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AN63539_E 04/12S
Application Note 563
Data Examples of Clinical Research Samples
Figure 4 shows the SRM chromatograms of MN and
NMN in an individual plasma sample. Figure 5 shows the
SRM chromatograms of MN and NMN in a CSS sample.
Conclusion
A fast, automated and analytically sensitive LC-MS/MS
method was developed to quantify plasma metanephrines for
clinical research purposes
4
. By using TurboFlow technology,
the sample preparation procedure was significantly simplified
compared to a previously reported offline IP-SPE method.
The presence of PFHA during the online sample preparation
was critical to the success of this method. A PGC column
was used for chromatographic separation of metanephrines.
The total online extraction and analytical LC runtime was
12 minutes. This method was linear from 6.3 to 455.4 pg/mL
for metanephrine and 12.6 to 954.5 pg/mL for
normetanephrine, with an accuracy of 80.6% to 93.5% and
80.9% to 101.7%, respectively. The lower limit of
quantitation was 6.3 pg/mL for metanephrine and 12.6 pg/mL
for normetanephrine. Inter-assay and intra-assay precision
for metanephrine and normetane-phrine at low and high
concentration level ranged from 2.0% to 10.5%.
Overall, the analytical performance achieved with this
automated online TurboFlow method is consistent with
the previously reported offline SPE method
2
. More
importantly, the online method significantly saved sample
preparation time by more than 50% and eliminated the
expense of SPE cartridges with an offline approach.
References
1.He, X. and Kozak, M. Quantitative Measurement of
Plasma Free Metanephrines by Ion-Pairing Solid Phase
Extraction and LC-MS/MS with Porous Graphitic
Carbon Column,
Thermo Scientific Application Note
:
AN539.
2.He, X.; Gabler, J.; Yuan, C.; Wang, S.; Shi, Y.; and
Kozak, M. Quantitative Measurement of Plasma Free
Metanephrines by Ion-pairing Solid Phase Extraction
and Liquid Chromatography-Tandem Mass
Spectrometry with Porous Graphitic Carbon Column,
J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.
2011
,
879
(23), 2355-2359.
3. Yuan, C; Kosewick, J; He, X; Kozak, M and Wang,
S. Sensitive Measurement of Serum1, 25-dihydroxy-
vitamin D by Liquid Chromatography/Tandem Mass
Spectrometry after Removing Interference with
Immunoaffinity Extraction,
Rapid Commun Mass
Spectrom.
2011
, 25(9), 1241-1249.
4.He, X and Kozak, M. Development of a Liquid
Chromatography–Tandem Mass Spectrometry Method
for Plasma-Free Metanephrines with Ion-pairing
Turbulent Flow Online Extraction,
Anal. Bioanal.
Chem.
2012
,
402(9)
, 3003-3010.
Figure 4. SRM chromatograms of MN and NMN in human
plasma sample
Figure 5. Representative SRM chromatograms of MN (31.0 pg/mL)
and NMN (61.0 pg/mL) in CSS sample
For Research Use Only. Not for use in diagnostic procedures.