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6 Confirmatory Determination of Buprenorphine and Norbuprenorphine in Urine Using A High-Throughput LC-HRAM-MS/MS Forensic Methodology
Results
Q E ti ™ xac ve
R i thi LC th d lti h l i MS lt d i
d-MS2 (t-MS2) unn ng s me o across mu -c anne s us ng one resu e n a
of 17 500 and sample cycle time of 1.07 minutes. The method was linear from 5 to 2000
,
l i li t ng/mL for both BUP and NBUP with correlation of coefficients (R2) > 0 996
us on s .
(1/ i hti i i i d) d li it f tit ti (LOQ) f 5 / L
ect t-MS2 x we g ng; or g n gnore an a m o quan a on o ng m .
ethod Figure 3 shows a representative extracted ion chromatogram (XIC) for the
analytes and their respective IS with a mass accuracy of 5 ppm at their LOQ
(5 ng/mL) Table 5 contains a 3 day summary of precision and accuracy in. -
the QCs across two LC channels.
t of settings. Matrix Effects
The absolute recoveries of BUP and NBUP were tested in urine obtained
from six different drug-free human donors as compared to the average in the
Species water samples (n=3) Absolute recoveries of BUP in these samples ranged.
from 30 0% to 84 1% and for NBUP from 28 9% to 57 5% at 100 ng/mL The. . . . .
l i i f BUP d NBUP i h h i i IS i h
H
re at ve recover es o an w t t e r respect ve n t ese
+ samples ranged from 90.2% to 111%.
+H
H d l i C t ly ro ys s on ro
+H As a result of enzymatic hydrolysis demonstrated here between 98.7% and
113% of expected NBUP was recovered from its glucuronide standard in the
+H t l ft h d l i d LC/MS/MS l i b d th ti f th i con ro a er y ro ys s an ana ys s ase on e ra o o e r
molecular weights (0.701).
ed Figure 3 Representative chromatogram of quan and qual XICs for . .
NBUP and its IS at left and for BUP and its IS at right at LOQ (5 ng/mL).
C:\Xcalibur\...\CH1a\SurCal-2-01 05/22/1418:30:18
5.0 cal
RT: 0.00 -0.30 SM: 7G
RT:0.05
AA:57527
NL: 2.46E4
m/z=187 07442 187 07630
RT: 0.00 -0.30 SM: 7G
RT:0.10
AA:5887590
NL: 2.53E6
m/z=187 07442 187 07630
80
100
dance
SN:177
. - .
F:FTMS+p ESIFullms2
50.00
[50.00-440.00] MS ICIS
SurCal-2-01 80
100
dance
SN:7485
. - .
F:FTMS+p ESIFullms2
50.00
[50.00-500.00] MS ICIS
SurCal-2-01
40
60
ativeAbund
40
60
ativeAbund
0
20
Relat RT:0.05
AA:27610
NL: 1.26E4
m/z=211.07430-211.07642
0
20
Relat RT:0.10
AA:8292683
NL: 3.62E6
m/z=400.24004-400.24404
80
100
ndance
SN:129 F:FTMS+p ESIFullms2
50.00
[50.00-440.00] MS ICIS
SurCal-2-01 80
100
ndance
SN:6705 F:FTMS+p ESIFullms2
50.00
[50.00-500.00] MS ICIS
SurCal-2-01
20
40
60
ativeAbun 20
40
60
ativeAbun
0
Rela RT:0.05
AA:5049640
SN 8743
NL: 2.31E6
m/z=187.07442-187.07630
F:FTMS+p ESIFullms2
0
Rela RT:0.09
AA:65604
SN 74
NL: 2.99E4
m/z=187.07442-187.07630
F:FTMS+p ESIFullms2
60
80
100
undance
: 50.00
[50.00-445.00] MS ICIS
SurCal-2-01
60
80
100
undance
: 50.00
[50.00-495.00] MS ICIS
SurCal-2-01
20
40
elativeAbu
20
40
elativeAbu 0
Re RT:0.05
AA:2509319
SN:4462
NL: 1.12E6
m/z=211.07430-211.07642
F:FTMS+p ESIFullms2
0
Re RT:0.10
AA:83356
SN:165
NL: 3.52E4
m/z=396.21495-396.21891
F:FTMS+p ESIFullms2
60
80
100
bundance
50.00
[50.00-445.00] MS ICIS
SurCal-2-01
60
80
100
bundance
50.00
[50.00-495.00] MS ICIS
SurCal-2-01
20
40
RelativeAb
20
40
RelativeAb
0.00 0.05 0.10 0.15 0.20 0.25 0.30
Time (min)
0 e
0.00 0.05 0.10 0.15 0.20 0.25 0.30
Time (min)
0 e
T bl 5 I t A (3 d ) P i i (%CV 5) d A a e . n er- ssay - ays rec s on , n= an ccuracy
(%Diff, n=5) obtained by staggered injection across Channel 1 (CH1)
and Channel 2 (CH2) of a Multi-Channel LC.
rties used QC L l CH1 CH1 CH2 CH2 . Analyte eve
(ng/mL) %CV %Diff %CV %Diff
BUP 10 8 7 7 4 9 7 6 7 . . . .
800 3 3 3 3 4 2 4 4 . . . .