The National Metrology Institute of Japan has issued a certified reference material of bioethanol (NMIJ CRM 8301-a) for the quantification of water, methanol, sulfur, and copper. This paper presents technical details for the characterization of the water in NMIJ CRM 8301-a. The characterization was performed using coulometric and volumetric Karl-Fischer (KF) titrations. To reduce moisture absorption, sample handling and KF titration were performed in a glove box under a dried nitrogen atmosphere. In addition, a rubber cap with a three-way valve was attached to the ampoule immediately after opening so as to minimize the influence of moisture. Sample aliquots were obtained using a gas-tight microsyringe through the valve, and injected into the KF cell as soon as possible. The certified value of water obtained from coulometric and volumetric KF titrations was 1.688 mg g -1 , and the expanded uncertainty (coverage factor, k = 2) was 0.028 mg g -1 . This CRM would be suitable for the monitoring of water in bioethanol and similar matrices.