In the diploma thesis, we tried to regenerate waste deuterated chloroform that is usually discarded after being used for NMR spectroscopy. First we did pre-experiments with nondeuterated chloroform, which directed us when purifying an actual, deuterated sample. There were several methods we used for separation: filtration, extraction, distillation and drying. Due to properties of substances, we used fractional distillation and did NMR analysis for individual fractions. The results obtained by NMR spectroscopy showed us succsessfulness of regeneration and exposed substances that stay in the chloroform and would need further separation procedures.