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

Sample Preparation Details

Part No:8E-S035-TGB-S
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 30 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:500uL plasma (Human plasma di sodium EDTA) diluted with 1 mL water to load
Dry:For 2 mins at 10" of Hg pressure
Final Prep and Analysis: Sample was reconstituted in 100 uL of 95/5 ACN/100mM ammonium formate pH 3.2 Inject: 30 µL on HPLC / Mass Spectrometer (MS) @ (ambient)

Analytes:

Name Spiked
Conc.
Log P pKa % Rec %RSC (n=0)
1 Metanephrine 100 
2 Metanephrine-d3 500 
3 Normetanephrine 100 
4 Normetanephrine-d3 500 

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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