New stationary phases for high performance liquid chromatography have been prepared by the reaction of Alizarin Red-S solution with either Amberlite anion exchange or Alumina resins. The FTIR elemental analyses have configured the attachment of Alizarin on the resin as has indicated by the physical appearance of the new resins. The resin capacities of newly prepared resins have calculated. The resin capacity for the Amberlite-Alizarin Red-S resin was found to average 6.65 meq./g which are approximately twice as those of the Amberlite resin, 3.05 meq./g. Similarly, the resin capacity for Alumina-Alizarin Red-S and Alumina resins were 4.39 meq./g and 1.68 meq./g respectively, which is approximately double value. The new stationary phases have been packed into stainless steel columns. This has been done by the special techniques. The chromatographic performance of the packed column was characterized. The number of plate numbers, height equivalent of a theoretical plates, capacity factors, selectivity factors, and peaks asymmetry, were measured by analyzing different analyses on the new columns using different mobile phase's compositions and f l ow r a t e s . These parameters were measured from analysis of some amino acids such as Histidine, phenylalanine, Tryptophane, and Tyrosine, and analysis of PAHs such as Anthracene, Fluorine, Naphthalene, Acenophthene, and Phenathrene. The analysis of amino acids using Ambulate-Alizarin Red-S column has performed with an aqueous mobile phase adjusted at pH 2.5. The average value of plate numbers N was 178.82. The plate heights H was value averaged 0.2483cm, capacity factors K, were ranged from 1.67-3.13, separation factors a, were ranged from 1.21-1.29, and peaks asymmetry value was 1.88. The analysis of the some amino acids on the some column but with a mobile phase djusted at pH 8.2, the chromatographic parameters were plate numbers N was 108.27. The plate heights H was 0.2483cm, capacity factors K, from 1.78-2.51, separation factors a, from 1.11-1.13, and peaks asymmetry value was 1.89. However, in the analysis of PAHs with the same column but conditioned at either pH 2.5 or 8.2. The plate numbers N was 11491.45 and 9986.53. The plate heights H was 0.0032 cm and 0.0035cm, capacity factors K, were ranged from 1.12-2.84 and 1.61-2.76, separation factors a, were ranged from 1.11 -1.49 and 1.12-1.19, and peaks asymmetry value was 1.89 and 1.25. The analysis of amino acids using Alumina-Alizarin Red-S column with mobile phase at pH 8.5 has given an average value of plate numbers N was 331.88. The plate heights H was 0.0793cm, capacity factors K, were ranged from 1.29-2.34, separation factors a, were ranged from 1.18- 1.30, a n d p e a k s a s ymme t r y v a l u e w a s 1 . 7 9 . However, in the analysis of PAHs using Alumina-Alizarin Red-S column conditioned at pH 8.5. The plate numbers N was 9524.76. The plate heights H was 0.0029cm, capacity factors K, were ranged from 2.16-3.56, separation factors a, were ranged from 1.11-1.16, and peaks asymmetry value was 2.24. Calibration curves for all analyzed compound were linear from their detection limits to at least 50 ppm with correlation coefficient ranged from 0.9990-0.9998. The detection limits were ranged from 0.05-0.1 ppm at signal to noise ratio of three or more.