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Chromatography of acid phytohormones on columns of Sephadex LH‐20 and insoluble poly‐N‐vinylpyrrolidone, and application to the analysis of conifer extracts
Author(s) -
Sandberg Göran,
Dunberg Arne,
Odén PerChrister
Publication year - 1981
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1981.tb04490.x
Subject(s) - chromatography , chemistry , sephadex , elution , abscisic acid , phenylacetic acid , acetic acid , fractionation , picea abies , column chromatography , gibberellin , indole 3 acetic acid , ethyl acetate , biochemistry , botany , auxin , biology , gene , enzyme
The chromatographic behaviour of abscisic acid (ABA), indole‐3‐acetic acid (IAA), phenylacetic acid (PAA), and gibberellins A 1 , A 4 , A 8 , A 9 , A 13 and A 20 on columns of Sephadex LH‐20 and insoluble poly‐ N ‐vinylpyrrolidone (PVP) eluted with buffers of different pH values is described. PVP shows considerable batch differences that must be carefully checked. Chromatography of acidic ethyl acetate‐soluble fractions of Scots pine ( Pinus sylvestris L.) extracts at pH 4.5 resulted in great losses of phytohormones, due to poor solubility of the extracts. If the extracts were applied to the column dissolved in buffer of pH 7.5, subsequent elution at pH 4.5 was possible with only small losses. The performance of the chromatographic column was strongly affected by the application volume. A combined PVP/Sephadex LH‐20 column eluted at pH 4.5 allows remarkable purification of pine and spruce ( Picea abies (L.) Karst.) extracts, collection of IAA in a fraction that can be directly analyzed by e.g. the indolo‐α‐pyrone method, and collection of another fraction containing ABA, PAA and probably most of the known C 19 ‐gibberellins; whereas the C 20 ‐gibberellin A 13 is eluted later (with IAA).