Since the application of genetic markers in forensic practise requires availability of suitable profile database, the allele frequency and forensic evaluation of four pentanucleotide markers: Penta B, C, D and E in central Poland population were established. Penta B, C, D and E show a high degree of the power of discrimination i.e. 0.974, 0,912, 0.928 and 0.978, respectively. The power of exclusion of Penta B, C, D and E markers in the investigated population were 0.755, 0.413, 0.695 and 0.826, respectively. Application of four investigated markers gives the total value of the power of discrimination of 0.999996 and the total value of the power of exclusion of 0.9924. It makes the markers, especially Penta E, B, and D, a very useful genetic tool in complex cases of kinship analysis.