The present invention relates to methods for the production and isolation of D-chiro-inositol (DCI) from kasugamycin. More specifically, the present invention relates to a method of producing DCI by hydrolysis of kasugamycin with aqueous trifluoroacetic acid in the presence of a strongly acidic ion exchange resin. The present invention further relates to methods of isolating DCI from an aqueous mixture, such as a hydrolysis mixture, either directly or by forming an organic derivative of DCI.