Various crosslinking polymers were synthesized by condensation polymerization of triethanolamine and a mixture of (triethanolamine : glycerol) with different diacid chlorides; such as glytarylchloride, adipoylchloride, suberoylchloride, sebacoylchloride and 1,10-dodecanedioyldichloride. And with different anhydride compounds like succinic anhydride, phthalic anhydride, maleic anhydride, and 1,8-naphthalic anhydride. The products were characterized by FTIR. The polymer produced from condensation of mixture of triethanolamine and glycerol with maleic anhydride has high rigidity, easy grinding, and high stability. Different percentage of triethanolamine and glycerol (25:75,50:50,75:25%) and reacted with maleic anhydride. The polymer produced from the mixture of (50:50%) triethanolamine and glycerol with maleic anhydride was used as a stationary phase for high performance liquid chromatography. This stationary phase has been packed into a stainless steel column. The chromatographic performance of the newly packed column was characterized. The number of theoretical plates (N), height equivalent to theoretical plates (H), capacity factors, and selectivity factors, were measured by analyzing different analytes on the new packed column using different mobile phase compositions and flow rates. Some of amino acids such as Histidine, Phenylalanine, Tryptophane, and Tyrosine we analyzed by this new column. The analyzed compounds have shown characteristics retention times, and different selectivities. This has allowed separation of Phenylalanine, Histidine, and Tryptophane with this column using 100% hexane at 1.5ml/min. as a mobile phase and detection wave length of 254nm. Some of poly aromatic compounds such as Fluorine, Phenathrene, Anthracene, Naphthalene, and Acenaphthylene were also analyzed by this column. We were able to separate some of these PAHs compounds using only distilled water as a mobile phase with flow rate of Iml/min. Calibration curves for all the analyzed compounds were linear from their detection limits to 25ppm with correlation coefficient ranged from 0.9982 to 0.9995. The detection limits were ranged from 0.05 to O.lppm at signal to noise ratio of three or more.