LC-MS/MS is a powerful tool for the analysis of Pharmaceuticals and Personal Care Products in environmental samples. The combination of high resolution LC separation and high sensitivity MS/MS is the most powerful tool to screen and quantify targeted compounds.

1. Stacy Tremintin and André Schreiber
AB SCIEX
Using LC-MS/MS and Advanced Software Tools
to Screen for Unknown and Non-targeted PPCP
in Environmental Samples

2. 2 © 2010 AB SCIEX
Outline
− Pharmaceuticals and Personal Care Products (PPCP), Endocrine
Disrupting Compounds (EDC), Pesticides
– Screening for PPCP in water samples
– Comprehensive screening using Multi-MRM methods
– Direct injection of water with LOD < 10ppt
– Hybrid triple quadrupole linear ion trap systems (QTRAP®)
− Compound Identification
– MRM ratio
– MS/MS library searching
– Multi-Target Screening (MTS) and Quantitation
− General Unknown Screening (GUS)
– Non-targeted peak finding algorithm
– Statistical data analysis (Principal Components Analysis PCA)
– Automatic data reporting

3. 3 © 2010 AB SCIEX
LC-MS/MS analysis of EDC and PPCP
− Screening for hundreds or possibly thousands of compounds covering a wide
range of chemical properties in environmental samples (water, soil, biota…)
− Minimum sample cleanup without time consuming derivatization and generic LC
setup
− LC-MS/MS ideal for analysis of medium polar, polar and ionic compounds in
trace levels
− Positive and negative ionization with different ion sources (ESI, APCI, APPI)
− Quantitation and identification at ng/L (ppt) levels
− Screening for unknown environmental pollutants

4. 4 © 2010 AB SCIEX
Screening strategies using LC-MS/MS
− Screening using full scan (high resolution) spectra lacks in selectivity and sensitivity
causing a high risk of false positive and false negative results
− Fragment ions are needed for identification
− 2 Multiple Reaction Monitoring transitions per compound
– MRM ratio for identification [1]
– Scheduled MRM for extended screening [3-4]
− Information Dependent Acquisition (IDA) of MS/MS
– Multi Target Screening: MRM to trigger MS/MS [2]
– Unknown Screening: Full scan MS to trigger MS/MS [5]
[1] Christopher Borton et al.: at ASMS conference (2006) Seattle, WA
[2] Jim Krol et al.: at AOAC annual meeting (2007) Anaheim, CA
[3] André Schreiber et al.: at ASMS conference (2008) Denver, CO
[4] Rolf Kern et al.: at Pittcon (2009) Chicago, IL
[5] André Schreiber et al.: at LC/MS/MS workshop (2010) Barcelona, Spain

5. 5 © 2010 AB SCIEX
Experimental
− Sample preparation
– Direct injection of drinking water samples
– SPE cleanup of waste water and sea water
− HPLC separation
– Shimadzu Prominence UFLC XR system
– Different RP columns with fast gradients of water/methanol and volatile
buffer, such as (ammonium acetate, formate, formic acid)
− MS/MS detection
– New QTRAP® 5500 LC-MS/MS system with Turbo V™ source and
Electrospray Ionization (ESI) probe
– MRM with Scheduled MRM™ algorithm for targeted quantitation
– Enhanced MS (EMS) for non-target screening
– Enhanced Product Ion (EPI) CE=35 with CES=15V for compound
identification using MS/MS library searching

6. 6 © 2010 AB SCIEX
Q1
Q3/LIT
Q2/Q2L
Q0
QJet
Q3L
Orifice
Curtain Plate
New Technology in the 5500 QTRAP® Systems
− TurboV™ source for highest ionization efficiency, ruggedness and
robustness with low carry-over
− Ceramic interface and QJet® 2 ion guide for best ion transmission
− Space saving design of collision cell with Linear Accelerator fields
− Faster electronics for fast MRM (2ms dwell time) and polarity
switching in 50ms without loss in S/N
− Linear Accelerator Q3 for fastest QTRAP® scanning (20,000Da/s)

7. 7 © 2010 AB SCIEX
Multi Target Screening and Quantitation

8. 8 © 2010 AB SCIEX
MRM offers highest Selectivity and Sensitivity
Ion filtering
Ion transport
Ion production Ion filtering
Fragmentation

