analysis of trace organic contaminants in water by lc · pdf file ·...
TRANSCRIPT
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Analysis of Trace Organic Contaminants in Water by LC-MS/MS
Tarun AnumolAgilent Environmental Workshop
March 18th, 2014
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Introduction
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Automation
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Au.to.mation
“the technique of making an apparatus, a process, or a system operate automatically”
Past Present Future ?
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Automation in the lab
Workbench for automated sample preparation
Robot for automated pipetting into 96 well plates
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Traditional Extraction Techniques
Analysis of TOrCs in Water
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Sample collection
Surrogate addition
Extraction (SPE)
Evaporation
Analysis
Conventional
Method
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Sample Transport
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Sample Volume
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Instrument Setup
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Instrument Setup
Agilent Flexcube connected to 6460 MS/MS
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Configuration
10
9
76
2
3
4
5
Binary Pump
SPE cartridge 1
Flex cube pump
Sample Injection
Loop
Analytical Column
SPE cartridge 2
1
Mass Spectrometer
Waste
Flexcube Pump Flow Path
Binary Pump Flow Path
Equilibrating
Loading
8
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Configuration
10
9
8
76
2
3
4
5
Binary Pump
SPE cartridge 1
Flex cube pump
Sample Injection
Loop
Analytical Column
SPE cartridge 2
1
Mass Spectrometer
Waste
Flexcube Pump Flow Path
Binary Pump Flow Path
Elution
Loading
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Trace Organic Chemicals
Atenolol (ß-blocker) Atrazine (Herbicide)
Bisphenol A (plasticizer) Caffeine (stimulant)
Benzophenone (UV-blocker)
Carbamazepine (Anti-seizure)
DEET (Insect-repellant) Dexamethasone (glucocorticoid) Trimethoprim (Antibiotic)
Estrone (Hormone) Fluoxetine (Anti-depressant) Gemfibrozil (Anti-cholesterol)
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Trace Organic Chemicals
Ibuprofen (Analgesic) Meprobamate (Anti-anxiety)
PFBS (Fluoro-surfactant) PFOA (Fluoro-surfactant)
Naproxen (Pain-reliever)
PFOS (Fluoro-surfactant)
Primidone (Anticonvulsant) Simazine (Herbicide) Sulfamethoxazole (Antibiotic)
TCPP (Flame-retardant) Triclocarban (Anti-microbial) Triclosan (Anti-microbial)
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Analysis of OSPE Cartridges
150 ng on column
<50%
50-60
%
60-70
%
70-80
%
80-90
%
90-10
0%
100-1
10%
110-1
20%
120-1
30%
>130
%
70-13
0%
0
4
8
12
16
20
24
28
SB-AQPLRP-sPhenyl-hexylCarbon-X
Absolute recovery range (%)
Num
ber
of c
ompo
unds
PLRP-s selected for further studies
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Loading Volume
PLRP-s
Flowrate: 1 mL/min
4 mL loading volume selected
for further studies
<50%
50-60
%
60-70
%
70-80
%
80-90
%
90-10
0%
100-1
10%
110-1
20%
120-1
30%
>130
%
70-13
0%
0
2
4
6
8
10
1215
18
21
24
27
30
2.0 mL2.5 mL3.0 mL3.5 mL4.0 mL5.0 mL
Absolute recovery range (%)
Num
ber
of c
ompo
unds
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Loading Volume
Atenolol
Caffein
e Trim
ethoprim
Prim
idone
Sulfameth
oxazo
