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Peaks | tR (min) | Precursor Ion | Product Ion | |
---|---|---|---|---|
1. | Succinic acid | 1.69 | 117.3 | 73.1 |
2. | Methyl-D3-malonic acid | 2.02 | 120.2 | 76.2 |
3. | Methylmalonic acid | 2.03 | 117.3 | 73.1 |
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) | ||||||||||||||||||||||||
Temp.: | 35 °C | ||||||||||||||||||||||||
Standard/Sample | |||||||||||||||||||||||||
Conc.: | 50 ng/mL methylmalonic acid in SeraFlx BIOMATRIX | ||||||||||||||||||||||||
Inj. Vol.: | 3 µL | ||||||||||||||||||||||||
Mobile Phase | |||||||||||||||||||||||||
A: | 0.5% Formic acid in water | ||||||||||||||||||||||||
B: | 0.5% Formic acid in methanol | ||||||||||||||||||||||||
|
Detector | MS/MS |
---|---|
Ion Mode: | ESI- |
Mode: | MRM |
Instrument | HPLC |
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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