LC/MS Analysis of Opioid Glucuronide Metabolites in Urine on Ascentis® Express F5 after Solid Phase Extraction (SPE) using Supel™-Select HLB

LC/MS Analysis of Opioid Glucuronide Metabolites in Urine on Ascentis® Express F5 after Solid Phase Extraction (SPE) using Supel™-Select HLB

Conditions

sample preparation Solid Phase Extraction
sample/matrix urine spiked with drugs and metabolites at 300 ng/mL
SPE well plate Supel-Select HLB 96-well plate, 60 mg/well (575662-U)
condition 1 mL methanol followed by 1 mL water
sample addition 1 mLhydrolyzed urine
washing 1 mL water followed by 1 mL 25% methanol
elution 1 mL acetonitrile
column Ascentis Express F5, 10 cm x 2.1 mm I.D., 2.7 µm particles (53569-U)
column temp. 35 C
mobile phase [A] 5 mM ammonium formate, pH 4.2 (adjusted with formic acid); [B] 5 mM ammonium formate in (95:5 acetonitrile:water) pH 5.2 (adjusted with formic acid)
gradient 15% B held for 1 min, to 70% B in 2 min, held at 70% B for 7 min
flow rate 0.4 ml/min
sample 300 ng/mL
injection 3 µl
detector ESI(+) TOF, full scan, m/z 100-1000

Description

Analysis Note The workflow for the LC/MS analysis of native drugs and associated metabolites from urine is shown here. The convenient liquid form of beta-glucuronidase from Sigma was used for hydrolysis of the glucuronide metabolites. SPE using Supel-Select HLB yielded clean extracts. The Ascentis Express F5 columns gave highly efficient, well-resolved peaks and rapid separation. Ultra high purity LC-MS solvents and additives provided trouble-free operation, and Cerilliant CRMs gave reliable quantification., Sample pretreatment: β-glucuronidase hydrolysis
Categories Analytical Chromatography, SPE Applications, Clinical, Forensics and Toxicology, catecholamines, Opiate, Opiates
Featured Industry Clinical
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 SPE, application for HPLC

Materials

     
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