Method for comprehensive extraction of cyclopeptide toxins and other metabolites

Inventors

Chialvo, Clare Scott • Ciesla, Lukasz

Assignees

University of Alabama at Birmingham UAB

Abstract

Methods for extracting metabolites from mushroom comprising dispersing a sample of the mushroom; contacting the sample with a first solvent system at a temperature of 80° C. or greater to extract at least a first portion of metabolites from the sample; and contacting the sample with a second solvent system at a temperature of 80° C. or greater to extract at least a second portion of metabolites from the sample, wherein the combined first portion of metabolites and the second portion of metabolites include primary and secondary metabolites from the mushroom. A high-throughput process for establishing a metabolic profile of a mushroom, such as Amanita mushroom, can comprise extracting metabolites from mushroom and analyzing the extracted metabolites by automatic peak recognition or peak integration. Assay for identifying and/or assessing candidate metabolites extracted from Amanita mushroom are also disclosed.

Core Innovation

The invention relates to an extraction framework for recovering comprehensive primary and secondary metabolite profiles from mushroom samples using two sequential solvent extraction steps. The method mixes a sample of the mushroom with sand and then performs contacting in an extraction cell with a first solvent system followed by a second solvent system. Both contacting steps occur at a temperature of 80 °C or greater for a sufficient time to obtain first and second portions of metabolites.

The first portion of metabolites and the second portion of metabolites include primary and secondary metabolites from the mushroom, so the sequential extraction supports recovery across metabolite classes. In related aspects, the framework is applied to extract cyclopeptide toxins from a mushroom, again using sand mixing and two sequential solvent extractions in an extraction cell at temperatures of 80 °C or greater to obtain first and second portions of cyclopeptide toxins.

The described framework is further associated with specific mushroom sources, where the mushroom can be selected from genera including Amanita, Agaricus, Boletus, Lactarius, Lepiota, Russula, Tricholoma, Coprinus, or Pluteus, and can include cyclopeptide-toxin-containing mushrooms. Additional disclosed refinements include defining an aqueous solvent system with a water and methanol volume ratio and constraining both extraction-cell temperatures to 80–160 °C, together with reporting yield targets for cyclopeptide toxins.

Claims Coverage

The partial claims include two independent claims, one directed to extracting metabolites from a mushroom and one directed to extracting cyclopeptide toxins from a mushroom. Across these independent claims, the core inventive structure is a two-step sequential solvent extraction in extraction cells at 80 °C or greater, starting from a mushroom sample mixed with sand.

Together, the independent claims cover sequential first and second solvent system contacting steps performed in extraction cells at 80 °C or greater after mixing the mushroom sample with sand. The metabolite claim covers recovery of primary and secondary metabolites, and the cyclopeptide-toxin claim specifically covers extraction of cyclopeptide toxins into first and second portions.

Stated Advantages

Documented Applications