Bovine spleen phosphodiesterase 2

o-Toluidine hydrochloride (T536215; purity, 97%) was obtained from Toronto Research Chemicals, Toronto, ON, Canada, p-toluidine (PT) hydrochloride (T0303; purity, >98%) was obtained from Tokyo Chemical Industry, Tokyo, Japan, and aniline (ANL) hydrochloride (017-04092; purity, 97%) was obtained from Wako Pure Chemical Industries, Osaka, Japan. AAOT (A1020; purity, >98%) and apocynin (H0261; purity, >98%) were obtained from the Tokyo Chemical Industry. Nuclease P1 and HPLC-grade acetonitrile were purchased from Wako Pure Chemical Industries, Ltd. Phosphodiesterase I was purchased from Worthington Biochem (Lakewood, NJ, USA). Bovine spleen Phosphodiesterase II, DNase I, and bacterial alkaline phosphatase Type III (Escherichia coli) were purchased from SigmaAldrich (St. Louis, MO, USA). All other chemicals used were of analytical grade and purchased from Wako Pure Chemical Industries, Ltd.

PhIP-HCl was purchased from the Nard Institute (Osaka, Japan), and its purity was confirmed to be >99% by HPLC. AA was obtained from Sigma Aldrich (St. Louis, MO, United States). Nuclease P1 was purchased from Wako Pure Chemical Industries (Osaka, Japan). Phosphodiesterase I was purchased from Worthington. Nicotinamide, bovine spleen Phosphodiesterase II, DNase I, and bacterial alkaline phosphatase type III (E. coli) were purchased from Sigma Aldrich (St. Louis, MO, United States). Difco™ Nutrient Broth and Cofactor-I were acquired from Becton, Dickinson and Company (Sparks, MD, United States), and Oriental Yeast Co., Ltd (Tokyo, Japan), respectively. The S9 mix was purchased from the IEDA Trading Corporation (Tokyo, Japan). All other chemicals used in the study were of analytical grade and were purchased from Wako Pure Chemical Industries.