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
材料
used together
相关产品
吗啡-3- β-D-葡糖醛酸 溶液
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100 μg/mL in methanol: water (2:8), ampule of 1 mL, certified reference material, Cerilliant®Hydromorphone-3-β-D-glucuronide
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100 μg/mL in methanol: water (1:1), ampule of 1 mL, certified reference material, Cerilliant®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 mL urine
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
detector
ESI(+) TOF, m/z 100-1000
injection
3 μL
sample
300 ng/mL
说明
分析说明
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.
法律信息
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany
Supel is a trademark of Sigma-Aldrich Co. LLC