A new stationary phase for high performance liquid Chromatography has been prepared from the reaction of Alizarin Red S solution with Amberlite anion exchange resin. The stability of the new attached functional group on the resin was studied against different solvents such as methanol, ethanol, benzene, petroleum
ether, chloroform, as well as NaOH and HC1 solutions. The new prepared resin was found stable and no depletion of the Alizarin moity from the resin was noticed with the above materials. The IR and CHN elemental analysis have confirmed the attachment of the Alizarin on the resin as has indicated by the physical appearance of the new resin. The new stationary phase has been packed into a stainless steel column. This has been done by pouring a water slurry of the resin manually into the column which has been placed in ultrasonic bath, and the packing was then pressurized using high pressure pump. The chromatographic performance of the packed column was characterized. The number of theoretical plates, height equivalent to theoretical plates, capacity factors, and selectivity .factors, were measured by analyzing different analyles on the new column using different mobile phase compositions and How rates. Some of polynuclear aromatic hydrocarbons such as Naphthalene, Anthracene, Phenanthrene, Fluorene, Acenaphthene, and Acenaphthylene were analyzed by this column. The analyzed compounds have shown characteristics retention times, and different selectivities. This has allowed separation of Naphthalene, Phenanthrene and Anthracene with this column using 1.3ml/min, 100% methanol as a mobile phase and detection wave length of 254nm. Three amino acid were also analyzed by .this column; Histidine, Tyrosine, and Phenylalanine. We were able to separate these amino acids on the new column using only distilled water as a mobile phase with (low rate of Iml/min. Calibration curves for all the analyzed compound were linear from their detection limits to l00ppm with correlation coefficient ranged from 0.9988-0.9997. The detection limits were ranged from O.()9ppm-0.3ppm at signal to noise ratio of three or more.