Supporting Information for
Ultrasonic Assisted Extraction Method for Simultaneous Determination of
Emerging Contaminants in Freshwater Sediments
Diana Nara Ribeiro de Sousa1, Guilherme Martins Grosseli1, Antonio Aparecido
Mozeto1, Renato Lajarim Carneiro2, Pedro Sergio Fadini1*
1
Laboratório de Biogeoquímica Ambiental,
1,2
2
Grupo de Quimiometria Aplicada,
Departamento de Química, Universidade Federal de São Carlos - UFSCar, Rodovia
Washington Luís km 235, São Carlos 13.565-905, SP, Brazil.
*Corresponding Author: Tel: +55 1633066432, Fax: +55 1633518065.
E-mail adresses: dnrsousa@yahoo.com.br (D. N. R. Sousa), grosseli@ufscar.br (G. M.
Grosseli), amozeto@ufscar.br (A. A. Mozeto), renato.lajarim@ufscar.br (R. L.
Carneiro), psfadini@ufscar.br (P. S. Fadini)*.
Table of Contents
Table S1. Characteristic of emerging contaminants studied.
Table S2. Monitored conditions for analysis of the target compounds by UPLCMS/MS.
Table S3. Characteristics of the analyzed sediment river samples.
Table S1. Molecular structure and physical-chemical properties of emerging contaminants studied.
Emerging
Water
Characteristics
CAS
Log
a
Compound
Contaminant
pKa
Solubility
number
Kowa
Use
(mg L-1)a
Atenolol
29122-68-7 0.16
9.6
13300
Basic
β-Blocker
Caffeine
Stimulant
58-08-2
-0.07
10.4
21600
Neutral
Carbamazepine Antiepileptic
3564-73-6
2.46
7.0
16.8
Neutral
Diclofenac
Antinflamatory
15307-86-5 4.51
4.15
2.37
Acidic
Ibuprofen
Analgesic and
15687-27-1 3.97
4.91
21.0
Acidic
Antinflamatory
Naproxen
Analgesic and
22204-53-1 3.18
4.15
15.9
Acidic
Antinflamatory
Propranolol
525-66-6
3.48
9.42
61.7
Basic
β-Blocker
Triclosan
Bactericide
3380-34-5
4.76
7.68
10
Acidic
Estrone
Hormone
53-16-7
3.13
10.3b
30
Basic
b
Hormone
50-28-2
4.01
10.3
3.6
Basic
17-β-Estradiol
b
17-αHormone
57-63-6
3.67
10.3
11.29
Basic
Ethinylestradiol
a
www.syrres.com
b
strongest acidic (www.drugbank.ca)
Table S2. Monitored conditions for analysis of the target compounds by UPLC-MS/MS.
Target compounds
Analyzed in negative mode
Naproxen
Naproxen-d3
Ibuprofen
Ibuprofen-d3
Diclofenac
Diclofenac-d4
Estrone
Estrone-d4
17-β-estradiol
17-β-estradiol-d5
17-α-ethinylestradiol
17-α-ethinylestradiol-d4
Triclosan
Triclosan-d3
Rt window (min)
1.55 - 2.05
1.80 - 2.40
2.80 - 3.20
3.15 - 4.00
Analyzed in positive mode
Caffeine
1.50 - 1.85
Caffeine-d3
Atenolol
1.85 - 2.25
Atenolol-d7
Carbamazepine
2.40 - 2.85
Carbamazepine-d10
Propranolol
3.20 - 3.70
Propranolol-d7
CV-CE: Cone Voltage-Collision Energy (eV).
Precursor ion
CV-CE
MRM1
229
232
205.1
208
294
298
269.1
273.2
271.2
276.2
295.2
299.2
286.9
292.6
15-15
12-8
15-8
15-8
20-12
20-12
53-38
53-35
53-35
53-40
55-40
55-40
18-10
18-10
170.1
188
161.1
164
250.1
254
145.1
147
145.1
147
145
147
35
35
195
198
267
274
237.1
247
260.1
267.2
35-20
35-22
32-30
32-28
33-20
33-25
34-24
34-25
138.1
138
145
145
194.1
204
72
72
CV-CE
MRM2
15-8
185.1
15-5
205.1
20-12
296>252
53-40
143
53-40
183.1
55-34
159
18-7
288.9>35
35-30
42
32-20
190
33-20
192
34-22
116
Table S3. Characteristics of the analyzed sediment river samples.
