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Sample pre-treatment
1ml human oral fluid was collected on cellulose pad of the applicator tip provided by the Intercept® i2
oral fluid device (OFC) . Saturated pad was placed into
transport tube containing buffer solution and allowed to
sit overnight. Centrifuge at 600g for 15mins to collect
supernatant.
SPE cartridge: Strata-X-C, 30 mg 96-Well Plate
Part No. 8E-S029-TGB
Step Procedure
Condition: 1 mL Methanol
Equilibrate: 1 mL DI Water
Load: Combine 0.5mL of pretreated sample spiked with internal standards and 1 mL 1% formic acid, mix/vortex 10-15 secs and load in Strata-X-C well plate.
Weak Wash: 1 mL DI Water
Strong Wash: 1 mL 50:50 Acetone/Water
Dry Down: 5 minutes at maximum vacuum (15" Hg or higher)
Elute: 2 x 500 µL Methanol/Acetonitrile/30% Ammonium Hydroxide (5:5:2)
Dry Down: Evaporate to dryness under gentle nitrogen and 45-50°C.
Reconstitute: 200 µL initial mobile phase