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Automated Extraction of Tricyclic Antidepressants by Perkin Elmer Multiprobe II

Sample Preparation Details

Load :Load 0.50 mL Sample diluted in 100 mM sodium acetate buffer (pH5.0)
Wash 1 :0.8 mL 100mM sodium acetate buffer (pH5.0) at 1 mL/min
Wash 2 :0.8 mL Methanol at 1 mL/min
Dry :8-10 mins at 10" of Hg under N2 at 45 degC
Elution 1 :0.4 mL Methanol/ACN(5/50) + 2% ammonium hydroxide at 1 mL/min
Evaporate: :Evaporate to dryness
Reconstitute :0.50 mL LC initial mobile phase A/B(60/40)
Notes :To do % absolute recovery, the IS was spiked in the initial mobile phase in the reconstitution step

Application Detail (App ID: 20937)

Part No:00B-4462-AN
Dimensions:50 x 2.1 mm ID
Elution Type:Gradient
Elution A:0.1% formic acid/water
Elution B:0.1% Formic acid/methanol
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 60 40
2 3.5 20 80
3 4 20 80
4 4.01 60 40
5 5 60 40

Flow Rate:0.4 mL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ amu (22°C)
Detector Info: AB Sciex API 4000™
Note:MS/MS Parameters
Compound Q1 Q3 Dwell time DP EP CE CXP
Protrip 264.2 191.1 35 60 10 35 10
Notrip 264.3 191.2 35 60 10 35 10
DM-Dox 266.2 107.1 25 60 10 35 10
Desip 267.2 208.2 35 60 10 35 10
Protrip-D3 267.2 191.1 35 60 10 35 10
Notrip-D3 267.3 191.2 35 60 10 35 10
Desip-D3 270.2 208.2 35 60 10 35 10
Amitrip 278.2 191.2 35 60 10 35 10
Dox 280.3 107.1 25 60 10 35 10
Imip 281.3 208.2 35 60 10 35 10
Imip-D3 284.2 208.1 35 60 10 35 10
DM-Clomip 301.2 242.2 35 60 10 35 10
Clomip 315.2 242.2 35 60 10 35 10
Clomip-D3 318.2 242.2 35 60 10 35 10

Ionization Parameters
ESI in positive polarity
Gas 1 50
Gas 2 50
CAD 8
Cur 20
IS 5500
Temp 600

Analyte Details

Analyte Detail not available

1
Doxepin
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2
DM-Doxepin

3
Imipramine-d3
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4
Imipramine
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5
Desipramine-d3
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6
Desipramine
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7
Nortriptyline-d3
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8
Nortriptyline
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9
Amitriptyline
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10
Protriptyline-d3

11
Protriptyline
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12
Clomipramine-d3
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13
Clomipramine
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14
DM-Clomipramine