CN104628736A - Method for extracting galanthamine from lycoris plant - Google Patents
Method for extracting galanthamine from lycoris plant Download PDFInfo
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- CN104628736A CN104628736A CN201510066820.5A CN201510066820A CN104628736A CN 104628736 A CN104628736 A CN 104628736A CN 201510066820 A CN201510066820 A CN 201510066820A CN 104628736 A CN104628736 A CN 104628736A
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- lycoremine
- lycoris
- obtains
- dry thing
- silica gel
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- ASUTZQLVASHGKV-JDFRZJQESA-N galanthamine Chemical compound O1C(=C23)C(OC)=CC=C2CN(C)CC[C@]23[C@@H]1C[C@@H](O)C=C2 ASUTZQLVASHGKV-JDFRZJQESA-N 0.000 title claims abstract description 120
- 229960003980 galantamine Drugs 0.000 title claims abstract description 60
- 241001633628 Lycoris Species 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 38
- HPOIPOPJGBKXIR-UHFFFAOYSA-N 3,6-dimethoxy-10-methyl-galantham-1-ene Natural products O1C(C(=CC=2)OC)=C3C=2CN(C)CCC23C1CC(OC)C=C2 HPOIPOPJGBKXIR-UHFFFAOYSA-N 0.000 title abstract 5
- LPCKPBWOSNVCEL-UHFFFAOYSA-N Chlidanthine Natural products O1C(C(=CC=2)O)=C3C=2CN(C)CCC23C1CC(OC)C=C2 LPCKPBWOSNVCEL-UHFFFAOYSA-N 0.000 title abstract 5
- BGLNUNCBNALFOZ-WMLDXEAASA-N galanthamine Natural products COc1ccc2CCCC[C@@]34C=CCC[C@@H]3Oc1c24 BGLNUNCBNALFOZ-WMLDXEAASA-N 0.000 title abstract 5
- ASUTZQLVASHGKV-UHFFFAOYSA-N galanthamine hydrochloride Natural products O1C(=C23)C(OC)=CC=C2CN(C)CCC23C1CC(O)C=C2 ASUTZQLVASHGKV-UHFFFAOYSA-N 0.000 title abstract 5
- IYVSXSLYJLAZAT-NOLJZWGESA-N lycoramine Natural products CN1CC[C@@]23CC[C@H](O)C[C@@H]2Oc4cccc(C1)c34 IYVSXSLYJLAZAT-NOLJZWGESA-N 0.000 title abstract 5
- 238000000605 extraction Methods 0.000 claims abstract description 27
- 108090000790 Enzymes Proteins 0.000 claims abstract description 17
- 102000004190 Enzymes Human genes 0.000 claims abstract description 17
- 229940088598 enzyme Drugs 0.000 claims abstract description 17
- 239000000843 powder Substances 0.000 claims abstract description 17
- 108010059892 Cellulase Proteins 0.000 claims abstract description 13
- 108010059820 Polygalacturonase Proteins 0.000 claims abstract description 13
- 229940106157 cellulase Drugs 0.000 claims abstract description 13
- 239000002904 solvent Substances 0.000 claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000010898 silica gel chromatography Methods 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 238000000638 solvent extraction Methods 0.000 claims abstract description 6
- 239000012043 crude product Substances 0.000 claims abstract description 5
- 238000000746 purification Methods 0.000 claims abstract description 5
- 238000001953 recrystallisation Methods 0.000 claims abstract description 5
- 238000001179 sorption measurement Methods 0.000 claims abstract description 5
- 238000001291 vacuum drying Methods 0.000 claims abstract description 5
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 30
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 20
- 238000011068 loading method Methods 0.000 claims description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 229960001701 chloroform Drugs 0.000 claims description 12
- 239000003480 eluent Substances 0.000 claims description 11
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 238000004587 chromatography analysis Methods 0.000 claims description 8
- 239000000945 filler Substances 0.000 claims description 8
- 238000004108 freeze drying Methods 0.000 claims description 8
- 239000000741 silica gel Substances 0.000 claims description 8
- 229910002027 silica gel Inorganic materials 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 7
- 241001654189 Lycoris aurea Species 0.000 claims description 6
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 4
- 125000002091 cationic group Chemical group 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000004440 column chromatography Methods 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 238000010828 elution Methods 0.000 claims description 4
- 238000001704 evaporation Methods 0.000 claims description 4
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000003921 oil Substances 0.000 claims description 4
- 238000002203 pretreatment Methods 0.000 claims description 4
- 239000011347 resin Substances 0.000 claims description 4
- 229920005989 resin Polymers 0.000 claims description 4
- 229920006395 saturated elastomer Polymers 0.000 claims description 4
