CN103614450A - Method for resolving DL-menthol through catalysis of lipase - Google Patents
Method for resolving DL-menthol through catalysis of lipase Download PDFInfo
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- CN103614450A CN103614450A CN201310703291.6A CN201310703291A CN103614450A CN 103614450 A CN103614450 A CN 103614450A CN 201310703291 A CN201310703291 A CN 201310703291A CN 103614450 A CN103614450 A CN 103614450A
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- menthol
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Abstract
The invention relates to a method for resolving DL-menthol through the catalysis of lipase, and belongs to the technical field of biological catalysis, solving the technical problems of very difficult acylating agent separation and cumbersome step of the traditional method for resolving the DL-menthol. The method comprises the following steps of 1, mixing the LD-menthol, porcine pancreatic lipase and succinic anhydride with ether, then shaking at the rotation speed of a shaking table, carrying out centrifugal filtration to remove the porcine pancreatic lipase, diluting by using diethyl ether, adding an NaCO3 solution, then extracting for three times by using the diethyl ether, then drying by using MgSO4, and then carrying out decompressing rotary evaporation to remove the diethyl ether to obtain D-menthol; 2, adding an NaOH solution to a water phase for stirring and hydrolysis, then extracting for three times by using the diethyl ether, drying by using MgSO4, and carrying out the decompressing rotary evaporation to remove the diethyl ether to obtain L-menthol. The method disclosed by the invention is used for obtaining the D-menthol and the L-menthol.
Description
Technical field
The invention belongs to biocatalysis technology field, relate to a kind of method of preparing MENTHOL; Be specifically related to the method for utilizing succinyl oxide to be catalyzer fractionation DL-menthol for acylating agent, lipase.
Background technology
Menthol claims again mentha camphor (menthacamphor), formal name used at school 1-methyl-4-sec.-propyl-hexamethylene-3-alcohol, English name DL-Menthol (± menthol); Molecular formula C
10h
20o is very important cyclic terpene alcohol chipal compounds.Owing to existing three unsymmetrical carbons in menthol molecule, so it can produce four pairs of different stereomeric racemic modifications.Wherein, that MENTHOL (-menthol) has is pure and fresh, brisk, elegant, with sweet irritating smell, and there is similar peppermint main note odor characteristics, and D-menthol has stimulation, assorted has wood smell mint flavored and be slightly with the peculiar smell of camphor.The refrigerant effect of MENTHOL makes it much more valuable than D-menthol (+menthol).At aspect MENTHOLs such as medical and health, foodstuffs industry and daily fine chemical products, have a wide range of applications, can be used as local anesthetic, sterilant, and it is medium to can be used for makeup, cigarette, refrigerant drink and other food flavor(ing).Therefore, how research is obtained efficiently optically active MENTHOL tool and is of great significance.
The common method that obtains menthol optically active substance mainly contains following several: chiral source method, the fractionation of dissymmetric synthesis and racemic modification.Certainly in whole fractionation or directed building-up process, can be the combination of two kinds of approach, some step be directed synthetic, and some step is to split or epimerization, thereby achieves the goal cost-effectively.In addition, some new technology are the numerous and confused preparation process for chiral drug also, as supercritical CO
2middle enzyme catalysis Chiral pesticide is synthetic, and Yu etc. have studied chirality building-up reactions lipase-catalyzed in supercutical fluid, Chrlsochoon etc. by supercritical technology be used in the enzyme process of Chiral pesticide synthetic on.
Preparation resolution of racemates process in, the method for utilizing biocatalysis to prepare optical purity enantiomorph just more and more demonstrates its importance.At present, for the enzyme majority of resolution of racemic compound, be lytic enzyme, and the overwhelming majority is lipase.Along with the development of non-aqueous media zymetology, for solving this difficult problem of fractionation of the fractionation, particularly racemic alcohol of racemic compound, brought hope.The research report of the application of relevant lipase in organic solvent is a lot, and A.M.Klibanov splits and prepared a large amount of optically active esters, acid in organic solvent.
