CN102850189B

CN102850189B - A kind of preparation method of high-purity saturation laccol and its formal derivatives

Info

Publication number: CN102850189B
Application number: CN201210310773.0A
Authority: CN
Inventors: 王成章; 何源峰
Original assignee: Institute of Chemical Industry of Forest Products of CAF
Current assignee: Institute of Chemical Industry of Forest Products of CAF
Priority date: 2012-08-29
Filing date: 2012-08-29
Publication date: 2018-03-20
Anticipated expiration: 2032-08-29
Also published as: CN102850189A

Abstract

This patent discloses the preparation method of a kind of high-purity saturation laccol and its formal derivatives, the acetone extract laccol of formal method modified raw lacquer is applied first, a kind of laccol derivative of new structure is obtained, while the process significantly reduces the polarity with catechol structure laccol monomer；Hydrogenation reaction is carried out by catalyst of palladium carbon, obtains saturation formalizing laccol；Positive post separation so that laccol monomer is easy to separate with polar impurity in polyurushiol and extract；Obtained saturation formal laccol obtains the saturation laccol of high-purity after sloughing formal group, and laccol and its derivative have strong bioactivity, has good market prospects.

Description

A kind of preparation method of high-purity saturation laccol and its formal derivatives

Technical field

This patent is related to a kind of preparation method of high-purity saturation laccol and its formal derivatives, is widely used in life The field such as thing pharmacy, cosmetics, fine chemistry industry.

Background technology

Raw lacquer is a kind of sumac bark secretion, is mainly used in coating industry at present, because raw lacquer coating has bloom Pool, it is corrosion-resistant, the characteristic of elegant and be known as the good reputation of " king of coating "；China's lacquer tree cultivated area is extensive, is distributed in me The areas such as Yunnan of state, Guizhou, Sichuan, Chongqing, Gansu, Shaanxi, Hubei, paint is often produced per year up to 5000 tons.The main component of raw lacquer It is laccol, the urushiol compound for presently mainly having three types, Chinese laccol, Vietnam's laccol and thitsiol, three kinds of laccols Main difference is that the length of the location of side chain and side chain, mainly by meta in Chinese laccol（Relative to No. 1 hydroxyl Base, similarly hereinafter）For the catechol derivatives composition of 15 carbon atoms, Vietnam's laccol is the laccol that meta contains 17 carbon atoms, Thitsiol is the side chain that contraposition is 17 carbon atoms.Laccol possesses good bioactivity, there is the record being used as medicine from ancient times；Closely Researcher has carried out substantial amounts of research to laccol bioactivity both at home and abroad over year：South Korea member of Imperial Academy university researcher has found laccol pair Helicobacter pylori has the inhibitory action arranged in detail, and research shows that optimal antibacterial pH is 6-9, to the minimum inhibitory concentration of bacterial strain For 0.064-0.256mg/mL；Taiwan's scholars separate 2 new chemical combination of hydroquinones from Rhus succedanea Rhus succedanea paint juice extractions Thing, its side chain are C17 diene or three allylic alkylations, have good anti-oxidant and anti-cancer effectiveness, find to cervical epithelium cancer HeLa, liver cancer Huh7, colorectal cancer HCT116 and adenocarcinoma of colon Lovo have obvious inhibitory action；Patent CN101805246A is public The cloth serial laccol salt derivative such as laccol and its acetylation, methyl-etherified, and the bioactivity of laccol is inquired into, tie Fruit shows that laccol and its salt derivative have splendid inhibition to breast cancer cell, esophageal cancer cell etc.；YUN-RU CHEN etc. have studied in raw lacquer regulation and control of the laccol to TYR enzyme so as to control the generation of melanin, it is contemplated that urushiol compound is being changed Whitening agent is used as in cosmetic, substitutes the catechol that tradition uses；Itokawa etc. separates long chain alkylphenol from gingko episperm Class（C13-17）There is good suppression to tumour cell S180, HCT-15, MCF-7, A-549, HT-1197 and SKOV-3.In The content of laccol is about 70% or so in the hair dam paint of state, and abundant yield provides the raw material of abundance for extraction laccol, is also paint Phenol has established solid foundation in cosmetics, bio-pharmaceuticals industry.

