CN104069150A - Preparation method for honeysuckle extract - Google Patents
Preparation method for honeysuckle extract Download PDFInfo
- Publication number
- CN104069150A CN104069150A CN201410342185.4A CN201410342185A CN104069150A CN 104069150 A CN104069150 A CN 104069150A CN 201410342185 A CN201410342185 A CN 201410342185A CN 104069150 A CN104069150 A CN 104069150A
- Authority
- CN
- China
- Prior art keywords
- flos lonicerae
- ethanol
- extract
- medical material
- macroporous resin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention relates to a preparation method for a honeysuckle extract. The method comprises the steps as follows: firstly, ethanol extracting; secondly, vacuum concentrating; thirdly, pH value regulating; fourthly, macroporous resin purifying; fifthly, concentrating, drying and the like. The technology of the method is simple and easy to operate; as the macroporous resin can be recycled, the method is low in manufacturing cost and small in environmental pollution; the ethanol solution, adopted as an extracting and eluting solvent, is high in safety, cheap and easy to obtain, and low in production cost; as the obtained honeysuckle extract and the honeysuckle have basically identical ingredients, namely compounds such as an organic acid, iridoid glycoside, and flavone, an active substance group is wholly transferred; under the situation that invalid substances are removed effectively, the recycling rate of the organic acid, the iridoid glycoside, and the flavone can reach higher than 80%, and the resemblance of the extract finger-print before and after the macroporous resin purification can reach over 0.91.
Description
Technical field
The preparation method that the present invention relates to a kind of Flos Lonicerae extract, belongs to the field of Chinese medicines.
Background technology
" multicomponent, many target spots, group effect " is the feature of Chinese medicine, its drug effect depends on the cooperative effect of a series of compound groups with relatively stable formation, but owing to there being certain physicochemical property difference between each composition, cause each composition response rate in separation and purification process to differ greatly, destroy its intrinsic composition, certainly will affect the performance of its effect.As macroporous adsorption resin technology, be considered at present the most promising Purification of traditional Chinese herbs technology, applied research in the field of Chinese medicines is increasing, but it is preferred to be mainly with one or several composition that index is carried out purification condition, due to the difference of constitutive property, effective ingredient retention rate difference of different nature, after resin purification, there is huge change in original component composition not only, destroy the globality of material group, and caused the loss of effective ingredient and the waste of resource.Fingerprint pattern technology is to evaluate at present the multi-component effective tool of Chinese medicine, if can carry out the refining effect of evaluating resin with finger printing, just can keep the globality of material group, keeps the concordance of drug effect.
Flos Lonicerae is the dry flower of caprifoliaceae plant Radix Ophiopogonis (Lonicera japonica Thunb.) or the flower that band is just opened, and has heat-clearing and toxic substances removing, effect of wind-heat dissipating.Its main active comprises the multiclass compounds such as the flavone such as iridoid glycoside and luteoloside such as the organic acid such as chlorogenic acid, disconnected loganin hemiacetal lactone, has antibacterial, antiviral, antiinflammatory, the pharmacological action such as antipyretic, is one of clinical the most frequently used medical material.At present, separation, purification for Flos Lonicerae active ingredient mainly do investigation index with single component, for example Chinese patent CN101837039A provides a kind of Flos Lonicerae extract and preparation method that does not contain chlorogenic acid, specifically comprise the steps such as alcohol reflux extracts, filtration is concentrated, macroporous resin column separation, concentrate drying, prepared the Flos Lonicerae extract of luteoloside content more than 40%, this extract is not containing chlorogenic acid.Chinese patent CN101530449 discloses a kind of method of preparing honeysuckle extract by applying membrane filtration technology, comprise first Flos Lonicerae is decocted with water to extraction, concentrating under reduced pressure is centrifugal, remove most of impurity by micro-filtration membrane, further refining with ultrafilter membrane again, the rate of transform of extract obtained chlorogenic acid reaches 90%, but the method is carried out separation and purification with micro-filtration membrane and ultrafilter membrane, the active ingredient beyond chlorogenic acid is not detected, in removing impurity, also removed other active ingredient.Liu Enli etc. are studied total organic acids technique in macroporous adsorbent resin separation and purification Flos Lonicerae, taking static saturated adsorption capacity and static eluting rate as investigating index, compared 6 kinds of macroporous resins separation, purification Flos Lonicerae total organic acids technique, result shows with HPD100 separation and purification total organic acids best results; But the method is only using organic acid as evaluation index, do not relate to extraction, the separation (Liu Enli of the total constituents of Flos Lonicerae, Li Qingshan. the research [J] of total organic acids in macroporous adsorbent resin separation and purification Flos Lonicerae. Chinese herbal medicine, 2006, (12): 1792-1796).The separation such as Ma Shuancheng have obtained 10 kinds of iridoid glycoside compounds in Flos Lonicerae, and adopt RP-HPLC method to measure the content of 4 kinds of iridoid glycosides in Chinese medicine honeysuckle, but the method is used and has used respectively petroleum ether, ether, ethyl acetate, n-butyl alcohol separates as eluting solvent with chloroform, purification, easily cause extract toxic dissolvent residual, operating procedure is various, be not suitable for large production, and the method is only with using iridoid glycoside constituents as evaluation index, other effective ingredient (Ma Shuancheng in cannot integrated survey Flos Lonicerae, Liu Yan, Bi Peixi, Deng. the quantitative study [J] of the iridoid glycoside constituents that in Chinese medicine honeysuckle, preventing respiratory viruses infects. pharmaceutical analysis magazine 26 (8): 1039-1043).
