11_PO-CON1751E.pdf - 第5页
5 Quantitation of plasma metanephrine and normetanephrine by derivatization using an integrated LC-MS/MS analyzer equipped with fully-automated sample preparation device Fig. 4 MRM chromatograms of NMN and NMN-d3 spiked …
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Quantitation of plasma metanephrine and normetanephrine
by derivatization using an integrated LC-MS/MS analyzer equipped
with fully-automated sample preparation device
Metanephrine (MN)
MN-d3
Normetanephrine (NMN)
NMN-d3
254.15
257.15
296.20
299.20
Precursor m/z
(derivatized)
197.23
200.23
183.20
186.20
MW
-14
-21
-17
-21
CE (V)
236.15
154.10
278.20
154.10
Product m/zCompound
Fig. 3 MRM chromatograms of MN and MN-d3 spiked in control plasma at 1 ng/mL concentration.
Shown below are the list of target compounds, their MRM transitions and the HPLC condition for LC/MS/MS analysis.
Analytical conditions
Results
MN
(run 1)
MN
(run 2)
MN-d3
(run 1)
MN-d3
(run 2)
Column : Shimpack GISS C18 (100 mm x 2.0 mm, 3 μm)
Mobile phase A : 0.1% formic acid in water
Mobile phase B : Methanol
Flow rate : 0.4 mL/min
Column temp. : 40 °C
Injection volume : 1 μL
8.09e4ISTD 257.15>154.10
A=171010
H=80171
RT =2.446
2.0 2.5 3.0
0.00
%
100.00
8.34e4ISTD 257.15>154.10
A=171613
H=81989
RT =2.443
2.0 2.5 3.0
0.00
%
100.00
7.27e4Q 254.15>236.15
A=147336
H=68771
RT =2.426
2.0 2.5 3.0
0.00
%
100.00
8.02e4Q 254.15>236.15
A=158391
H=76320
RT =2.430
2.0 2.5 3.0
0.00
%
100.00
5
Quantitation of plasma metanephrine and normetanephrine
by derivatization using an integrated LC-MS/MS analyzer equipped
with fully-automated sample preparation device
Fig. 4 MRM chromatograms of NMN and NMN-d3 spiked in control plasma at 1 ng/mL concentration.
NMN
(run 1)
NMN
(run 2)
NMN-d3
(run 1)
NMN-d3
(run 2)
1.54e6ISTD 299.20>154.10
A=5958581
H=1534491
RT =3.313
2.5 3.0 3.5
0.00
%
100.00
1.57e6ISTD 299.20>154.10
A=6077891
H=1574332
RT =3.310
2.5 3.0 3.5
0.00
%
100.00
8.42e6Q 296.20>278.20
A=31185517
H=8346933
RT =3.322
2.5 3.0 3.5
0.00
%
100.00
8.56e6Q 296.20>278.20
A=31474446
H=8478530
RT =3.317
2.5 3.0 3.5
0.00
%
100.00
Two consecutive measurement of MN and NMN were made, each starting fresh sample pretreatment. High intensity
signal was detected demonstrating that direct one-pot derivatization in plasma was successful, and that there was
minimal difference between the two runs. Under the given condition, NMN showed higher signal; derivatization
condition was likely more favorable for NMN than for MN.
Quantitation of plasma metanephrine and normetanephrine
by derivatization using an integrated LC-MS/MS analyzer equipped
with fully-automated sample preparation device
First Edition: June, 2017
© Shimadzu Corporation, 2017
Conclusion
• Although still preliminary, the experiment clearly demonstrated that complex sample pretreatment such as derivatization
was greatly facilitated by automation by CLAM-2000.
• Due to sequential pretreatment scheme, the incubation times and vortex conditions were kept constant, thus
contributing to exceptional inter-run reproducibility for a derivatization method.
• In-depth evaluation and optimization requires the use of synthetic standards for the derivatized compounds – for future
work.
Disclaimer: The products and applications in this presentation are intended for Research Use Only (RUO). Not for
use in diagnostic procedures. Not available in China.