Laccol is a kind of derivative of catechol, and its oxidation activity is high, and laccol begins to occur to gather from raw lacquer collection Close reaction, research shows to take the raw lacquer of fresh collection to use alcoholic extraction laccol immediately, wherein the content of polymer and reaches 7%, Long-time storage, even if using deepfreeze raw lacquer also progressively there is milky to be changed into dark brown, and polyurushiol and laccol list It is difficult to separate after body mixing, and the laccol separated is easy to oxidative deformation without easy to maintain；Meanwhile raw lacquer has strong sensitization Property, the main reason for corresponding research shows, the double bond in the catechol structure and pendant alkyl group structure of laccol is sensitization.From knot From the point of view of on structure, laccol is serial different saturation catechol derivatives, while identical saturation degree has homology similar in structure Thing, the separation of such laccol is very difficult, and the laccol of different saturation can be separated in C18 posts using methanol-water solution, Occur simultaneously using LC-MS analysis shows laccol monomer and its dimer, therefore external generally use gel chromatography is to sample Pre-separation is carried out, the polymer removed in raw material obtains series monomers, then carries out follow-up Separation Research.

Prepared by the separation on laccol, the method that presently mainly application prepares liquid phase, and this method requires high to instrument and equipment, Solvent consumption is big, while the yield of laccol is extremely low；Chen Lianxi etc. is directly carried out hydrogenation using raw lacquer acetone extract as raw material and begged for By the hydrogenation principle of laccol, this method can make unsaturated laccol be changed into saturation laccol；But apply liquid-phase chromatographic analysis hydrogenation products It is to find that composition is not single, it is preliminary to infer wherein be that different chain length laccol and polyurushiol are mingled with, thus direct hydrogenation Can not solve the problems, such as the difficult separation of laccol；Foreign literature is reported in before the separation of laccol first to laccol sample application gel color Spectrum separation, but because laccol easily aoxidizes, during gel column is crossed easily chemical bond and and cause extremely to adsorb, post is filled out The injury of material is big, even if while finding directly to apply C in experimentation₁₈Liquid phase post analyzes the acetone extract of raw lacquer, liquid phase Chromatographic column post effect and rapidly reduction, the cost of separation is undoubtedly added by the way of laccol is directly separated；The neighbour of laccol Benzenediol hydroxyl is its polar group, has identical retention time with the polymer or oxidation product of laccol, anti-in formal Catechol reacts during answering, and the phenolic hydroxyl group with polar character disappears, and causes polarity to significantly decrease, on the other hand Oxidation or polymerizate due to laccol react to form benzylalcohol structure, so possess the monomer of catechol structure in formal The difference for changing polarity after reacting becomes big, while can also reduce the sensitization of raw lacquer, thin-layer chromatography and the production of liquid phase analysis formal Thing, it is found that the laccol polarity of part diminishes, Rf values are about 1 under the conditions of original thin layer, and regulation chromatographic solution is 99.7% petroleum ether With 0.3% ethyl acetate, discovery Rf values 0.8-0.95 component, while formal reaction product appearance time hysteresis in the liquid phase. So easily this moieties can be separated from system.The material application H-NMR separated is analyzed into it is Urushiol-Formaldehyde product, carry out the experiment of next step.

Using the methylene above Boron tribromide removing formal laccol, Boron tribromide is a kind of thing to moisture-sensitive Matter, the removing of system moisture is should be noted that during operation, configure the dichloromethane solution of Boron tribromide, dichloromethane need to be first dehydrated Processing, to ensure requirement of the reaction to moisture；Formal laccol adds N in reaction bulb₂Protection, by the dichloromethane of boron trifluoride Solution is slowly dropped into reaction system, reacts as low-temp reaction device, controls -40--20 DEG C of temperature, reaction time 1-2h.

