Method for comprehensive extraction of cyclopeptide toxins and other metabolites
View Patent ↗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.
1. A method for extracting metabolites from mushroom comprising:
a) mixing a sample of the mushroom with sand;
b) contacting the sample with a first solvent system in an extraction cell at a temperature of 80° C. or greater for a sufficient time to extract at least a first portion of metabolites from the sample; and
c) contacting the sample with a second solvent system in an extraction cell at a temperature of 80° C. or greater for a sufficient time to extract at least a second portion of metabolites from the sample;
wherein the first portion of metabolites and the second portion of metabolites include primary and secondary metabolites from the mushroom.
2. The method of claim 1 , wherein the mushroom is selected from any mushroom containing cyclopeptide toxins.
3. The method of claim 1 , wherein the mushroom is selected from a species occurring in Amanita, Agaricus, Boletus, Lactarius, Lepiota, Russula, Tricholoma, Coprinus , or Pluteus genus.
4. The method of claim 1 , wherein the first solvent system comprising water and a polar organic solvent.
5. The method of claim 4 , wherein the polar organic solvent comprises an alcohol selected from methanol, ethanol, propanol, or a mixture thereof.
6. The method of claim 5 , wherein the aqueous solvent system comprises water and methanol present in a volume ratio of from 1:5 to 5:1.
7. The method of claim 1 , wherein the second solvent system comprises a polar organic solvent, a non-polar organic solvent, or a mixture thereof.
8. The method of claim 1 , wherein the second solvent system consists essentially of an alcohol.
9. The method of claim 1 , wherein step b) and step c) are carried out at a temperature from 80° C. to 160° C.
10. The method of claim 1 , having a yield of at least 80% by weight cyclopeptide toxins, based on the total amount of cyclopeptide toxins from the mushroom.
11. A method for extracting cyclopeptide toxins from a mushroom comprising:
a) mixing a sample of the mushroom with sand;
b) contacting the sample with a first solvent system in an extraction cell at a temperature of 80° C. or greater for a sufficient time to extract at least a first portion of cyclopeptide toxins from the sample; and
c) contacting the sample with a second solvent system in an extraction cell at a temperature of 80° C. or greater for a sufficient time to extract at least a second portion of cyclopeptide toxins from the sample.
12. The method of claim 11 , wherein the biological sample is obtained from a mushroom.