9. 9 © 2010 AB SCIEX
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0
Time, min
0.0
5.0e5
1.0e6
1.5e6
1.7e6
0.58
cycle
time
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0
Time, min
0.0
5.0e5
1.0e6
1.5e6
1.7e6
0.58
cycle
time
2.9 3.0 3.1 3.2 3
Time min
0.0
2.0e4
4.0e4
6.0e4
8.0e4
9.1e4
3.1
cycle
time
2.9 3.0 3.1 3.2 3
Time min
0.0
2.0e4
4.0e4
6.0e4
8.0e4
9.1e4
3.1
cycle
time
MRM in Multi-Target Analysis
Many MRM transitions with long cycle time – insufficient number of data points
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0
Time, min
0.0
5.0e5
1.0e6
1.5e6
1.7e6
0.58
cycle
time
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0
Time, min
0.0
5.0e5
1.0e6
1.5e6
1.7e6
0.58
cycle
time
2.9 3.0 3.1 3.2 3.
Time, min
0.00
2.00e4
4.00e4
6.00e4
8.00e4
1.00e5
1.11e5
3.1
cycle
time
2.9 3.0 3.1 3.2 3.
Time, min
0.00
2.00e4
4.00e4
6.00e4
8.00e4
1.00e5
1.11e5
3.1
cycle
time
Many MRM transitions with short dwell time – for better qualitative data

10. 10 © 2010 AB SCIEX
The Key for Multi-Target Analysis – Smart Software
− Adjusts detection windows automatically depending on retention time
− Optimizes dwell times for each analyte and cycle time
→ Allows detecting many more MRM transitions
→ Allows using UHPLC
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0
Time, min
0.0
5.0e5
1.0e6
1.5e6
1.7e6
0.58
0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0
Time, min
0.0
5.0e5
1.0e6
1.5e6
1.7e6
0.58
Scheduled MRM™ Algorithm

11. 11 © 2010 AB SCIEX
Maintaining Quantitative Accuracy
Maximizing Effective Duty Cycle
− Poor peak sampling makes robust peak integration very difficult
(left)
− Proper cycle time provides good peak sampling and increases
assay reproducibility
8.4% CV 5.3% CV
12% CV 7.8% CV
300 Normal MRMs 300 Scheduled MRMs

12. 12 © 2010 AB SCIEX
XIC of +MRM (158 pairs): 415.1/178.1 amu Expected RT: 5.2 ID: Diltiazem 1 from Samp... Max. 8.2e6 cps
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Time, min
0.0
1.0e6
2.0e6
3.0e6
4.0e6
5.0e6
6.0e6
7.0e6
8.0e6
5.1
LC-MS/MS Screening – Direct Injection of Water
− 158 Scheduled MRM™ transitions to screen for PPCP
(EPA 1694)
− Injection of 100µL into QTRAP® 5500 LC/MS/MS system
− Phenomenex Luna C18 HST (100x3mm 2.5µm) and
gradient of water/acetonitrile with 0.1% formic acid
1μg/L
(100μL injected)

13. 13 © 2010 AB SCIEX
XIC of +MRM (158 pairs): 216.1/96.1 amu Expected RT: 6.9 ID: Atrazine 2 from Sample 37 (Farmland Drai... Max. 1.1e5 cps.
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Time, min
0.00
5000.00
1.00e4
1.50e4
2.00e4
2.50e4
3.00e4
3.50e4
4.00e4
4.50e4
5.00e4
5.50e4
6.00e4
6.50e4
7.00e4
7.50e4
8.00e4
8.50e4
9.00e4
9.50e4
1.00e5
1.05e5
1.10e5
1.13e5
6.9
70ppt Atrazine
6ppt Metolachlor
2ppt
Sulfadimethoxazine
2ppt
Propazine
XIC of +MRM (158 pairs): 216.1/96.1 amu Expected RT: 6.9 ID: Atrazine 2 from Sample 37 (Farmland Drai... Max. 1.1e5 cps.
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Time, min
0.00
5000.00
1.00e4
1.50e4
2.00e4
2.50e4
3.00e4
3.50e4
4.00e4
4.50e4
5.00e4
5.50e4
6.00e4
6.50e4
7.00e4
7.50e4
8.00e4
8.50e4
9.00e4
9.50e4
1.00e5
1.05e5
1.10e5
1.13e5
6.9
70ppt Atrazine
6ppt Metolachlor
2ppt
Sulfadimethoxazine
2ppt
Propazine
LC-MS/MS Screening – Farmland Drain Sample
LOD of PPCP <10ppt

