Background: Decellularization methods using anionic detergents have been applied to engineered tissues for eliminating tissue immunogenicity. Sodium deoxycholate is a water soluble ionic detergent commonly used in applications ranging from decellularisation, liposome preparation and isolation of membrane proteins and lipids. Sodium deoxycholate has been reported to form many reactive intermediates which bind to tissue and exert toxic effects. Methods: This work reports on methods to detect and estimate residual sodium deoxycholate in decellularized saphenous vein. The method reported is based on the extraction of anionic detergent (sodium deoxycholate)-Methylene blue complex in chloroform which can be subsequently quantified by measuring the optical density at 653 nm. Results and conclusion: This assay may be used for detecting sodium deoxycholate levels in decellularized tissue before implantation. © 2010 Indian Association of Cardiovascular-Thoracic Surgeons.