CN102060824A - Method for preparing coumarin - Google Patents

Abstract

The invention discloses a method for preparing coumarin by extraction. The method comprises the following steps of: weighing coumarin and crushing; adding the crushed coumarin into water in an amount which accounts for 3 to 12 times the weight part of the crushed coumarin; performing reflux extraction for 2 to 3 times, wherein the extraction is performed for 30 to 60 minutes every time at a solid-liquid ratio of 1 to 40 and the extraction temperature of between 40 and 95 DEG C; mixing extract and filtering to remove insoluble impurities; distilling and concentrating the extract under reduced pressure; adding ethanol into concentrate with stirring to guarantee that alcohol content reaches 80 percent; standing; filtering; and collecting precipitates. The coumarin has the effects of protecting liver, reducing glutamic-pyruvic transaminase, enhancing immunity, calming, lowering blood sugar, preventing tumor and the like, is widely applied to Chinese patent medicine, health-care foods and beatifying medicated foods, and has broader development prospect.

Description

A kind of preparation method of tonka bean camphor

Technical field

The invention belongs to technical field of traditional Chinese medicine pharmacy, relate to the preparation method of tonka bean camphor.

Background technology

The coumarins medicine is the oral anticoagulant of a class.Their common structure is a 4 hydroxy coumarin.Simultaneously, temparin can also be used to tackle the plague of rats.People caused the temparin found in the lethal process of internal hemorrhage the pasture livestock because of anticoagulation originally, recognized the anticoagulation of this class material, after having caused to the research of coumarins medicine and synthetic, thereby provide many a kind of important blood-clotting agents for medical circle.Common coumarins medicine has temparin (dicoumarol), warfarin (warfarin, neodicoumarin) and Acenocoumarol (acenocoumarol, Syncumar).

The effect of coumarins medicine is the anticoagulant factor synthesizing at liver.The structural similitude of coumarins medicine and vitamin b6 usp K.The coumarins medicine combines with vitamin b6 usp K epoxide reductase at liver, suppresses vitamin K and is transformed to hydroquinone type by epoxide, and the circulation of vitamin K is suppressed.We can say that the coumarins medicine is the vitamin K antagonist, or competitive inhibitor (referring to enzyme).Contain prothrombin, VII, the IX of glutaminic acid residue, the carboxylation of X is suppressed, and its precursor does not have blood coagulation activity, so coagulation process is suppressed.But it is invalid to established thrombin.。Show that (1) have antitumour activity in vivo, and can promote the immunologic function of human body; Contained tonka bean camphor can reach 96.88% to the inhibition efficient of sarcoma S-180.(2) preserved material has diuretic properties to normal rabbit as abdominal injection; Ether or ethanol extraction can make isolated frog heart shrink reinforcement, to tame rabbit blood glucose rising are then reduced.(3) fushen's decoction has sedative effect to mouse, and the sedative effect of Poria cocos is inferior to fushen.[property of medicine] sweet, light, equal, return lung, stomach, kidney channel.Anticancer, sharp aquation drink, the peaceful heart of invigorating the spleen relieve internal heat.Promoting diuresis to eliminate damp pathogen, invigorating the spleen is reduced phlegm, tranquillizing by calming the heart.

Tonka bean camphor (Pachymaran) derives from polyporaceae fungi Poria cocos (Poriacocos Wolf).There is invigorating the spleen to be good for effects such as kidney, tranquillizing by calming the heart.The modern pharmacology effect shows that tonka bean camphor can strengthen immune function of human body, improves the human body resistance against diseases, plays diseases prevention, function in delaying senility.Select to be fit to the extracting method of Poria cocos, the effective constituent of extracting Poria cocos to greatest extent is the basis of research Poria cocos pharmacological action.Existing two kinds of up-to-date extracting method can make the extraction yield of water-soluble polysaccharide be significantly improved.

Summary of the invention

The technical solution used in the present invention is as follows:

A kind of preparation method of tonka bean camphor is characterized in that, prepares with following method:

(1) take by weighing Poria cocos section back and pulverize, add the water of 3-12 times of parts by weight, refluxing extraction 2-3 time solid-liquid ratio 1: 40, is extracted 40-95 ℃ of temperature, extracts 30-60min/ united extraction liquid filtration, removes insoluble impurities;

(2) get the extracting solution underpressure distillation and concentrate, concentrated solution adds ethanol while stirring, makes to contain the alcohol amount and reach 80%, leaves standstill, and filters collecting precipitation.

Preparation method of the present invention, wherein extracting temperature is 40 ℃.

Preparation method of the present invention, wherein extraction time is 30min/ time.

Preparation method of the present invention wherein adds 95% ethanol.

The objective of the invention is experimental design and optimize the extracting method and the processing condition of tonka bean camphor.Method utilizes experiment of single factor to optimize the extracting method of tonka bean camphor, utilizes the response surface experimental design to investigate the influence to the polysaccharide extracted amount of alkali concn in the alkaline process leaching process, extraction time, solid-liquid ratio on the basis of experiment of single factor.The optimum extracting method of polysaccharide is that alkaline process extracts in the tonka bean camphor as a result, and its extraction conditions can be described with regression equation Y=7.89+15.249X1-35.4X12+8.375X2-0.885X22+7.333X1X2.The conclusion optimum extracting method is an alkaline process, and the optimum extraction process condition is: NaOH concentration 0.84mol/L, time 8.89h, solid-liquid ratio 1: 150.

Equipment

1, A key instrument UV752 type visible ultraviolet spectrophotometer; JJ series electronic balance; Sai Duolisi BS series electronic balance; HHS type electric-heated thermostatic water bath; HS10260D ultrasonic extraction device; The XYJ802 centrifugal precipitation mechanism.

The Tuckahoe peel dry product that the B tonka bean camphor Yunnan white piece of Simao Diqu processing Poria cocos is cut is pulverized standby.

The C main agents vitriol oil, sodium hydroxide, phenol, dextrose anhydrous are homemade analytical pure.Phenol solution: take by weighing phenol 6g, add water to 100ml promptly.The glucose reference liquid: precision takes by weighing the glucose 100.3mg adding distil water dissolving of 105 ℃ of following dry constant weights and is settled to 100ml.

2, method and result

The A tonka bean camphor is measured the method that adopts sulfuric acid phenol and is measured.

The B water extraction takes by weighing a certain amount of Tuckahoe peel, under the certain condition of solid-liquid ratio, investigates the relation of extraction time, extraction time and extraction temperature and tonka bean camphor extracted amount.

The C ultrasonic extraction takes by weighing a certain amount of Tuckahoe peel, under the certain condition of solid-liquid ratio, investigates the relation of supersound extraction time and extraction temperature and polysaccharide extracted amount.

The D alkaline process extracts polysaccharide and takes by weighing a certain amount of tonka bean camphor, and under the certain condition of solid-liquid ratio, room temperature is extracted 30min, investigates the relation of alkali concn and polysaccharide extracted amount.

The E water extraction

(1) investigation of extraction time takes by weighing a certain amount of tonka bean camphor, solid-liquid ratio 1: 40, extracts 95 ℃ of temperature, extracts under 30min/ time the condition relation of investigation extraction time and tonka bean camphor extracted amount.The polysaccharide amount that Tuckahoe peel extracts increases with the increase of accumulation extraction time, but since the polysaccharide amount extracted for the 2nd time and reduce rapidly, takes all factors into consideration factors such as energy consumption in the leaching process, water consumption, should adopt once extraction.

(2) investigation of extraction time takes by weighing a certain amount of Tuckahoe peel, solid-liquid ratio 1: 40, extracts 95 ℃ of temperature, extracts under the condition once, investigates the relation of different extraction times and polysaccharide extracted amount.Between 10～20min, the extracted amount of tonka bean camphor polysaccharide increases fast with the increase of extraction time, then changes after the 20min and relaxes, so extraction time is advisable with 20～30min.

(3) investigation of extracting temperature takes by weighing a certain amount of Tuckahoe peel, and solid-liquid ratio 1: 40, extraction time 30min under the lixiviate condition once, investigated the relation of extracting temperature and polysaccharide extracted amount.The polysaccharide extracted amount increases with the increase of extracting temperature between 20～40 ℃ in the Tuckahoe peel, it is then on a declining curve after temperature is higher than 40 ℃, change little between 60～80 ℃, raw material is in continuous kinestate under boiling situation (95 ℃), its polysaccharide extracted amount rises to some extent, prompting is stirred and is made the continuous motion of Tuckahoe peel help the extraction of polysaccharide, so should select for use 40 ℃ of stirrings to extract.

The F ultrasonic extraction

(1) investigation of supersound extraction time takes by weighing a certain amount of Tuckahoe peel, and solid-liquid ratio 1: 40, ultrasonic frequency 40kHz under the supersound extraction condition once, investigated the relation of supersound extraction time and polysaccharide extracted amount., Tuckahoe peel polysaccharide extracted amount increases with the increase of ultrasonic time, and after extraction time reached 30min, the influence of extraction time to extracted amount reduced, and it is milder that curve becomes.So the time of a supersound extraction is advisable with 30min.

(2) investigation of supersound extraction number of times takes by weighing a certain amount of Tuckahoe peel, and solid-liquid ratio 1: 40, ultrasonic frequency 40kHz under the condition that supersound extraction is 30min/ time, investigated the relation of supersound extraction number of times and polysaccharide extracted amount.

The G alkaline process extracts and to take by weighing a certain amount of Tuckahoe peel, solid-liquid ratio 1: 200, under the room temperature lixiviate 30min condition, investigates the relation of alkali concn and polysaccharide extracted amount.The result shows that Tuckahoe peel polysaccharide extracted amount is in rising trend between NaOH concentration 0.25～0.5mol/L, and is later on a declining curve greater than 0.5mol/L when concentration, is that 0.5mol/L is that central point carries out secondary rotating orthogonal combination experiment so should select NaOH concentration.

H secondary rotating orthogonal combination experiment is put forward in the process each factor interaction to the influence of Tuckahoe peel leaching process for multianalysis alkali, under 1: 150 condition of solid-liquid ratio, this research adopts 23 rotation response surface experimental designs to carry out, factor of influence is NaOH concentration, extraction time, and index is the polysaccharide extracted amount of Tuckahoe peel as a result.Experimental result adopts quadratic regression analysis and variance analysis, draws suitable alkali according to regression equation and puies forward processing condition.

2 results carry out regression analysis and variance analysis with two factor secondary rotating orthogonal combination experiment statistics program his-and-hers watches, obtaining with the polysaccharide extracted amount is index, with NaOH concentration X1 (mol/L) and extraction time X2 is the quadratic regression equation and the The results of analysis of variance (seeing Table 3) thereof of variable, the result shows in the lixiviate of Tuckahoe peel polysaccharide alkali, on α=0.05 level, the polysaccharide extracted amount is influenced the direct action that significant factor is X1, X2, the secondary action of X2 and the interaction of X1X2 (P＜0.05).

The coumarins medicine is the oral anticoagulant of a class.Their common structure is 4 hydroxy coumarinSimultaneously, temparin can also be used to tackle the plague of rats.People caused the temparin found in the lethal process of internal hemorrhage the pasture livestock because of anticoagulation originally, recognized the anticoagulation of this class material, after having caused to the research of coumarins medicine and synthetic, thereby provide many a kind of important blood-clotting agents for medical circle.

Common coumarins medicine have temparin (dicoumarol), Warfarin(warfarin, neodicoumarin) and Acenocoumarol(acenocoumarol, Syncumar).

The effect of coumarins medicine is to suppress Thrombin LiverSynthetic.The structural similitude of coumarins medicine and vitamin b6 usp K.The coumarins medicine combines with vitamin b6 usp K epoxide reductase at liver, suppresses vitamin K and is transformed to hydroquinone type by epoxide, and the circulation of vitamin K is suppressed.We can say that the coumarins medicine is The vitamin K antagonist, or Competitive inhibitor(referring to enzyme).Contain L-glutamic acidThe carboxylation of the prothrombin of residue, VII, IX, X is suppressed, and its precursor does not have blood coagulation activity, therefore Blood coagulationProcess is suppressed.But it is invalid to established thrombin.In the polysaccharide alkali leaching process, direct action, secondary action, interaction that NaOH concentration X1, extraction time are carried X2 to the mathematical model of polysaccharide extracted amount influence are: Y=7.8899+15.248 8X1-35.3996X12+8.372 6X2-0.884 7X22+7.3333 X1X2.The relation conefficient 0.967 7 of this mathematical model, standard error 3.792 1, The results of analysis of variance is (P＜0.05 sees Table 3) significantly, can be used for predicting the polysaccharide extracted amount in the Tuckahoe peel polysaccharide alkali leaching process.Figure is replied on the surface of Tuckahoe peel polysaccharide extracted amount to each factor and isogram is seen Fig. 8.

3 conclusions

With the tonka bean camphor extracted amount is index, compares water extraction, ultrasonic extraction and alkali extraction method, and the extraction of tonka bean camphor should be extracted with alkaline process in the Tuckahoe peel.In the Tuckahoe peel alkali leaching process, NaOH concentration X1 and extraction time X2 to the mathematical model of polysaccharide extracted amount influence are: Y=7.8899+15.248 8X1-35.399 6X12+8.372 6X2-0.884 7X22+7.333 3X1X2.Obtaining best alkali extracting technology condition by prediction equation is: NaOH concentration X1:0.84mol/L, time X2:8.89h.Tonka bean camphor polysaccharide extracted amount reaches 54.99% under this condition.

Embodiment

Following examples are with helping understand the present invention, and are not used in and also should be interpreted as the restriction to inventing in the listed claim by any way.The method preparation that wherein raw material can reference example or buying on the market.

Embodiment 1

(1) take by weighing tonka bean camphor 500g section back and pulverize, add the water of 12 times of parts by weight, refluxing extraction 3 times solid-liquid ratio 1: 40, is extracted 40 ℃ of temperature, extracts 30min/ united extraction liquid filtration, removes insoluble impurities;

(2) get the extracting solution underpressure distillation and concentrate, concentrated solution adds 95% ethanol while stirring, makes to contain the alcohol amount and reach 80%, leaves standstill, and filters, and collecting precipitation obtains tonka bean camphor, and the polysaccharide extracted amount reaches 54.99% in the Tuckahoe peel.

Embodiment 2

(1) take by weighing tonka bean camphor 500g section back and pulverize, add the water of 10 times of parts by weight, refluxing extraction 2 times solid-liquid ratio 1: 40, is extracted 80 ℃ of temperature, extracts 60min/ united extraction liquid filtration, removes insoluble impurities;

(2) get the extracting solution underpressure distillation and concentrate, concentrated solution adds 95% ethanol while stirring, makes to contain the alcohol amount and reach 80%, leaves standstill, and filters, and collecting precipitation obtains tonka bean camphor, and the polysaccharide extracted amount reaches 55.9% in the Tuckahoe peel.

Embodiment 3

In per 1000 milliliters of injection liquids, add tonka bean camphor 500mg, tween-80 30g, surplus is the sodium-chlor isotonic solution.Add the small amount of activated heated and stirred, press filtration, filtrate is through filtering with microporous membrane, and filtrate is supplied water for injection, mixing, embedding, sterilization is made into injection liquid.

Embodiment 4

In per 1000 milliliters of injection liquids, add tonka bean camphor 150mg, tween-80 10g, surplus is oozed Klorvess Liquid for waiting.Add the small amount of activated heated and stirred, press filtration, filtrate is through filtering with microporous membrane, and filtrate is supplied water for injection, mixing, embedding, sterilization is made into injection liquid.

Embodiment 5

In per 1000 milliliters of injection liquids, add tonka bean camphor 200mg, tween-80 15g, surplus is for waiting magnesium chloride osmometer solution.Add the small amount of activated heated and stirred, press filtration, filtrate is through filtering with microporous membrane, and filtrate is supplied water for injection, mixing, embedding, sterilization is made into injection liquid.

Embodiment 6

In per 1000 milliliters of injection liquids, add tonka bean camphor 280mg, tween-80 25g, surplus is a water for injection.Add the small amount of activated heated and stirred, press filtration, filtrate is through filtering with microporous membrane, and filtrate is supplied water for injection, mixing, embedding, sterilization is made into injection liquid.

Embodiment 7

In per 1000 milliliters of injection liquids, add tonka bean camphor 300mg, tween-80 18g, surplus is the Sodium.alpha.-hydroxypropionate isotonic solution.Add the small amount of activated heated and stirred, press filtration, filtrate is through filtering with microporous membrane, and filtrate is supplied water for injection, mixing, embedding, sterilization is made into injection liquid.

Embodiment 8

In per 1000 milliliters of injection liquids, add tonka bean camphor 400mg, tween-80 28g, surplus is a water for injection.Add the small amount of activated heated and stirred, press filtration, filtrate is through filtering with microporous membrane, and filtrate is supplied water for injection, mixing, embedding, sterilization is made into injection liquid.

Claims (4)

1. the preparation method of a tonka bean camphor is characterized in that, prepares with following method:

(1) take by weighing tonka bean camphor and pulverize, add the water of 3-12 times of parts by weight, refluxing extraction 2-3 time solid-liquid ratio 1: 40, is extracted 40-95 ℃ of temperature, extracts 30-60min/ united extraction liquid filtration, removes insoluble impurities;

(2) get the extracting solution underpressure distillation and concentrate, concentrated solution adds ethanol while stirring, makes to contain the alcohol amount and reach 80%, leaves standstill, and filters collecting precipitation.

2. preparation method as claimed in claim 1, wherein extracting temperature is 40 ℃.

3. preparation method as claimed in claim 1, wherein extraction time is 30min/ time.

4. preparation method as claimed in claim 1 wherein adds 95% ethanol.