Extraction of Metanephrine and Normetanephrine from Plasma On strata X-CW Using Luna HILIC
LC Conditions
Column
Luna 3 µm HILIC 200 Å, LC Column 50 x 2 mm, Ea
Brand
Luna
Part No
00B-4449-B0
Phase Name
HILIC
Elution Type
Gradient
Mobile Phase
A: 95/5 ACN/100 mM ammonium formate, pH 3.2
B: 50/45/5 ACN/Water/100 mM ammonium formate, pH 3.2
Gradient Profile
Step No.
Time(min)
%A
%B
1
0
100
0
2
2.5
100
0
3
2.51
0
100
4
3
0
100
5
3.01
100
0
Flow Rate
0.40 mL/min
Temperature
22°C
Detection
Mass Spectrometer (MS) @ (22°C)
Detection Info
Sample Notes
LC/MS/MS & Chromatographic Condition MS Detector: API 3000 LC/MS/MS with ESI (TurboIonSpray®) (www.Appliedbiosystems.com), Positive Ion Mode; MRM; heater gas flow 7000 cc/min; heater temperature 550 °C; Column: LUNA 3µm HILIC, 50x2.0mm; Flow rate: 0.4ml/min ESI Source Conditions: NEB: 9.00 CUR: 10.00 IS: 3500.00 TEM: 550.00 CAD: 11.00 CE 23.00 Normetanephrine CE 25.00 Metanephrine Analyte Q1 Q3 Time DP FP CE CXP NMN 166.4 134.2 150 61 370 23 8 MN 180.4 148.2 150 71 370 25 8
Sample Preparation Details
Pretreatment Note
Sample was reconstituted in 100 uL of 95/5 ACN/100mM ammonium formate pH 3.2
Condition 1
400 µL Methanol:ACN (50/50) at 1.0 mL/min
Condition 2
400 µL Water at 1.0 mL/min
Load
Load 500.00 µL Sample diluted in Water
Wash 1
800 µL Water at 1.0 mL/min
Wash 2
2000 µL Methanol:ACN (50/50) at 1.0 mL/min
Dry
For 2 mins at 10 of Hg pressure
Elution 1
400 µL 5% formic acid in Methanol:ACN (50/50) in 2 aliquots of 200 uL at 1.0 mL/min
Evaporation
Evaporate to dryness
Reconstitute
100 µL Mobile Phase
Order Items Used in This Application
- 1. Metanephrine
- 2. Metanephrine-d3
- 3. Normetanephrine
- 4. Normetanephrine-d3
-OEChem-06211003162D<br><br> 29 29 0 1 0 0 0 0 0999 V2000<br> 3.7320 1.6550 0.0000 O 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.8660 -1.8450 0.0000 O 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 -2.8450 0.0000 O 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 2.6550 0.0000 N 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 1.1550 0.0000 C 0 0 3 0 0 0 0 0 0 0 0 0<br> 4.5981 0.1550 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 1.6550 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.7320 -0.3450 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 -0.3450 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.7320 -1.3450 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.4641 -1.3450 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 -1.8450 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.3301 3.1550 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.0000 -1.3450 0.0000 C 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.5981 2.0050 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.6762 1.0724 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.0747 1.7627 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.1951 -0.0350 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.0010 -0.0350 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 4.9272 2.9650 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.0010 -1.6550 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 3.7320 2.2750 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.6401 2.6181 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.8671 3.4650 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 6.0201 3.6919 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 5.1350 -3.1550 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 2.3100 -0.8081 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1.4631 -1.0350 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1.6900 -1.8819 0.0000 H 0 0 0 0 0 0 0 0 0 0 0 0<br> 1 5 1 0 0 0 0<br> 1 22 1 0 0 0 0<br> 2 10 1 0 0 0 0<br> 2 14 1 0 0 0 0<br> 3 12 1 0 0 0 0<br> 3 26 1 0 0 0 0<br> 4 7 1 0 0 0 0<br> 4 13 1 0 0 0 0<br> 4 20 1 0 0 0 0<br> 5 6 1 0 0 0 0<br> 5 7 1 0 0 0 0<br> 5 15 1 0 0 0 0<br> 6 8 2 0 0 0 0<br> 6 9 1 0 0 0 0<br> 7 16 1 0 0 0 0<br> 7 17 1 0 0 0 0<br> 8 10 1 0 0 0 0<br> 8 18 1 0 0 0 0<br> 9 11 2 0 0 0 0<br> 9 19 1 0 0 0 0<br> 10 12 2 0 0 0 0<br> 11 12 1 0 0 0 0<br> 11 21 1 0 0 0 0<br> 13 23 1 0 0 0 0<br> 13 24 1 0 0 0 0<br> 13 25 1 0 0 0 0<br> 14 27 1 0 0 0 0<br> 14 28 1 0 0 0 0<br> 14 29 1 0 0 0 0<br>M END
Compound Name
Metanephrine
CID
21100
Molecular Formula
C10H15NO3
Molecular Weight
197
No. Hydrogen Bond Acceptors
4
No. Hydrogen Bond Donors
3
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