The versatile characterization of organic materials in Anatolian black pine (Pinus nigra) especially the significant organic acids is great importance. A rapid, simple, and sensitive HPLC-DAD method was developed and presented for a simultaneous detection of six organic acids. This method was thoroughly validated to determine the most abundant organic acids present in Anatolian black pine and their distribution in branches, leaves and cones. Separation of six organic acids were developed and achieved on ODS C-18 column (250mmx4.6mm, 5 mu m particle size), using a mixture of (A) water (pH: 2.0, adjusted by H2SO4) and (B) acetonitrile in the ratio of (95:5, v/v) at wavelength of 210 nm with a flow rate of 0.8 mL min(-1) within 14 minutes. The dynamic range was between 5 to 1000 mg L-1 with relative standard deviation less than 0.73%, (n=4). Limits of detection and the recoveries for oxalic acid, tartaric acid, formic acid, lactic acid, acetic acid and citric acid were ranged 1.85 mg L-1 (87.75-99.36%), 3.61 mg L-1 (100.89-119.55%), 4.01 mg L-1 (83.57-119.64%), 1.16 mg L-1 (82.29-112.19%), 1.19 mg L-1 (80.86-110.42%) and 2.87 mg L-1 (96.96-117.74%), respectively.