Process for extracting constituents from organic material
The present invention relates to a process for extracting constituents from organic material, comprising the step of extracting with a monophasic mixture of 50% by volume to 90% by volume of methanol and 50 to 10% by volume of water and optionally 0% by volume to 40% by volume of a further solvent or solvent mixture.
1. A process for extracting one or more metabolites from an organic material, comprising an extraction step, comprising
(A) extracting an organic material wherein the extracting comprises two extractions with different solvents or solvent mixtures comprising
(a) extracting the organic material with a monophasic mixture of 50% by volume to 90% by volume of methanol and 50% by volume to 10% by volume of water and, optionally, 0% by volume to 40% by volume of a further solvent or solvent mixture; and
(b) extracting the organic material with a monophasic unpolar solvent or solvent mixture; and
(B) combining the extracts obtained from step (a) and step (b) to form one phase;
wherein the extraction of step (a) and the extraction of step (b) are carried out in parallel or in succession.
2. The process of claim 1 , wherein step (a) is carried out with a mixture of 80% by volume of methanol and 20% by volume of water.
3. The process of claim 1 , wherein step (b) is carried out with a mixture of 30% by volume to 60% by volume of C 1 - to C 6 -alkyl alcohol, acetone, acetonitrile, ethyl acetate, dimethyl sulfoxide or N,N-dimethylformamide and 40% by volume to 70% by volume of chloroform, dichloromethane, pentane, hexane, heptane, cyclohexane or carbon tetrachloride.
4. The process of claim 1 , wherein step (b) is carried out with a mixture of 30% by volume to 70% by volume of methanol and 40% by volume to 60% by volume of dichloromethane.
5. The process of claim 1 , wherein, in one or more further steps, the one or more metabolites are derivatized, chromatographed and/or analyzed.
6. The process of claim 1 , wherein the process comprises at least one of the following further steps:
i) freezing the organic material;
ii) freeze-drying the organic material;
iii) homogenizing and/or dispersing the organic material;
iv) evaporating the extract or the phase to dryness;
v) carrying out an esterification/transesterification in the unpolar phase;
vi) carrying out an oxime formation in the unpolar phase;
vii) carrying out an oxime formation in the polar phase;
viii) carrying out, in the unpolar phase, a trialkylsilylation where each alkyl radical contains 1 to 6 carbon atoms; or
ix) carrying out, in the polar phase, a trialkylsilylation where each alkyl radical contains 1 to 6 carbon atoms.
7. The process of claim 6 , wherein the esterification/transesterification is carried out with a volatile acid as catalyst.
8. The process of claim 6 , wherein one or more standard substances are added before carrying out step v), vi), vii), viii) or ix).
9. The process of claim 8 , wherein the standard substance is an internal standard or a chromatographic standard.
10. The process of claim 1 , wherein extracting is carried out at a temperature of from 0° C. to 200° C. and at a pressure of from 40 bar to 200 bar.
11. The process of claim 1 , wherein the solvent or solvent mixture used in step (a) and/or step (b) for the extraction additionally comprise up to 5% by weight of buffer salts, acids and/or bases.
12. The process of claim 1 , wherein the phase is analyzed via LC, GC, MS, LC/MS, GC/MS, and/or LC/MS/MS analysis.
13. The process of claim 1 , wherein the organic material is composed of plant material.
14. The process of claim 1 , wherein the process is part of another process for analyzing a metabolic profile.
15. The process of claim 14 , further comprising analyzing the resulting data by automatic peak recognition and peak integration.
16. The process of claim 1 , wherein step (a) comprises plural extraction steps.
17. The process of claim 16 , wherein the extraction of step (a) and the extraction of step (b) are carried out in succession, wherein the extraction of step (a) is carried out followed by the extraction of step (b), or the extraction of step (b) is carried out followed by the extraction of step (a).
18. The process of claim 16 , wherein the extraction of step (a) and the extraction of step (b) are carried out in parallel by dividing the organic material and carrying out the extraction of step (a) and step (b) separately.
19. The process of claim 1 , wherein step (b) comprises plural extraction steps.
20. The process of claim 19 , wherein the extraction of step (a) and the extraction of step (b) are carried out in succession, wherein the extraction of step (a) is carried out followed by the extraction of step (b), or the extraction of step (b) is carried out followed by the extraction of step (a).
21. The process of claim 19 , wherein the extraction of step (a) and the extraction of step (b) are carried out in parallel by dividing the organic material and carrying out the extraction of step (a) and step (b) separately.
22. The process of claim 1 , which step (a) and step (b) comprise plural extraction steps.
23. The process of claim 22 , wherein the extraction of step (a) and the extraction of step (b) are carried out in succession, wherein the extraction of step (a) is carried out followed by the extraction of step (b), or the extraction of step (b) is carried out followed by the extraction of step (a).
24. The process of claim 22 , wherein the extraction of step (a) and the extraction of step (b) are carried out in parallel by dividing the organic material and carrying out the extraction of step (a) and step (b) separately.
25. The process of claim 1 , wherein the extraction of step (a) and the extraction of step (b) are carried out in succession, wherein the extraction of step (a) is carried out followed by the extraction of step (b), or the extraction of step (b) is carried out followed by the extraction of step (a).
26. The process of claim 1 , wherein the extraction of step (a) and the extraction of step (b) are carried out in parallel by dividing the organic material and carrying out the extraction of step (a) and step (b) separately.
27. The process of claim 1 , further comprising treating the combined extracts, obtained from combining the extracts of step (a) and step (b), with one or more analytical standards.
28. The process of claim 27 , wherein the analytical standard is an internal standard or a chromatographic standard.
29. The process of claim 1 , wherein the analytical standard is an isotope-labeled, radiolabeled, or fluorescence-labeled substance.
30. The process of claim 1 , further comprising treating the combined extracts, obtained from combining the extracts of step (a) and step (b), with water.