Modification of the formalizing method applied to catechol structure, synthesis pepper base class medicine, agricultural chemicals etc. is important to organise The intermediate of compound.At present, catechol formal has two big synthetic routes, and a kind of is using excessive dichloromethane and adjacent benzene Diphenol reacts in head type reactor mesohigh, but this method conversion ratio is relatively low, while excessive dichloromethane also increases conjunction Into cost, in the basic conditions, equimolar amounts dichloromethane and catechol, which carry out synthesis under normal pressure, can both reach higher Conversion ratio.

Using the Urushiol-Formaldehyde thing and application process reduction laccol monomer polarity of formalizing reaction synthesizing new So that monomer is easily isolated, then by hydrogenation reaction, de- methylene is come to prepare high-purity saturation laccol be invention Innovative point.

The content of the invention

Object of the present invention is to provide the preparation method of a kind of high-purity formal laccol and saturation laccol, the method The formal laccol and saturation laccol of high-purity can be prepared by succinct technological process, is that one kind prepares saturation laccol And its short-cut method of formal derivatives.

A kind of preparation method of high-purity saturation laccol and its formal derivatives, its key step are as follows：

The first step：The extraction of laccol

Raw lacquer is laccol, 20% or so the water by 70%, 7% urushipolysaccharide and albumen and laccase composition, using organic solvent pair Active ingredient-laccol of raw lacquer is extracted, and is extracted respectively from methanol, ethanol, acetone, ethyl acetate, petroleum ether, excellent Methanol, ethanol, petroleum ether are selected, raw lacquer is 1 with organic solvent mass volume ratio:10-1:100, filtering, filter residue repeated washing, directly It is rendered as colourless, is extracted using ultrasonic assistant, extraction time 1s-300s to filtrate, Extracting temperature is 0 DEG C -40 DEG C；Gained Filtrate merges cryogenic vacuum concentration, the laccol of as serial different saturation.

Second step：Urushiol-Formaldehydeization is reacted

By volume 1：1~1：4 prepare dichloromethane-dimethyl sulphoxide solution, the laccol mixture that the first step is extracted With 1-5 times of dmso solution, laccol lysate and 10%-50% sodium hydrate aqueous solutions synchronously add dichloromethane-diformazan Base sulfoxide solution reaction system, 80 DEG C -130 DEG C, reaction time 5h of reaction temperature, reaction terminate vacuum concentration and obtain yellowish-brown oil Shape thing, puts plate analysis reaction effect, and products therefrom polarity substantially reduces.

3rd step：Laccol isolates and purifies

Product obtained by second step is separated, Urushiol-Formaldehyde thing separated using positive phase system, preferably silicon Glue uses silica gel as 100-200 mesh, sample is 1 with silica gel mass ratio as filler:20, with acetone, petroleum ether, acetic acid second Ester, n-hexane, normal heptane gradient elution, preferably petroleum ether and ethyl acetate are as eluant, eluent, 0.1% increase gradient to acetic acid second Ester is 10% stopping, and each gradient elution volume is the column volume of 3-10 times of bed volume, preferably 5 times, and capture range is 0.5-3%'s Component；

4th step：The hydrogenation reaction of laccol

It is catalyst using palladium carbon, hydrogenation reaction is carried out to Urushiol-Formaldehyde compound, reacts the solvent used for methanol, second A kind of in alcohol, petroleum ether, ethyl acetate or acetone, ethyl acetate and acetone, palladium-carbon catalyst dosage are 0.01-5%, Hydrogenation time is 0.5-8h, hydrogenation pressure 0.05-1.5MPa, and hydrogenation is concentrated in vacuo reactant after terminating, obtains slightly yellow oil Shape thing, room temperature place crystallizable solidification.

5th step：The de- methylene reaction of formal laccol

De- methylene reaction is carried out to the formal laccol of the 4th step separation, formal laccol is in -80 DEG C -0 DEG C and tribromo Change the dichloromethane solution mixing of boron, carry out the reaction of anhydrous and oxygen-free, stir 0.5-15h, obtain saturation laccol, purity 94- 98%。

Raw lacquer is a kind of water-in-oil emulsion, and wherein contains urushipolysaccharide and protein matter, is reached using organic solvent The effect of demulsification, while alcohols material can make the species precipitate of polysaccharide and protide, laccol and the polysaccharide and albumen of oily obtain To separation, organic solvent used includes petroleum ether, ethyl acetate, acetone, ethanol and methanol, preferred alcohol, third in experiment Ketone and methanol are as extracts reagent.

