LC-MS/MS Chromatogram for Amlodipine Enantiomers and Internal Standards

LC Conditions

Column

Lux 3 µm Cellulose-4, LC Column 150 x 2 mm, Ea

Brand

Lux

Part No

00F-4490-B0

Phase Name

Cellulose-4

Elution Type

Gradient

Mobile Phase

A: 0.05% Ethanolamine in Acetonitrile and Isopropyl Alcohol (96:4 v/v)

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

100

2

4

100

Flow Rate

0.30 mL/min

Temperature

25°C

System

Sciex SCIEX 4500

Detection

LC/MS/MS @

Detection Info

Sample Preparation Details

Description

Strata-X 33 µm Polymeric Reversed Phase, 30 mg / 1 mL, Tubes, 100/Pk

Part No

8B-S100-TAK

Pretreatment Note

5 µL of internal standard working solution was added to 100 µL spiked human plasma (spiked at 0.050, 0.150, 20.0, 40.0, and 50.0 ng/mL) and mixed, 500 µL of 0.2% (v/v) Ethanolamine in water was added to sample and vortexed.

Condition 1

1 mL Methanol

Load

Spiked Plasma

Wash 1

1 mL Water

Wash 2

1 mL 20% Methanol in Water

Elution 1

1 mL of 0.1% Formic Acid in Methanol

Evaporation

Evaporate to dryness

Reconstitute

100 µL of Mobile Phase

Steps

0

Condition

1 mL Methanol

1

Equilibrate

1 mL Water

2

Load

Spiked Plasma

3

Wash

1 mL Water

4

Wash

1 mL 20% Methanol in Water

5

Elute

1 mL of 0.1% Formic Acid in Methanol

6

Dry Down

Take to dryness under a gentle stream of Nitrogen at 50 °C

7

Reconstitute

100 µL of Mobile Phase