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Automated Extraction of Tricyclic Antidepressants by Perkin Elmer Multiprobe II

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Sample Preparation Details

Part Number :8E-S128-TGB

Description :Strata-X-Drug B 33 µm Polymeric Strong Cation, 30 mg / well, 96-Well Plates, 2/Pk

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)

Column: Kinetex® 2.6 µm C18 100 Å, LC Column 50 x 2.1 mm, Ea

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: 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

Doxepin

DM-Doxepin

Imipramine-d3

Imipramine

Desipramine-d3

Desipramine

Nortriptyline-d3

Nortriptyline

Amitriptyline

Protriptyline-d3

Protriptyline

Clomipramine-d3

Clomipramine

DM-Clomipramine