14. 14 © 2010 AB SCIEX
XIC of +MRM (158 pairs): 216.1/96.1 amu Expected RT: 6.9 ID: Atrazine 2 from Sample 36 (Urban) of 200... Max. 1.9e4 cps.
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Time, min
0.00
5000.00
1.00e4
1.50e4
2.00e4
2.50e4
3.00e4
3.50e4
4.00e4
4.50e4
5.00e4
5.50e4
6.00e4
6.50e4
7.00e4
7.50e4
8.00e4
8.50e4
9.00e4
9.50e4
1.00e5
1.05e5
1.10e5
1.15e5
6.9
7.26.96.6
10ppt Atrazine
200ppt Cotinine
20ppt
Diuron
100ppt Metformin
5ppt Metolachlor
2ppt Simazine1ppt
Thiabendazole
420ppt Caffeine
XIC of +MRM (158 pairs): 216.1/96.1 amu Expected RT: 6.9 ID: Atrazine 2 from Sample 36 (Urban) of 200... Max. 1.9e4 cps.
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Time, min
0.00
5000.00
1.00e4
1.50e4
2.00e4
2.50e4
3.00e4
3.50e4
4.00e4
4.50e4
5.00e4
5.50e4
6.00e4
6.50e4
7.00e4
7.50e4
8.00e4
8.50e4
9.00e4
9.50e4
1.00e5
1.05e5
1.10e5
1.15e5
6.9
7.26.96.6
10ppt Atrazine
200ppt Cotinine
20ppt
Diuron
100ppt Metformin
5ppt Metolachlor
2ppt Simazine1ppt
Thiabendazole
420ppt Caffeine
LC-MS/MS Screening – Urban Water Sample
LOD of PPCP <10ppt

15. 15 © 2010 AB SCIEX
Multi-Target Screening with
MS/MS Library Searching

16. 16 © 2010 AB SCIEX
An Alternative to MRM Ratios – Full Scan MS/MS
Ion transport
Ion production
Q3 of a QTRAP® System can be used as quadrupole or as linear ion trap
Ion trapping
Ion filtering Ion filtering
Fragmentation

17. 17 © 2010 AB SCIEX
Trapping and Scanning in a Linear Ion Trap (Q3)
Exit lens
(1) Trapping
Entrance lens
(2) Scanning

18. 18 © 2010 AB SCIEX
Multi-Target Screening with MS/MS Library Search
intensity threshold
297
159
69 173
255
176
20181 141
109 115
Threshold
Library Search
Scheduled MRM™ Algorithm
EPI

19. 19 © 2010 AB SCIEX
XIC of +MRM (79 pairs): Exp 1, 130.1/71.1 amu Expected RT: 0.6 ID: Metformin from S... Max. 9.2e4 cps
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Time, min
0.0
2.0e4
4.0e4
6.0e4
8.0e4
9.2e4
0.5
0.8
+EPI (311.30) Charge (+0) CE ... Max. 1.2e5 cps.
100 150 200 250 300 350 400
m/z, Da
0.00
5.00e4
1.00e5
1.16e5
311.0
108.0
156.0
92.0 292.9
+EPI (216.10) Charge (+0) CE ... Max. 1.1e6 cps
100 150 200 250 300 350 400
m/z, Da
0.00
5.00e5
1.00e6
1.15e6
Intensity,cps
216.1
174.1
103.767.9
146.0
78.6
+EPI (230.20) Charge (+0) CE ... Max. 1.9e5 cps.
100 150 200 250 300 350 400
m/z, Da
0.0
5.0e4
1.0e5
1.5e5
1.9e5
230.2
146.0
79.0 195.1 212.2
+EPI (284.20) Charge (+0) CE ... Max. 7.9e4 cps
100 150 200 250 300 350 400
m/z, Da
0.0
2.0e4
4.0e4
6.0e4
7.9e4
Intensity,cps
252.1
284.2
176.2
133.9 146.373.0 184.0 266.5
LC-MS/MS Screening – Farmland Drain Sample
Scheduled MRM™ – EPI
Sulfadimethoxazine
FIT 83.7%
Atrazine
FIT 98.3%
Propazine
FIT = 98.8%
Metolachlor
FIT=91.8%