le
Carbam
ezap
ine TCEP
Atrazin
e Tes
tostero
ne PFOA
Benzo
phenone
Ibuprofen
PFOS
Gemfib
rozil
Tric
losan
0
25
50
75
100
125
150
175
200
2.0 mL2.5 mL3.0 mL3.5 mL4.0 mL5.0 mL
Abso
lute
rec
over
y (%
)
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Loading Rate
Atenolol
Caffein
e
Trimeth
oprim
Primidone
Norges
trel
Mepro
bamate
Sulfameth
oxazo
le
Benzo
triaz
ole
Hydro
corti
sone
Diphenhyd
ramine
Simaz
ine Ditia
zem
Carbam
ezap
ine TCEP
Fluoxetin
e Atra
zine
DEET PFHxA
Bisphen
ol A
Propylp
araben
Testoste
rone
Clofibric
Acid
Benzo
phenone
Napro
xen
PFOA TCPP
Diclofen
ac
Ibuprofen
Gemfib
rozil
PFOS
Tricloca
rban
Tric
losan
0
25
50
75
100
125
150
175
0.5 mL/min1.0 mL/min1.5 mL/min
Increasing Retention Time
Abus
olut
e R
ecov
ery
(%)
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Analytical Method
Ate
nolo
l
Caf
fein
eTr
imet
hopr
im
Prim
idon
e
Mep
roba
mat
e
Diphenhydramine
Dilt
iaze
m
TCE
PD
EE
TTe
stos
tero
neN
apro
xen
PFO
A TCP
PB
enzo
phen
one
Dic
lofe
nac
Ibup
rofe
n Gem
fibro
zil
PFO
STr
iclo
carb
an
Tric
losa
n
Sim
azin
e
Pre
dnis
one
Ben
zotri
azol
e
Atra
zine
Simultaneous analysis of 36 TOrCsin positive and negative ESI
Injection Volume: 1.7 mL
Cycle time (Extraction + Analysis): 14.5 min
Analytical Column: Poroshell 120 EC, 2.1x50 mm
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Method Validation
Atenolol
Atrazin
e
Benzo
phenone
Bisphen
ol ACaff
eine
Carbam
ezap
ineDEET
Diphenhyd
ramine
Ditiaze
mGem
fibro
zil
Hydro
chloro
thiazide
Ibuprofen
Mepro
bamate
Napro
xen
PFOAPFOS
Primidone
Sulfameth
oxazo
leTCPP
Testoste
rone
Triclosa
nTrim
ethoprim
0
25
50
75
100
125
150Ultrapure Water (30 ng/L)Ultrapure Water (100 ng/L)
Rec
over
y (%
)
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Method Validation
Atenolol
Atrazin
e
Benzo
phenone
Bisphen
ol ACaff
eine
Carbam
ezap
ineDEET
Diphenhyd
ramine
Ditiaze
mFluoxe
tine
Gemfib
rozil
Hydro
chloro
thiazide
Ibuprofen
Mepro
bamate
PFOAPFOS
Primidone
Sulfameth
oxazo
leTCPP
Testoste
rone
Triclosa
nTrim
ethoprim
0
25
50
75
100
125
150Surface Water (30 ng/L)Surface Water (100 ng/L)
Rec
over
y (%
)
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Method Validation
Atenolol
Atrazin
e
Benzo
phenone
Benzo
triaz
ole
Bisphen
ol ACaff
eine
Carbam
ezap
ine
Clofibric
Acid
DEETDicl
ofenac
Diphenhyd
ramine
Ditiaze
mFluoxe
tine
Gemfib
rozil
Hydro
chloro
thiazide
Hydro
corti
sone
Ibuprofen
Mepro
bamate
Napro
xen
Norges
trel
PFHxAPFOAPFOS
Primidone
Propran
olol
Propylp
araben
Simaz
ine
Sulfameth
oxazo
leTCEPTCPP
Testoste
rone
Tricloca
rban
Triclosa
n
Trimeth
oprim
0
25
50
75
100
125
150
1:5 Diluted Waste Water (100 ng/L)
Rec
over
y (%
)
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Method Validation
Atenolol
Atrazin
e
Benzo
phenone
Benzo
triaz
ole
Bisphen
ol ACaff
eine
Carbam
ezap
ine
Clofibric
Acid
DEETDicl
ofenac
Diphenhyd
ramine
Ditiaze
mFluoxe
tine
Gemfib
rozil
Hydro
chloro
thiazide
Hydro
corti
sone
Ibuprofen
Mepro
bamate
Napro
xen
Norges
trel
PFHxAPFOAPFOS
Primidone
Propran
olol
Propylp
araben
Simaz
ine
Sulfameth
oxazo
leTCEPTCPP
Testoste
rone
Tricloca
rban
Triclosa
n
Trimeth
oprim
0
3
6
9
12
15
Intra-day variability (n=4)Inter-day variability (n=4)
Rel
ativ
e St
anda
rd D
evia
tion
(%)
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Method Detection Limit
8 replicates using Glaser et al. method
Atenolol
Atrazin
e
Benzo
phenone
Benzo
triaz
ole
Bisphen
ol ACaff
eine
Carbam
ezap
ine
Clofibric
Acid
DEETDicl
