Removal of Phospholipids from Milk Sample Using Strata-X PRO With a Kinetex 2.6 µm C18 50 x 2.1 mm

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 in Water

B: 0.1%Formic acid in Methanol

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

60

40

2

0.5

5

95

3

11.5

5

95

4

11.51

60

40

5

13

60

40

Flow Rate

0.40 mL/min

Temperature

45°C

System

Agilent Technologies 1260 Infinity SCIEX 4500

Detection

LC/MS/MS @ (30°C)

Detection Info

Sample Notes

2% liquid Milk (ALTA DENA DairyPure) utilized for sample prep extraction

Sample Preparation Details

Description

Strata-X PRO, 60mg/3mL, 50/Pk

Part No

8B-S536-UBJ

Sample Pretreatment

To 1 mL of milk add 3 mL of 0.2% formic acid in Acetonitrile /Methanol (90:10). Mix/vortex for 15-20 secs. Centrifuge for 5 min at 10,000 rpm. Collect supernatant to perform SPE in next step.

Load

Supernatant (from the pre-treatment) through cartridge and collect

Evaporation

Evaporate to dryness

Reconstitute

Dried sample in 1 mL of 0.1% Formic acid in Water/ 0.1% Formic acid in Acetonitrile (95:5)

Steps

0

Load

Supernatant (from the pre-treatment) through cartridge and collect

1

Dry Down

Sample under a gentle stream of Nitrogen

2

Reconstitute

Dried sample in 1 mL of 0.1% Formic acid in Water/ 0.1% Formic acid in Acetonitrile (95:5)