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|Precursor Ion||Product Ion|
|Column||Raptor Biphenyl (cat.# 9309A52)|
|Dimensions:||50 mm x 2.1 mm ID|
|Particle Size:||2.7 µm|
|Pore Size:||90 Å|
|Conc.:||0.5 ng/mL Fortified SeraFlx BIOMATRIX sample|
|Inj. Vol.:||5 µL|
|A:||Water, 0.1% formic acid, 5 mM ammonium formate|
|B:||Methanol, 0.1% formic acid|
|Sample Preparation||A 500 µL aliquot of fortified sample (SeraFlx BIOMATRIX) was mixed with 5 µL of internal standard solution (K1-d7, MK4-d7, and MK7-d7 at 100 ng/mL in methanol) and 1.5 mL of acetonitrile followed by vortexing for 20 seconds at 3000 rpm. After centrifugation at 4300 rpm for 10 minutes, the supernatant was loaded onto a Biotage ISOLUTE PLD+ 96-well plate (50 mg) and vacuum was applied to collect the eluate. The eluate was then evaporated to dryness at 50 °C under a gentle stream of nitrogen. The dried extract was reconstituted with 100 μL of diluent and 5 μL of sample was injected for analysis.|