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After filtration, the filtrate is evaporated in vacuo. The resulting viscous liquid is loaded on 5 g of DEAE-Sephadex (A-50-120; Sigma, St. Louis, MO) pretreated with 10 mM of ammonium acetate. 20 M solution to obtain the product). 2 M ammonium acetate. 2%) of p-hydroxyphenacyl diammonium phosphate (16). 18, IR (KBr) 3350, 3088, 2950, 1679, 1594, 1570, 1440, 1268, 1090 cm-1; UV-vis (CH3CN/H20) Ama~(e)220 (8400), 282 (14,000). 68; N, 10,52. 33. Coupling of p-Hydroxyphenacyl Phosphate with ADP p-Hydroxyphenacyl Adenosine 5'-Triphosphate, Triammonium Salt (17).
33. Coupling of p-Hydroxyphenacyl Phosphate with ADP p-Hydroxyphenacyl Adenosine 5'-Triphosphate, Triammonium Salt (17). 27 The coupling reaction is modeled after the method of Hoard and Ott. , Phillipsburg, NJ) (10 g) in a sintered glass funnel is treated with 5% hydrochloric acid (20 ml x 2) and washed with water until the filtrate is neutral. The resin is then treated with 20% aqueous pyridine solution and washed with water until the filtrate is neutral. A solution of adenosine 5'-diphosphate (ADP, potassium salt, 39 C.
Chem. , 18-20 (1955). 37(a) M. Kulbaand A. Zwierzak,Ann. Soc. Chim. Polonorum48,1603(1974);(b) A. Zwierzak and M. Kulba, Tetrahedron22, 3163-3170 (1971);and (c) E. P. Serebryakov,L. M. Suslova, and V. F. Kucherov, Tetrahedron34, 345-351 (1978). 5)] is stirred for 2 hr. Evaporation of acetone and H20 on a Rotavapor gives a concentrated aqueous solution of the salt. 66. 70 mmol), and 20 ml of benzene is placed in a 50-ml round-bottom flask and refluxed with efficient stirring for 10 hr. The solution is cooled to room temperature and extracted with the HzO/ethyl acetate solution.