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|Peaks||tR (min)||Precursor Ion||Product Ion|
|Column||Force C18 (cat.# 963431E)|
|Dimensions:||100 mm x 3.0 mm ID|
|Particle Size:||3 µm|
|Pore Size:||100 Å|
|Guard Column:||Force C18 EXP guard column cartridge 3 mm ID (cat.# 963450253)|
|Conc.:||50 ng/mL methylmalonic acid in SeraFlx BIOMATRIX|
|Inj. Vol.:||3 µL|
|A:||0.5% Formic acid in water|
|B:||0.5% Formic acid in methanol|
|Sample Preparation||100 μL of sample (SeraFlx BIOMATRIX) was aliquoted for extraction. The sample was fortified at 50 ng/mL with methylmalonic acid. It was also fortified with 5 μL of succinic acid (20 μg/mL in water, for validation purposes only). 5 μL of the internal standard (2500 ng/mL MMA-D3 in water) was added to the sample. The sample was precipitated using 300 μL of 0.5% formic acid in methanol followed by a 10 second vortex at 3000 rpm. The sample was then centrifuged at 4000 rpm for 10 minutes at 10 °C. 250 μL of the supernatant was filtered using a Thomson SINGLE StEP standard filter vial (PVDF, 0.2 μm, Restek cat.# 25895) prior to analysis.|
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