LC/MS (TOF) Analysis of Opioid Drugs and Glucuronide Metabolites in Urine on Ascentis® Express HILIC after SPE using Supel™-Select HLB, β-Glucuronidase Enzyme Digestion

LC/MS (TOF) Analysis of Opioid Drugs and Glucuronide Metabolites in Urine on Ascentis® Express HILIC after SPE using Supel™-Select HLB, β-Glucuronidase Enzyme Digestion

Conditions

sample preparation SPE (Solid Phase Extraction)
sample/matrix urine spiked at 300 ng/mL
SPE well plate Supel-Select HLB 96-well plate, 60 mg/well (575662-U)
condition 1 mL methanol, 1 mL water
sample addition 1.0 mLurine
washing 1 mL water, 1 mL 25% methanol
elution 1 mL acetonitrile
column Ascentis Express HILIC, 10 cm x 2.1 mm x 2.7 µm particles (53939-U)
mobile phase [A]: 5 mM ammonium formate pH 4.2 [B]: 5 mM ammonium formate (95:5 acetonitrile:water) pH 5.2
gradient 100% B for 3 min, to 80% B in 7 min, held for 2 min
flow rate 0.6 mL/min
sample 300 ng/mL
injection 3 µL
detector ESI(+) TOF, m/z 100-1000

Description

Analysis Note The glucuronide metabolites were converted back to the parent drug using β-glucuronidase, then processed using SPE prior to LC/MS analysis using HILIC mode chromatography. The Supel-Select HLB provided clean extracts and the Ascentis Express HILIC provided rapid resolution of parent drugs and metabolites. Cerilliant CRMs provided reliable quantitation.
Categories Analytical Chromatography, SPE Applications, Opiate, Opiates
Featured Industry Forensics and Toxicology
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, Supel is a trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC

Materials

     
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