Laccol is easy to aoxidize, and laccol hydroxyl loses hydrogen ion and is changed into negative oxygen ion in the presence of Laccase Catalyzed agent, and oxygen is born The double bond of ion and phenyl ring is conjugated to form metastable conjugated system, the ortho para position of such laccol and the oxygen of phenolic hydroxyl group bear from Son turns into the site of reflection and forms the polymer of C-C, C-O structure, and catechol structure is consumed, while laccol conjugated double bond D-A can occur and react the laccol polymer to form side side structure, exist by the polymer and monomer of liquid matter analysis shows laccol Flowed out simultaneously in reverse phase silica gel, the reaction of this patent application formal, because laccol monomer has catechol structure, polarity hair Life significantly changes, thus the screening that the method can be to there is the laccol monomer of catechol structure to carry out selectivity, laccol contracting first Aldehyde is reacted using dichloromethane, and in the presence of sodium hydroxide, laccol forms laccol sodium, and the carbon atom in dichloromethane carries Extremely strong electropositive, both are prone to necleophilic reaction and form the laccol of formal；Laccol negative oxygen ion while attack laccol Benzene ring structure, dichloromethane can also react with the sodium hydroxide of surplus, thus during should pass through regulation both relative drop The reverse side of acceleration one make it that laccol is suitable with the amount of sodium hydroxide material, on the other hand controls laccol concentration, reduces laccol molecule The chance of copolymerization, laccol are 1 with solvent DMSO volume ratios:1-1:10, it is 0.5-6d/s, preferably 0.5-1d/ to control rate of addition s；10-50% sodium hydroxide solutions, control rate of addition 0.05-0.6d/s, preferably 0.05-0.3d/s.

The polarity of formal laccol is third there occurs normal phase column separation, optional organic solvent is carried out after significantly changing Ketone, petroleum ether, ethyl acetate, n-hexane, normal heptane, preferably gradient elution, petroleum ether-ethyl acetate system；With 0.1% gradient Increase gradient, collect 0-5% components, tracking merges collection liquid, is concentrated to give the formal laccol of high-purity.

It is mainly that side chain is C in Chinese hair dam raw lacquer₁₅Laccol, to formal laccol carry out hydrogenation reaction i.e. obtain saturation Urushiol compound, unsaturated laccol is mainly the chain alkyls that side chain contains three step saturated bonds in hair dam raw lacquer, by phase Close document report to understand, when using palladium carbon as catalyst, hydrogenation reaction both can be smoothly carried out under room temperature normal pressure, reaction uses Solvent be a kind of in methanol, ethanol, petroleum ether, ethyl acetate or acetone, ethyl acetate and acetone, palladium-carbon catalyst Dosage is 0.01-5%, hydrogenation time 0.5-8h, hydrogenation pressure 0.05-1.5MPa；Reaction temperature is 0-40 DEG C, is preferably 20-30℃；Hydrogenation finishes concentration, and due to losing for double bond, room temperature cools down to obtain white crystalline saturation formal laccol, and purity can Up to 95-98%.

Last this patent provides the method for the Deprotection of formal laccol to prepare saturation laccol, the side of Deprotection Method is relatively more, with reference to laccol chemical property it is active the characteristics of, select Boron tribromide Deprotection method, reaction temperature is -80- 0 DEG C, preferably -40--20 DEG C, both material ratios are 1:1-1:10, preferably 1:2-1:4, reaction time 0.5-15h, preferably 0.5- 2h.The reaction be anhydrous and oxygen-free reaction, during should strictly control the moisture of reaction system and the content of oxygen, gained saturation Laccol purity is 94-96%.

Beneficial effects of the present invention are shown as：

1. being reacted by formalizing, the serial formal urushiol compound with new structure is prepared first；

2. first Application formalizing reacts, by the laccol class material polarity with catechol structure and other types thing Matter polarity produces difference, and laccol monomer can both have been separated in positive silica gel；Using atmospheric hydrogenation by unsaturated Partial Conversion For saturation formal laccol, the yield of raising saturation formal laccol.