Below be all about the extraction of Chlorogenic Acid of Flos Lonicerae, luteoloside, total organic acids or total iridoid methods of glycosides, the research of separating technology, but very few about evaluate on the whole the literature research of the each extracts active ingredients of Flos Lonicerae, purification effect taking finger printing as instrument.Although chlorogenic acid, luteoloside, total organic acids or total iridoid glycoside etc. are one of active ingredient of Flos Lonicerae, only using single component as index, can not the existing Flos Lonicerae extract drug action of holohedron.
In view of the deficiencies in the prior art, be necessary to invent a kind of new Flos Lonicerae active substance group purifying process, make it to simplify the operation course, to the full extent few with or without hazardous solvent, be applicable to industrialized great production, improve the content of multiple active ingredient in extract, the whole machine balancing that can reach again medical material active substance group shifts, and reaches the object being consistent with former medicine.
Summary of the invention
Technical problem to be solved by this invention is the deficiency that overcomes existing extraction, purifying process, and a kind of method of simply, effectively preparing Flos Lonicerae extract is provided, and the method can keep the globality of Flos Lonicerae material group, keeps the concordance of drug effect.
The object of the invention is to realize in the following manner.
The preparation method of Flos Lonicerae extract of the present invention comprises: ethanol extraction, concentrating under reduced pressure, adjusting pH value, purification by macroporous resin, concentrated and dry etc.
Further, the preparation method of a kind of Flos Lonicerae extract involved in the present invention comprises the following steps:
1) extracting honeysuckle medical material, alcohol reflux, filters, merging filtrate, concentrating under reduced pressure reclaims ethanol, obtains concentrated solution;
2) step 1) concentrated solution is diluted with water to and is equivalent to 2-5 times of medical material weight, filter, it is 2-6 that filtrate regulates pH, be added in the macroporous resin column that pretreatment is good and adsorb, first use the pure water eluting of 2-4BV with the flow velocity of 1-3BV/h, discard, then use respectively the ethanol elution of 2-4BV30%-50% ethanol and 2-4BV60%-80% with the flow velocity of 1-3BV/h, collect ethanol elution;
3) combining step 2) ethanol elution, concentrated, dry, obtain Flos Lonicerae extract.
Preferably, step 1) described reflux, extract, is 60%-80% with the percentage composition of ethanol, and extracting alcohol adding amount is 8-12 times of medical material weight, and extraction time is 3 times, each 1 hour.
Preferably, step 2) described concentrated solution is diluted to respect to 3 times of medical material weight, and regulating pH value is 3.0, and applied sample amount is equivalent to medical material: macroporous resin is 0.8-1.0:1.
Preferably, step 2) model of described macroporous resin is the one in LX-17, LX-207, HPD-100, HPD-450, XDA-8 and AB-8, optimum is HPD-100.
Preferably, step 2) described eluting is respectively 50% and 80% by the concentration of alcoholic solution.
Compared with the prior art, Flos Lonicerae extract preparation method of the present invention has following significant progress:
1, simple to operate, condition is controlled, the repeatability that tool is good, and big pore resin can reuse, and preparation cost is low and free from environmental pollution, is applicable to suitability for industrialized production;
2, this method is in the situation that removing invalid components, and the compounds such as organic acid, iridoid glycoside, flavone have obtained the higher response rate, are all greater than 80%, and extraction used and eluting solvent are water or ethanol, cheap and easy to get, extract obtained residual without hazardous solvent, drug safety is high;
3, with finger printing contrast preferred resin, loading and elution requirement, gained Flos Lonicerae extract and Flos Lonicerae composition are basically identical, the total peak of HPLC finger printing is no less than 15, and before and after purification by macroporous resin, the similarity of extract finger printing is that similarity is greater than more than 0.91;
4, compare with existing effective ingredient of honeysuckle preparation method, in the Flos Lonicerae extract of preparing by the inventive method, organic acid chlorogenic acid, 3,5-Dicaffeoylquinic acid, 4,5-Dicaffeoylquinic acid sum are greater than 30%, and disconnected loganin hemiacetal lactone content is greater than 1.0%;
5, extract obtained organic acid, the iridoid glycoside of not only containing, also contain the compounds such as flavonoid, realize the global transfer of active substance group, can at utmost ensure the former powerful of Chinese medicine honeysuckle, and reduce dose, be beneficial to preparation and improved the compliance of patient's medication.
Brief description of the drawings
Fig. 1, reference substance and sample HPLC figure, wherein:
S1. mix reference substance; S2. Flos Lonicerae extractive solution; S3. sample 1; S4. sample 2; S5. sample 3;
1.5-O-CQA；2.3-O-CQA；3.4-O-CQA；4.vogeloside；5.luteoloside；6.3,4-DCQA；7.3,5-DCQA；8.4,5-DCQA。
Fig. 2, ethanol elution HPLC figure, wherein: A.20% ethanol; B.40% ethanol; C.60% ethanol; D.80% ethanol
Detailed description of the invention
The invention will be further described to contrast detailed description of the invention prepared by preferred elution requirement, Flos Lonicerae extract fingerprint and Flos Lonicerae extract by Flos Lonicerae extract anti-influenza A H 1 N 1 virus test, finger printing below, but this is not limitation of the present invention, those skilled in the art are according to basic thought of the present invention, can make various amendments or improvement, but only otherwise depart from basic thought of the present invention, all within the scope of the present invention.
