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Vitamin B 12 was separated from the protein by high performance liquid chromatography (HPLC) (1200, Agilent) using a Zorbax SB300-C8 reverse phase chromatography column, (4.6 mm x 150 mm, Agilent, 863973–906) with a gradient elution consisting of mobile phase A: 0.1% trifluoroacetic acid in LC/MS grade water (34978, Fluka) and mobile phase B: 0.08% trifluoroacetic acid (28901, Thermo) in LC/MS grade acetonitrile (34967, Fluka)
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