Extraction of Steroids Using Strata-X on Kinetex 2.6µm C18 50 x 2.1mm ID by LCMS

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%FA in H2O

B: 0.1%FA in Acetonitrile

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

80

20

2

2.5

20

80

3

2.51

5

95

4

2.8

5

95

5

2.81

80

20

6

4.6

80

20

Flow Rate

450.00 mL/min

Temperature

0°C

Detection

Mass Spectrometer (MS) @ (22°C)

Detection Info

Sample Preparation Details

Description

Strata-X 33 µm Polymeric Reversed Phase, 30 mg / 3 mL, Tubes, 50/Pk

Part No

8B-S100-TBJ

Pretreatment Note

Blown down samples (after evaporation) were reconstituted in 100 uL of initial mobile phase (A/B=80/20).

Condition 1

1 mL Methanol at 2 mL/min

Condition 2

1 mL 0.1M Acetic acid at 2 mL/min

Load

Load 0.50 mL Sample diluted in 0.1M Acetic acid

Wash 1

1 mL 0.1M Acetic acid at 2 mL/min

Wash 2

1 mL 40% Methanol/water at 2 mL/min

Dry

2 mins at 10 of Hg pressure

Elution 1

0.5 mL Methanol at 1 mL/min

Elution 2

0.5 mL Methanol at 1 mL/min

Evaporation

Evaporate to dryness

Reconstitute

100 µL Initial mobile phase (A/B=80/20)