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 K1 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.
Martinson
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Institute of Marine Science, University of Alaska at Fairbanks 99775, USA.
Plumley
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D000440:Butanols; D002851:Chromatography, High Pressure Liquid; D008055:Lipids / Q000302:isolation & purification*; D010860:Pigments, Biological / Q000302:isolation & purification*; D010944:Plants / Q000737:chemistry
D000440:Butanols; D008055:Lipids; D010860:Pigments, Biological