1. Liquid Chromatograph Mass Spectrometer
SSI-LCMS-061
Liquid Chromatography Mass Spectrometry
Simultaneous Ultrafast Analysis for
Research of Anti-Cancer Drugs
Summary: Three anti-cancer drugs were
prepared in plasma using a Biotage® SLE
96-well plate and analyzed on an LCMS-
8050 .
Background: Anti-cancer research requires
large volumes of samples covering a wide
range of drugs to be analyzed. This can
strain laboratory resources and impede the
generation of analytical data necessary to
accomplish research goals. The LCMS-8050
triple quadrupole mass spectrometer which
combines industry leading UHPLC, scanning
speeds, and detection limits was evaluated.
Method:
A 100 µl aliquot of plasma was extracted
using a Biotage® SLE 96-well plate. Samples
were diluted with H2O and eluted using
ethyl acetate. The eluent was then dried
down on an SPE Dry 96 evaporation system
and reconstituted in mobile phase, all in a
96-well plate. Samples were analyzed on an
LCMS-8050 in MRM mode with a Nexera
UHPLC front end. Drugs were separated
using a Raptor ARC C18 column (2.0 x 100-
2.7 µm particle) from Restek. All analytes
were eluted in under 1.4 minutes.
Figure 1. Chromatogram of all analytes eluted in under 1.4 minutes.
0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 1.3 1.4 1.5 min
0
50000
100000
150000
200000
250000
300000
Gemcitabine
Crizotinib
Cyclophosphamide

2. SSI-LCMS-061
Figure 2. Calibration curves and chromatograms at the LLOQ for representative analytes.
Name
Concentration
(ng/mL)
Calculated. Con.
(ng/mL)
%CV Accuracy
Gemcitabine
5 4.97 11.3 99.3
100 98.80 1.6 98.8
600 587.24 2.8 97.9
Crizotinib
5 4.93 10.3 98.6
100 96.63 1.9 96.6
600 580.10 2.8 96.7
Cyclophosphamide
5 5.03 10.9 100.6
100 108.17 3.4 108.2
600 632..37 3.1 105.4
Table 1. Quality control samples at n=5 for all three analytes.
1.25 1.50
0
250
500
750
1000
261.00>140.10(+)
0.75 1.00
0
100
200
300
400
500
264.20>112.20(+)
1.25 1.50
0
1000
2000
3000
4000
450.30>260.20(+)
Crizotinib
Gemcitabine
Cyclophosphamide
0 100 200 300 400 500 600 700 800 900 Conc. Ratio
0.0
2.5
5.0
7.5
10.0
Area Ratio
0 100 200 300 400 500 600 700 800 900 Conc. Ratio
0
1
2
3
4
5
6
7
Area Ratio
0 100 200 300 400 500 600 700 800 900 Conc. Ratio
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
Area Ratio
R2=0.998
R2=0.996
R2=0.996
O
P
O
N
H
N
Cl
Cl
O
NN
N
NH
NH2
Cl
F
Cl
N
O
N
NH2
O
OH
OH
F
F

3. SSI-LCMS-061
Results and Discussion: Three anti-cancer
drugs from different classes were extracted
from plasma. Coupling a fast analysis time
of under 1.4 minutes with SLE extraction in
a 96-well format provides a solution to the
need for ultrafast analysis of anti-cancer
drugs for high throughput research labs.
The LCMS-8050’s unrivaled scan speed and
sensitivity allows for consistent, robust, and
accurate high throughput analysis in
complex matrices.
Figure 1. shows representative
chromatograms of all analytes and their
reference ions. As demonstrated in Figure
2. and Table 1., excellent accuracy and
linearity was achieved. The accuracy of the
quality controls ranged from 96.6% to
108.2% and the R2 value for all curves was
>0.996.
Conclusion: The accurate and sensitive
detection of 3 anti-cancer drugs in under
1.4 minutes was accomplished using an
LCMS-8050 in conjunction with fast
chromatography. In addition, the
incorporation of 96-well plates for sample
preparation demonstrates compatibility
with high throughput workflows as a
complete high speed solution.