LC/MS Analysis of Amphetamine Enantiomers on Astec CHIROBIOTIC® V2 in Urine after Solid Phase Extraction (SPE) using Supel™-Select SCX
Materials
analytical column
SPE tube or plate
eluent
standard
(±)-Amphetamine solution
1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
sample/matrix
1 mL of synthetic urine spiked with 50 ng/mL of each enantiomer
SPE well plate
Supel-Select SCX, 96-well plate, 30 mg/well (575664-U)
condition
1 mL 1% formic acid in acetonitrile followed by 1 mL water
sample addition
1 mL spiked urine sample
washing
2 mL water followed by 1 mL 5 mM ammonium phosphate dibasic in 50% methanol followed by 1 mL 25% v/v methanol in water
elution
1 mL 10% w/v ammonium hydroxide in acetonitrile
eluate post-treatment
evaporate to dryness under nitrogen at 40 °C, reconstitute in 1 mL mobile phase
column
Astec CHIROBIOTIC V2, 15 cm x 4.6 mm I.D., 5 μm particles (15023AST)
mobile phase
[A] methanol; [B] water; [C] acetic acid; [D] ammonium hydroxide; (95:5:0.1:0.02, A:B:C:D)
flow rate
1 mL/min
pressure
1220 psi (84 bar)
column temp.
20 °C
detector
MS, ESI(+),SIR m/z 150.1
injection
2 μL
Description
Analysis Note
Shown here is the chiral separation of amphetamine enantiomers under MS-compatible conditions on Astec CHIROBIOTIC V2 after extraction from urine using Supel-Select SCX SPE 96-well plates. The highest grade mobile phase solvents and additives were used to supply low background interference and low particulate contaminants for robust, trouble-free operation. Cerilliant CRMs provided reliable identification and quantification.
Legal Information
CHIROBIOTIC is a registered trademark of Sigma-Aldrich Co. LLC
Supel is a trademark of Sigma-Aldrich Co. LLC