Embodiment 1 Flos Lonicerae extract anti-influenza A H 1 N 1 virus Experiment on Function
1 test objective
Adopt H 1 N 1 influenza A virus infection murine pneumonia model to inquire into Flos Lonicerae extract pharmacodynamic action.
2 test materials
2.1 tested medicines
Flos Lonicerae extract, lot number: 20111927 (with the Flos Lonicerae extracts of the embodiment of the present invention 13 gained), brown-black powder, gas fragrance hardship, quality are clear-cut, water-soluble and ethanol.
2.2 positive control drug YINHUANG KOUFUYE: produced by Lunan Pharmaceutical Co., Ltd.; The accurate word of traditional Chinese medicines: Z10870001; Product batch number: 09122001; Date of manufacture: 091203; Valid until 201211; Character: the supernatant liquid of rufous, sweet, the micro-hardship of taste; Function cures mainly: cleaning heat and expelling wind, liyan jiedu.For dry pharynx, pharyngalgia, pharyngeal tonsils enlargement, thirsty, heating due to affection due to external wind and heat, exuberant lung-stomach heat; Acute and chronic tonsillitis, acute/chronic pharyngitis, upper respiratory tract infection are shown in above-mentioned patient.
2.3 animals: CD-1 (ICR) mice, SPF/VAF rank, body weight 14 ± 1g, totally 30, female.Purchased from Beijing HFK Bio-Technology Co., Ltd..Animal licence numbering: SCXK (capital) 2009-0004.
2.4 Strain: influenza A H1N1 influenza virus Mus lung adapted strain (FM/1/47 strain), purchased from CDC virosis prevention and control institute, gone down to posterity by this institute ABSL-2 test chamber ,-80 DEG C save backup.
2.5 test apparatuses: ether, Beijing reagent company product, lot number: 20090707; A2 type Biohazard Safety Equipment, model: MSC1.8, German Thermo product; The weight of animals electronic balance, model: YP/0001, Shanghai Yue Ping scientific instrument company limited product; Animal viscera electronic balance, model: AR1140, U.S. Chaus product; IVC mice rearging cage, Suzhou teaching Long Ju factory product.
3 methods and result
3.1 dosage designs: according to the anxious poison test of each medicine mice gained LD50.Using the 1/7-1/10 of LD50 as middle dosage, using 2 times and 1/2 of middle dosage respectively as heavy dose of and low dose of.
3.1.1 Flos Lonicerae extract median lethal dose(LD 50) is 4.053g/kg Mouse Weight, and the 1/10 middle dosage as tested medicine of this experimental selection LD50 adopts 1.0g/kg, 0.5g/kg, tri-dosage of 0.25g/kg when test.
3.1.2 YINHUANG KOUFUYE: according to body surface area dose,equivalent transformation approach, people's consumption is scaled to mice dosage, the dosage of mice is 11 times with dosage of people.The dose,equivalent of mice is 11ml/kg/d.When test, each medicine holds dense 0.2ml/10g intraperitoneal injection and the gastric infusion such as not by waiting.
3.2 test methods: animal is divided into following group at random according to body weight: dosage group (0.5g/kg), Flos Lonicerae small dose group (0.25g/kg) in Normal group, model control group, positive drug group (YINHUANG KOUFUYE group), the heavy dose of group of Flos Lonicerae (1.0g/kg), Flos Lonicerae.Each administration component is gastric infusion and drug administration by injection, every group of 5 mices.Except Normal group, mice is slightly anaesthetized with ether, infect every 35ul with 15 LD50 influenza virus drop noses.Infect and started at every turn the same day by 0.2ml/10g body weight gastric infusion, every day 1 time, continuous 4 days, Normal group and model control group be distilled water gavage under equal conditions.After within the 5th day, weighing, dissect and claim to calculate lung index and lung index by lung weight.Between result employing group, relatively statistical procedures is carried out in t inspection.The results are shown in Table 1
Lung index=lung weight in wet base (g)/body weight (g)
3.3 medicines preparations principle Flos Lonicerae extract gastric infusions: adopt maximum dosage-feeding 24.0g/kg in early stage 1/10 as middle dosage, when test, adopt 4.8g/kg, 2.4g/kg, tri-dosage of 1.2g/kg.
4 results
Table 1 infected by influenza infects the effect (lung index) of normal mouse pulmonary inflammation model
Note: with relatively ##P<0.01 of normal group; With model control group comparison, * * P<0.01, * P<0.05
5 brief summaries
5.1 Flos Lonicerae extracts show the effect of significant inhibition influenza virus mice pneumonia.
5.2 Flos Lonicerae extracts are in the time of the dosage of 1/20 maximum dosage-feeding, and gastric infusion has obvious effect, and suppression ratio is 51.92% to the maximum.
The foundation of embodiment 2 Flos Lonicerae extract finger printing
1 instrument and reagent
Agilent1100 high performance liquid chromatograph (Anjelen Sci. & Tech. Inc), AG285 analytical balance (Mei Teletuo benefit company); PHS-25 type acidometer (Shanghai great achievement instrument plant); Glass chromatography column (lcm × 50cm).