Lipase majority derives from fungus and bacterium, and the different business-like lipase ，Qie in nearly kind more than 30 source scientific research institution constantly finds novel lipase now, for selecting a kind of efficient stereoselectivity lipase to give Maximum Space.
Enzymatic Resolution chiral material is mainly the stereoselectivity of utilizing enzyme, whole reaction process is exactly the active centre of 2 enantiomorph competition enzymes of racemic substrate, because both speed of reaction are different, produce selectivity, thereby make reaction product or residue substrate there is single optical activity.The optical selective of enzyme is exactly usually said enantioselectivity rate (enantioselectivity, E).
In the nonaqueous phase enzymatic esterification of chiral alcohol, the affect highly significant of the selection of acylating agent on enzymatic activity and enantioselectivity.In acyl group shift reaction, nucleophilic reagent as alcohol, ester etc. be limited.Thereby transesterification reaction is usually reversible; this will cause the decline of speed of response and the generation of side reaction; can adopt excessive acry radical donor for this reason; but this method costs dearly; best bet is to select special acry radical donor; to produce an irreversible reaction, acid anhydrides is also conventional acry radical donor, can form irreversible reaction.Its speed of response and selectivity are suitable with carboxylic acid alkene ester, and it can not produce aldehyde material, can not cause the inactivation of enzyme.Ethyl acetate, methoxyacetic acid ethyl ester, ethoxy ethyl acetate are all to study at present more acylating agent, and synthetic after product need to be by loaded down with trivial details column chromatography for separation, complex steps.
Summary of the invention
The object of this invention is to provide the method that organic phase system enzyme-catalyzed change splits DL-menthol, the method acylating agent that the invention solves existing fractionation DL-menthol is difficult to the technical problem of separated and complex steps.Method of the present invention improves the selectivity of resolution reaction, more than D-menthol enantiomeric excess can reach 95ee%, more than MENTHOL enantiomeric excess can reach 93ee%, DL-menthol transformation efficiency can reach more than 87%, and reaction conditions is gentle, the method reaction conditions is safe, gentle, easy, efficient, does not need lengthy and tedious chromatographic separation.
For solving the problems of the technologies described above, the method for lipase-catalyzed fractionation DL-menthol of the present invention completes by following step:
Be in step 1,1～20mmolDL-menthol, 1000～2000U/mmol porcine pancreatic lipase (PPL), 10～100mmol succinyl oxide to be mixed with 1～200mL ether.
Ether described in step 1 is by ether.
Described in step 1, the temperature of shaking table is 20～35 ℃.
Described in step 1, the rotating speed of shaking table is 100～300r/min.
Described in step 1, the jolting time is 6～72h.
The usage quantity of diluting the ether of use described in step 1 is 5～100mL.
NaCO described in step 1
3the usage quantity of solution is 2～100mL, described NaCO
3solution volumetric molar concentration is 0.1～10mol/L, and in step 1, each usage quantity of extracting the ether of use is 10～50mL.
Described in step 1, the usage quantity of NaOH solution is 5～100mL, and the volumetric molar concentration of NaOH solution is 0.1～10mol/L, and hydrolysis time is 1～10h.
Described in step 2, the usage quantity of NaOH solution is 5～100mL, and the volumetric molar concentration of NaOH solution is 0.1～10mol/L, and hydrolysis time is 1～10h, and in step 2, each usage quantity of extracting the ether of use is 50～150mL.
Reactional equation is as follows:
The highly-solid selectively of this research and utilization lipase splits DL-menthol and obtains MENTHOL, thereby has avoided the drawbacks such as chiral induction body that chemosynthesis MENTHOL reactions steps is many and use is expensive.Using succinyl oxide as acylating reagent, and one of product is hemisuccinic acid, forms sodium salt be easy to from organic phase separated with sodium bicarbonate.Avoided miscellaneous column chromatography for separation unreacting alcohol and product, improved simultaneously and reflected body overrate and transformation efficiency.
