The invention provides a novel culture medium using marine aspergiolide glaucus to manufacture a novel antineoplastic active compound. Per liter of the novel culture medium comprises 20 to 100g of tragantine, 10 to 100g of cane sugar, 20 to 100g of maltose, 0.5 to 6g of peptone, 0.3 to 6g of yeast extract, 0.2 to 4g of soybean flour, 0.5 to 10g of sodium glutamate, 0.01 to 1g of heptahydrate magnesium sulfate, 0.01 to 2g of monopotassium phosphate and the balance being artificial seawater. A method for preparing the novel culture medium is also provided. The novel culture medium using the marine aspergiolide glaucus to manufacture the novel antineoplastic active compound has the advantages of easily received raw materials, low cost and simple preparing method, can obviously improve the yield of griseoviridin A produced and fermented by the marine aspergiolide glaucus HB1-19, improve the yield of the griseoviridin A from the prior 15mg/L to 71mg/L, is advantageous to industrial production in large scale, and has significance for the fermentation investigation of the marine aspergiolide glaucus HB1-19 and the medicine and pharmacology investigation of the griseoviridin A.