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Precision and accuracy of indole‐3‐acetic acid analyses performed with the 2‐methylindolo‐α‐pyrone fluorescence assay and with a high performance liquid chromatography technique with spectrofluorimetric detection, exemplified on pine tissue ( Pinus sylvestris L.)
Author(s) -
Sandberg Göran,
Dunberg Arne
Publication year - 1982
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.1399-3054.1982.tb00298.x
Subject(s) - chromatography , high performance liquid chromatography , fluorescence , chemistry , accuracy and precision , acetic acid , indole 3 acetic acid , analytical chemistry (journal) , biochemistry , mathematics , auxin , gene , physics , statistics , quantum mechanics
The precision and accuracy of two versions of the 2‐methylindolo‐α‐pyrone (2‐MIP) assay and of a high performance liquid chromatography (HPLC) technique was assessed when applied to analysis of indole‐3‐acetic acid (IAA) in pine ( Pinus sylvestris ) tissue. In one version of the 2‐MIP assay total fluorescence at wavelengths > 450 nm was recorded, whereas in the other version fluorescence spectra were recorded and analyzed. Such analysis revealed the occurrence in some samples of fluorescent contaminants that impaired both precision and accuracy. The precision and accuracy of the 2‐MIP assay could be improved by additional sample purification, and the over‐all accuracy of the assay was finally verified, at a high level of sample purity, by successive approximation. The precision of the assay was then 14%. Successive approximation also verified that the HPLC technique was accurate at a less advanced stage of sample purity than the 2‐MIP assay, and had a precision of 14%.