Procedures for the extraction, separation and estimation of the major fat‐soluble pigments of hay

New methods are described for the extraction, chromatographic separation and spectro‐photometric identification and estimation of chlorophyll‐a, chlorophyll‐b, phaeophytin‐a, phaeophytin‐b, carotene and xanthophyll in hay. The most suitable solvent for extraction was 85% acetone. The chlorophylls were separated chromatographically from the phaeophytins, carotene and xanthophyll on a sucrose‐sodium sulphate mixture. The chlorophyll‐a and chlorophyll‐b were estimated simultaneously in the green solution, and phaeophytin‐a and phaeophytin‐b were estimated simultaneously in the yellow solution, by spectrophotometry. Carotene and, subsequently, xanthophyll were separated chromato‐graphically from the yellow solution on magnesium oxide, and were estimated separately.