Bleaching of syringes has been advocated to prevent HIV-1 transmission among injection drug users (IDUs). Bleach is frequently distributed by needle exchange, outreach, and educational programs targeting IDUs. We applied a sensitive HIV-1 microculture assay to determine the effectiveness of bleach in disinfecting syringes contaminated with HIV-1. This study demonstrates that in a laboratory environment designed to replicate injection behaviors, undiluted bleach is highly effective in reducing the viability of HIV-1 even after minimal contact time. However, it did not reduce the HIV-1 recovery to zero. Furthermore, three washes with water were nearly as effective as a single rinse with undiluted bleach in reducing the likelihood that contaminated syringes harbored viable HIV-1. Given the reality that IDUs share syringes and may not have access to a new, sterile syringe for each injection, the results suggest that they should be encouraged through harm reduction interventions to clean their syringes, preferably with undiluted bleach.