ONE-STEP EXTRACTION AND CONCENTRATION OF PIGMENTS AND ACYL LIPIDS BY SEC-BUTANOL FROM IN-VITRO AND IN-VIVO SAMPLES

Photosynthetic pigments and acyl lipids were simultaneously extracted and concentrated by sec-butanol. Pigments extracted with sec-butanol were indistinguishable from those extracted using acetone as determined by quantitative and qualitative HPLC, Use of sec-butanol has several advantages over conventional extraction solvents: (1) pigments are extracted directly from polyacrylamide gel slices without an elution step; (2) pigments in dilute, isolated pigment-protein complexes are extracted and concentrated without first concentrating the sample; (3) when necessary, the concentration factor is readily increased by addition of water; (4) sec-butanol extracts acyl lipids and vitamin K-1 as effectively, but much quicker, than chloroform:methanol; (5) sec-butanol rapidly extracts and concentrates pigments from thylakoids of all plant species tested and even directly from many algal/higher plant cells, facilitating analysis of pigment biosynthetic pathways using radioactive substrates; and (6) pigments are stable in sec-butanol for several days at room temperature in the dark or for many weeks if stored at -20 degrees C in darkness. Finally, sec-butanol is preferable to ether for concentrating pigments extracted with acetone. (C) 1995 Academic Press, Inc.