Amphetamines by LC/MS/MS using a Kinetex C8 2.6µm, 50x2.1mm

LC Conditions

Column

Kinetex 2.6 µm C8 100 Å, LC Column 50 x 2.1 mm, Ea

Brand

Kinetex

Part No

00B-4497-AN

Phase Name

C8

Elution Type

Gradient

Mobile Phase

A: 5mM Ammonium formate with 0.1% FA

B: Methanol with 0.1% FA

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

90

10

2

1

30

70

3

3

30

70

4

3

30

70

Flow Rate

400.00 mL/min

Temperature

22°C

Detection

Mass Spectrometer (MS) @ (22°C)

Detection Info

Sample Notes

SecurityGuard™ ULTRA Guard Cartridge System extends column lifetime. - SecurityGuard ULTRA Cartridges, UHPLC C8 for 2.1mm ID Columns, 3/Pk Part No.: AJ0-8784 - Holder Part No.: AJ0-9000

Sample Preparation Details

Pretreatment Note

After eluting add 300µL of 0.5 N methanolic hydrochloride to each sample. Samples can then be blown down under a stream of N2 gas at < 40ºC until dry. Reconstitute samples with 1mL of 10% methanol in 0.1% formic acid. Before injecting onto the LC/MS dilute the samples by a factor of 20 to bring the concentration into a suitable range for analysis.

Load

Load Sample

Wash 1

2 mL of 100 mM sodium acetate buffer (pH 5.0)

Wash 2

2 mL methanol

Dry

10 min at 10 in. of Hg

Elution 1

2mLs of (7:2:1) Ethyl Acetate: Isopropanol:NH4OH mixture

Evaporation

Evaporate to dryness

Reconstitute

1 mL 1mL of 10% methanol in 0.1% formic acid