Page 129 - CRFTApplication Compendium_090613

3 Quality Controls
Quality control samples analyzed in this study showed
good recovery and reproducibility. Table 4 shows
validation statistics for quality controls analyzed in this
study. Imprecisions, as given by %CV, were also better
than those given in the package insert for all compounds
and levels tested except for those for everolimus, which
still compared favorably to the test kit (data not shown).
Table 4. Mean %bias and %CV of quality controls
Method Comparison Samples
Donor samples previously analyzed with a validated
method utilizing a triple-stage quadrupole mass
spectrometer were reanalyzed on the Exactive Plus MS.
A total of 114 samples containing tacrolimus, 34 containing
sirolimus, and 32 containing cyclosporine A were
analyzed. No donor sample values were available for
everolimus. Figure 3 shows the correlation between the
two methods. All slopes were greater than 0.9, indicating
good agreement between the two methods. R-squared
values were also greater than 0.99 for tacrolimus and
cyclosporine A and greater than 0.94 for sirolimus.
*Concentration of (tacrolimus, sirolimus, everolimus)/cyclosporine A in ng/mL
y = 0.9796x - 0.391
R = 0.99057
1
10
100
1 10 100
Triple Stage Quadrupole
Exactive Plus
Tacrolimus (n=114)
y = 0.987x - 1.5167
R = 0.99591
10
100
1000
10000
10 100 1000 10000
Triple Stage Quadrupole
Exactive Plus
Cyclosporine A (n=32)
y = 0.9132x - 1.0846
R = 0.94422
1
10
100
1 10 100
Triple Stage Quadrupole
Exactive Plus
Sirolimus (n=34)
Figure 3. Correlation of results from Exactive Plus and triple-stage
quadrupole instruments
Analyte Control 1
(3/30)*
Control 2
(12/125)*
Control 3
(25/375)*
Control 4
(0/700)*
Tacrolimus -3.20/6.60 -4.60/2.22 -1.85/4.17 NA
Sirolimus 0.691/12.9 -10.1/7.56 -14.7/4.66 NA
Everolimus -9.17/18.6 -10.9/9.49 -5.18/8.12 NA
Cyclosporine A 5.12/7.07 -3.20/4.94 1.71/3.98 8.10/3.78