Pigment Extraction from Plant Leaves for Plant Physiology Studies

Abstract

Pigments are organic compounds that to be synthesized by plants for various functions. Moreover, it uses to photosynthesis. The major pigments founded in plants, such as chlorophyll (including chlorophyll a and chlorophyll b), carotenoids, and anthocyanins. Chlorophyll is green pigment which absorbs the sunlight energy and transfer to other receptors. Finally, sunlight energy becomes to carbohydrate and water. Carotenoids absorb other wavelength that chlorophyll can’t be absorbed and then transfer to chlorophyll for photosynthesis. Carotenoids can also protect chlorophyll from photooxidation by photoprotection. With anthocyanin can absorb green light that reflected from sun plant leaves. After that, green light transferred to chlorophyll for shade or shade tolerant plant photosynthesis. Therefore, both carotenoids and anthocyanins are called “accessory pigments” because play roles to be promote chlorophyll in photosynthesis. Using the N,N-dimethylformamide (DMF) solution is solvent for chlorophyll extraction, it has efficient, save time, and tools, with decreasing chlorophyll loss more than acetone (80% or 100% acetone) extraction when low chlorophyll in leaf samples. Anthocyanin extraction using 6M HCl:H₂O:MeOH (7:23:70) solution is solvent anthocyanin and chlorophyll out of leaf tissue. Then separate chlorophyll by chloroform solvent will get pure anthocyanin but mixed chlorophyll. Afterward, extract the pigments; bring the pigment solution to absorbance measurement for pigment content calculation by spectrophotometer.