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Thermo Scienti c Poster Note
•
PN63785_E 03/13S
TABLE 1. Method Range, Linearity and Recovery
TABLE 2. Intraday Accuracy and Precision
Preparation Liquid
that reduces solvent
then traditional systems.
, LC/MS/MS, Multiplexing
erone, the immunosuppressant
, the chemotherapeutic drugs
were validated using a Prelude
rument maintenance, down time,
ons which require sample clean-
syringe pumps designed to
analysis with a single push of
ar on pump seals and check
ps can move several hundred if
enance required on traditional
lves; therefore, syringe pumps
PLC Systemʼs also have
phases possible. The time
ed resulting in shorter run times
several LC/MS methods that are
ccessful validation of such a wide
he Prelude SPLC offers a viable
volumes resulted in methods that
run on a conventional HPLC and
tion.
olution and centrifuged.
tainers for LC-MS/MS analysis.
c HTLC-C18 XL TurboFlow
H-D
2
, 25-OH-D
3
,
sterone using a 50x2.1mm, 2.6
lumn. The detector was a TSQ
nization probe in positive mode.
(B) 10 mM ammonium formate in
All run times were 4 minutes or
uced to 2 minutes per sample.
ood with cell lysis and protein
internal standard. Testosterone
an plasma. Chemotherapeutics
thetic urine but was validated
tisol
ystem verification from three
fferent days . The interday and
-OH-D
2
and 25-OH-D
3
at a
testosterone was 0.02-10 ng/mL.
in ranges from 1-2000 ng/mL.
nM). The method precision had
Additionally, accuracy was ±15%
oefficient values for all the
oughout all concentrations and
ty, autosampler stability, and
Compound Name
Intraday Accuracy Range
(% Difference from Theoretical)
Intraday Precession Range (%RSD)
Low QC Mid QC High QC Low QC Mid QC High QC
Cyclosporin A
2.38 – 12.4 3.61 – 10.9 2.11 – 9.72
1.7 – 4.2
1.1 – 2.9
1.4 – 2.7
Sirolimus
1.78 – 16.5 2.33 – 14.9 0.11 – 13.6
7.5 -10.6
1.8 – 2.8
4.7 – 7.6
Everolimus
1.98 – 18.9 2.66 – 13.4 0.81 – 10.2
5.4 – 8.3
1.7 – 3.5
1.6 – 4.1
Tacrolimus
1.09 – 13.3 0.87 – 5.32 0.34 – 8.38
4.8 – 6.0
1.3 – 2.6
1.4 – 2.3
Testosterone
0.18 – 11.4 0.15 – 5.24 1.63 – 4.84
3.4 – 3.6
1.5 – 2.6
0.8 – 1.2
Cortisol
1.6 - 9.3
0.76 – 12.0 0.03 – 15.1
4.0 – 6.3
2.3 – 3.9
2.6 – 5.1
Busulfan
0.56 - 16.5 0.17 – 8.17 0.22 – 5.83
1.1 – 10.9
1.8 – 3.3
1.6 – 4.2
Docetaxel
0.37 – 11.9 0.14 – 5.61 0.26 – 6.98
1.6 – 9.4
1.1 – 3.7
0.9 – 3.4
Imatinib
1.0 - 9.5
0.3 – 9.8
0.0 – 11.7
1.0 – 1.9
1.1 – 7.4
1.3 – 6.2
Methotrexate
0.13 – 18.5 0.12 – 9.74 0.10 – 10.5
3.3 – 7.5
0.6 – 5.9
2.8 – 7.8
25-hydroxy Vit D2
0.5 – 14.8
0.09 – 12.5
0.3 – 11.2
5.0 – 11.5
2.9 – 6.6
1.9 – 5.1
25-hydroxy Vit D3
1.0 – 17.8
0.3 – 12.9
0.9 – 13.3
6.3 – 6.8
2.3 – 3.9
2.0 – 3.2
TABLE 3. Interday Accuracy and Precision
Compound Name
Interday Accuracy
(% Difference from Theoretical)
Interday Precession (%RSD)
Low QC Mid QC High QC Low QC Mid QC High QC
Cyclosporin A
2.00
0.75
3.06
12.2
9.7
12.2
Sirolimus
2.00
4.00
3.75
7.8
8.1
1.8
Everolimus
2.35
3.11
2.98
9.7
5.4
4.6
Tacrolimus
1.67
0.50
3.75
5.1
3.2
2.9
Testosterone
5.00
0.32
3.12
3.5
1.3
0.15
Cortisol
1.10
1.72
3.50
3.3
3.8
2.7
Busulfan
4.76
0.35
3.85
5.6
5.4
3.9
Docetaxel
2.66
1.51
1.28
4.2
4.4
3.1
Imatinib
11.0
1.33
3.74
4.0
2.0
5.9
Methotrexate
2.33
2.80
0.48
5.5
2.8
7.5
25-hydroxy Vit D2
4.83
2.52
2.87
3.9
4.0
4.8
25-hydroxy Vit D3
5.33
2.53
0.00
3.4
3.1
3.9
FIGURE 1. Standard Curves for Eac
LC/MS/MS System
Compound Name Method Range (ng/mL)
Linearity (r
2
)
Recovery
Cyclosporin A
10 – 2000
0.992 – 0.998
87.3 – 93.9
Sirolimus
1 – 50
0.998 – 0.999
86.9 – 93.9
Everolimus
1 – 50
0.992 – 0.998
88.5 – 95.2
Tacrolimus
1 – 50
0.998 – 0.999
87.3 – 97.9
Testosterone
0.020 -10.0
0.994 – 0.999
99.9 – 103.5
Cortisol
3.62 - 362
0.997 – 0.999
88.3 – 114.1
Busulfan
20 - 2000
0.995 – 0.998
89.4 – 93.5
Docetaxel
10 - 1000
0.993 – 0.999
96.6 – 102.1
Imibitib
10 - 2000
0.991 – 0.998
92.0 – 110.2
Methotrexate
10 - 750
0.992 – 0.998
102 – 111.8
25-hydroxy Vit D2
2.0 - 100
0.992 – 0.998
92.2 – 94.5
25-hydroxy Vit D3
2.0 - 100
0.992 – 0.996
95.0 - 98.9
0
200 400 600 800 1000 1200 1400 1600 1800
0
10
20
30
40
50
Cyclosporin A
Concentration (ng/mL)
Peak Area Ratio (Sample/IS)
0
10
20
30
40
0.0
0.5
1.0
1.5
2.0
2.5
Everolimus
Concentration (ng/mL)
Peak Area Ratio (Sample/IS)
0
2000
4000
6000
8000
0
10
20
30
40
50
60
70
80
Testosterone
Concentration (pg/mL)
Peak Area Ratio (Sample/IS)
0
500
1000
1500
0
5
10
15
20
25
30
Busulfan
Concentration (ng/mL)
Peak Area Ratio (Sample/IS)
0
500
1000
1500
0
10
20
30
40
50
60
70
Imatinib
Concentration (ng/mL)
Peak Area Ratio (Sample/IS)
0
20
40
60
80
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
25-Hydroxy Vitamin D2
Concentration (ng/mL)
Peak Area Ratio (Sample/IS)