Catecholamines and Metabolites in Human Urine on Raptor Biphenyl by LC-MS/MS

LC_CF0703

	Peaks	t_R (min)	Precursor Ion	Product Ion
1.	Norepinephrine-d6	1.14	158.16	111.26
2.	Norepinephrine	1.14	152.07	106.91
3.	Epinephrine-d6	1.32	172.16	111.86
4.	Epinephrine	1.33	166.07	135.00
5.	Normetanephrine-d3	1.51	169.00	136.96
6.	Normetanephrine	1.52	166.00	148.99

	Peaks	t_R (min)	Precursor Ion	Product Ion
7.	Dopamine-d4	1.55	141.16	94.69
8.	Dopamine	1.56	136.97	64.66
9.	Metanephrine-d3	1.94	183.00	151.15
10.	Metanephrine	1.96	179.94	165.01
11.	3-Methoxytyramine-d4	2.42	155.07	122.93
12.	3-Methoxytyramine	2.45	151.00	91.02

Column

Raptor Biphenyl (cat.# 9309A62)

Dimensions:

150 mm x 2.1 mm ID

Particle Size:

2.7 µm

Pore Size:

90 Å

Temp.:

30 °C

Standard/Sample

Conc.:

Endogenous level

Inj. Vol.:

2 µL

Mobile Phase

A:

Water, 0.2% formic acid

B:

Methanol, 0.2% formic acid

Time (min)	Flow (mL/min)	%A	%B
0.00	0.3	95	5
2.50	0.3	75	25
3.00	0.3	5	95
3.01	0.3	95	5
5.00	0.3	95	5

Detector	MS/MS
Ion Mode:	ESI+
Mode:	MRM
Instrument	UHPLC
Sample Preparation	A 200 µL aliquot of urine sample (Bio-Rad Lyphochek quantitative urine control, normal level) was mixed with 10 µL (1 µg/mL in methanol) internal standard solution and 600 μL of 250 mM ammonium acetate solution. The mixture was loaded onto the EVOLUTE EXPRESS WCX 96-well plate (30 mg), washed with 1 mL water and 1 mL methanol:acetonitrile (60:40), eluted with 200 µL of 5% formic acid in water:methanol (95:5), and injected (2 μL) for analysis.

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