CN1923829B - Preparation method and use of separating and purifying lotus leaf flavone and lotus leaf alkaloid by employing polyamide - Google Patents
Preparation method and use of separating and purifying lotus leaf flavone and lotus leaf alkaloid by employing polyamide Download PDFInfo
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Abstract
The invention discloses a preparing method and application of lotus leaf flavone and lotus leaf alkaloid, which comprises the following steps: adopting rough-extract of lotus leaf, film separating impurity-removing lotus leaf extract or resin separating purifying lotus leaf extract as raw material; adding water to stir into suspension; adsorbing lotus leaf flavone through polyamide; decompressing non-adorbed part to recycle; condensing solvent into lotus leaf alkaloid in the vacuum; washing lotus leaf flavone on the polyamide; removing polar impurity and pigment; eluting through polar organic solvent; decompressing eluent; recycling solvent; condensing into lotus leaf flavone in the vacuum; fitting for prepare fat-reducing, antihyperglycemic, antiviral or oxidation resistance oral agent.
Description
Technical field
The present invention relates to a kind of from plant the Green Chemistry preparation method of separation and purification active pharmaceutical ingredients, more specifically relate to a kind of preparation method and purposes that adopts polyamide separation and purification lotus flavone and lotus leaf alkaloid.
Background technology
In China, the lotus leaf aboundresources, cheap and easy to get, drug effect is clear and definite, for rare can intensive processing and large natural medicine-food dual purpose plant resource of comprehensive development and utilization.A large amount of lotus leaves are used as waste and are not used appropriately, and cause the significant wastage of resource, also bring environmental pollution to a certain degree but at source.
Nymphaeaceae plant lotus (Nelumbo nucifera Gaertn), be a kind of medicine food dual purpose plant, at China's the South and the North plantation is arranged all, plantation history reaches more than 3000 year, abound with in Fujian at present, province such as Jiangxi, Hubei, Hunan and Zhejiang, especially white lotus of producing with Jianning, Fujian (building lotus) and the Hunan Dongting Lake Area red lotus (Hunan lotus seeds) of producing is known far and wide at home and abroad.
Lotus leaf (Nelumbo nucifera leaf) is the dry leave of lotus, have another name called lotus leaf or lotus root leaf, the plant register of selected ministry of Health of China " being food; be again medicine ", food and the medicine of being widely used as among the people, also often be used for the treatment of obesity and hyperlipidaemia on the clinical medicine with rough machined compound preparation, curative effect is obvious, but less to furtheing investigate such as effective part groups such as lotus flavone and lotus leaf alkaloids.
Traditional medicine thinks, the lotus leaf nature and flavor are cold, has clearing summer-heat and damp, Weight-reducing and lipid-lowering, clears away heart-fire and reduce phlegm and internal heat, stop blooding the activity of Li Shui, can develop protective foodss such as making teabag, beverage, electuary, capsule.Modern pharmacological research shows that the biological activity of compositions such as flavones that lotus leaf is contained and alkaloid is higher, is active pharmaceutical ingredients main in the lotus leaf, and content can reach 1.5% and 0.3% respectively.Lotus flavone and lotus leaf alkaloid have tangible fat-reducing, lipopenicillinase, antibacterial, antiviral, detoxifcation, drug rehabilitation, function [Kong Wenqi etc., " traditional Chinese medicine research and information ", 2005,7 (6): 22-24] such as anti-oxidant.These achievements in research will provide scientific basis for the application of lotus leaf in food, medicine.The preparation method and the purposes of research lotus flavone and lotus leaf alkaloid have certain directive significance for the intensive processing and the series product development of lotus leaf.
At present, the main method of extraction lotus flavone has: water extraction method [Chen Haiguang etc., " Food science ", 2002,23 (1): 69-72], adopt macroporous resin enrichment absorption method [Cai Weirong etc., " Jiangsu food and fermentation " behind the water extraction, 2002,24 (4): 1-5; Zhao Jun etc., " Chinese medicinal materials ", 2005,28 (9): 832-834] etc.Extracting lotus leaf alkaloid method commonly used has: acidic aqueous solution extraction method [Chen Haiguang etc., " food and machinery ", 2001,17 (5): 16-17; Jiang Yihong, " journal of Zhejiang university (agricultural and life science version) " 2004,30 (5): 519-523; ], adopt macroporous resin enrichment absorption method [Zhao Jun etc., " Chinese medicinal materials ", 2003,26 (9): 669-670] behind the acidic water extract.
Because lotus leaf complex chemical composition, pure dissolubility active pharmaceutical ingredientss such as the alkaloid that is soluble in ethanol isopolarity solvent, flavones had both been contained, also contain water-soluble biological small molecular weight impurities such as the water-soluble biological macromole impurity such as protein, enzyme, polysaccharide of soluble in water or damping fluid and tannin, organic acid, oligose, even also contain oil-soluble impurities compositions such as the chlorophyll that is soluble in non-polar solvents such as sherwood oil, lipid, natural gum.
Adopt the Lotus Leafextract of water extracting method preparation, though contain main active pharmaceutical ingredients such as lotus leaf alkaloid and lotus flavone, but also there are impurity components such as amounts of protein, polysaccharide, tannin, chlorophyll and organic acid to exist, defectives such as the purity that obviously there is easy moisture absorption in product, preserve difficulty, alkaloid and flavones is not high, drug effect is not obvious, the quality of product and market competitiveness deficiency.
Adopt the macroporous resin enrichment absorption method behind the water extraction, though help the purity of alkaloid and flavones in the Lotus Leafextract to improve, but because the protein, polysaccharide and other biomacromolecule impurity that remain in the Lotus Leafextract, separation and purification efficient and the unit resin saturated extent of adsorption of macroporous adsorbent resin to lotus leaf alkaloid and lotus flavone will be had a strong impact on, even cause the non-renewable of macroporous adsorbent resin, to have a strong impact on the work-ing life of macroporous adsorbent resin, cause that the Lotus Leafextract product yield is lower, preparation cost is high.
Because polymeric amide has the effect of selective adsorption to the Flavonoid substances that is rich in hydroxyl, and the adsorbed material hydroxyl what its adsorptive power depends on, so become the Flavonoid substances good adsorbent.Utilize polyamide separation and purification flavones and alkaloid, can obtain the common macroporous resin product purity that is beyond one's reach.
The present invention utilizes the specific adsorption characteristics of polymeric amide to flavones, carry out the further separation and purification of lotus flavone and lotus leaf alkaloid, prepare highly purified lotus flavone and lotus leaf alkaloid product respectively, has tangible novelty, a kind of equipment is simple, separation and purification is effective, product yield is high, production cost is low lotus flavone and lotus leaf alkaloid preparation method can be provided, not see bibliographical information both at home and abroad.
Summary of the invention
One of purpose of the present invention is to provide a kind of low cost, high yield, high-quality, the preparation method that is suitable for industrial lotus flavone and lotus leaf alkaloid.
Two of purpose of the present invention is prepared lotus leaf crude extract, the Lotus Leafextract behind the membrane separation purification, the Lotus Leafextract behind the resin separation purification, lotus flavone and the lotus leaf alkaloid after the polyamide separation and purification, further make any medicinal preparation for oral administration applicatory clinically with fat-reducing, reducing blood-fat, antibacterial, antiviral or antioxygenation, as: capsule, granule, tablet, oral liquid, syrup, microcapsule, powder, pill or pill etc.
The lotus flavone of employing polyamide separation and purification of the present invention and the preparation method of lotus leaf alkaloid are achieved in that the polar organic solvent that comprises the lotus leaf crude extract extracts or the water extraction step, the membrane separation for removing impurities step of lotus leaf crude extract, the macroporous adsorbent resin purification procedures of lotus leaf crude extract, it is characterized in that: with the lotus leaf crude extract, Lotus Leafextract behind the membrane separation purification or the Lotus Leafextract behind the resin separation purification are raw material, add suitable quantity of water, it is stirred into suspension, adopt polymeric amide enrichment absorption lotus flavone then, not by the polymeric amide absorbed portion, through decompression and solvent recovery, vacuum concentration becomes the lotus leaf alkaloid product.The lotus flavone that is adsorbed in the above-mentioned steps on the polymeric amide is reclaimed, wash depolarization impurity and pigment earlier with water, use polar organic solvent aqueous solution wash-out lotus flavone again, elutriant becomes the lotus flavone product through decompression and solvent recovery, vacuum concentration.
The preparation method of the lotus leaf crude extract that described polar organic solvent extracts is: be raw material with the lotus leaf, adopt 30～65% the polar organic solvent aqueous solution of 10～20 times of lotus leaf dry weights (W/W), under 50～80 ℃ of conditions, extract 1～3 time, each 1～3 hour, united extraction liquid, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.
The preparation method of the lotus leaf crude extract of described water extraction is: be raw material with the lotus leaf, adopt the water of 10～30 times of lotus leaf dry weights (W/W), under 50～80 ℃ of conditions, extract 1～3 time, each 1～3 hour, united extraction liquid, filter, filtrate-0.06～-0.095MPa, 50～80 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.
Described membrane separation for removing impurities method is: the lotus leaf crude extract with polar organic solvent extraction or water extraction is a raw material, add water, the lotus leaf crude extract is stirred into suspension, make suspension proportion reach 1.05～1.30, under 0.2～1.0MPa,, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification through membrane separation for removing impurities.
Described macroporous adsorbent resin separation purification method is: with the Lotus Leafextract behind lotus leaf crude extract or the membrane separation purification is raw material, add suitable quantity of water, it is stirred into suspension, make suspension proportion reach 1.1～1.2, it is saturated to adopt the macroporous adsorbent resin enrichment to be adsorbed to then, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 30～60% the polar organic solvent aqueous solution of using 4～8 times of amount of resin again with 1～3 times of amount of resin/hour flow velocity (V/V) wash-out target substance, filtrate-0.05～-0.09MPa, under 50～65 ℃ of conditions, through decompression and solvent recovery, Lotus Leafextract after the separation and purification of vacuum concentration resin.
The preparation method of described polyamide separation and purification lotus leaf alkaloid is: with Lotus Leafextract behind lotus leaf crude extract, the membrane separation purification or the Lotus Leafextract behind the resin separation purification is raw material, add suitable quantity of water, it is stirred into suspension, make suspension proportion reach 1.1～1.2, adopt polymeric amide enrichment absorption lotus flavone then, not by the polymeric amide absorbed portion, filtrate-0.06～-0.095MPa, 50～80 ℃ of conditions under, become the lotus leaf alkaloid product through decompression and solvent recovery, vacuum concentration.
The preparation method of described polyamide separation and purification lotus flavone is: with the lotus leaf crude extract, Lotus Leafextract behind the membrane separation purification or the Lotus Leafextract behind the resin separation purification are raw material, add suitable quantity of water, it is stirred into suspension, make suspension proportion reach 1.1～1.2, adopt polymeric amide enrichment absorption lotus flavone then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, use again 6～10 times of amount of resin 40～70% final concentrations the polar organic solvent aqueous solution with 1～3 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, filtrate-0.05～-0.09MPa, under 50～65 ℃ of conditions, through decompression and solvent recovery, vacuum concentration becomes the lotus flavone product.
Above-mentioned polar organic solvent can be one or more mixed solvents in methyl alcohol, ethanol, propyl alcohol, Virahol, propyl carbinol or the acetone, preferentially selects methyl alcohol or acetone for use.Mainly the extraction effect based on methyl alcohol, ethanol, propyl alcohol, Virahol, propyl carbinol or acetone isopolarity organic solvent is suitable for this, but for other polar organic solvent, the boiling point of methyl alcohol or acetone is lower, energy consumption is littler, can significantly reduce the preparation cost of product.
Above-mentioned membrane separation for removing impurities process can be selected the separatory membrane of 10000 molecular weight cut-offs～0.22 μ m membrane pore size for use.
The separation membrane material of above-mentioned membrane separation for removing impurities process can be one or more hybrid films materials in inorganic ceramic film, polysulfone membrane or the polyvinylidene fluoride film, preferentially selects polysulfone membrane or polyvinylidene fluoride film for use.
Above-mentioned macroporous adsorbent resin can be one or more hybrid resins among HZ801, AB-8, HZ806, D101, DK110, D113, HZ802, HZ803, HZ816, the HZ818, preferentially selects HZ801, HZ802, HZ803, HZ806, HZ816, HZ818 for use.
One of major advantage of preparation technology of the present invention is: combination adopts polar organic solvent to extract, membrane separation for removing impurities, the method of macroporous adsorbent resin enrichment and polyamide separation and purification, can not only the suitability for industrialized production different purity and other Lotus Leafextract of level, and the further suitability for industrialized production higher degree of energy and other lotus flavone of level and lotus leaf alkaloid product, be used for follow-up fat-reducing, the preparation of lipid-reducing function product, safety more veritably, easy, rationally, realize the total composition comprehensive utilization of lotus leaf and the suitability for industrialized production of series product economically, demonstrate fully the theory of Green Chemistry production and the strategy of recycling economy, it is theoretical novel to have, reasonable in technology, operational safety, technology is easy, economically feasible, advantages of environment protection.
Two of preparation technology's of the present invention major advantage is: adopt modernization of Chinese medicine production technology to extract various purity and other Lotus Leafextract of level that the main active pharmaceutical ingredients of lotus leaf is made the current international practice, and higher degree and other lotus flavone of level and lotus leaf alkaloid product, " three height ", " three is little " and advantages such as " three just " with modern Chinese herbal medicine product, energy adapts to and satisfies modern Chinese medicine clinical application demand, meets the modernization of Chinese medicine and international industry policy that country advocates.
Embodiment
The present invention adopts the preparation method of the lotus flavone and the lotus leaf alkaloid of polyamide separation and purification, the polar organic solvent that comprises the lotus leaf crude extract extracts or the water extraction step, the membrane separation for removing impurities step of lotus leaf crude extract, the macroporous adsorbent resin purification procedures of lotus leaf crude extract, it is characterized in that: with the lotus leaf crude extract, Lotus Leafextract behind the membrane separation purification or the Lotus Leafextract behind the resin separation purification are raw material, add suitable quantity of water, it is stirred into suspension, adopt polymeric amide enrichment absorption lotus flavone then, not by the polymeric amide absorbed portion, through decompression and solvent recovery, vacuum concentration becomes the lotus leaf alkaloid product.The lotus flavone that is adsorbed in the above-mentioned steps on the polymeric amide is reclaimed, wash depolarization impurity and pigment earlier with water, use polar organic solvent aqueous solution wash-out lotus flavone again, elutriant becomes the lotus flavone product through decompression and solvent recovery, vacuum concentration.
The preparation method of the lotus leaf crude extract that polar organic solvent extracts is: be raw material with the lotus leaf, adopt 30～65% the polar organic solvent aqueous solution of 10～20 times of lotus leaf dry weights (W/W), under 50～80 ℃ of conditions, extract 1～3 time, each 1～3 hour, united extraction liquid, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.
Above-mentioned polar organic solvent can be one or more mixed solvents in methyl alcohol, ethanol, propyl alcohol, Virahol, propyl carbinol or the acetone, preferentially selects methyl alcohol or acetone for use.Mainly the extraction effect based on methyl alcohol, ethanol, propyl alcohol, Virahol, propyl carbinol or acetone isopolarity organic solvent is suitable for this, but for other polar organic solvent, the boiling point of methyl alcohol or acetone is lower, energy consumption is littler, can significantly reduce the preparation cost of product.
The preparation method of the lotus leaf crude extract of water extraction is: be raw material with the lotus leaf, adopt the water of 10～30 times of lotus leaf dry weights (W/W), under 50～80 ℃ of conditions, extract 1～3 time, each 1～3 hour, united extraction liquid, filter, filtrate-0.06～-0.095MPa, 50～80 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.
The membrane separation for removing impurities method is: the lotus leaf crude extract with polar organic solvent extraction or water extraction is a raw material, add water, the lotus leaf crude extract is stirred into suspension, make suspension proportion reach 1.05～1.30, under 0.2～1.0MPa,, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification through membrane separation for removing impurities.
Above-mentioned membrane separation for removing impurities process can be selected the separatory membrane of 10000 molecular weight cut-offs～0.22 μ m membrane pore size for use.
The separation membrane material of above-mentioned membrane separation for removing impurities process can be one or more hybrid films materials in inorganic ceramic film, polysulfone membrane or the polyvinylidene fluoride film, preferentially selects polysulfone membrane or polyvinylidene fluoride film for use.
The macroporous adsorbent resin separation purification method is: with the Lotus Leafextract behind lotus leaf crude extract or the membrane separation purification is raw material, add suitable quantity of water, it is stirred into suspension, make suspension proportion reach 1.1～1.2, it is saturated to adopt the macroporous adsorbent resin enrichment to be adsorbed to then, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 30～60% the polar organic solvent aqueous solution of using 4～8 times of amount of resin again with 1～3 times of amount of resin/hour flow velocity (V/V) wash-out target substance, filtrate-0.05～-0.09MPa, under 50～65 ℃ of conditions, through decompression and solvent recovery, Lotus Leafextract after the separation and purification of vacuum concentration resin.
Above-mentioned macroporous adsorbent resin can be one or more hybrid resins among HZ801, AB-8, HZ806, D101, DK110, D113, HZ802, HZ803, HZ816, the HZ818, preferentially selects HZ801, HZ802, HZ803, HZ806, HZ816, HZ818 for use.
The preparation method of polyamide separation and purification lotus leaf alkaloid is: with Lotus Leafextract behind lotus leaf crude extract, the membrane separation purification or the Lotus Leafextract behind the resin separation purification is raw material, add suitable quantity of water, it is stirred into suspension, make suspension proportion reach 1.1～1.2, adopt polymeric amide enrichment absorption lotus flavone then, not by the polymeric amide absorbed portion, filtrate-0.06～-0.095MPa, 50～80 ℃ of conditions under, become the lotus leaf alkaloid product through decompression and solvent recovery, vacuum concentration.
The preparation method of polyamide separation and purification lotus flavone is: with the lotus leaf crude extract, Lotus Leafextract behind the membrane separation purification or the Lotus Leafextract behind the resin separation purification are raw material, add suitable quantity of water, it is stirred into suspension, make suspension proportion reach 1.1～1.2, adopt polymeric amide enrichment absorption lotus flavone then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, use again 6～10 times of amount of resin 40～70% final concentrations the polar organic solvent aqueous solution with 1～3 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, filtrate-0.05～-0.09MPa, under 50～65 ℃ of conditions, through decompression and solvent recovery, vacuum concentration becomes the lotus flavone product.
The purposes of the Lotus Leafextract behind lotus leaf crude extract of the present invention, the membrane separation purification, the Lotus Leafextract behind the resin separation purification, the lotus flavone after the polyamide separation and purification and lotus leaf alkaloid is: the medicinal preparation for oral administration that can have fat-reducing, lipopenicillinase, antibacterial, detoxifcation, drug rehabilitation, antiviral or antioxygenation according to the preparation of the formulation method of routine.Described medicinal preparation for oral administration is: capsule, granule, tablet, oral liquid, syrup, microcapsule, powder, pill or pill etc.
The physical and chemical parameter measuring method of each prepared product of the present invention is as follows:
(1) assay of lotus flavone: adopting high performance liquid chromatography, is contrast with the Quercetin.Chromatographic condition: chromatographic column is selected Waters Nova-Pak C18 post (Φ 3.9 * 15mm, 4 μ m) for use, and pre-column C18 (3.9mm * 15mm), moving phase is methyl alcohol: 0.2% phosphate aqueous solution (45: 55), flow velocity 1.0mL/min, 34 ℃ of column temperatures, detect wavelength 365nm, sample size 20 μ L.The mensuration of sample: get liquid 4mL to be measured (or determinand powder 0.1g, be dissolved in the 4mL solvent), add methyl alcohol 6mL and 12mol/L hydrochloric acid 1mL, shake up rearmounted 80 ℃ of water-bath back hydrolysis 1h.Then hydrolyzed solution is changed in the 50mL volumetric flask, be diluted to scale, as HPLC liquid to be measured with 50% methyl alcohol.Adopting external standard method quantitative in the HPLC mensuration process, is contrast with the Quercetin, and the content of each aglycon after the calculating hydrolysis is unified with the factor 2.51 calculating lotus flavone content.
(2) qualitative judgement of lotus flavone: adopting aluminium salt colorimetry, is contrast with the rutin.Sample solution is diluted to certain multiple, gets 1mL in test tube, add 0.1mL 5%NaNO
2Solution shakes up, and adds 0.1mL 10%Al (NO behind the 5min
3)
3Solution adds 1mL 1mol/L NaOH solution again behind the 6min, mixing, behind the 10min if producing redness or red-purple shows and has lotus flavone, with reagent as blank.
(3) assay of lotus leaf alkaloid; Adopting high performance liquid chromatography, is contrast with the Nuciferine.Chromatographic condition is: chromatographic column is selected Waters Nova-Pak C for use
18Post (Φ 3.9 * 15mm, 4 μ m), pre-column C
18(3.9mm * 15mm), moving phase is methyl alcohol: H
2O: diethylamine (75: 25: 0.01, pH value 9.0～10.5), flow velocity 1.0mL/min, 34 ℃ of column temperatures detect wavelength 270nm, sample size 20 μ L.
(4) lotus leaf alkaloid qualitative judgement: adopting the IKI colorimetry, is contrast with the Nuciferine.Sample solution is diluted to certain multiple, gets 1mL in test tube, add 1～2 Wagner's reagent solution, shake up, place 5min, if produce brown or the brown precipitation shows and has lotus leaf alkaloid, with reagent as blank.
(5) measurement of the polysaccharide content: adopting the phenolsulfuric acid method, is contrast with glucose or D-semi-lactosi.
(6) mensuration of protein content: adopting FoLin-phenol method to measure, is contrast with the bSA.
(7) water content adopts quick moisture content tester to measure.
Medicinal preparation for oral administration preparation method of the present invention is as follows:
(1) the spraying drying condition is: feeding liquid concentration 10～20 degree Beaume (60 ℃), 160～250 ℃ of PG-5 type spray-drier inlet temperatures, 60～110 ℃ of temperature outs, centrifugal head operating pressure 1.6～3.0kgf/cm
2
(2) lyophilize condition is: drying temperature-10～-60 ℃, 35～70 ℃ of sublimation temperatures, pressure 0.05～0.18mbar, time of drying 20～40h.
(3) vacuum-drying condition: 45～75 ℃ of drying temperatures, pressure-0.06～-0.095MPa, time of drying 15～50h.
(4) capsule preparation: according to the formulation method of routine, select No. 1 hard capsule, the about 0.30g of the tolerant loadings of every intragranular, on the full-automatic hard capsule filler of NJP-1000B type, carry out the medicinal powder filling, on the flat Autoblisterpackagingmachine of JPB-25IIB type, carry out the aluminium foil Blister Package, can be made into clinical suitable Lotus Leafextract oral capsule.
(5) granule preparation: according to the formulation method of routine, granulation on SHK-220 type efficient wet mixer-granulator is packed on JXD-30A type particle automatic packing machine, can be made into clinical suitable Lotus Leafextract oral granular formulation.
(6) pill preparation: according to the formulation method of routine, adjust the temperature controlling system of the general dripping pill machine of TZDW-1 type, make the water dropper temperature remain on 50～85 ℃, the temperature of refrigerants such as whiteruss remains on-4～40 ℃; Get the glycogelatin of Lotus Leafextract and 1～9 times of weight thereof, place well heater, suspension is made in heating under agitation condition, be placed in the water dropper jar of dripping pill machine, splash in the refrigerant by water dropper, the dripping pill that will be shunk moulding by the outlet of dripping pill machine takes out, and removes the refrigerant on surface, is dried to the agent of Lotus Leafextract oral drip-pill.
Preparation method's of the present invention embodiment is presented below:
Embodiment 1
Producing lotus leaf with Jianning, Fujian Province is raw material, with new lotus leaf washing, dry (air-dry, all can dry or dry), pulverize, cross 16～20 mesh sieves and get Lotus Leaf, the 3.0kg Lotus Leaf is put into extractor, with 40% methanol aqueous solution of 10 times of dry weights of Lotus Leaf (W/W), under 63 ℃ of conditions, extract 3 times each 2 hours, united extraction liquid, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 96.7%, and the extraction yield of lotus leaf alkaloid is 95.9%.
Then, earlier the lotus leaf crude extract is added suitable quantity of water and stir into suspension, make suspension proportion reach 1.05, adopting molecular weight cut-off then is 10000 polysulfone membrane, under 1.0MPa, carry out membrane separation for removing impurities, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 95.2%, and the lotus leaf alkaloid rate of recovery is 95.6%, and the protein removal rate is 96.1%, and the polysaccharide clearance is 96.9%.
To the Lotus Leafextract behind the membrane separation purification, adopt the HZ806 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, use 60% aqueous ethanolic solution of 8 times of amount of resin again, with 3 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 96.2% of lotus flavone, the rate of recovery 95.8% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.19, adopt 200 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, 60% aqueous ethanolic solution of using 10 times of amount of resin again with 3 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after spray-dried.After measured, lotus flavone product water content 4.6%, lotus flavone purity 64.0%, lotus flavone total recovery 68.8%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting spray drying treatment in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 3.8%, lotus leaf alkaloid purity 85.5%, lotus leaf alkaloid total recovery 78.5%.
Embodiment 2
16～20 order Lotus Leaf 2.5kg are put into extractor, 30% methanol aqueous solution with 20 times of dry weights of Lotus Leaf (W/W), under 50 ℃ of conditions, extract 2 times, each 3 hours, united extraction liquid, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 93.4%, and the extraction yield of lotus leaf alkaloid is 92.3%.
The lotus leaf crude extract is added suitable quantity of water stir into suspension, make suspension proportion reach 1.18, adopting molecular weight cut-off then is 60000 polyvinylidene fluoride film, under 0.6MPa, carry out membrane separation for removing impurities, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 95.8%, and the lotus leaf alkaloid rate of recovery is 95.9%, and the protein removal rate is 94.6%, and the polysaccharide clearance is 96.2%.
To the Lotus Leafextract behind the membrane separation purification, adopt the HZ801 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 45% aqueous ethanolic solution of using 6 times of amount of resin again with 1 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 97.2% of lotus flavone, the rate of recovery 98.3% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.13, adopt 120 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, 50% aqueous ethanolic solution of using 6 times of amount of resin again with 1 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after lyophilize.After measured, lotus flavone product water content 5.9%, lotus flavone purity 63.7%, lotus flavone total recovery 64.1%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting lyophilize to handle in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 5.8%, lotus leaf alkaloid purity 84.1%, lotus leaf alkaloid total recovery 78.7%.
Embodiment 3
16～20 order Lotus Leaf 2.5kg are put into extractor, 65% methanol aqueous solution with 17 times of dry weights of Lotus Leaf (W/W), refluxing extraction is 1 hour under 80 ℃ of conditions, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 92.7%, and the extraction yield of lotus leaf alkaloid is 91.2%.
The lotus leaf crude extract is added suitable quantity of water stir into suspension, make suspension proportion reach 1.30, adopting membrane pore size then is the inorganic ceramic film of 0.22 μ m, carries out membrane separation for removing impurities under 0.2MPa, removes the Lotus Leafextract that promptly gets behind the macromole impurity such as protein and polysaccharide behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 96.2%, and the rate of recovery of lotus leaf alkaloid is 96.4%, and the protein removal rate is 93.1%, and the polysaccharide clearance is 95.4%.
To the Lotus Leafextract behind the membrane separation purification, adopt the HZ816 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 30% aqueous ethanolic solution of using 8 times of amount of resin again with 2 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 95.4% of lotus flavone, the rate of recovery 95.9% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.12, adopt 60 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, use again 9 times of amount of resin 40% final concentration aqueous ethanolic solution with 2 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after vacuum-drying.After measured, lotus flavone product water content 5.1%, lotus flavone purity 63.1%, lotus flavone total recovery 64.4%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting the vacuum-drying mode to handle in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 4.2%, lotus leaf alkaloid purity 85.0%, lotus leaf alkaloid total recovery 79.3%.
Embodiment 4
Producing lotus leaf with Jianning, Fujian Province is raw material, with new lotus leaf washing, dry (air-dry, all can dry or dry), pulverize, cross 16～20 mesh sieves and get Lotus Leaf, the 3.0kg Lotus Leaf is put into extractor, 65% aqueous ethanolic solution with 20 times of dry weights of Lotus Leaf (W/W), under 80 ℃ of conditions, extracted 1 hour, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 92.1%, and the extraction yield of lotus leaf alkaloid is 90.4%.
Earlier the lotus leaf crude extract is added suitable quantity of water and stir into suspension, make suspension proportion reach 1.15, adopting molecular weight cut-off then is 80000 polyvinylidene fluoride films, under 0.4MPa, carry out membrane separation for removing impurities, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 96.0%, and the lotus leaf alkaloid rate of recovery is 96.1%, and the protein removal rate is 94.1%, and the polysaccharide clearance is 95.8%.
To the Lotus Leafextract behind the membrane separation purification, adopt the D101 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 40% methanol aqueous solution of using 4 times of amount of resin again with 2 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 96.3% of lotus flavone, the rate of recovery 96.4% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.14, adopt 120 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, 45% methanol aqueous solution of using 8 times of amount of resin again with 2 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after lyophilize.After measured, lotus flavone product water content 6.1%, lotus flavone purity 63.5%, lotus flavone total recovery 67.7%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting the lyophilize mode to handle in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 5.8%, lotus leaf alkaloid purity 84.2%, lotus leaf alkaloid total recovery 82.0%.
Embodiment 5
16～20 order Lotus Leaf 2.5kg are put into extractor, tap water with 30 times of dry weights of Lotus Leaf (W/W), under 50 ℃ of conditions, extract 2 times, each 2 hours, united extraction liquid, filter, filtrate-0.06～-0.095MPa, 50～80 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 92.1%, and the extraction yield of lotus leaf alkaloid is 90.5%.
The lotus leaf crude extract is added suitable quantity of water stir into suspension, make suspension proportion reach 1.30, adopting membrane pore size then is the inorganic ceramic film of 0.22 μ m, under 0.2MPa, carry out membrane separation for removing impurities, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 96.3%, and the rate of recovery of lotus leaf alkaloid is 96.2%, and the protein removal rate is 93.4%, and the polysaccharide clearance is 95.3%.
To the Lotus Leafextract behind the membrane separation purification, adopt the DK110 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 30% methanol aqueous solution of using 6 times of amount of resin again with 3 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 93.9% of lotus flavone, the rate of recovery 94.7% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.13, adopt 30 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, 40% methanol aqueous solution of using 10 times of amount of resin again with 3 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after vacuum-drying.After measured, lotus flavone product water content 4.9%, lotus flavone purity 63.4%, lotus flavone total recovery 64.5%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting the vacuum-drying mode to handle in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 4.8%, lotus leaf alkaloid purity 84.9%, lotus leaf alkaloid total recovery 78.3%.
Embodiment 6
16～20 order Lotus Leaf 2.5kg are put into extractor, 38% aqueous ethanolic solution with 10 times of dry weights of Lotus Leaf (W/W), under 50 ℃ of conditions, extract 3 times, each 3 hours, united extraction liquid, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 94.7%, and the extraction yield of lotus leaf alkaloid is 95.1%.
The lotus leaf crude extract is added suitable quantity of water stir into suspension, make suspension proportion reach 1.05, adopting molecular weight cut-off then is 20000 polysulfone membrane, carries out membrane separation for removing impurities under 0.8MPa, removes the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 95.4%, and the lotus leaf alkaloid rate of recovery is 95.9%, and the protein removal rate is 94.8%, and the polysaccharide clearance is 96.4%.
To the Lotus Leafextract behind the membrane separation purification, adopt the AB-8 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 60% methanol aqueous solution of using 6 times of amount of resin again with 1 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 95.7% of lotus flavone, the rate of recovery 96.4% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.14, adopt 200 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, 60% methanol aqueous solution of using 6 times of amount of resin again with 1 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after spray-dried.After measured, lotus flavone product water content 3.6%, lotus flavone purity 63.4%, lotus flavone total recovery 70.3%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting the spraying drying mode to handle in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 3.4%, lotus leaf alkaloid purity 85.4%, lotus leaf alkaloid total recovery 78.7%.
Embodiment 7
Producing lotus leaf with Jianning, Fujian Province is raw material, with new lotus leaf washing, dry (air-dry, all can dry or dry), pulverize, cross 16～20 mesh sieves and get Lotus Leaf, the 3.0kg Lotus Leaf is put into extractor, with 30% aqueous acetone solution of 14 times of dry weights of Lotus Leaf (W/W), under 50 ℃ of conditions, extract 3 times each 3 hours, united extraction liquid, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 94.6%, and the extraction yield of lotus leaf alkaloid is 93.8%.
The lotus leaf crude extract is added suitable quantity of water stir into suspension, make suspension proportion reach 1.30, adopting membrane pore size then is the polyvinylidene fluoride film of 0.22 μ m, under 0.3MPa, carry out membrane separation for removing impurities, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 96.2%, and the lotus leaf alkaloid rate of recovery is 96.3%, and the protein removal rate is 93.6%, and the polysaccharide clearance is 95.1%.
To the Lotus Leafextract behind the membrane separation purification, adopt the D113 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 30% aqueous acetone solution of using 4 times of amount of resin again with 1 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 95.8% of lotus flavone, the rate of recovery 96.1% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.13, adopt 30 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, 40% aqueous acetone solution of using 6 times of amount of resin again with 1 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after vacuum-drying.After measured, lotus flavone product water content 5.2%, lotus flavone purity 63.1%, lotus flavone total recovery 64.0%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting the vacuum-drying mode to handle in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 4.5%, lotus leaf alkaloid purity 84.7%, lotus leaf alkaloid total recovery 77.5%.
Embodiment 8
16～20 order Lotus Leaf 2.5kg are put into extractor, 65% aqueous acetone solution with 10 times of dry weights of Lotus Leaf (W/W), refluxing extraction is 2 times under 80 ℃ of conditions, each 1 hour, united extraction liquid, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 93.2%, and the extraction yield of lotus leaf alkaloid is 92.1%.
The lotus leaf crude extract is added suitable quantity of water stir into suspension, make suspension proportion reach 1.21, adopting molecular weight cut-off then is 50000 polysulfone membrane, carries out membrane separation for removing impurities under 0.5MPa, removes the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 95.2%, and the lotus leaf alkaloid rate of recovery is 96.2%, and the protein removal rate is 95.0%, and the polysaccharide clearance is 96.1%.
To the Lotus Leafextract behind the membrane separation purification, adopt the HZ802 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 60% aqueous acetone solution of using 4 times of amount of resin again with 2 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 93.6% of lotus flavone, the rate of recovery 94.3% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.13, adopt 200 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, 60% aqueous acetone solution of using 8 times of amount of resin again with 2 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after spray-dried.After measured, lotus flavone product water content 3.9%, lotus flavone purity 63.6%, lotus flavone total recovery 68.4%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting spray drying treatment in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 3.5%, lotus leaf alkaloid purity 85.8%, lotus leaf alkaloid total recovery 77.0%.
Embodiment 9
16～20 order Lotus Leaf 2.5kg are put into extractor, with the tap water of 20 times of dry weights of Lotus Leaf (W/W), under 80 ℃ of conditions, extracted 2 hours, filter, filtrate-0.06～-0.095MPa, 50～80 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration.After measured, the extraction yield of lotus flavone is 90.8%, and the extraction yield of lotus leaf alkaloid is 90.1%.
The lotus leaf crude extract is added suitable quantity of water stir into suspension, make suspension proportion reach 1.16, adopting molecular weight cut-off then is 100000 inorganic ceramic film, under 0.3MPa, carry out membrane separation for removing impurities, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification.After measured, the lotus flavone rate of recovery is 96.0%, and the lotus leaf alkaloid rate of recovery is 96.2%, and the protein removal rate is 94.1%, and the polysaccharide clearance is 95.7%.
To the Lotus Leafextract behind the membrane separation purification, adopt the HZ803 macroporous adsorbent resin, enrichment is adsorbed to saturated, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 45% aqueous acetone solution of using 8 times of amount of resin again with 3 times of amount of resin/hour flow velocity (V/V) wash-out target substance, reclaim the Lotus Leafextract that promptly gets behind the solvent behind the resin separation purification.After measured, the rate of recovery 93.8% of lotus flavone, the rate of recovery 94.2% of lotus leaf alkaloid.
Add suitable quantity of water, Lotus Leafextract behind the resin separation purification is stirred into suspension, make suspension proportion reach 1.15, adopt 120 order polymeric amide enrichments absorption lotus flavone to saturated then, earlier with water (V/V) the flush away polar impurity and the pigment that are no less than 4 times of amount of resin, 50% aqueous acetone solution of using 10 times of amount of resin again with 3 times of amount of resin/hour flow velocity (V/V) wash-out lotus flavone, vacuum concentration reclaims solvent, promptly gets the lotus flavone product after lyophilize.After measured, lotus flavone product water content 5.9%, lotus flavone purity 63.8%, lotus flavone total recovery 64.2%.
Not by the polymeric amide absorbed portion, after vacuum concentration reclaims solvent, promptly get the lotus leaf alkaloid product after further adopting spraying drying, lyophilize, air stream drying, microwave drying or vacuum-drying mode to handle in the above-mentioned steps.After measured, lotus leaf alkaloid product water content 4.7%, lotus leaf alkaloid purity 84.6%, lotus leaf alkaloid total recovery 76.0%.
Explanation about Lotus Leafextract purposes of the present invention
The lotus leaf crude extract of embodiment 1 preparation, the Lotus Leafextract behind the membrane separation purification, Lotus Leafextract, lotus flavone and the lotus leaf alkaloid behind the resin separation purification are carried out spraying drying respectively, the oral applying jianfeijiangzhi capsule agent of clinical use is made in direct then filling, carries out the evaluation of animal pharmacodynamics.
1 material
(body weight 14～16g) is provided by Medical University Of Fujian's animal center female mice.Basal feed is available from Medical University Of Fujian's animal center.High lipid food (basal feed 92.5%, salt 2%, yolk powder 2%, lard 3%, cholesterol 0.5%), this institute makes by oneself.Total cholesterol (TCH) test kit (CHOD-PAP method), Eastern Europe, Zhejiang biotechnology company limited.Triglyceride level (TG) test kit (GPO-PAP method), Eastern Europe, Zhejiang biotechnology company limited.Mda (MDA) test kit (TBA method), bio-engineering research institute is built up in Nanjing.Cholesterol, Shanghai are tried a chemical reagent company limited.
2 experimental techniques
70 mouse are raised with basal feed be divided into seven groups of A, B, C, D, E, F, G, 10 every group, mouse ad lib and drinking-water after 5 days at random.Wherein administration group A, B, C, D and E are respectively Lotus Leafextract, the Lotus Leafextract behind the resin separation purification, the lotus flavone after the polyamide separation and purification and the lotus leaf alkaloid behind lotus leaf crude extract, the membrane separation purification, the F group is as high fat positive controls, and the G group is as normal negative control group.A～F group employing high lipid food modeling, the G group is raised with basal feed, and each is organized mouse and all begins administration after raising 7 days, and each medicine all is made into 1g/mL, and dosage is only sky of 0.6ml/, and wherein F group and G group make physiological saline into.Administration is 42 days respectively, plucks eyeball behind the fasting 12h and gets blood and dissection, carries out the mensuration of every biochemical indicator.TCH, TG and MDA all measure with the reagent corresponding box and in strict accordance with specification sheets.Data are carried out with SPSS software, adopt Independent-Sample T Test method that each group data is carried out average, standard error calculating and test of significance.
3 lipopenicillinase animal pharmacodynamic result
Different extracts see Table 1 to the influence of mice plasma total cholesterol, triglyceride level and MDA content.
After the administration 42 days, the cholesterol of high fat positive controls mice plasma has significantly improved 96.7% than normal negative control group, and mouse hypercholesterolemia model modeling success under this experiment condition is described.Find simultaneously, the mice plasma cholesterol level of administration group A, B, C, D and E all significantly is lower than high fat positive controls, best with A group effect, cholesterol level has significantly reduced 34.3%, B, C, D and E group cholesterol level also significantly reduce more than 25%, illustrate that the lotus flavone of different purity and lotus leaf alkaloid sample all have tangible reducing blood-fat, decreasing cholesterol effect.
(X ± SD) influences n=10 to mice plasma total cholesterol, triglyceride level and mda content for the lotus flavone of table 1 different purity and lotus leaf alkaloid
A, B, C, D and E administration group are respectively Lotus Leafextract, the Lotus Leafextract behind the resin separation purification, lotus flavone and the lotus leaf alkaloid sample behind lotus leaf crude extract, the membrane separation purification; F: high fat positive controls; G: normal negative control group.Compare with high fat positive controls
*: P＜0.05;
*: P＜0.01;
* *: P＜0.001.
Above embodiment is intended to further describe for example the present invention, rather than limits the present invention by any way.
The present invention is novel, technology is simple, the extraction efficiency height, production cost is low, and drug effect is obvious, and made lotus flavone and lotus leaf alkaloid, obviously have " three height " of modern Chinese herbal medicine product, the advantage of " three is little " and " three just ", can adapt to and satisfy modern Chinese medicine clinical application demand, meet the modernization of Chinese medicine and international industry policy that country advocates, have bigger promotional value, can produce remarkable economic efficiency and social benefit.
Claims (6)
1. preparation method who adopts polyamide separation and purification lotus flavone and lotus leaf alkaloid, the polar organic solvent that comprises the lotus leaf crude extract extracts or the water extraction step, the membrane separation for removing impurities step of lotus leaf crude extract, the macroporous adsorbent resin purification procedures of lotus leaf crude extract, it is characterized in that: with the Lotus Leafextract behind the resin separation purification is raw material, add suitable quantity of water, it is stirred into suspension, adopt polymeric amide enrichment absorption lotus flavone then, not by the polymeric amide absorbed portion, become the lotus leaf alkaloid product through decompression and solvent recovery, vacuum concentration;
The polar organic solvent extraction step of described lotus leaf crude extract is: be raw material with the lotus leaf, adopt 30～65% the polar organic solvent aqueous solution of 10～20 times of lotus leaf dry weights (W/W), under 50～80 ℃ of conditions, extract 1～3 time, each 1～3 hour, united extraction liquid, filter, filtrate-0.05～-0.09MPa, 50～65 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration;
The water extraction step of described lotus leaf crude extract is: be raw material with the lotus leaf, adopt the water of 10～30 times of lotus leaf dry weights (W/W), under 50～80 ℃ of conditions, extract 1～3 time, each 1～3 hour, united extraction liquid, filter, filtrate-0.06～-0.095MPa, 50～80 ℃ of conditions under, become the lotus leaf crude extract through decompression and solvent recovery, vacuum concentration;
Described membrane separation for removing impurities method is: the lotus leaf crude extract with polar organic solvent extraction or water extraction is a raw material, add water, the lotus leaf crude extract is stirred into suspension, make suspension proportion reach 1.05～1.30, under 0.2～1.0MPa,, remove the Lotus Leafextract that promptly gets behind protein and polysaccharide and other macromole impurity behind the membrane separation purification through membrane separation for removing impurities;
Described macroporous adsorbent resin separation purification method is: with the Lotus Leafextract behind the membrane separation purification is raw material, add suitable quantity of water, it is stirred into suspension, make suspension proportion reach 1.1～1.2, it is saturated to adopt the macroporous adsorbent resin enrichment to be adsorbed to then, earlier with water (V/V) the flush away polar impurity that is no less than 2 times of amount of resin, 30～60% the polar organic solvent aqueous solution of using 4～8 times of amount of resin again with 1～3 times of (V/V) amount of resin/hour flow velocity wash-out target substance, filtrate-0.05～-0.09MPa, under 50～65 ℃ of conditions, through decompression and solvent recovery, Lotus Leafextract after the separation and purification of vacuum concentration resin;
The preparation method of described polyamide separation and purification lotus leaf alkaloid is: with the Lotus Leafextract behind the resin separation purification is raw material, add suitable quantity of water, it is stirred into suspension, make suspension proportion reach 1.1～1.2, adopt polymeric amide enrichment absorption lotus flavone then, not by the polymeric amide absorbed portion, filtrate-0.06～-0.095MPa, 50～80 ℃ of conditions under, become the lotus leaf alkaloid product through decompression and solvent recovery, vacuum concentration;
The lotus flavone that is adsorbed in the above-mentioned steps on the polymeric amide is reclaimed, wash depolarization impurity and pigment earlier with water, use polar organic solvent aqueous solution wash-out lotus flavone again, elutriant becomes the lotus flavone product through decompression and solvent recovery, vacuum concentration.
2. the preparation method of employing polyamide separation and purification lotus flavone according to claim 1 and lotus leaf alkaloid is characterized in that: it is one or more mixed solvents in methyl alcohol, ethanol, propyl alcohol, Virahol, propyl carbinol or the acetone that described lotus leaf is slightly carried the polar organic solvent that adopts in the process.
3. the preparation method of employing polyamide separation and purification lotus flavone according to claim 1 and lotus leaf alkaloid is characterized in that: the membrane separation for removing impurities process is selected the separatory membrane of 10000 molecular weight cut-offs～0.22 μ m membrane pore size for use.
4. the preparation method of employing polyamide separation and purification lotus flavone according to claim 1 and lotus leaf alkaloid is characterized in that: the separation membrane material of described membrane separation for removing impurities process is one or more hybrid films materials in inorganic ceramic film, polysulfone membrane or the polyvinylidene fluoride film.
5. the preparation method of employing polyamide separation and purification lotus flavone according to claim 1 and lotus leaf alkaloid is characterized in that: macroporous adsorbent resin is one or more hybrid resins among HZ801, AB-8, HZ806, D101, DK110, D113, HZ802, HZ803, HZ816, the HZ818.
6. the preparation method of employing polyamide separation and purification lotus flavone as claimed in claim 1 and lotus leaf alkaloid is characterized in that: Lotus Leafextract, lotus flavone and the lotus leaf alkaloid of described preparation method's preparation have be used to prepare have fat-reducing, the purposes of the medicinal preparation for oral administration of lipopenicillinase, antibacterial, detoxifcation, drug rehabilitation, antiviral or antioxygenation.
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| CN101402627B (en) * | 2008-10-15 | 2011-08-31 | 南昌大学 | Synchronous separation, extraction and purification method for flavone, alkaloid and polysaccharide in lotus leaf |
| CN101423508B (en) * | 2008-11-26 | 2012-08-01 | 南昌大学 | Method for preparing flavonoid from lotus leaf and drug application thereof |
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| CN103285089A (en) * | 2013-05-13 | 2013-09-11 | 张娟 | Preparation process flow of lotus leaf extract |
| CN103479730B (en) * | 2013-09-27 | 2014-11-26 | 周爱琴 | Method for synchronously separating lotus leaf flavone and chlorophyll from fresh lotus leaves |
| CN107805257A (en) * | 2017-09-12 | 2018-03-16 | 烟台大学 | A kind of capsule of weeping forsythia alkaloid extract and its preparation method and application |
| CN113444043A (en) * | 2020-03-27 | 2021-09-28 | 福建中医药大学 | Lotus leaf alkaloid and extraction and purification method thereof |
| CN116063265A (en) * | 2023-03-16 | 2023-05-05 | 常州大学 | Method for separating peony seed meal flavone by using organic silicon/PA composite membrane |
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| CN1482125A (en) * | 2002-09-13 | 2004-03-17 | 中国科学院武汉植物研究所 | Process for extracting lotus leaf flavone |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1374280A (en) * | 2002-03-26 | 2002-10-16 | 贵州省生化工程中心 | Technological process of eliminating alkaloid from plant extractive |
| CN1482125A (en) * | 2002-09-13 | 2004-03-17 | 中国科学院武汉植物研究所 | Process for extracting lotus leaf flavone |
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