Tech Note: Rapid and Comprehensive Screening for ... - AB Sciex

Rapid and Comprehensive Screening for Pharmaceuticals
and Personal Care Products using LC-MS/MS with Fast
Polarity Switching
André Schreiber 1 , Tania Sasaki 2
1 AB SCIEX, Concord, Ontario, Canada; 2 AB SCIEX, Foster City, California, USA
Introduction
There is an emerging environmental concern that
pharmaceuticals and personal care products (PPCP) are
entering and contaminating the drinking water supply. Chemicals
like hormones and antibiotics are especially of interest because
of proven endocrine disrupting effects and a possible
development of bacterial resistance. Powerful screening
methods are required to detect and quantify the presence of
these compounds in our environment.
LC-MS/MS is the technology of choice to monitor numerous
compounds in environmental samples. Multiple Reaction
Monitoring mode (MRM) is typically used because of its excellent
sensitivity, selectivity, and speed. Because PPCP span such a
wide variety of compound classes and chemical properties, it is
necessary to employ both positive and negative Electrospray
Ionization (ESI) for complete analysis. It is also desirable to
obtain the maximum amount of information in the shortest
amount of time.
The novel AB SCIEX QTRAP ® 5500 Systems incorporates the
proven technology of the Turbo V source and the Curtain
Gas interface for ultimate sensitivity and robustness. The
advanced eQ electronics and the new Qurved LINAC ®
collision cell was designed for unparalleled speed of MRM
detection and fast polarity switching for multi-component
analysis. In addition, the new Linear Accelerator Trap
technology allows the acquisition of fast and highly sensitive
Enhanced Product Ion (EPI) spectra for compound confirmation
with highest confidence.
The ability of the QTRAP ® 5500 System to detect a large panel
of PPCP while performing fast positive/negative switching,
resulting in the maximum amount of information from a single
LC-MS/MS injection is demonstrated.
Method Details
• Ultra High Pressure Liquid Chromatography using a
Shimadzu UFLCXR system with a Zorbax SB-C18 column (1.8
μm) and a gradient of water and methanol with 0.1% formic
acid
• AB SCIEX QTRAP ® 5500 System with Turbo V Source and
ESI probe
• Experiment 1: dedicated positive mode to monitor 78 MRM
transitions (5 ms dwell time and 3 ms pause time)
• Experiment 2: dedicated negative mode to monitor 33 MRM
transitions (5 ms dwell time and 3 ms pause time)
• Experiment 3: positive/negative switching to monitor all 111
MRM transitions (3 ms dwell time, 3 ms pause time, and
settling time of 50 ms)
• Experiment 4: positive/negative switching to monitor all 111
MRM transitions with Information Dependent Acquisition (IDA)
of EPI spectra with CE = +/- 35V and CES = 15V
p 1

Table 1. Details comparing parameters of the various experiments: Chromatographic peak widths were on the order of 8-10 seconds base-to-base.
Cycle times were adjusted to allow a minimum of 10 scans across the chromatographic peak.
Experiment MRM Transitions Dwell Time (ms) Cycle Time (ms)
Dedicated positive 78 5 624
Dedicated negative 33 5 264
Polarity switching 111 3 766
Results
Intensity, cps
Intensity, cps
XIC of +MRM (78 pairs): 235.2/86.1 Da ID: Lidocaine_1 from Sample 3 of Dedicated Posit... Max. 1.2e6 cps.
1.20e6
1.00e6
8.00e5
6.00e5
4.00e5
2.00e5
0.00
0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0
Time, min
10.0 11.0 12.0 13.0 14.0 15.0 16.0
XIC of +MRM (78 pairs): Exp 1, 235.2/86.1 Da ID: Lidocaine_1 from Sample 3 of Pos Neg... Max. 1.1e6 cps.
1.20e6
1.00e6
8.00e5
6.00e5
4.00e5
2.00e5
Dedicated
positive polarity
Positive
negative switching
Figure 1. LC-MS/MS analysis in positive polarity: Comparison of a data
acquired using a dedicated positive mode acquisition method (top) versus
a method that utilized positive/negative switching (bottom)
Intensity, cps
Intensity, cps
Figure 1a. Zoomed in view of Figure 1
6.5
6.5
0.00
0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0
Time, min
10.0 11.0 12.0 13.0 14.0 15.0 16.0
XIC of +MRM (78 pairs): 235.2/86.1 Da ID: Lidocaine_1 from Sample 3 of Dedicated Posit... Max. 1.2e6 cps.
4.0e5
3.0e5
2.0e5
1.0e5
0.0
2.0 3.0 4.0 5.0 6.0 7.0 8.0
Time, min
9.0 10.0 11.0 12.0 13.0 14.0
XIC of +MRM (78 pairs): Exp 1, 235.2/86.1 Da ID: Lidocaine_1 from Sample 3 of Pos Neg... Max. 1.1e6 cps.
4.0e5
3.0e5
2.0e5
1.0e5
0.0
Dedicated
positive polarity
Positive
negative switching
2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0
Time, min
Intensity, cps
Intensity, cps
XIC of -MRM (33 pairs): 307.0/160.9 Da ID: Warfarin_1 from Sample 7 of Dedicated Nega... Max. 3.0e6 cps.
2.5e6
2.0e6
1.5e6
1.0e6
5.0e5
0.0
2 4 6 8
Time, min
10 12 14 16
XIC of -MRM (33 pairs): Exp 2, 307.0/160.9 Da ID: Warfarin_1 from Sample 7 of Pos Neg... Max. 2.7e6 cps.
2.7e6
2.5e6
2.0e6
1.5e6
1.0e6
5.0e5
0.0
Dedicated
negative polarity
Positive
negative switching
2 4 6 8 10 12 14 16
Time, min
Figure 2. LC-MS/MS analysis in negative polarity: Comparison of a data
acquired using a dedicated negative mode acquisition method (top)
versus a method that utilized positive/ negative switching (bottom)
Intensity, cps
Intensity, cps
XIC of -MRM (33 pairs): 307.0/160.9 Da ID: Warfarin_1 from Sample 7 of Dedicated Nega... Max. 3.0e6 cps.
3.0e5
2.5e5
2.0e5
1.5e5
1.0e5
5.0e4
0.0
7.0 7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0
Time, min
11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0
XIC of -MRM (33 pairs): Exp 2, 307.0/160.9 Da ID: Warfarin_1 from Sample 7 of Pos Neg... Max. 2.7e6 cps.
3.0e5
2.5e5
2.0e5
1.5e5
1.0e5
5.0e4
0.0
Dedicated
negative polarity
Positive
negative switching
7.0 7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0
Time, min
Figure 2a. Zoomed in view of Figure 2
11.8
p 2

