This study aimed to determine the amount of vitamin C from vegetable & fruit juice by high performance liquid chromatograhy (HPLC). Components for estimation of measurement uncertainty associated with the analysis of vitamin C, such as standard weight, purity, molecular weight, dilution of standard solution, calibration curve, recovery, and precision, were importantly applied. The estimation of uncertainty obtained with systematic and random error based on the GUM (Guide to the expression of uncertainty in measurement) and EURACHEM document with mathematical calculation and statistical analysis. The components, evaluated ty either Type A or Type B methods, were combined to produce an overall value of uncertainty known as the combined standard uncertainty. An expanded uncertainty was obtained by multiplying the combined standard uncertainty with a coverage factor (k) calculated from the effective degree of freedom. The content of vitamin C from vegetable and fruit juice was 27.53 mg/100g and the expanded uncertainty by multiplying by the coverage factor (k, 2.06) was 0.63 mg/100g at a 95% confidence level. It was concluded that the main sources were, in order of recovery and precision, weight and purity of the reference material, dilution of the standard solution, and calibration curve. Careful experiments on other higher uncertainties is further needed in addition to better personal proficiency in sample analysis in terms of accuracy and precision.