Flos Lonicerae, purchased from nine Jian Peng Flos Lonicerae Specialty Co-operative Organizations; Luteoloside (luteoloside, purity >=98%, lot number 111720-201307, National Institute for Food and Drugs Control); Chlorogenic acid (3-O-CQA, purity >=98%, lot number 110753-200413, National Institute for Food and Drugs Control); Neochlorogenic acid, 4-dicaffeoylquinic acid (lot number is respectively MUST-10091501 for 5-O-CQA, 4-O-CQA, purity >=98%, MUST-10091811, Man Site bio tech ltd, Chengdu); The two caffeoyl quinic acids, 3 of 4,5-O-, the two caffeoyl quinic acids, 3 of 5-O-, 4-O-two caffeoyl quinic acid (4,5-DCQA, 3,5-DCQA, 3,4-DCQA, purity >=98%, lot number is respectively 12022804,12040507, D100624, Chengdu Purification Technology Development Co., Ltd.), disconnected loganin hemiacetal lactone (vogeloside, purity >=98%, self-control); HPD100, HPD450, AB-8 macroporous adsorbent resin (Cangzhou, Hebei Bao En Chemical Co., Ltd.); LX207, LX17, XDA-8 macroporous adsorbent resin (Lan Xiao Science and Technology Co., Ltd.); Methanol is chromatographically pure (Merk company), and water is redistilled water, the equal analytical pure of all the other reagent.
2 methods and result
2.1 Flos Lonicerae extractive solutions prepare extracting honeysuckle 500g, with 60% alcohol reflux of 8 times of amounts 3 times, each 1 hour, backflow decompression recycling ethanol, extractum adds water to 1500ml, absorbent cotton filters, filtrate is for subsequent use.
The foundation of 2.2 finger printing
2.2.1 chromatographic condition Kromasil C18 chromatographic column (250mm × 4.6mm, 5 μ m); 35 DEG C of column temperatures; Detecting wavelength 238nm flow velocity is 1.0mlmin
-1; Sample size 5 μ L; Number of theoretical plate calculates and should be not less than 6000 by chlorogenic acid peak.Mobile phase acetonitrile (A)-0.05% trifluoroacetic acid solution (B) gradient elution: 0～25min, 10%～15%A; 25～60min, 15%～35%A.
2.2.2 mix reference substance preparation and take respectively a certain amount of luteoloside (luteoloside), vogeloside (disconnected loganin hemiacetal lactone), 3-O-CQA (chlorogenic acid), 5-O-CQA (neochlorogenic acid), 4-O-CQA (silver-colored chlorogenic acid), 4; 5-DCQA (4; the two caffeoyl quinic acids of 5-O-), 3; 5-DCQA (3; the two caffeoyl quinic acids of 5-O-), 3; 4-DCQA (3; the two caffeoyl quinic acids of 4-O-) reference substance, add 50% Methanol and must mix reference substance solution.
2.2.3 reference fingerprint is set up precision and is measured Flos Lonicerae extractive solution 2ml, is settled to 10ml, and filtering with microporous membrane, as need testing solution, is set up reference fingerprint under above-mentioned chromatographic condition.Separately get mixing reference substance, Criterion product chromatograph collection of illustrative plates under above-mentioned chromatographic condition, wherein peak 1-8 5-O-CQA, 3-O-CQA, 4-O-CQA, vogeloside, luteoloside, 3 respectively, 4-DCQA, 3,5-DCQA, 4,5-DCQA, the results are shown in Figure 1.
2.2.4 precision is investigated and is got same need testing solution, and METHOD FOR CONTINUOUS DETERMINATION 6 times is measured RSD≤2.11% of 15 main peaks retention times of collection of illustrative plates 6 times, RSD≤2.83% of peak area, and instrument precision is good.
2.2.5 study on the stability is got same need testing solution, measures in 0,2,4,6,10,12h, measures RSD≤2.78% of 15 main peaks retention times of collection of illustrative plates, RSD≤2.92% of peak area, and test sample is good at 12h internal stability.
2.2.6 repeatability is investigated and is got 6 parts of same batch samples, by the preparation method operation of need testing solution, measures, and measures RSD≤2.08% of 15 main peaks retention times of collection of illustrative plates, RSD≤2.28% of peak area, and the repeatability of method of proof is good.
2.3 assays are the accurate 3-O-CQA, 3 that draws respectively, 5-DCQA, 4, and 5-DCQA reference substance is appropriate, puts in measuring bottle, and 50% methanol is diluted to scale, makes the gmL containing 3-O-CQA79.2 μ
-1, 3,5-DCQA56.1 μ gmL
-1with 3,4-DCQA53.0 μ gmL
-1solution.Measure absworption peak area at 327nm place, Criterion working curve, obtain 3-O-CQA, 3,5-DCQA, 4,5-DCQ regression equation is respectively Y=2513.1X-1.910, Y=2661.1X-6.652, Y=2649.6X-8.317, r is respectively 0.9999,0.9992,0.9991, good in 0.0792～0.792,0.0561～0.561,0.0530～0.530 μ g scope internal linear relation respectively.Through precision, repeatability, study on the stability, RSD is all less than 2.86%, and precision, repeatability, stability are better; Three kinds of composition response rate of average recovery test are all less than 2.79% at 98.6～102.4%, RSD.
