METHOD FOR QUALITATIVE DETERMINATION OF 7-OXA 6-OXOBRASSINOSTEROID

摘要

The invention relates to immunochemical analysis, namely to qualitative determination of plant's growth regulators, 7-oxa-6 oxobrassinosteroids – brassinolid, 24-epibrassinolid, 28-homobrassinolid, 28-norbrassinolid etc, and is used for evaluation of said brassinosteroid content in plant objects, control of the substance production and measurement concentration thereof in agro preparations based thereon and can also be used for evaluating the quality of agricultural production grown therewith, determination residual amounts in soil, waste waters etc. The present invention is aimed at providing a method for qualitative determination of 7-oxa-6-oxobrassinosteroids and a composition thereof which are specified by specificity, high sensitivity, ecological safety, simplicity, reliability and produce ability. The proposed result is achieved firstly by using the inventive method comprising mixing brassinosteroid samples to be analyzed with a conjugate of 28-norbrassinolid-horseradish peroxidase in polymer holes with immobilized polyclonal antibodies on the inner surface to 28-norbrassinolid, subsequent incubation, adding to bound form of chromogen-substrate mixture for determining peroxidase activity by which calculate the amount of determined 7-oxa-6-oxobrassinosteroid; and secondly a composition therefor comprising antibodies to 28-norbrassinolid immobilized on polymer holes; the conjugate of 28-norbrassinolid with horseradish peroxidase; buffer solution for dissolving conjugate; washing buffer solution; choromogen-substrate mixture-solution of 3.3',5,5'-tetramethylbenzidine in a substrate buffer with hydrogen peroxide; stop-reagent -5% HSOsolution.