Method for profiling phytohormone levels in plant tissue

摘要

The present invention provides a method for profiling phytohormone levels in plant tissue or tissue of other plastid containing organisms, i.e. a method for the simultaneous determination of a multitude of phytohormone levels in plant tissue or tissue of other plastid containing organisms.

主权项

1. A method for profiling phytohormone levels in tissue of a plastid containing organism, the method comprising,
i. extracting particulate tissue material of the plastid containing organism with a liquid extractant, which is a mixture of at least one water miscible neutral organic solvent having from 1 to 3 carbon atoms and 1 heteroatom selected from O and N with water containing from 0.1 to 5% by weight, based on the extractant, of at least one acid, whereby a first liquid extract is obtained; ii. contacting the liquid extract obtained in step i. with a solid absorbent having a hydrophobically modified surface and removing the solid absorbent to obtain a second liquid extract; iii. evaporating the solvent from the second extract to obtain a residue and then re-dissolving the obtained residue in a solvent mixture of at least one water miscible neutral organic solvent having from 1 to 3 carbon atoms and 1 heteroatom selected from O and N with water containing from 0.1 to 5% by weight, based on the solvent mixture, of at least one acid to obtain a reconstituted extract; iv. purifying the reconstituted extract by passing it through a bed of a solid absorbent having a hydrophobically modified surface to obtain a purified reconstituted extract; and v. determining relative concentrations of at least two plant hormones in the purified reconstituted extract obtained in step iv. by directly subjecting the purified reconstituted extract to an analyzing unit comprising a separation unit for separation of the phytohormones and an analyzer for identifying the phytohormones which separation unit is coupled to the analyzer; wherein step iv. is performed as a solid phase extraction by using a solid phase extraction cartridge containing the solid absorbent; wherein the solid phase extraction cartridge is directly coupled to a column of a reverse phase LC-MS; and wherein the solid absorbent of step iv. is selected from the group consisting of C6-C20-alkyl modified silica particles and polymer particles comprising homo or copolymers of divinylbenzene with neutral monoethylenically unsaturated monomers.