Embodiment 3 purification by macroporous resin technical studies
2.4 purification by macroporous resin researchs
2.4.1 resin pretreatment by resin with packing glass chromatography column into after soaked in absolute ethyl alcohol 24h, rinse and add deionized water to effluent and be not muddy with the flow velocity of 2BV/h with dehydrated alcohol, again with deionized water rinsing to without alcohol taste, then after rinsing with 2BV/h flow velocity with 2BV5% dilute hydrochloric acid solution, be neutral with deionized water rinsing resin to effluent pH value again, after finally rinsing with 2BV/h flow velocity with 2BV2% sodium hydroxide solution, be neutral with deionized water rinsing resin to effluent pH value again, sucking filtration, removes the liquid that anhydrates for subsequent use.
2.4.2 the resin 5.0g that it is good that Resin sieving selection accurately takes pretreatment packs in tool plug conical flask, accurately add Flos Lonicerae extractive solution 40mL, putting vibration on constant temperature oscillator adsorbs after 24h, resin is packed in glass chromatography column, with deionized water 40mL flushing resin, then rinse resin with 50% ethanol 30mL, rinse resin with 70% ethanol 60mL again, receive ethanol elution, be settled to 100mL, measure finger printing, calculate 3-O-CQA, Vogeloside, Luteoloside, 3,5-DCQA, 4, the 5-DCQA response rate, the results are shown in Table 1.Result shows, HPD-100 is the highest to the each main component response rate of Flos Lonicerae, determines that HPD-100 is as adsorbing material.
The impact of table 1 resin model on the main component response rate
2.4.3 competitive Adsorption impact accurately takes pretreatment good resin 5.0g packs in tool plug conical flask, accurately add respectively and regulate pH value to the Flos Lonicerae extractive solution 40mL of different acidity and do not regulate pH value Flos Lonicerae extractive solution 20mL, putting vibration on constant temperature oscillator adsorbs after 24h, resin is packed in glass chromatography column, with deionized water 40mL flushing resin, then rinse resin with 50% ethanol 30mL, rinse resin with 80% ethanol 60mL again, receive ethanol elution, be settled to 100mL, measure finger printing, calculate 3-O-CQA, Vogeloside, 3, 5-DCQA, 4, the 5-DCQA response rate, the results are shown in Table 2.Show that pH value reduces, 3-O-CQA adsorption competitiveness strengthens, and the response rate improves, and the vogeloside response rate reduces; PH value is lower than 3.5 o'clock, 3,5-DCQA, 4, and the 5-DCQA response rate all declines, and considers that to select pH value be 3.0; Loading liquid measure reduces by half, and each composition response rate is all significantly increased, and wherein 3-O-CQA improves the most remarkable because competition reduces.
The impact of table 2 competitive adsorption
2.4.4 applied sample amount investigation accurately takes pretreatment, good resin 10.0g packs in chromatographic column, accurately add respectively the Flos Lonicerae extractive solution 44mL that regulates pH value to 3.5, 40mL, 36mL and 32mL, be equivalent to medical material: resin is 1.1:1, 1.0:1, 0.9:1 and 0.8:1, pass through after resin bed with flow velocity 0.5mL/min, with 40mL deionized water rinsing resin bed, then rinse resin with 50% ethanol 30mL, rinse resin with 80% ethanol 60mL again, receive ethanol elution, be settled to 100mL, measure, calculate absorption front and back collection of illustrative plates similarity and 3-O-CQA, Vogeloside, 3, 5-DCQA, 4, the 5-DCQA response rate, the results are shown in Table 3.When result shows that applied sample amount is 0.9:1, each component recovery is more balanced, and similarity is the highest.
The impact of table 3 applied sample amount
2.4.5 eluting solvent screening accurately takes pretreatment, good resin 10.0g packs in chromatographic column, add the Flos Lonicerae extractive solution 36mL that regulates pH value to 3.0, pass through after resin bed with flow velocity 0.5mL/min, with 40mL deionized water rinsing resin bed, then with 20%, 40%, 60%, 80% ethanol 40mL eluting, collect respectively, be settled to 100mL, measure finger printing, calculate 3-O-CQA, Vogeloside, 3,5-DCQA, 4, the 5-DCQA response rate, the results are shown in Figure 2, show that 80% ethanol is more complete to each composition eluting.
2.4.6 the resin 10.0g that it is good that process certification accurately takes pretreatment packs in chromatographic column, add the Flos Lonicerae extractive solution 36mL that regulates pH value to 3.0, pass through after resin bed with flow velocity 0.5mL/min, with 60mL deionized water rinsing resin bed, then with 50% ethanol 40mL and 70% ethanol 40mL eluting, collect ethanol elution, be settled to 100mL, measure, finger printing is shown in Fig. 1." similarity evaluation 2004A version " computed in software and the Flos Lonicerae extractive solution fingerprint similarity that adopts Chinese Pharmacopoeia committee to publish.Measure respectively 3-O-CQA, 3,5-DCQA, 4,5-DCQA content and paste-forming rate, the results are shown in Table 4.
3 batches of verification sample results of table 4
Flos Lonicerae 60% ethanol extraction paste-forming rate can reach 35%, and after Optimization Technology purification, paste-forming rate is only 12%, and each main component response rate, 80～99%, realizes the synchronous transfer of composition substantially, and fingerprint similarity 0.98 has kept the concordance of component.
