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An electrochemical deposition method was used to fabricate a gold nanoflower (AuNF) and carbon nanoparticle (CNP) modified carbon paper (CP) sensor (AuNFs-CNPs/CP) for the low-cost detection of 5-methyltetrahydrofolate (5-mTHF) in egg yolk. AuNF morphology and structures were characterized by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS), revealing nanoflower sizes in the 50 to 200 nm range. AuNFs formed on the sensor were in the Au0. We evaluated 5-mTHF assay performance using cyclic voltammetry, differe