analysis of trace organic contaminants in water by lc · pdf file ·...

Analysis of Trace Organic Contaminants in Water by LC-MS/MS

Tarun AnumolAgilent Environmental Workshop

March 18th, 2014

Introduction

Automation

Au.to.mation

“the technique of making an apparatus, a process, or a system operate automatically”

Past Present Future ?

Automation in the lab

Workbench for automated sample preparation

Robot for automated pipetting into 96 well plates

Traditional Extraction Techniques

Analysis of TOrCs in Water

Sample collection

Surrogate addition

Extraction (SPE)

Evaporation

Analysis

Conventional

Method

Sample Transport

Sample Volume

Instrument Setup

Instrument Setup

Agilent Flexcube connected to 6460 MS/MS

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

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

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)

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)

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

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

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 (%

)

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

(%)

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

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 (%

)

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 (%

)

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 (%

)

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

(%)

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)

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

(%)

Ion Suppression: Online SPE

Ultrapure water

Wastewater effluent (0.2 um filtered)

Carbamazepine_d10Concentration: 100 pptInjection Vol: 1.5 mLMass: 150 pg

Ion Suppression: Conventional SPE

Ultrapure water

Wastewater effluent (0.2 um filtered)

Carbamazepine_d10Concentration: 50 ppbInjection Vol: 3 µLMass: 150 pg

Ion Suppression: Direct Injection

Ultrapure water

Wastewater effluent (0.2 um filtered)

Carbamazepine_d10Concentration: 3 ppbInjection Vol: 50 µLMass: 150 pg

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

(%)

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)

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)

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

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.

Acknowledgements

Dr. Shane Snyder

Snyder Research Lab Dr. Sylvain Merel Dr. Sonia Dagnino

Agilent Technologies Joe Weitzel Dr. Sheher Mohsin

Questions

Contact: tanumol@email.arizona.edu