20. 20 © 2010 AB SCIEX
XIC of +MRM (79 pairs): Exp 1, 216.1/174.1 amu Expected RT: 6.9 ID: Atrazine from Sample 15 (Urban) ... Max. 3174.0 cps.
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Time, min
0.0
1.0e4
2.0e4
3.0e4
4.0e4
5.0e4
5.7e4
7.0
+EPI (130.10) Cha... Max. 2.0e5 cps.
100 150 200
m/z, Da
0.0
5.0e4
1.0e5
1.5e5
2.0e5
130.1
71.0
88.8 112.1
60.8
102.0
+EPI (177.20) Cha... Max. 3.1e5 cps.
100 150 200
m/z, Da
0.0
5.0e4
1.0e5
1.5e5
2.0e5
2.5e5
3.0e5
Intensity,cps
177.1
80.0
149.3 159.0118.172.8
97.9
+EPI (202.10) Cha... Max. 1.4e5 cps.
100 150 200 250
m/z, Da
0.0
2.0e4
4.0e4
6.0e4
8.0e4
1.0e5
1.2e5
1.4e5
Intensity,cps
202.1
184.1
175.0
131.0
139.1
156.070.9 90.8
+EPI (202.10) Cha... Max. 5.7e4 cps.
100 150 200 250
m/z, Da
0.0
1.0e4
2.0e4
3.0e4
4.0e4
5.0e4
5.7e4 202.1
130.1114.8
105.0 127.9 167.273.8
+EPI (216.10) Cha... Max. 7.4e4 cps.
100 150 200 250 300
m/z, Da
0.0
2.0e4
4.0e4
6.0e4
7.4e4
Intensity,cps
216.2
170.0
133.9
188.2
76.9
129.1 154.0 198.2
96.1
+EPI (284.20) Cha... Max. 8.2e4 cps.
100 150 200 250 300 350
m/z, Da
0.0
2.0e4
4.0e4
6.0e4
8.0e4
Intensity,cps
284.3
252.0
176.0
134.0
163.1105.0 266.2
Metformin
Fit 57.8%
Cotinine
Fit 76.8%
Simazine
Fit 83.8%
Metolachlor
Fit 87.9%
Thiabendazole
Fit 94.2%
Atrazine
Fit 94.9%
XIC of +MRM (79 pairs): Exp 1, 216.1/174.1 amu Expected RT: 6.9 ID: Atrazine from Sample 15 (Urban) ... Max. 3174.0 cps.
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0
Time, min
0.0
1.0e4
2.0e4
3.0e4
4.0e4
5.0e4
5.7e4
7.0
+EPI (130.10) Cha... Max. 2.0e5 cps.
100 150 200
m/z, Da
0.0
5.0e4
1.0e5
1.5e5
2.0e5
130.1
71.0
88.8 112.1
60.8
102.0
+EPI (177.20) Cha... Max. 3.1e5 cps.
100 150 200
m/z, Da
0.0
5.0e4
1.0e5
1.5e5
2.0e5
2.5e5
3.0e5
Intensity,cps
177.1
80.0
149.3 159.0118.172.8
97.9
+EPI (202.10) Cha... Max. 1.4e5 cps.
100 150 200 250
m/z, Da
0.0
2.0e4
4.0e4
6.0e4
8.0e4
1.0e5
1.2e5
1.4e5
Intensity,cps
202.1
184.1
175.0
131.0
139.1
156.070.9 90.8
+EPI (202.10) Cha... Max. 5.7e4 cps.
100 150 200 250
m/z, Da
0.0
1.0e4
2.0e4
3.0e4
4.0e4
5.0e4
5.7e4 202.1
130.1114.8
105.0 127.9 167.273.8
+EPI (216.10) Cha... Max. 7.4e4 cps.
100 150 200 250 300
m/z, Da
0.0
2.0e4
4.0e4
6.0e4
7.4e4
Intensity,cps
216.2
170.0
133.9
188.2
76.9
129.1 154.0 198.2
96.1
+EPI (284.20) Cha... Max. 8.2e4 cps.
100 150 200 250 300 350
m/z, Da
0.0
2.0e4
4.0e4
6.0e4
8.0e4
Intensity,cps
284.3
252.0
176.0
134.0
163.1105.0 266.2
Metformin
Fit 57.8%
Cotinine
Fit 76.8%
Simazine
Fit 83.8%
Metolachlor
Fit 87.9%
Thiabendazole
Fit 94.2%
Atrazine
Fit 94.9%
LC-MS/MS Screening – Urban Water Sample
Scheduled MRM™ – EPI
?

