In this research, operational parameters were assessed to optimise the production of volatile fatty acids (VFA) from artisanal fish waste (FW) using mixed-culture fermentation. The experiments were performed using a randomised block design with pH and Inoculum/Substrate (I/S) ratio as factors with three pH levels (5, 7 and 9) and five I/S ratios levels (0.20, 0.15, 0.10, 0.05, 0.00) with VFA production, total ammonia nitrogen (TAN), biogas and soluble chemical oxygen demand (sCOD) as response variables. The results indicated that pH and I/S ratio significantly influenced VFA production and the other variables evaluated. The highest VFA concentration (54.5 g/L) was obtained at pH 7 and with an I/S ratio of 0.05. These conditions were also optimal for TAN and sCOD. External pH control was proved as unnecessary as the highest VFA production (70.3 g VFA/L) was obtained without pH control. In these conditions, the fermentation broth was able to maintain its pH between 6.6 and 7.7. The VFA profile had acetic (35%) and butyric (32%) acids as the dominant species and propionic, iso-valeric and iso-butyric were minor species (< 10%). An economic assessment evaluating the establishment of a FW VFA biorefinery in Tumaco, Colombia was performed including capital, operational and labour cost, discounted payback period and cost curves. These results evidenced the potential of artisanal FW as a raw material for producing high concentrations of VFA and smooth the path for the establishment of a VFA and biogas biorefinery from FW.