Accompanying drawing explanation
Fig. 1 is the gas chromatogram of embodiment one product, No. 1 peak MENTHOL, No. 2 peak D-menthols; Fig. 2 is the nmr spectrum of DL-menthol; Fig. 3 is 4-((1S, 2R, 5S)-2-isopropy-5-methylcyclohexyloxy)-4-oxobutanoic acid(mono succinate menthyl ester) nucleus magnetic resonance figure.
Embodiment
Embodiment one: in present embodiment, the method for lipase-catalyzed fractionation DL-menthol completes by following step: step 1,5mmolDL-menthol, 1777U/mmol porcine pancreatic lipase (PPL), 30mmol succinyl oxide and 50mL ether are mixed in 250ml triangular flask, then at 30 ℃ of shaking tables with 200r/min rotating speed jolting 48h, filter and remove porcine pancreatic lipase, with 20mL ether dilution, then to add 20mL volumetric molar concentration be the NaCO of 1mol/L
3solution, then uses ether extraction three times, and the consumption that at every turn extracts ether is 20mL, uses MgSO
4after dry, decompression rotary evaporation is removed ether, obtains D-menthol; D-menthol enantiomeric excess rate can reach 95ee%, and it is as follows that transformation efficiency can reach 87%(calculating formula).
Product is carried out to GC analysis:
Agilent 6890N gas chromatograph is selected in gas chromatographic analysis.CYCLOSILB chiral column (0.25 μ m * 30m * 0.25mm), flame ionization ditector, carrier gas is nitrogen.Sample introduction and detector temperature are respectively 200 ℃ and 250 ℃.Heating schedule is as follows: 90 ℃ of initial temperatures insulation 10min, 2 ℃/min is warming up to 150 ℃, then 5 ℃/min is warming up to 165 ℃ and be incubated 5min.Chromatogram result is processed by Agilent look general workstation, and sample retention time is respectively MENTHOL 4.026min, D-menthol 4,556min.
Fig. 1 has illustrated the optical purity of product.Fig. 2 characterizes resolved product.
With microsyringe, getting 2 μ l samples analyzes for gas-chromatography (GC).Wherein reaction conditions take ether as solvent, substrate molar ratio as 1.5:1, reaction times 48h, under the condition of 30 ℃ of temperature of reaction, and porcine pancreatic lipase is the best results that catalyzer splits DL-menthol.
(1) result is calculated
For chirality is synthetic, should consider chemical yield, consider again optical yields.Optical yields represents by enantio-selectivity, uses.For enzymic catalytic reaction being had to a comprehensive evaluation, consider transformation efficiency and the selectivity of enzymic catalytic reaction simultaneously, enantiomeric excess (enantiomeric excess, ee) can quantitative description the enantio-selectivity of reaction, thereby draw enantioselectivity rate E (enantiomeric ratio).Calculation formula is as follows:
Wherein c represents transformation efficiency, and ee represents enantiomeric excess rate, and S represents unreacted substrate, P
land P
drepresent respectively L-and D-product.
Claims (10)
1. the method for lipase-catalyzed fractionation DL-menthol, is characterized in that the method for lipase-catalyzed fractionation DL-menthol completes by following step:
Step 1, DL-menthol, porcine pancreatic lipase, succinyl oxide are mixed with ether, then shaking speed jolting, porcine pancreatic lipase is removed in centrifuging, with ether dilution, adds NaCO
3solution, then uses ether extraction three times, uses MgSO
4dry, then the rotary evaporation removal ether that reduces pressure, D-menthol obtained;
Step 2, step 1 is extracted after remaining solution with stirring hydrolysis under NaOH solution room temperature, then use ether extraction three times, use MgSO
4dry, decompression rotary evaporation is removed ether, obtains MENTHOL.
2. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, is characterized in that in step 1,1 ~ 20mmolDL-menthol, 1000 ~ 2000U/mmol porcine pancreatic lipase, 10 ~ 100mmol succinyl oxide being mixed with 1 ~ 200mL ether.
3. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, is characterized in that the ether described in step 1 is by ether.
4. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, is characterized in that the temperature of shaking table is 20 ~ 35 ℃ described in step 1.
5. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, is characterized in that the rotating speed of shaking table is 100 ~ 300r/min described in step 1.
6. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, is characterized in that described in step 1 that the jolting time is 6 ~ 72h.
7. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, the usage quantity that it is characterized in that diluting described in step 1 the ether of use is 5 ~ 100mL.
8. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, is characterized in that NaCO described in step 1
3the usage quantity of solution is 2 ~ 100mL, described NaCO
3solution volumetric molar concentration is 0.1 ~ 10mol/L, and in step 1, each usage quantity of extracting the ether of use is 10 ~ 50mL.
9. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, is characterized in that the usage quantity of NaOH solution is 5 ~ 100mL described in step 1, and the volumetric molar concentration of NaOH solution is 0.1 ~ 10mol/L, and hydrolysis time is 1 ~ 10h.
10. the method for lipase-catalyzed fractionation DL-menthol according to claim 1, it is characterized in that the usage quantity of NaOH solution is 5 ~ 100mL described in step 2, the volumetric molar concentration of NaOH solution is 0.1 ~ 10mol/L, hydrolysis time is 1 ~ 10h, and in step 2, each usage quantity of extracting the ether of use is 50 ~ 150mL.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109651437A (en) * | 2018-12-14 | 2019-04-19 | 万华化学集团股份有限公司 | A kind of chiral nitrogen phosphorus ligand and preparation method thereof and a kind of method for splitting racemization menthol |
| CN115286507A (en) * | 2022-07-18 | 2022-11-04 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Menthol hapten, artificial antigen, preparation methods of menthol hapten and artificial antigen, antibody and application of menthol hapten and artificial antigen |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1471584A (en) * | 2000-11-02 | 2004-01-28 | Csir | Process for preparing (-)-menthol and the like compounds |
| CN1914190A (en) * | 2004-01-29 | 2007-02-14 | 巴斯福股份公司 | Method for producing enantiomer-pure aminoalcohols |
| CN101045936A (en) * | 2007-03-27 | 2007-10-03 | 吉林大学 | Process of preparing chiral fatty alcohol with acid anhydride as acry radical donor |
-
2013
- 2013-12-19 CN CN201310703291.6A patent/CN103614450A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1471584A (en) * | 2000-11-02 | 2004-01-28 | Csir | Process for preparing (-)-menthol and the like compounds |
| CN1914190A (en) * | 2004-01-29 | 2007-02-14 | 巴斯福股份公司 | Method for producing enantiomer-pure aminoalcohols |
| CN101045936A (en) * | 2007-03-27 | 2007-10-03 | 吉林大学 | Process of preparing chiral fatty alcohol with acid anhydride as acry radical donor |
Non-Patent Citations (1)
| Title |
|---|
| 周舒扬等: "脂肪酶催化拆分外消旋薄荷醇的研究进展", 《黑龙江科学》, vol. 4, no. 6, 30 June 2013 (2013-06-30), pages 63 - 66 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109651437A (en) * | 2018-12-14 | 2019-04-19 | 万华化学集团股份有限公司 | A kind of chiral nitrogen phosphorus ligand and preparation method thereof and a kind of method for splitting racemization menthol |
| CN109651437B (en) * | 2018-12-14 | 2021-03-09 | 万华化学集团股份有限公司 | Chiral nitrogen-phosphorus ligand, preparation method thereof and method for resolving racemic menthol |
| CN115286507A (en) * | 2022-07-18 | 2022-11-04 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Menthol hapten, artificial antigen, preparation methods of menthol hapten and artificial antigen, antibody and application of menthol hapten and artificial antigen |
| CN115286507B (en) * | 2022-07-18 | 2023-11-17 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Menthol hapten and artificial antigen as well as preparation methods, antibodies and applications thereof |
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Application publication date: 20140305 |