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Objective Uncertainty evaluation of determination of vitamin B6 content in oyster soybean peptide carnitine oral liquid by high performance liquid chromatography (HPLC assay). Methods HPLC was adopted for the determination of vitamin B6 content. Mathematical model was established for the evaluation of uncertainty to quantize influential factors of uncertainty and evaluate the uncertainty factors further. Results The content of vitamin B6 in sample oral liquid was 0.498 mg/100mL, in which, the relative uncertainty of vitamin B6 content was 0.024 9, the standard uncertainty was 0.0124 mg/100 mL, the expanded uncertainty was 0.024 8 mg/100 mL(k=2). Conclusion The calculation method is practical and reliable, being access to the uncertainty evaluation of content of vitamin B6 in oyster soybean peptide carnitine oral liquid by high performance liquid chromatography. Upon the evaluation, it is found that the quantitative repetitive of the HPLC is the major impact. [ABSTRACT FROM AUTHOR]