Brief description of the drawings：

The composition of laccol in the raw lacquer of accompanying drawing 1

The Urushiol-Formaldehyde of accompanying drawing 2 reacts

The laccol of accompanying drawing 3 and hydrogenation laccol HPLC collection of illustrative plates

The thin-layer chromatographic analysis laccol of accompanying drawing 4（a）, formal laccol（b）And saturation formal laccol（c）

The infrared spectrogram of the formal laccol of accompanying drawing 5 and laccol

The mass spectrogram of the saturation laccol of accompanying drawing 6

Embodiment

Following examples are some citings of the present invention, should not be seen as limitation of the invention.

Case study on implementation

The extraction of the raw lacquer laccol of case study on implementation 1

10g hairs dam raw lacquer is taken, 1-20 times of mass volume ratio acetone extraction, the acetone of preferably 7-15 times mass volume ratio, is filled Divide stirring, ultrasonic wave added 10-300s, filter out coating removal albumen, polysaccharide etc., 50ml acetone washing filter residue is taken for several times, until filtrate Become colorless（Fresh raw lacquer sample, the filter residue of gained are changed into pure white）, 20-35 DEG C of vacuum concentration obtain 7g yellowish-brown oilies Thing, it is the unsaturated laccol of series.

The formal reaction of the laccol of case study on implementation 2

Take gained laccol mixture to be dissolved in 1-10 times of volume dimethyl sulfoxide (DMSO), preferably 2-5 times of volume, take 1-4 times of mole 10-50% sodium hydrate aqueous solutions, prepare the solution of 2-10ml dichloromethane（Solvent is dimethyl sulfoxide (DMSO)）In tri- mouthfuls of burnings of 100ml Adjust 80-130 DEG C of oil bath temperature in bottle, preferably 100-110 DEG C, while by the DMSO solution of the aqueous solution of sodium hydroxide and laccol Reaction bulb is instilled, rate of addition is controlled, 1-5h, preferably 2-3 hours is persistently added dropwise, waits that supplement 2ml dichloromethane, which is added dropwise, to be held Continuous backflow 2h terminates to react, and the deionized water that 0.2-2 times of volume is added in system is fully washed to be produced during elimination reaction Salinity, 1-10 times of petroleum ether extraction be concentrated to give 5.2g yellow oils, the as formal production of series of saturation laccol Thing.

The hydrogenation reaction of the formal laccol of case study on implementation 3

The Urushiol-Formaldehyde product of gained in case 2 is taken to carry out hydrogenation reaction, using acetone, methanol, petroleum ether, acetic acid second Ester dissolves the formal thing of laccol, preferably acetone and petroleum ether, carries out hydrogenation reaction, volume ratio 1:1-1:5 times of acetone solution paints Phenol formal product, add 0.01-1% palladium-carbon catalysts, 25 DEG C, hydrogenation reaction 0.5-6h, preferably 4-5h under 0.1MPa, reaction After the completion of remove solvent, obtain light yellow oil, room temperature cools down to obtain white crystals thing.

The purifying of the formal laccol of case study on implementation 4

Formal laccol purification on normal-phase silica gel separation after concentration, using petroleum ether-ethyl acetate gradient elution, every 0.1% increase Gradient to the content of ethyl acetate is 4%, takes 0.2%-2.0% ethyl acetate component to concentrate, obtains white plates crystal 3.8g, i.e., For the formal product of saturation laccol, the purity of gained formal product is 95-98%.

The preparation of the saturation laccol of case study on implementation 5

Selection Boron tribromide sloughs the methylene group of formal laccol, prepares Boron tribromide dichloromethane solution, dichloro For methane through drying and dehydrating, both volume ratios are 1:1-1:5, take the saturation formal laccol of the gained of case 4 to insert anhydrous three mouthfuls of burnings Bottle carries out low-temp reaction, -40 DEG C of dropwise addition Boron tribromides of controlling reaction temperature, and stirring 3h stops, 5 times of deionized water cleanings, removing Solvent and the Boron tribromide for having neither part nor lot in reaction, white waxy solid is obtained, as saturation urushiol compound, reaction conversion ratio reaches 95%, the purity of saturation laccol is 94-98%.

