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Extraction of Metanephrine and Normetanephrine from Plasma On strata X-CW Using Luna HILIC

Sample Preparation Details

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
Evaporate: :Evaporate to dryness
Reconstitute :100 µL Mobile Phase
Notes :Sample was reconstituted in 100 uL of 95/5 ACN/100mM ammonium formate pH 3.2

Application Detail (App ID: 18462)

Part No:00B-4449-B0
Dimensions:50 x 2 mm ID
Elution Type:Gradient
Elution A:95/5 ACN/100 mM ammonium formate, pH 3.2
Elution B:50/45/5 ACN/Water/100 mM ammonium formate, pH 3.2
Gradient Profile:
Step No. Time (min) Pct A Pct 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:0.4 mL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ (22°C)
Note: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

Analyte Details

Analyte Detail not available

1
Metanephrine
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2
Metanephrine-d3
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3
Normetanephrine
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4
Normetanephrine-d3
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