Separation of Ribos E-5-Phosphate, Ribose-L-Phosphate, Deoxyribose-1-Phosphate By High Performance Liquid Chromatography and Spectrophotometric Determination Using 2-Cyanoacetamide

A simple procedure for separation of ribose-5-phosphate, deoxyribose-1-phosphate and ribose-l-phosphate is based on high performance liquid chromatography using reversed phase 4 x 300 mm“pBondapak/NH2” column. The column is equilibrated with 0.13 M borate buffer (pH 7.5) followed by gradient elution of ribose-5-phosphate, deoxyribose-l-phosphate and ribose-l-phosphate using water, 0.05 M borate buffer containing 0.1 M MgCl (pH 9.6) and 0.05 M sodium acetate-acetic acid buffer containing 0.1 M MgCI (pH 5.0) as eluants respectively. Eluates of borate complex “μ Bondapak/NH₂” column are brought to pH 9.6 by the addition of 1 N KOH and enzymatically hydrolysed with alkaline phosphatase (EC 3.1.3.1) to.