Chinese medicine material group is the material base of Chinese medicine performance effect, has the feature [10] of globality, in Purification of traditional Chinese herbs process, realizes the synchronous transfer of active component, is to keep the consistent prerequisite of the property of medicine.Evaluate the result of purification with effective ingredient or effective site, lay particular emphasis on pursuit purity, isolate contacting of purification technique and Chinese medical theory, fingerprint pattern technology is Chinese medicine multicomponent quality control effective means, can more comprehensively evaluate the overall picture of component, evaluate purge process with finger printing, the concordance of component before and after can keeping, the unification that keeps drug effect.
The preparation of embodiment 4 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 6 times of amount ethanol extractions of 80% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 2 times of medical material weight, regulating pH is 2.0, be added in the LX-17 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 0.8:1 (volume/volume), first use the pure water eluting of 2BV with the flow velocity of 1BV/h, discard, again with the flow velocity of 1BV/h respectively with the ethanol elution with 2BV30% ethanol and 2BV60%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.846kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
" the Flos Lonicerae extract finger printing " set up with embodiment 2 similarity of " similarity evaluation 2004A version " computed in software Flos Lonicerae extract finger printing that adopts Chinese Pharmacopoeia committee to publish, obtaining the similarity of finger printing before and after purification by macroporous resin is 0.917.
The preparation of embodiment 5 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 12 times of amount ethanol extractions of 80% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 5 times of medical material weight, regulating pH is 6.0, be added in the LX-207 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 1.0:1 (volume/volume), first use the pure water eluting of 4BV with the flow velocity of 3BV/h, discard, again with the flow velocity of 3BV/h respectively with the ethanol elution with 4BV50% ethanol and 4BV80%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.768kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.935.
The preparation of embodiment 6 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 11 times of amount ethanol extractions of 70% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 4 times of medical material weight, regulating pH is 5.0, be added in the HPD-100 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 0.9:1 (volume/volume), first use the pure water eluting of 3BV with the flow velocity of 2BV/h, discard, again with the flow velocity of 2BV/h respectively with the ethanol elution with 3BV40% ethanol and 3BV70%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.582kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.951.
The preparation of embodiment 7 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 10 times of amount ethanol extractions of 80% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 3 times of medical material weight, regulating pH is 4.0, be added in the XDA-8 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 0.8:1 (volume/volume), first use the pure water eluting of 3BV with the flow velocity of 3BV/h, discard, again with the flow velocity of 2BV/h respectively with the ethanol elution with 3BV50% ethanol and 4BV60%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.660kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.911.
The preparation of embodiment 8 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 9 times of amount ethanol extractions of 70% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 4 times of medical material weight, regulating pH is 3.5, be added in the HPD-100 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 0.9:1 (volume/volume), first use the pure water eluting of 2BV with the flow velocity of 2BV/h, discard, again with the flow velocity of 2BV/h respectively with the ethanol elution with 2BV40% ethanol and 2BV80%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.722kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.946.
The preparation of embodiment 9 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 9 times of amount ethanol extractions of 60% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 2 times of medical material weight, regulating pH is 3.0, be added in the HPD-450 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 0.9:1 (volume/volume), first use the pure water eluting of 4BV with the flow velocity of 1BV/h, discard, again with the flow velocity of 1BV/h respectively with the ethanol elution with 4BV30% ethanol and 2BV80%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.492kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.920.
The preparation of embodiment 10 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 8 times of amount ethanol extractions of 60% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 3 times of medical material weight, regulating pH is 3.0, be added in the HPD-100 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 1.0:1 (volume/volume), first use the pure water eluting of 2BV with the flow velocity of 2BV/h, discard, again with the flow velocity of 2BV/h respectively with the ethanol elution with 2BV50% ethanol and 2BV80%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.558kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.936.
The preparation of embodiment 11 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 10 times of amount ethanol extractions of 70% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 5 times of medical material weight, regulating pH is 6.0, be added in the LX-17 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 1.0:1 (volume/volume), first use the pure water eluting of 2BV with the flow velocity of 3BV/h, discard, again with the flow velocity of 2BV/h respectively with the ethanol elution with 3BV40% ethanol and 3BV80%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.612kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.924.
The preparation of embodiment 12 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 8 times of amount ethanol extractions of 80% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 4 times of medical material weight, regulating pH is 2.0, be added in the LX-207 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 1.0:1 (volume/volume), first use the pure water eluting of 4BV with the flow velocity of 1BV/h, discard, again with the flow velocity of 3BV/h respectively with the ethanol elution with 2BV30% ethanol and 3BV60%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.822kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.947.
The preparation (patent most preferred embodiment) of embodiment 13 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 10 times of amount ethanol extractions of 60% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 3 times of medical material weight, regulating pH is 3.0, be added in the HPD-100 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 1.1:1 (volume/volume), first use the pure water eluting of 2BV with the flow velocity of 2BV/h, discard, again with the flow velocity of 2BV/h respectively with the ethanol elution with 2BV50% ethanol and 2BV80%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.695kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.971.
The preparation of embodiment 14 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 11 times of amount ethanol extractions of 80% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 4 times of medical material weight, regulating pH is 2.0, be added in the XDA-8 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 0.8:1 (volume/volume), first use the pure water eluting of 3BV with the flow velocity of 2BV/h, discard, again with the flow velocity of 3BV/h respectively with the ethanol elution with 2BV40% ethanol and 2BV70%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.870kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.955.
The preparation of embodiment 15 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 12 times of amount ethanol extractions of 70% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 3 times of medical material weight, regulating pH is 5.0, be added in the AB-8 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 0.9:1 (volume/volume), first use the pure water eluting of 4BV with the flow velocity of 1BV/h, discard, again with the flow velocity of 3BV/h respectively with the ethanol elution with 2BV50% ethanol and 2BV70%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.522kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.934.