ofenac
Diphenhyd
ramine
Ditiaze
mFluoxe
tine
Gemfib
rozil
Hydrac
ortiso
ne
Hydro
chloro
thiazide
Ibuprofen
Mepro
bamate
Napro
xen
Norges
trel
PFHxAPFOAPFOS
Primidone
Propran
olol
Propylp
araben
Simaz
ine
Sulfameth
oxazo
leTCEPTCPP
Testoste
rone
Tricloca
rban
Triclosa
n
Trimeth
oprim
0.0625
0.125
0.25
0.5
1
2
4
8
16
MDL (ng/L)
Con
cent
ratio
n (n
g/L)
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Matrix Effects
7
Atenolol d
5
Atrazin
e d
10
Benzo
phenone d
4
Benzo
triaz
ole d 12C13
Bisphen
ol A
3C13
Caffein
e
10
Carbam
ezap
ine d
6
DEET d
6C13
Diclofen
ac
5
Diphenhyd
ramine d
3
Ditiaze
m d
5
Fluoxetin
e d
6
Gemfib
rozil
dIbupro
fen d3 3
Mepro
bamate
d 3 d1C13
Napro
xen
2 C13
PFHxA
4 C13
PFOA
4 C13
PFOS
5
Primidone d
4
Propylp
araben
d
6C13
Sulfameth
oxazo
le
12
TCEP d
6 C13
Tricloca
rban
12C13
Triclosa
n
3
Trimeth
oprim d
Averag
e-30
-20
-10
0
10
20
30
40
50
60
70
80
90
100SW WWE (1:5 dil) WWE
Mat
rix E
ffect
(%)
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Ion Suppression: Online SPE
Ultrapure water
Wastewater effluent (0.2 um filtered)
Carbamazepine_d10Concentration: 100 pptInjection Vol: 1.5 mLMass: 150 pg
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Ion Suppression: Conventional SPE
Ultrapure water
Wastewater effluent (0.2 um filtered)
Carbamazepine_d10Concentration: 50 ppbInjection Vol: 3 µLMass: 150 pg
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Ion Suppression: Direct Injection
Ultrapure water
Wastewater effluent (0.2 um filtered)
Carbamazepine_d10Concentration: 3 ppbInjection Vol: 50 µLMass: 150 pg
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Ion Suppression Effects
Comparison of 3 methods
Atenolol d
7
Sulfameth
oxazo
le 13
C3Carb
amaz
epine d
10
DEET d6
TCEP d12Gem
fibro
zil d6
0
20
40
60
80
100
Conventional SPE (500 fold)Online SPEDirect Injection
Surr
ogat
e R
ecov
ery
(%)
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Ion Suppression Effects
Atenolol
Mepro
bamate
Sulfameth
oxazo
le
Carbam
azep
ine
DEET
TCEP
Gemfib
rozil
0.1
1
10
100
1000
CSPEOSPELVI
Met
hod
Det
ectio
n Li
mit
(ng/
L)
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Ion Suppression Effects
Atenolol
Mepro
bamate
Sulfameth
oxazo
le
Carbam
azep
ine
DEET
TCEP
Gemfib
rozil
0.1
1
10
100
1000
CSPE-MRL
OSPE-MRL
LVI-MRL
Met
hod
Rep
ortin
g Li
mit
(ng/
L)
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Analysis of Emerging Contaminants in Water
Conventional SPE Method Online SPE Method Direct Injection Method
1 L sample 1.5 mL sample 0.1 mL sample
0.01-10 ng/L
36 CECs
5 hours
0.1-20 ng/L
33 CECs
30 min
10-100 ng/L
21 CECs
15 min
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Conclusions Online SPE allows sensitive analysis of trace organics in water while
allowing significant time and labor savings.
Method has been proven to be robust in several different water matrixes.
Online SPE is significantly less affected by ion suppression compared with conventional offline extraction techniques.
LVI is currently not sensitive enough to attain desired MRLs but offers promise with rapid increase in sensitivity of newer mass spectrometers.
Use of online SPE with accurate mass detectors (ToF & Q-ToF) could allow for real-time analysis of trace unknowns in water.
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Acknowledgements
Dr. Shane Snyder
Snyder Research Lab Dr. Sylvain Merel Dr. Sonia Dagnino
Agilent Technologies Joe Weitzel Dr. Sheher Mohsin