Spiculisporic acid and its sodium salts are frequently studied as the soil flushing agents. The biosurfactants are environmentally friendly when used in soil remediation. However, they are very expensive compared to the synthetic surfactants. The recovery of the costly surfactant is highly recommended in an economic point of view in order to reuse it after regeneration. In this work, the trisodium salt of spiculisporic acid was recovered from the model waste solution containing heavy metal ions and an organic substance. To regenerate the spiculisporic acid from the waste solution, it was acid-treated to exclude the contaminants. The resulting mixture in slurry phase was separated by the successive membrane filtrations. Spiculisporic acid of an open-ring structure was recovered around 90% yield from the retentate by the multiple operations of acid treatment and microfiltration.