A method for determining chlorophyll in intact etiolated cucumber cotyledons using N,N-dimethylformamide (DMF) is described. The method involves direct immersion of the cotyledons into DMF, which is more efficient than traditional extraction methods in preserving pigment and reducing loss. The specific absorption coefficient of chlorophyll a in DMF was determined. The study compared DMF with acetone for extracting chlorophyll, finding that DMF was more efficient, especially when pigment concentrations were low. Extraction with DMF was simpler and resulted in comparable outcomes to acetone. The method allowed for the extraction of chlorophyll from intact tissues without grinding, which is particularly useful when material is limited. The absorption spectra of chlorophyll a in DMF and acetone were compared, showing slight differences in peak positions and intensity. The maxima of chlorophyll a in DMF were found at 626.0 nm, and the specific absorption coefficient of chlorophyll a in DMF at 664.5 nm was calculated to be 83.89 g⁻¹l⁻¹. The results showed that DMF extracts remained stable during storage and that the time between extraction and spectrophotometric analysis was not critical. The study highlights the advantages of using DMF for chlorophyll determination in intact tissues, including simplicity, efficiency, and minimal pigment loss.A method for determining chlorophyll in intact etiolated cucumber cotyledons using N,N-dimethylformamide (DMF) is described. The method involves direct immersion of the cotyledons into DMF, which is more efficient than traditional extraction methods in preserving pigment and reducing loss. The specific absorption coefficient of chlorophyll a in DMF was determined. The study compared DMF with acetone for extracting chlorophyll, finding that DMF was more efficient, especially when pigment concentrations were low. Extraction with DMF was simpler and resulted in comparable outcomes to acetone. The method allowed for the extraction of chlorophyll from intact tissues without grinding, which is particularly useful when material is limited. The absorption spectra of chlorophyll a in DMF and acetone were compared, showing slight differences in peak positions and intensity. The maxima of chlorophyll a in DMF were found at 626.0 nm, and the specific absorption coefficient of chlorophyll a in DMF at 664.5 nm was calculated to be 83.89 g⁻¹l⁻¹. The results showed that DMF extracts remained stable during storage and that the time between extraction and spectrophotometric analysis was not critical. The study highlights the advantages of using DMF for chlorophyll determination in intact tissues, including simplicity, efficiency, and minimal pigment loss.