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Therapeutic Drug Monitoring of 9 New Anticancer Agents by High-Performance Liquid Chromatography-Tandem Mass Spectrometry
DASA
SUNI
ccuracy
Precision
Accuracy
Precision
106.9
16.2
119.8
20.0
101.5
8.2
99.7
6.3
98.3
11.9
97.6
7.0
97.8
6.0
93.9
11.0
97.2
7.7
90.7
5.1
98.4
5.6
91.3
4.0
102.2
3.0
105.8
2.1
IMAT
LAPA
Accuracy
Precision
Accuracy
Precision
93.4
12.8
105.5
7.7
98.1
9.9
96.8
6.6
107.6
7.8
109.2
6.1
98.3
5.9
90.5
7.8
100.2
5.1
96.8
6.1
99.2
3.8
99.6
4.8
100.8
2.3
101.9
2.7
SORA
VAND
ccuracy
Precision
Accuracy
Precision
113.1
5.4
86.5
16.2
98.9
3.3
91.7
11.0
108.2
5.8
111.2
17.4
91.8
5.7
103.8
10.7
91.2
8.4
108.5
5.0
98.3
2.7
101.3
2.4
105.3
3.3
96.0
3.3
and 4 controls samples are given in Table
cted to reflect low, medium and high range
y were chosen to encompass the clinically
lasma. The mean intra-assay precision was
and lower than 8.2 %. Overall, the mean
ithin 5.3 and 13.8%. The intra-assay and
ntrations of QCs for each considered TKI
113.5 %. Ratios of ion transitions were
ation for all of them below 25%.
Extraction recovery and matrix effect
The assessment of matrix effects and extraction recoveries is reported in Table 5. A
value above or below 100% for the matrix effects indicates an ionization
enhancement or suppression, respectively. Matrix effects and extraction yields were
ranged from 84.6 to 109 % and 84.0 to 101.2% respectively. Overall recoveries
were ranged from 77.8 to 93.3 % for lower concentrations, 78.6 to 98.4% for
medium concentrations and from 79.8 to 105.6 % for higher concentrations. The
extraction recovery of D8-imatinib was 93.7%. There was no effect of
hyperbilirubinemia, hyperlipemia and haemolysis on matrix effect as evaluated in
medium CQs.
Stability
The stability of TKIs in human plasma samples was studied with low and high QC
samples left at room temperature up to 48h. The variations are contained within
±
15% of starting concentrations indicating that TKIs can be considered stable at RT
excepted for lapatinib which decreases of -36% at RT after 24h and of -76% after
48h. It has been demonstrated that lapatinib was stable at RT for 6 hours. Sunitinib
is sensitive to light and decreases by -15% after 48h even light protection. By
contrast, all TKIs in plasma samples left during the same period of time at +4
°
C
were found stable.
QC samples prepared in human plasma undergoing three freeze-thaw cycles
showed no significant degradation (variation < 8.2 %) for all analytes.
Long-term stability studies indicated that all analytes were stable in human plasma
when stored at -70
°
C for 150 days (ratios between 96.0 to 100.5%, degradation <
7.9%).
The stability of stock solutions held at -70
°
C and left in the dark for 10 months
showed decrease less than 6% for each analyte.
In neutral extracts, all analytes were stable up to 7h when left in the autosampler
without any degradation allowing more than 40 samples to be analyzed
simultaneously within a single chromatographic batch.
External quality controls
The external quality controls (low and high concentrations) for imatinib (18
laboratories), nilotinib and dasatinib (9 laboratories) showed a good accuracy (97.2
to 101.4%) in comparison to data obtained from others laboratories.
Application to biological samples
We applied the assay to the analysis of samples obtained from patients receiving
imatinib, nilotinib, dasatinib, sunitinib or sorafenib.
DASA, IMAT and NILO were frequently detected in patients with chronic myeloid
leukemia (n=75). In 71 patients treated with 400 (84%) or 600 mg imatinib daily,
detected though concentrations were around 871 ng/mL (median: 789 ng/mL).
Among these 71 patients, 45 % of them presented a major molecular response
associated with a trough concentration higher than 1,000 ng/mL such as
recommended [50].
We applied the assay to samples provided from an obese patient treated with 50
mg sunitinib for a renal carcinoma. The profile of SUNI concentrations measured in
this obese woman showed no difference with AUC (1592
±
41 ngh/ml) observed in
patients without obesity.
Conclusion
In overall, the method that has been developed is precise, accurate and sensitive. It
concerns nine inhibitors of tyrosine kinase acquired in a single run Confirmation is
performed using confirmation/quantification ion ratios criteria. The method is very
simple and therefore used in a routine environment for clinical studies; it is also
possible to add new TKIs that could potentially have an interest in clinical practices
and performed a partial analytical validation. The dynamic range of the
concentrations allow to carry out some pharmacokinetics studies.
ion samples for BORT, DASA, SUNI and of the high
RA, VAND in human plasma (n=20)
were observed at the drugs retention time
valuated. The endogeneous responses in
of the signal at the LLOQ of 2 ng/mL for
e others. The endogeneous responses in
ients were always less than 7.1% of the
cts of others concomitant treatments (40
in, 25 mg/l of vancomycin, ceftazidime,
rphine, 3 mg/l of docetaxel, 5 mg/l of
nd fluconazole).
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