Extraction of Amphetamines-NIDA5 Using Strata X-C on Kinetex 2.6µm C18 50 x 2.1mm 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-5mM NH4Ac in H2O

B: 0.1%FA-5mM NH4Ac in Methanol

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

90

10

2

1

80

20

3

3

70

30

4

3.01

5

95

5

3.3

5

95

6

3.31

90

10

7

5.1

90

10

Flow Rate

450.00 mL/min

Temperature

0°C

Detection

Mass Spectrometer (MS) @ (22°C)

Detection Info

Sample Preparation Details

Description

Strata-X-C 33 µm Polymeric Strong Cation, 30 mg / 3 mL, Tubes, 50/Pk

Part No

8B-S029-TBJ

Pretreatment Note

Blown down samples (after evaporation) were reconstituted in 200 uL of initial mobile phase (A/B=90/10).

Condition 1

1 mL Methanol at 2 mL/min

Condition 2

1 mL Water at 2 mL/min

Load

Load 1.00 mL Sample diluted in 100 mM KH2PO4 (pH 6)

Wash 1

1 mL 0.1N HCl at 2 mL/min

Wash 2

1 mL Methanol at 2 mL/min

Dry

2 mins @ 10 of Hg pressure

Elution 1

0.5 mL 5% ammonium hydroxide in methanol at 1 mL/min

Elution 2

0.5 mL 5% ammonium hydroxide in methanol at 1 mL/min

Evaporation

Evaporate to dryness

Reconstitute

200 µL Initial mobile phase (A/B=90/10)