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|Column||Raptor EtG/EtS (cat.# 9325A12)|
|Dimensions:||100 mm x 2.1 mm ID|
|Particle Size:||2.7 µm|
|Pore Size:||90 Å|
|Guard Column:||UltraShield UHPLC precolumn filter, 0.2 μm frit (cat.# 25809)|
|Ethyl sulfate-d5 sodium salt (EtS-d5) (cat.# 34104)|
|Ethyl sulfate sodium salt (EtS) (cat.# 34103)|
|Diluent:||0.01% Formic acid in water|
|Conc.:||500 ng/mL (40x acetonitrile precipitation sample prep)|
|Inj. Vol.:||10 µL|
|A:||0.01% Formic acid in water|
|B:||0.1% Formic acid in acetonitrile|
|Sample Preparation||A 500 ng/mL standard was prepared in urine. A 50 μL aliquot was mixed with 10 μL of internal standard (20 μg/mL EtG-d5 and 5 μg/mL EtS-d5 in water) and 150 µL of acetonitrile by vortexing at 3000 rpm for 10 seconds and centrifuged at 4300 rpm for 10 minutes at 10 °C. After centrifugation, 100 µL of the supernatant was diluted with 900 µL (40x dilution) of 0.01% formic acid in water. The sample was then vortexed at 3000 rpm for 10 seconds and injected for LC-MS/MS analysis.|