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Thermo Scientific Poster Note
•
PN-64108-ASMS-EN-0614S
nants of emerging
ECs in samples.
by high
Orbitrap MS).
esented. Targeted
es of CECs
.g., DEET,
r degradation
eted CECs, e.g.,
, as well as their
mple preparation
d has been
strial biomes
n the CECs in
d where
n HRMS data
ducts for possible
ilable with other
were used in the
1L-amber bottles
alysis. The same
ed to observe the
-Aldrich (Oakville,
hased from CDN
ries (Andover, MA,
luting intermediate
ol (CH
3
OH) were
rity water used for
passing reverse
water purification
ed through a 200
xtraction. Sample
es, 20 mL of the
:30 (v/v), 1 mM
gates. The tubes
in and centrifuged
ss centrifuge tube
, 50:50 (v/v)). The
min at 5000 rpm
e dissolved in 100
nalysis.
High Pressure Liquid Chromatography Separation
Sample analysis was achieved on a Thermo
Scientific™ Dionex™
UltiMate
™
3000 HPLC
consisting of a HRG-3400RS binary pump, WPS-3000 autosampler, and a TCC-3400
column compartment. Separation was made by injecting 5 mL extracts into a Thermo
Scientific™
Betasil
™
and a Thermo
Scientific™
Hypersil
™
Gold, 2.1x100 mm column,
respectively, for positive and negative mode Orbitrap MS analysis. Three HPLC
separations were used for the analysis of PPCPs and their by-products.
TABLE 1. HPLC mobile phase and gradient used in the analysis
FIGURE 1.
Column oven temperature: 35
°
C; Flow rate: 450 mL/min
Mobile phase (Positive)
A: 5 mM HCOONH
4
/0.1% HCOOH in 10:90/CH
3
OH:H
2
O
B: 90:10/CH
3
OH:H
2
O
Mobile phase (Negative I) A: 10:90/CH
3
CN:H2O, pH 6.95
±
0.3
B: CH
3
CN
Mobile phase (Negative II) A: 5 mM CH
3
COONH
4
in 10:90/CH
3
CN:H2O, pH 6.95
±
0.3
B: CH
3
CN
HPLC Gradient
Time (min)
% A
% B
Curve
0.0
95
5
5
2.0
25
75
5
10.0
5
95
7
15.0
5
95
5
15.2
95
5
5
TABLE 2. Met
detection limit
deviation; RE
Current extracti
compounds. Ta
Quantitative D
Quantitative de
compounds, i.e
at the high ppb
Table 4 show
Mass Spectrometry
The HPLC was interfaced to a Thermo Scientific™
Exactive Plus
™
Orbitrap
™
MS using
a heated electrospray ionization (HESI) interface. The Orbitrap MS system was tuned and
calibrated in positive and negative modes by infusion of standard mixtures of MSCAL5
and MSCAL6. High purity nitrogen (>99%) was used in the ESI source (35 L/min). Spray
voltages used were 2500 and
−3200
V for positive and negative modes, respectively.
Mass spectrometric data was acquired at a resolving power of 140,000 (full-width-at-half-
maximum , at
m/z
200, R
FWHM
), resulting a scanning rate of > 1.5 scans/sec when using
automatic gain control target of 1.0x10
6
and a C-trap inject time of 100 msec.
Data Analysis
Thermo
Scientific™
TraceFinder
™
software were used to perform quantitative analysis
for 56 PPCPs. The same software was also used to perform non-targeted screening
along with a database of 312 compounds consisting of PPCPs and their metabolites,
steroids, hormones, perfluorohydrocarbons, surfactants, and organophosphorus flame
retardants. Quantitative analysis identified targeted compounds by retention time (RT)
obtained from extracted ion chromatogram (XIC) using a mass extraction window (MEW)
of 5 ppm. Non-targeted screening searched compounds listed in a database using
(M+H)
+
, (M+NH
4
)
+
and (M+Na)
+
adduct ions in the positive mode and (M-H)
−
quasi-
molecular ion in the negative mode, and created XICs for each compound. Those non-
targeted analytes with area counts larger than 200,000 (approximately 25
–
50 pg/mL
depending on compound), had a 5 ppm mass accuracy for the mono-isotopic mass (M)
and two isotopic peaks ((M+1) and (M+2)), and a relative intensity of 90%
±
10% from the
theoretical values were considered to be identified. Results obtained from TraceFinder
software were also exported to Thermo
Scientific™
SIEVE
TM
software to carry out a
ChemSpider
™
search.
Results
Method Performance
Figure 1 shows extraction method parameters with 100% CH
3
CN, CH
3
CN:H
2
O (0.1%
acetic acid in H2O, 70:30 (v/v), 1 mM EDTA), 100% CH
3
OH and CH
3
OH:H
2
O (0.1%
acetic acid in H2O, 70:30 (v/v). Both acetone and methanol extraction showed similar
recovery. Acetone was used in place of methanol to facilitate the evaporation step used
during the sample preparation.
Compound
19-Norethisterone
Acetamidophenol
a
-Estradiol
a
-Ethynyl Estradio
Atenolol
b
-Estradiol
Bisphenol A
Caffeine
Carbadox
Carbamazepine
Chloramphenicol
Chlorotetracycline
Ciprofloxacin
Clofibric acid
DEET
Diazepam
Diclofenac sodium
Doxycycline HCl
Enrofloxacin
Equilin
Esterone
Estriol
Gemfibrozil
Glipizide