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sample preparation	Solid Phase Extraction (SPE)
sample/matrix	human urine
extraction process	urine samples were spiked at 200 ng/mL each analyte and digested with 10kU of beta-glucuronidase enzyme at 60 °C for 120 min
SPE tube/cartridge	Supel-Select SCX, 60 mg/3 mL (54241-U)
condition	1 mL of 1% formic acid in acetonitrile followed by 1 mL water
sample addition	1.4 mLurine
washing	2 mL water followed by 1 mL of water:methanol (75:25)
elution	1 mL of 10% ammonium hydroxide in acetonitrile
eluate post-treatment	evaporate to dryness and reconstitute in 1 mL water
column	Titan C18, 5 cm x 2.1 mm I.D., 1.9 µm particles (577122-u)
column temp.	35 °C
mobile phase	[A] 5 mM ammonium formate; [B] 5 mM ammonium formate in acetonitrile:water (95:5)
gradient	5 to 90% B in 2.5 min, held for 0.5 min, to 5% B in 0.1 min, held for 0.5 min
flow rate	0.3 mL/min
injection	2 µL
detector	ESI(+) TOF, m/z 100-1000

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Analysis Note	Sample pretreatment: urine samples were spiked at 200 ng/mL each analyte and digested with 10kU of beta-glucuronidase enzyme at 60 °C for 120 min, A set of compounds representing five drug classes that are routinely tested in urine were extracted from urine and analyzed using LC/MS (TOF). The entire workflow, from enzymatic digestion to hydrolyze the glucuronide metabolites to analysis using a Titan UHPLC column is presented here. Cerilliant CRMS provided reliable quantification.
Categories	Analytical Chromatography, Clinical, Forensics and Toxicology, SPE Applications, steroids, Beta-blockers, Opiate, Anticonvulsants, anticonvulsants, Drugs of Abuse, Opiates, Drugs of abuse, Steroids, beta-blockers, analgesics, Analgesics, Anticonvulsant/antiepiletic/antiseizure, opiate, drugs of abuse
Featured Industry	Clinical Research \r\nForensics and Toxicology
suitability	application for SPE, application for HPLC

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