Metanephrine, Normetanephrine, and 3-Methoxytyramine in Human Plasma on Raptor HILIC-Si / Restek Searchable Chromatogram Library / Restek.com

	Peaks	t_R (min)	Conc. (pg/mL)	Precursor Ion	Product Ion
1.	3-Methoxytyramine-d4 (IS)	1.80	400	155.07	122.93
2.	3-Methoxytyramine	1.80	50	151.00	119.00
3.	Metanephrine-d3 (IS)	2.03	200	183.00	151.15
4.	Metanephrine	2.03	50	179.94	148.22
5.	Normetanephrine-d3 (IS)	2.11	400	169.00	136.96
6.	Normetanephrine	2.11	50	166.00	134.02

LC_CF0689

Column

Raptor HILIC-Si (cat.# 9310A52)

Dimensions:

50 mm x 2.1 mm ID

Particle Size:

2.7 µm

Pore Size:

90 Å

Temp.:

30 °C

Sample

Diluent:

Mobile phase A:mobile phase B (10:90)

Inj. Vol.:

10 µL

Mobile Phase

Water, 100 mM ammonium formate, pH 3.0

Acetonitrile

Time (min)	Flow (mL/min)	%A	%B
0.00	0.3	10	90
5.00	0.3	10	90

Detector

MS/MS

Ion Mode:

ESI+

Mode:

MRM

Instrument

UHPLC

Notes

Solid Phase Extraction Procedure:
Charcoal stripped human plasma was fortified with analytes at 50 pg/mL (0.25, 0.27, and 0.30 nmol/L for metanephrine, normetanephrine, and 3-methoxytyramine, respectively). Internal standard (IS) was prepared at 4 ng/mL for metanephrine-d3 and at 8 ng/mL for normetanephrine-d3 and 3-methoxytyramine-d4 in methanol. The plasma sample (200 μL) was mixed with 10 μL of IS solution and 600 μL of 50 mM ammonium acetate solution. The mixture was loaded in an EVOLUTE EXPRESS WCX 96-well plate (30 mg) and washed with 1 mL water and 1 mL methanol:acetonitrile (50:50). The elution was performed twice with 0.9 mL of 5% formic acid in methanol:acetonitrile (50:50) and evaporated to dryness at 55 °C under a gentle stream of nitrogen. Dried extract was reconstituted with 100 μL of diluent and injected (10 μL) for analysis.

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Metanephrine, Normetanephrine, and 3-Methoxytyramine in Human Plasma on Raptor HILIC-Si

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