The preparation of embodiment 16 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 11 times of amount ethanol extractions of 60% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 2 times of medical material weight, regulating pH is 4.0, be added in the AB-8 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 0.9:1 (volume/volume), first use the pure water eluting of 2BV with the flow velocity of 2BV/h, discard, again with the flow velocity of 3BV/h respectively with the ethanol elution with 2BV50% ethanol and 4BV60%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.873kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.974.
The preparation of embodiment 17 Flos Lonicerae extracts
1) preparation of Flos Lonicerae extract
Extracting honeysuckle medical material 6kg, with 9 times of amount ethanol extractions of 70% 3 times, each 1 hour, filter, merging filtrate, being evaporated to density is 1.15-1.25g/ml, let cool, concentrated solution is diluted with water to and is equivalent to 3 times of medical material weight, regulating pH is 2.0, be added in the HPD-100 type macroporous resin column that pretreatment is good with 2BV/h flow velocity, applied sample amount is equivalent to medical material: resin is 1.0:1 (volume/volume), first use the pure water eluting of 4BV with the flow velocity of 1BV/h, discard, again with the flow velocity of 3BV/h respectively with the ethanol elution with 4BV30% ethanol and 3BV70%, collect, merge ethanol elution, being evaporated to relative density is the extractum of 1.15～1.25 (70 DEG C), vacuum drying, obtain Flos Lonicerae extract 0.534kg.
2) investigation of Flos Lonicerae extract fingerprint similarity
Press fingerprint similarity evaluation methodology described in embodiment 4, calculating the similarity of extract finger printing before and after purification by macroporous resin is 0.943.
Claims (6)
1. a preparation method for Flos Lonicerae extract, is characterized in that, comprises the steps:
1) extracting honeysuckle medical material, alcohol reflux, filters, merging filtrate, concentrating under reduced pressure reclaims ethanol, obtains concentrated solution;
2) step 1) concentrated solution is diluted with water to and is equivalent to 2-5 times of medical material weight, filter, it is 2-6 that filtrate regulates pH, be added in the macroporous resin column that pretreatment is good and adsorb, first use the pure water eluting of 2-4BV with the flow velocity of 1-3BV/h, discard, then use respectively the ethanol elution of 2-4BV30%-50% ethanol and 2-4BV60%-80% with the flow velocity of 1-3BV/h, collect ethanol elution;
3) combining step 2) ethanol elution, concentrated, dry, obtain Flos Lonicerae extract.
2. the method for claim 1, is characterized in that step 1) described reflux, extract, is 60%-80% with ethanol percentage composition, and extracting alcohol adding amount is 8-12 times of medical material weight, and extraction time is 3 times, each 1 hour.
3. the method for claim 1, is characterized in that step 2) described concentrated solution is diluted to respect to 3 times of medical material weight, and regulating pH value is 3.0, and applied sample amount is equivalent to medical material: macroporous resin is 0.8-1.0:1.
4. the method for claim 1, is characterized in that step 2) model of described macroporous resin is the one in LX-17, LX-207, HPD-100, HPD-450, XDA-8 and AB-8.
5. method as claimed in claim 4, is characterized in that, the model of described macroporous resin is HPD-100.
6. the method as described in any one in claim 1-5, is characterized in that step 2) concentration of described eluting alcoholic solution is respectively 50% and 80%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410342185.4A CN104069150B (en) | 2014-07-18 | 2014-07-18 | Preparation method of honeysuckle extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410342185.4A CN104069150B (en) | 2014-07-18 | 2014-07-18 | Preparation method of honeysuckle extract |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104069150A true CN104069150A (en) | 2014-10-01 |
| CN104069150B CN104069150B (en) | 2020-02-14 |
Family
ID=51591016
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410342185.4A Active CN104069150B (en) | 2014-07-18 | 2014-07-18 | Preparation method of honeysuckle extract |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104069150B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104387273A (en) * | 2014-10-20 | 2015-03-04 | 湖北大别山药业有限公司 | Method for extracting chlorogenic acid from honeysuckle |
| CN104856032A (en) * | 2015-06-01 | 2015-08-26 | 巨鹿县三丰银杞饮品有限公司 | Method for preparing honeysuckle beverage concentrate through recycled solvent |
| CN106117284A (en) * | 2016-07-11 | 2016-11-16 | 山东省分析测试中心 | The method of six kinds of iridoid glycoside constituents in Extraction and enrichment Flos Lonicerae while of a kind of |
| CN106975021A (en) * | 2016-12-14 | 2017-07-25 | 江苏海王健康生物科技有限公司 | A kind of preparation method and applications of enriching yin Lonicera and Forsythia soup side antivirus extract |
| CN108645944A (en) * | 2018-05-10 | 2018-10-12 | 重庆医药高等专科学校 | A kind of quality evaluating method in honeysuckle general flavone extraction process |
| CN112691184A (en) * | 2020-12-31 | 2021-04-23 | 西藏自治区农牧科学院水产科学研究所 | Bacteriostatic composition for inhibiting saprolegnia and preparation method thereof |
