obtained by on-line SPE–HPLC and direct injection

Figure 3. Chromatograms obtained with various sample introduction techniques. A – direct injection of 20 µl of the sample; B – on-
line desorption with ACN–water (40:60 v:v) eluent; C – on-line desorption with SW at 175°C; D – on-line desorption with 100% ACN.
The total amount of each phenol was 0.1 µg (20 µl of 5 mg ml−1 solution for direct injection; 10 ml of 10 ng ml−1 solution for SPE
procedures). Peak labels are as follows: 1 – phenol, 2 – 4-nitrophenol, 3 – 2-chlorophenol, 4 – 2,4-dinitrophenol, 5 – 2-nitrophenol,
6 – 2,4-dimethylphenol, 7 – 4-chloro-3-methylphenol, 8 – 2,4-dichlorophenol, 9 – 2-methyl-4,6-dinitrophenol.
1984 D. R. BORISOVA ET AL.

Table 1. Analyte peak half-widths for direct injection and on- Table 2. Limits of detection (LODs) and enhancement factors.
line SPE–HPLC coupling(confidence limits were calculated for LOD, µg l−1,
3 replicates and P = 0.95). LOD, µg on-line SPE-
Peak half-width, seconds ml−1, direct HPLC via SW Enhancement
Analyte injection desorption factor*
On-line HPLC
Phenol 0.1 1 100
On-line SW eluent Direct
4-Nitrophenol 0.2 2 100
Analyte desorption desorption injection 2-Chlorophenol 0.2 3 67
Phenol 10 ± 1.4 26 ± 3.2 15 ± 1.8 2-Nitrophenol 0.1 1 100
4-Nitrophenol 14 ± 1.8 19 ± 2.0 18 ± 2.0 2,4-Dimethylphenol 0.2 3 67
2-Chlorophenol 14 ± 1.5 19 ± 1.9 21 ± 2.7 4-Chloro-3-methylphenol 0.4 4 100
2,4-Dinitrophenol 14 ± 1.1 14 ± 2.0 24 ± 2.4 2,4-Dichlorophenol 0.3 5 60
2-Nitrophenol 19 ± 2.2 24 ± 1.8 23 ± 3.4
* Enhancement factors were calculated as ratio of LODs for direct injection
2,4-Dimethylphenol 20 ± 2.9 32 ± 2.7 26 ± 3.7
4-Chloro-3-methylphenol 19 ± 2.7 33 ± 4.5 30 ± 3.2 and SPE–HPLC analysis.
2,4-Dichlorophenol 22 ± 3.3 45 ± 4.8 34 ± 2.4

accuracy of the proposed technique was tested by the

HPLC. Analytes were extracted from 10 ml of deio- analysis of spiked samples of tap, bottled mineral and
nized water spiked with 10 μg l−1 of each phenol; 20 μl river water; see Table S1 for results and Fig. S6 for
of ACN solution of analytes (5 mg l−1 each) was used chromatogram of spiked river water. Each sample was
for direct injection HPLC, the resultant peak widths are spiked with 10 ng ml−1 of each phenol. No analytes
given in Table 1. were detected in non-spiked (native) samples.
Comparison of peak widths for three different pro-
cedures (on-line SW desorption, on-line HPLC eluent
desorption and direct injection) reveals that desorption Conclusions
with HPLC eluent (ACN:water 40:60 v:v, in our case)
An approach to the on-line coupling of SPE on PGC
results in peaks that are 20–30% broader than for direct
column and HPLC separation on ODS column was
injection of 20 µl of sample. However, for some ana-
proposed. The approach includes extraction of phenols
lytes (2ClPh, 24NPh) peak widths are comparable. On
from 10 ml of water sample on PGC column; deso-
the other side, on-line SW desorption resulted in
rption with 5 ml of SW at 175°C, injection of the eluate
20–50% peak width reduction compared to direct
to the HPLC ODS column and separation of at an
injection.
ambient temperature with ACN–water eluent. LODs
Peak broadening reduces separation efficiency, for
for UV detection of phenols were 1–5 μg l−1, and
example peaks of 2NPh and 24MPh are poorly sepa-
analyte peaks were 20–50% narrower than for direct
rated when desorption is performed with the HPLC
injection of 20 μl of sample.
eluent (Fig. 3B, peaks 5 and 6, Rs = 1.07). Separation
is better (Fig. 3C, peaks 5 and 6, Rs = 1.56) for these
two peaks when on-line SW desorption is utilized.
Declaration of interest
Authors have declared no conflict of interest.

Analytical performance
Calibration plots for on-line SPE–HPLC coupling via
SW desorption were constructed for the concentration
of analytes ranging from 2.5 to 15 μg l−1. LODs were
calculated from the dispersion of the linear calibration
parameters according to Hubaux and Vos.[21] LODs for
on-line SPE–HPLC and direct injection HPLC are pre-
sented in Table 2. Application of the proposed SPE
procedure resulted in a 60–100-fold decrease of LODs.
The obtained LOD values are comparable to those
reviewed in Puig and Barceló.[22] For example, LODs
for on-line SPE–HPLC determination of phenols with
UV detection were in the range of 0.7–1 μg l−1,[22] but
the analytes were extracted from 100 ml of sample. We
have performed extraction from 10 ml of sample, but
obtained slightly higher LOD values (1–5 μg l−1). The
Funding
Authors are grateful to Russian Scientific Foundation for
financial support, grant 14-23-00012.
References
[1] Pan, J.; Zhang, C.; Zhang, Z.; Li, G. (2014) Review of
online coupling of sample preparation techniques
with liquid chromatography. Analytica Chimica
Acta, 815: 1–15.
[2] Hennion, M.-C. (1999) Solid-phase extraction: method
development, sorbents, and coupling with liquid chro-
matography. Journal of Chromatography A, 856: 3–54.
[3] Hennion, M.-C. (2000) Graphitized carbons for
solid-phase extraction. Journal of Chromatography
A, 885: 73–95.

