Eremothecium ashbyii NRRL 1363 was used in this investigation which is a highly flavinogenic strain. Studies on riboflavin production was conducted in shake culture using inexpensive and abundantly available raw materials. Among the stimulants rice bran oil at 1 kg m−3 stimulated the highest and promoted the maximum riboflavin yield followed by Eriodendron anfractuosum, yeast hydrolysate and corn steep liquor. The fermented broth (an enriched riboflavin concentrate) was stable for two months at 4 °C, to be used as feed grade riboflavin. To upgrade to pharmaceutical grade further investigations are required.