| CN116440184A (en) * | 2023-04-19 | 2023-07-18 | 山东中医药大学 | Total iridoid glycoside extract of honeysuckle and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101985421A (en) * | 2010-10-26 | 2011-03-16 | 西北农林科技大学 | Method for simultaneously preparing chlorogenic acid and luteoloside from honeysuckle flower |
| CN102391115A (en) * | 2011-09-26 | 2012-03-28 | 广东太阳神集团有限公司 | Method for preparing honeysuckle flower extract by jointly adopting membrane separation and column chromatography |
-
2014
- 2014-07-18 CN CN201410342185.4A patent/CN104069150B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101985421A (en) * | 2010-10-26 | 2011-03-16 | 西北农林科技大学 | Method for simultaneously preparing chlorogenic acid and luteoloside from honeysuckle flower |
| CN102391115A (en) * | 2011-09-26 | 2012-03-28 | 广东太阳神集团有限公司 | Method for preparing honeysuckle flower extract by jointly adopting membrane separation and column chromatography |
Non-Patent Citations (1)
| Title |
|---|
| 汪海鸿等: "中药制剂中间提取物制备工艺路线设计思路探讨――通塞脉微丸中间提取物制备工艺比较研究", 《世界科学技术(中医药现代化)》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104387273A (en) * | 2014-10-20 | 2015-03-04 | 湖北大别山药业有限公司 | Method for extracting chlorogenic acid from honeysuckle |
| CN104856032A (en) * | 2015-06-01 | 2015-08-26 | 巨鹿县三丰银杞饮品有限公司 | Method for preparing honeysuckle beverage concentrate through recycled solvent |
| CN106117284A (en) * | 2016-07-11 | 2016-11-16 | 山东省分析测试中心 | The method of six kinds of iridoid glycoside constituents in Extraction and enrichment Flos Lonicerae while of a kind of |
| CN106117284B (en) * | 2016-07-11 | 2019-01-11 | 山东省分析测试中心 | A kind of method of six kinds of iridoid glycoside constituents in while Extraction and enrichment honeysuckle |
| CN106975021A (en) * | 2016-12-14 | 2017-07-25 | 江苏海王健康生物科技有限公司 | A kind of preparation method and applications of enriching yin Lonicera and Forsythia soup side antivirus extract |
| CN108645944A (en) * | 2018-05-10 | 2018-10-12 | 重庆医药高等专科学校 | A kind of quality evaluating method in honeysuckle general flavone extraction process |
| CN108645944B (en) * | 2018-05-10 | 2020-08-28 | 重庆医药高等专科学校 | Quality evaluation method in honeysuckle total flavone extraction process |
| CN112691184A (en) * | 2020-12-31 | 2021-04-23 | 西藏自治区农牧科学院水产科学研究所 | Bacteriostatic composition for inhibiting saprolegnia and preparation method thereof |
| CN116440184A (en) * | 2023-04-19 | 2023-07-18 | 山东中医药大学 | Total iridoid glycoside extract of honeysuckle and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104069150B (en) | 2020-02-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104069150A (en) | Preparation method for honeysuckle extract | |
| CN105130796A (en) | Diterpenoid compound and preparing method and application thereof | |
| CN104592341A (en) | Method for extracting asiaticoside and madecassoside from centella | |
| Yu et al. | Purification of high-purity glycyrrhizin from licorice using hydrophilic interaction solid phase extraction coupled with preparative reversed-phase liquid chromatography | |
| CN107998212A (en) | A kind of preparation method of glutinous rehmannia cyclic olefine ether terpinyl side like extractive | |
| CN105294628A (en) | Method for preparing flavonoid component by separating wild chrysanthemum flower | |
| CN104327127A (en) | Method for preparing angroside C, aucubin and harpagide through separation and purification by high-speed countercurrent chromatography | |
| CN109879919B (en) | Method for separating and preparing three flavonoid glycosides from spina date seeds | |
| CN103830306A (en) | Preparation method of honey suckle leaf effective extract product | |
| CN104447910A (en) | Method for preparation of loganin from traditional Chinese medicine Cornus officinalis | |
| CN108440617A (en) | A method of extracting glucoside compound from the coptis | |
| CN103083392B (en) | A kind of method at tool isoamylene radical chromocor position in separation and concentration Radix Sophorae Tonkinensis | |
| CN102988480B (en) | Method for preparing bile and patchouli rhinitis tablets and application | |
| CN104069145A (en) | Application of lindley eupatorium herb in preparing anti-hepatitis B virus medicaments | |
| CN104027403B (en) | A kind of preparation method of Radix Polygalae sugar ester effective site | |
| CN103880895B (en) | A kind of method of utilizing high speed adverse current chromatogram separation and purification to prepare harpagoside and Wyrmslayer glycosides A | |
| CN102503996A (en) | Method for extracting active constituent from Swertia mussotii plant | |
| CN103044253B (en) | Extraction separation method of rosmarinic acid in salvia castabea diels f. tomentosa stib | |
| CN102988614B (en) | Preparation method and application of antiphlogosis tablet | |
| CN105646638B (en) | The preparation method of pedunculoside | |
| CN107721857A (en) | A kind of method that high-purity chlorogenic acid is prepared from Gynura procumbens (Lour.) Merr | |
| CN104725449A (en) | Paeoniflorin extracting method | |
| CN103923138B (en) | A kind of preparation method of Nicotifiorin and application thereof | |
| CN103833806A (en) | Preparation method of traditional Chinese medicine chemical component | |
| CN103919885B (en) | A kind of preparation method of soft capsule for nasosinusitis and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |