Clean-Up of Phospholipids in Human Plasma Comparing PPT VS. Strata-X PRO SPE on 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

13.5

60

40

Flow Rate

400.00 mL/min

Temperature

40°C

System

Agilent Technologies SCIEX 4500

Detection

LC/MS/MS @ (40°C)

Detection Info

Sample Notes

Analyte dilution not needed (Direct injection of the extracted sample)

Sample Preparation Details

Description

Strata-X PRO, 96-Well Plate, 30mg/well, Ea

Part No

8E-S536-TGA

Condition 1

1 mL of ACN

Sample Pretreatment

Sample Pre-treatment: • Add 100 µL Serum to tube • Add 900 µL of chilled (~0°C) ACN to the serum while vortexing • Centrifuge samples at 6000 rpm for 5 minutes and transfer supernatant

Load

supernatant (collected from pre-treatment)

Evaporation

Evaporate to dryness

Reconstitute

100 µL of initial mobile phase

Steps

0

Condition

1 mL of ACN

1

Load

supernatant (collected from pre-treatment)

2

Vacuum

Apply and collect extract

3

Dry Down

Sample under N2 and heat at 45 °C

4

Reconstitute

100 µL of initial mobile phase