The panna cotta is one of the Italian desserts served cold. The row materials for this dessert are cream and milk that are cooked with other ingredients such as sugar and gelatine. The characteristics of panna cotta resemble pudding, but the gel nature of this dessert is softer and can melt in the mouth when eaten. This study was purposed to determine the quality characteristics of panna cotta after the use of konjac flour (Amorphophallus konjac) as the substitution of gelatine. This research was conducted using a non factorial completely randomized design with 4 replications, with konjac substitution to gelatine were 100%, 75%, 50%, 25%, and 0%. Based on the tests carried out, it was found that the substitution of konjac flour to gelatine had a very significant effect on texture, vitamin C levels, and syneresis. The resulting panna cotta of texture ranges from 0.2751 mm/g to 0.2870 mm / g. The resulting panna cotta of vitamin C levels ranges from 8.3745 mg/100g to 11.5922 mg/100g. The resulting panna cotta of syneresis ranges from 6.4199% to 4.2715%. In addition, the substitution had no significant effect on the colour (°Hue), pH value, and organoleptic (colour, flavour, taste, and texture).