[4] Brouwer, E.R.; Kofman, S.; Brinkman, U.A.T. (1995)
Selected procedures for the monitoring of polar pesti-
cides and related microcontaminants in aquatic sam-
ples. Journal of Chromatography A, 703: 167–190.
[5] Kataoka, H.; Lord, H.L.; Yamamoto, S.; Narimatsu, S.;
Pawliszyn, J. (2000) Development of automated in-tube
SPME/LC/MS method for drug analysis. Journal of
Microcolumn Separation, 12: 493–500.
[6] Wissiack, R.; Rosenberg, E.; Grasserbauer, M. (2000)
Comparison of different sorbent materials for on-line
solid-phase extraction with liquid chromatography–
atmospheric pressure chemical ionization mass spec-
trometry of phenols. Journal of Chromatography A,
896: 159–170.
[7] Asperger, A.; Efer, J.; Koal, T.; Engewald, W. (2002)
Trace determination of priority pesticides in water by
means of high-speed on-line solid-phase extraction–
liquid chromatography–tandem mass spectrometry
using turbulent-flow chromatography columns for
enrichment and a short monolithic column for fast
liquid chromatographic separation. Journal of
Chromatography A, 960: 109–119.
[8] Hyötyläinen, T. (2007) Principles, developments and
applications of on-line coupling of extraction with
chromatography. Journal of Chromatography A,
1153: 14–28.
[9] Lamm, L.J.; Yang, Y. (2003) Off-line coupling of sub-
critical water extraction with subcritical water chroma-
tography via a sorbent trap and thermal desorption.
Analytical Chemistry, 75: 2237–2242.
[10] Tajuddin, R.; Smith, R.M. (2002) On-line coupled
superheated water extraction (SWE) and superheated
water chromatography (SWC). Analyst, 127: 883–885.
[11] Tajuddin, R.; Smith, R.M. (2005) On-line coupled
extraction and separation using superheated water for
the analysis of triazine herbicides in spiked compost
samples. Journal of Chromatography A, 1084: 194–200.
[12] Teutenberg, T. (2010) High-temperature Liquid
Chromatography: A User’s Guide for Method
SEPARATION SCIENCE AND TECHNOLOGY 1985
Development. Cambridge: Royal Society of Chemistry,
220 p.
[13] Welch, C.J.; Wu, N.; Biba, M.; Hartman, R.; Brkovic,
T.; Gong, X.; Helmy, R.; Schafer, W.; Cuff, J.; Pirzada,
Z.; et al. (2010) Greening analytical chromatography.
Trac-Trends Analytical Chemistry, 29: 667–680.
[14] Smith, R.M.; Burgess, R.J. (1997) Superheated water as an
eluent for reversed-phase high-performance liquid chro-
matography. Journal of Chromatography A, 785: 49–55.
[15] Hawthorne, S.B.; Yang, Y.; Miller, D.J. (1994)
Extraction of organic pollutants from environmental
solids with sub- and supercritical water. Analytical
Chemistry, 66: 2912–2920.
[16] Li, B.; Yang, Y.; Gan, Y.; Eaton, C.D.; He, P.; Jones, A.
D. (2000) On-line coupling of subcritical water extrac-
tion with high-performance liquid chromatography via
solid-phase trapping. Journal of Chromatography A,
873: 175–184.
[17] Yang, Y.; Jones, A.D.; Eaton, C.D. (1999) Retention beha-
vior of phenols, anilines, and alkylbenzenes in liquid
chromatographic separations using subcritical water as
the mobile phase. Analytical Chemistry, 71: 3808–3813.
[18] Kondo, T.; Yang, Y.; Lamm, L. (2002) Separation of
polar and non-polar analytes using dimethyl sulfoxide-
modified subcritical water. Analytica Chimica Acta,
460: 185–191.
[19] Yan, B.; Zhao, J.; Brown, J.S.; Blackwell, J.; Carr, P.W.
(2000) High-temperature ultrafast liquid chromatogra-
phy. Analytical Chemistry, 72: 1253–1262.
[20] Poole, C.F.; Gunatilleka, A.D.; Sethuraman, R. (2000)
Contributions of theory to method development in
solid-phase extraction. Journal of Chromatography A,
885: 17–39.
[21] Hubaux, A.; Vos, G. (1970) Decision and detection
limits for calibration curves. Analytical Chemistry, 42:
849–855.
[22] Puig, D.; Barceló, D. (1996) Determination of phenolic
compounds in water and waste water. TRAC Trends in
Analytical Chemistry, 15: 362–375.