Rapid Sample Extraction of Chloroquine, Hydroxychloroquine, and Desethylchloroquine from Serum for LC-MS/MS Analysis

LC Conditions

Column

Kinetex 2.6 µm F5 100 Å, LC Column 50 x 2.1 mm, Ea

Brand

Kinetex

Part No

00B-4723-AN

Phase Name

F5

Elution Type

Gradient

Mobile Phase

A: 0.1 % Formic acid in water

B: Methanol

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

98

2

2

2

93

7

3

3

85

15

4

3.2

60

40

5

4.2

5

95

6

6

98

2

Flow Rate

0.50 mL/min

Temperature

20°C

System

Agilent Technologies Applied Biosystems LC/MS Model: API 4000,  Mass Analyzer:  QQQ, Interface(s): ES/APCI, PI

Detection

LC/MS/MS

Detection Info

Sample Preparation Details

Description

Impact Protein Precipitation, 2mL Square Well Filter Plate, 2/Pk

Part No

CE0-7565

Load

150ul of Serum

Evaporation

Evaporate to dryness

Steps

0

Dispense

450ul of Methanol

1

Load

150ul of Serum

2

Vortex

2 minutes

3

Vacuum

Place an empty collection plate below the Impact protein precipitation plate and pass sample through by applying vacuum, positive pressure, or centrifuge

4

Add

Dilute filtrate 1:5 with aqueous 0.1 % Formic acid