21. 21 © 2010 AB SCIEX
XIC of +MRM (158 pairs): 130.1... Max. 1.8e5 cps.
2 4 6 8 10
Time, min
0.0
5.0e4
1.0e5
1.5e5
1.8e5
0.6
+EPI (130.10) Charge (+0) CE ... Max. 1.2e6 cps
80 100 120 140 160 180 200
m/z, Da
0.00
2.00e5
4.00e5
6.00e5
8.00e5
1.00e6
1.20e6
Intensity,cps
88.9
70.9
68.9
112.0
83.9
75.8
130.2
XIC of +MRM (158 pairs): 130.1... Max. 2.7e4 cps.
2 4 6 8 10
Time, min
0.0
1.0e4
2.0e4
3.0e4
4.0e4
5.0e4
0.7
+EPI (130.00) CE (20): Exp 1, ... Max. 1.1e6 cps
80 100 120 140 160 180 200
m/z, Da
0.00
2.00e5
4.00e5
6.00e5
8.00e5
1.00e6
1.14e6
Intensity,cps
71.0
130.2
85.168.0
113.0
66.4 132.0
LC-MS/MS Screening – MRM vs. EPI Identification
Standard 1μg/L
MRM ratio 0.083
Water sample 1μg/L
MRM ratio 0.082
Library spectrum
(Metformin)
Unknown spectrum
?

22. 22 © 2010 AB SCIEX
General Unknown Screening
and Quantitation

23. 23 © 2010 AB SCIEX
4 . 7
9 . 2
7 . 2
8 . 5
9 . 7
1 4 . 46 . 2
0 . 8
1 2 . 59 . 9
1 1 . 1
3 . 9
1 3 . 1
1 3 . 9
1 0 . 5
6 . 6
8 . 2
2 . 9 4 . 1
0 . 5 7 . 4
5 . 8
2 . 2
5 . 2
1 . 8
4 . 7
9 . 2
7 . 2
8 . 5
9 . 7
1 4 . 46 . 2
0 . 8
1 2 . 59 . 9
1 1 . 1
3 . 9
1 3 . 1
1 3 . 9
1 0 . 5
6 . 6
8 . 2
2 . 9 4 . 1
0 . 5 7 . 4
5 . 8
2 . 2
5 . 2
1 . 8
4 . 7
9 . 2
7 . 2
8 . 5
9 . 7
1 4 . 46 . 2
0 . 8
1 2 . 59 . 9
1 1 . 1
3 . 9
1 3 . 1
1 3 . 9
1 0 . 5
6 . 6
8 . 2
2 . 9 4 . 1
0 . 5 7 . 4
5 . 8
2 . 2
5 . 2
1 . 8
intensity threshold, DBS
297
159
69 173
255
176
20181 141
109 115
Threshold
Library Search
EMS survey
EPI
General Unknown Screening using a Non-Target
Peak Finding Algorithm in Cliquid® Software

