Automated Extraction of Tricyclic Antidepressants by Perkin Elmer Multiprobe II

LC Conditions

Column

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

Brand

Kinetex

Part No

00B-4462-AN

Phase Name

C18

Elution Type

Gradient

Mobile Phase

A: 0.1% formic acid/water

B: 0.1% Formic acid/methanol

Gradient Profile

Step No.

Time(min)

%A

%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

0.40 mL/min

Temperature

22°C

Detection

Mass Spectrometer (MS) @ (22°C)

Detection Info

Sample Notes

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

Sample Preparation Details

Description

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

Part No

8E-S128-TGB

Pretreatment Note

To do % absolute recovery, the IS was spiked in the initial mobile phase in the reconstitution step

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

Evaporation

Evaporate to dryness

Reconstitute

0.50 mL LC initial mobile phase A/B(60/40)