Data from the dedicated positive and negative experiment were
compared to data from the polarity switching experiment. Peak
areas and signal-to-noise (S/N) were compared, as well as %CV
of the peak areas.
Intensity, cps
Intensity, cps
Intensity, cps
Intensity, cps
XIC of +MRM (78 pairs): 195.5/13... Max. 2.3e4 cps.
2.0e4
1.0e4
(+) Caffeine
6.9
1.9e4
(+/-)Caffeine
S/N = 24.7 1.0e4 S/N = 20.0
0.0
5.0 5.5 6.0 6.5 7.0 7.5
Time, min
8.0 8.5
XIC of +MRM (78 pairs): 235.2/86... Max. 1.1e6 cps.
1.00e6
5.00e5
(+) Lidocaine
6.5
1.0e6
(+/-) Lidocaine
S/N = 705
5.0e5
S/N = 922
0.00
4.5 5.0 5.5 6.0 6.5 7.0
Time, min
7.5 8.0 8.5
XIC of +MRM (78 pairs): 213.1/72... Max. 9.1e4 cps.
9.1e4
5.0e4
0.0
9.0 9.5 10.0 10.5 11.0
Time, min
11.5 12.0 12.5
XIC of -MRM (33 pairs): 312.9/15... Max. 2.8e5 cps.
2.8e5
2.0e5
1.0e5
0.0
Variations between the sets of data were minimal, demonstrating
that no significant loss of data quality was observed when a
positive/negative switching method was used versus a dedicated
method (see Figure 1-4).
Dedicated polarity Positive negative switching
XIC of +MRM (78 pairs): Exp 1, 1... Max. 1.9e4 cps.
0.0
5.0 5.5 6.0 6.5 7.0 7.5
Time, min
8.0 8.5
XIC of +MRM (78 pairs): Exp 1, 2... Max. 1.0e6 cps.
Figure 3. Comparison of S/N for representative analytes: Data from the dedicated positive or negative experiment is shown on the left and data acquired
using a positive/negative switching experiment is shown on the right. No significant degradation of data quality was observed when using the polarity
switching experiment
Intensity, cps
Intensity, cps
6.9
6.5
0.0
4.5 5.0 5.5 6.0 6.5 7.0
Time, min
7.5 8.0 8.5
XIC of +MRM (78 pairs): Exp 1, 2... Max. 9.1e4 cps.
10.7
(+) Chlorotoluron
9.1e4
10.7
(+/-) Chlorotoluron
S/N = 52.6 5.0e4 S/N = 48.8
11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0
Time, min
Intensity, cps
0.0
9.0 9.5 10.0 10.5 11.0
Time, min
11.5 12.0 12.5
XIC of -MRM (33 pairs): Exp 2, 3... Max. 1.9e5 cps.
(-) Triclocarban
13.3
1.9e5
13.3
(+/-) Triclocarban
S/N = 869
1.0e5
S/N = 592
Intensity, cps
0.0
11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0
Time, min
p 3

Percent of compounds
Percent of compounds
60
50
40
30
20
10
0
90
80
70
60
50
40
30
20
10
0
Figure 4. Comparison of reproducibility: Peak areas of all analytes
monitored across five replicate injections and the %CV for each analyte
were determined. The charts compare the percent of compounds having
coefficients of variation in the various ranges. %CV from the dedicated
positive or negative experiments were compared with the %CV from a
positive/negative switching experiment. Even with increased acquisition
cycle times in the switching experiment, %CV of