Site
S1
S2
S3
S4
S5
S6
TOC (%) ± SD
3.80 ± 0.38
0.49 ± 0.04
1.63 ± 0.04
0.88 ± 0.04
0.98 ± 0.04
2.34 ± 0.05
Clay (%)
30
22
35
30
18
37
Silt (%)
10
5
16
8
3
10
Sand (%)
60
73
49
62
79
53
TOC – Total Organic Carbon
UPLC-MS/MS Analysis
The chromatographic separation was performed with a reverse phase column Acquity
UPLC BEH C18 (1.7 microns, 2.1 x 50 mm) equipped with a guard column of the same
material, both obtained from Waters. Ultrapure water (0.05% NH4OH) and methanol were
used as mobile phases in gradient elution mode, a flow rate of 0.45 mL min-1 and 10 μL of
injection volume. Analysis were performed in positive and negative modes with capillary
voltage 3000 V and 2500 V, respectively. The temperatures used were 500 °C for the
desolvation gas and 150 ºC for the source block.
Method Validation
Samples were quantified by internal standard calibration with a mixture of
water:methanol (75:25, v/v). A minimum eight-point calibration curves (0.1 – 400 μg L-1)
were obtained using linear regression analysis, corresponding at concentration of 0.05 – 200
ng g-1. The concentration of each IS was fixed at 100 μg L-1 (50 ng g-1). The linearity was
qualified by coefficient of determination (r2) in the linear regression. The recoveries of each
compound were used to correct the final concentration sample. Analysis and data processing
was performed using MassLynx 4.1 software with two transitions for each compound in the
Multiple Reaction Monitoring mode (MRM).
Recoveries were calculated based on the peak areas of the analytes in the spiked
sediment at different steps of extraction. For calculation of the total recovery (Rec T), a
sediment sample was spiked with a mixture of the target compounds and IS, both at 50 ng g-1,
in triplicate (X). The spiked matrices were mixed in vortex for 30 s and keeping at -20 °C for
24 h to equilibration time. After that, the extraction was carried out according to procedure
described in 2.3. In another tube, a sediment sample was extracted without spiking. At this
sample, the mixture of the target compounds and IS was added just in reconstitution step (Z).
RecT was obtained through the ration between peak areas for X and Z (Equation 1). SPE step
recoveries (RecSPE) was calculated from an extract obtained from UAE step (without spiking)
diluted in 250 mL of ultrapure water. In this solution was added a mixture of same
compounds and IS (Y) and then carried out the SPE extraction. RecSPE was obtained through
the ration between peak areas for Y and Z (Equation 2). UAE recovery (RecUAE) was
calculated as the difference between RecT and RecUAE (Equation 3). These relationships are
presented in Equations (1) to (3):
𝑋
𝑅𝑒𝑐𝑇 = ( ) × 100
𝑍
𝑌
𝑅𝑒𝑐𝑆𝑃𝐸 = ( ) × 100
𝑍
(1)
𝑅𝑒𝑐𝑈𝐴𝐸 = 100 − (𝑅𝑒𝑐𝑆𝑃𝐸 − 𝑅𝑒𝑐𝑇 )
(3)
(2)
The matrix effect (ME) was evaluated in terms of the suppression or increasing of
signal according to Stahnke et al. (2011) [1]. This effect was calculated using Equation (4),
where A corresponds to the peak area for standard solutions in solvent and B for the spiked
sample after the total extraction from the sediments. Negative ME values represent analyte
signal suppression and positive values denote matrix-induced enhancements. ME values close
to zero are desirables, since this represents the non-occurrence of the ME.
𝐵
𝑀𝐸(%) = ( − 1) × 100
𝐴
(4)
The overall precision of the method was evaluated at the level concentration of 50 ng
g-1 of analyte and IS for each compound. These samples were extracted for three consecutive
days, and for each day the extractions was carried out in triplicate. This sample is a mixture
of different sampling days at same site, with different sampling condition. The ruggedness of
the method can also be confirmed by this heterogeneous and complex sample. The precision
was expressed as the relative standard deviation (RSD) of replicates measurements.
The method limits of detection and quantification (MDL and MQL) were calculated
using spiked samples that were submitted to UAE-SPE. These calculations were made
according to Baker et al. (2011) [2], in equations (5) and (6), total recovery (RecT) and the
concentration factor used in the SPE extraction (CF), were considered. Firstly, the
instrumental detection limit and instrumental quantification limit (IDL and IQL) were
calculated using the signal-to-noise ratio, from a sample that was submitted to UAE-SPE and
spiked during the reconstitution. For the IDL we considered a signal-to-noise over than 3, and
for the IQL a signal-to-noise over 10 was established for the first transition and over 3 for the
second transition. CF used in this study was 250 and the recovery values at the concentration
level of 50 ng g-1.
𝑀𝐷𝐿 =
𝐼𝐷𝐿 × 100
𝑅𝑒𝑐𝑇 × 𝐶𝐹
(5)
𝑀𝑄𝐿 =
𝐼𝑄𝐿 × 100
𝑅𝑒𝑐𝑇 × 𝐶𝐹
(6)
References
[1] Stahnke, H., Kittlaus, S., Kempe, G., Alder, L., Anal. Chem. 2012, 84, 1474-1482.
[2] Baker, D. R., Kasprzyk-Hordern, B., J. Chromatogr. A 2011, 1218, 1620-1631.