24. 24 © 2010 AB SCIEX
Comparison of chromatograms and mass
spectra
TIC of +MRM (53 pairs): Exp 1, from Sample 9 (EDC River 2 Influent) of Samples POS mode_1EPI.wiff (Turbo Spray) Max. 5.2e5 cps.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
Time, min
0.0
5.0e4
1.0e5
1.5e5
2.0e5
2.5e5
3.0e5
3.5e5
4.0e5
4.5e5
5.0e5
8.17
6.42 7.626.24
1.74 5.46
10.06
7.14 8.39
10.58
TIC of +MRM (53 pairs): Exp 1, from Sample 10 (EDC River 2 Effluent) of Samples POS mode_1EPI.wiff (Turbo Spray) Max. 9.9e5 cps.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
Time, min
0.0
1.0e5
2.0e5
3.0e5
4.0e5
5.0e5
6.0e5
7.0e5
8.0e5
9.0e5
9.9e5
8.18
5.45
7.64
1.73
5.27 6.25
4.58 10.07
1.90 6.45
7.174.13
2.36
+EMS: Exp 1, 0.010 to 19.238 min from Sample 1 (GUS Testmix in urine extract (150ng/mL)) of Data Forensic samples ST Boniface_08.... Max. 8.1e5 cps.
100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600
m/z, amu
1.0e5
2.0e5
3.0e5
4.0e5
5.0e5
6.0e5
7.0e5
8.0e5
195.1
149.2
114.1
153.1
432.2
304.2
138.1
374.3
470.2211.2 416.3
171.1 287.2235.2 309.1 335.2120.1
251.2 346.3177.1156.1 311.2 429.2301.2123.1 227.2 448.2379.2193.2 475.1273.1 365.2355.2144.1 185.2
315.2298.2 413.3130.1 449.3385.2 445.2242.3 494.4267.2 467.2 540.2 563.3 573.2
+EMS: Exp 1, 8.024 to 29.929 min from Sample 1 (GUS Testmix in urine extract (150ng/mL)) of Data Forensic samples ST Boniface_08.... Max. 1.6e6 cps.
100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600
m/z, amu
2.0e5
4.0e5
6.0e5
8.0e5
1.0e6
1.2e6
1.4e6
1.6e6
149.0
332.4
432.2360.4153.1 305.2
287.2 374.3195.1 470.2416.3
363.2124.1 338.4330.2 429.2251.2211.2 391.3311.2167.2 284.3201.1 475.1427.3369.2155.2 522.5357.2 536.2 583.2105.1 423.2 485.3
2 unknown samples
Chromatograms (TIC) Mass spectra

25. 25 © 2010 AB SCIEX
PCA Explanation 1
x
y
2 dimensional plot

26. 26 © 2010 AB SCIEX
PCA Explanation 2
x
y
z
3 dimensional plot

27. 27 © 2010 AB SCIEX
PCA Explanation 3
x
y
z
(0, 0, 0)
PC1
PC1 Score
PC1 = 0.1 x + 0.3 y +0.95 z
PC1

28. 28 © 2010 AB SCIEX
Analysis of “Street Cocaine” Samples
Cocaine
Procaine
Diltiazem
Benzoylecgonine
Sample 1
Sample 2
Sample 3
Cocaine
Procaine
Diltiazem
Benzoylecgonine
Sample 1
Sample 2
Sample 3
Principal Components Analysis (PCA) – scores and loadings plot

29. 29 © 2010 AB SCIEX
Samples from wilderness areas
with extensive recreational use
Increasing urbanity of samples
Drinking water produced from
river bank filtrate during flood
DEET
Screening for Unexpected PPCP
Distribution of the insect repellent DEET
3.3µg/L
2.4µg/L
2.5µg/L
3.6µg/L
2.6µg/L

30. 30 © 2010 AB SCIEX
Quantitation of Benzoylecgonine (Metabolite of
Cocaine) in 70 Water Samples
0.000
0.025
0.050
0.075
0.100
0.125
0.150
0.175
0.200
Sam ple
BEConcentration(µg/
Std. 0.1µg/L
Drinking water
Rivers
Creeks
Zermatt
Berlin
Philadelphia
RiodeJaneiro
Rome
Vienna
SantaFe
Toronto
Dresden
Canoe Lake (Algonquin)
Lake Zürich
Lakes
0.000
0.025
0.050
0.075
0.100
0.125
0.150
0.175
0.200
Sam ple
BEConcentration(µg/
Std. 0.1µg/L
Drinking water
Rivers
Creeks
Zermatt
Berlin
Philadelphia
RiodeJaneiro
Rome
Vienna
SantaFe
Toronto
Dresden
Canoe Lake (Algonquin)
Lake Zürich
Lakes

31. 31 © 2010 AB SCIEX
Cliquid® – General Unknown Screening

32. 32 © 2010 AB SCIEX
Summary
− LC-MS/MS is a powerful tool for the analysis of Pharmaceuticals and Personal Care
Products in environmental samples
– The combination of high resolution LC separation and high sensitivity MS/MS is
the most powerful tool to screen and quantify targeted compounds.
– Direct injection of water samples
– MS/MS fragmentation required to identify compounds
– MRM transitions to quantify targeted contaminants
– MRM ratios or MRM to trigger MS/MS spectra (EPI) for compound identification
with high confidence
– GUS coupled with library searching and PCA is a powerful research tool for any
laboratory

33. 33 © 2010 AB SCIEX
Trademarks/Licensing
For Research Use Only. Not for use in diagnostic procedures.
The trademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their
respective owners.
© 2010 AB SCIEX.

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