Claims

1. the preparation method of a kind of high-purity saturation laccol and its formal derivatives, it is characterised in that by following step group Into：

The first step：The extraction of laccol, raw lacquer are 1 with organic solvent mass volume ratio (g/ml):10-1:100, extraction time is 1s-300s, Extracting temperature are 0 DEG C -40 DEG C；Extract solution filters, and filter residue with organic solvent washing until filter residue is white, filter by merging Liquid, concentrated in vacuum and low temperature, obtain the laccol mixture of serial different saturation；

Second step：Urushiol-Formaldehyde reacts, and by volume 1：1~1：4 prepare dichloromethane-dimethyl sulphoxide solution, by first It is synchronous with 1-5 times of dmso solution, laccol lysate and 10%-50% sodium hydrate aqueous solutions to walk the laccol mixture prepared Add dichloromethane-dimethyl sulphoxide solution reaction system, 80 DEG C -130 DEG C of reaction temperature, reaction time 1-7h；

3rd step：The separation of formal laccol, after second step reaction terminates, room temperature is cooled to, is extracted in equal volume with non-polar solven Extract reaction solution, extract 2-5 times, combining extraction liquid, be concentrated in vacuo, obtain yellowish-brown oily formal laccol；

4th step：Hydrogenation reaction, the formal laccol acetone solution that the 3rd step is separated, it is hydrogenated with palladium-carbon catalyst effect Reaction, catalyst amount be formal laccol quality 0.01%-5%, 0-40 DEG C of reaction temperature, pressure 0.05-1.5MPa, the time 0.5-8h；

5th step：Saturation formal laccol purifies, and takes hydrogenation reaction product, first filters, and filtrate carries out low-temperature reduced-pressure concentration, concentration The mass ratio of thing progress positive post separation, concentrate and positive column packing is 1：10~40, eluant, eluent is n-hexane-ethyl acetate Gradient elution, TLC and HPLC detections, collect saturation formalizing laccol collection point, are concentrated in vacuo, concentrate is saturation formal paint Phenol, purity 95-98%；

6th step：The removing of saturation formal laccol methylene, by saturation formal laccol and Boron tribromide in mass ratio 1:1- 1:5 add reactor, carry out removing methylene reaction at -80 DEG C -0 DEG C, react normal pressure, 0.5~5h of reaction time；

7th step：Prepared by high-purity saturation laccol, wash the product obtained by the 6th step using the deionized water of 1-10 times of volume, very Solvent in sky concentration removal system is the saturation laccol that can obtain high-purity, and the conversion ratio of reaction is more than 95%, and saturation laccol is pure Spend for 94-98%.

2. a kind of preparation method of high-purity saturation laccol and its formal derivatives according to claim 1, it is main special Organic solvent is any of methanol, ethanol or acetone used by sign is the first step.

3. the preparation method of a kind of high-purity saturation laccol according to claim 1 and its formal derivatives, its is main It is characterised by that first step vacuum and low temperature concentration condition is：Vacuum 500mmHg-750mmHg, temperature are 0-40 DEG C.

4. the preparation method of a kind of high-purity saturation laccol according to claim 1 and its formal derivatives, its is main Nonpolar solvent extraction agent is any of petroleum ether, ethyl acetate or acetone used by being characterised by the 3rd step.

5. the preparation method of a kind of high-purity saturation laccol according to claim 1 and its formal derivatives, its is main It is characterised by that using gradient elution agent in the 5th step be acetone-petroleum ether, n-hexane-ethyl acetate, normal heptane-ethyl acetate, Any one in petroleum ether-ethyl acetate or chlorofonn-ethylacetate.

6. the preparation method of a kind of high-purity saturation laccol according to claim 1 and its formal derivatives, its is main It is characterised by the 5th step that positive phase filling is any of silica gel, aluminum oxide.