Flash co140080 0

Twenty liters of the co-cultured supernatant was extracted three times with ethyl acetate (EtOAc). 2.5 g of crude EtOAc extract was concentrated under reduced pressure and purified on a silica gel (200–300 mesh) column and then eluted with petroleum ether/ethyl acetate and chloroform/methanol system to yield ten fractions (Song et al., 2014 (link)). Eight hundred and fifty milligrams of fraction 6 and 7 from the chloroform/methanol elution (20:1 and 10:1, v/v) was used to a medium pressure liquid chromatography (Flash CO140080-0, Agela Technologies, China) and eluted with methanol/water (the elution gradient was 10–90% methanol in 65 min) to generate a mixture of 216 mg. This mixture was purified on a silica gel (200–300 mesh, Qingdao Haiyang Chem. Ind. Co. Ltd. China) column by eluting with petroleum ether/ethyl acetate/ methanol (3:3:1, v/v) and then separated on a preparative column (Venusil XBP C18, Agela Technologies, China) by eluting with methanol/water (the elution gradient was 10–50% methanol in 40 min and flow rate was 8 mL/min) to obtain 10 mg of compound 1.

The mycelia in the culture broth (15 L) were filtered and the filtrate was extracted three times with Ethyl acetate (EtOAc). The crude EtOAc extract (1.1 g) was concentrated under reduced pressure and then purified on a silica gel (200–300 mesh) column, eluting stepwise with petroleum ether/ethyl acetate and chloroform/methanol system to yield eight fractions12 (link). Fraction 5 (93 mg) from the chloroform/methanol elution (10:1, v/v) was applied to a Medium Pressure Liquid Chromatography (Flash CO140080-0, Agela Technologies, China) and eluted with methanol/water (20–50% MeOH in 40 min) to yield a mixture (30 mg). This mixture was purified on a silica gel (200–300 mesh, Qingdao Haiyang Chem. Ind. Co. Ltd. China) column by eluting with chloroform/methanol (30:1, v/v) and then separated on a preparation column (Venusil XBP C18 (2), Agela Technologies, China) by eluting with methanol/water (5–30% MeOH in 40 min, 8 mL/min) to yield compound 2 (7.5 mg).