- 238000000967 suction filtration Methods 0.000 claims description 4
- 238000002525 ultrasonication Methods 0.000 claims description 4
- 241000319062 Lycoris radiata Species 0.000 claims description 2
- 241000220317 Rosa Species 0.000 claims description 2
- 239000010902 straw Substances 0.000 claims description 2
- 238000009210 therapy by ultrasound Methods 0.000 abstract description 2
- 108010093305 exopolygalacturonase Proteins 0.000 abstract 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 abstract 1
- 239000003729 cation exchange resin Substances 0.000 abstract 1
- 238000001914 filtration Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 abstract 1
- 238000005303 weighing Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 11
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 230000002255 enzymatic effect Effects 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 230000002195 synergetic effect Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- -1 phosphoric acid salt Chemical class 0.000 description 2
- 238000002137 ultrasound extraction Methods 0.000 description 2
- 229930013930 alkaloid Natural products 0.000 description 1
- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
- C07D491/10—Spiro-condensed systems
- C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for extracting galanthamine from a lycoris plant. The method comprises the following steps: (1) extraction: weighing the treated lycoris powder, adding the lycoris powder into water with pH of 4.5 at a feed-liquid ratio of 1:10 and a unit proportion of g/mL; adding the mixed enzyme of cellulase accounting for 1.0-3.0% of the mass of the lycoris powder and pectinase, wherein the mass ratio of the cellulase to pectinase is 1:1; performing enzymolysis for 20-100min at 50 DEG C; performing ultrasonic treatment for 5-25min and centrifuging for 20min at 5,000r/min; and fetching the supernate to obtain galanthamine extract; and (2) purification: solvent extraction; adsorption by cation exchange resin; C18 reverse phase silica gel column chromatography; normal-phase silica gel column chromatography; recrystallization: dissolving the dried product III obtained in the previous step with ethanol, crystallizing to obtain a crude product of galanthamine, filtering and performing vacuum drying at 50-60 DEG C to obtain galanthamine. The method disclosed by the invention has the advantages of easiness in operation, short extraction time, high extraction rate, small dosage of solvent and relatively low cost; and the purifying process is environment-friendly and the purity is high and can reach 99.2%.
Description
Technical field
The present invention relates to a kind of extracting method of lycoremine, especially relate to a kind of method extracting lycoremine from lycoris plants.
Background technology
In short-tube lycoris, the alkaloidal chemical synthesis process such as lycoremine has report, but from lycoris plants, extract the large important sources being still such material, in prior art, the extracting method of maryllidaceous alkaloid mainly contains the methods such as solvent-extraction process, ultrasonic extraction, microwave loss mechanisms, supercritical fluid extraction, and these methods generally all exist the shortcomings such as environmental pollution is serious, production cost is high.
Summary of the invention
Technical problem to be solved by this invention is, overcomes the above-mentioned defect that prior art exists, provides a kind of method extracting lycoremine from lycoris plants that extraction time is short, solvent load is few, extraction yield is high, cost is low.
The technical solution adopted for the present invention to solve the technical problems is, a kind of method extracting lycoremine from lycoris plants, comprises the following steps:
(1) extract: take treated short-tube lycoris powder, with solid-liquid ratio 1:10, unit ratio is g/mL, adding pH is in the water of 4.5, add short-tube lycoris powder quality 1.0%-3.0% (preferred 2.0%-2.8%, more preferably 2.5%) cellulase and the mixed enzyme of polygalacturonase, the mass ratio of described cellulase and polygalacturonase is 1:1, enzymolysis 20-100min (preferred 40-80min at 50 DEG C, more preferably 60min), then ultrasonication 5-25min (preferred 15min-20min, more preferably 15min), the centrifugal 20min of 5000r/min, get supernatant liquor and namely obtain lycoremine extracting solution,
(2) purifying
1. solvent extraction: lycoremine extracting solution NaOH step (1) extracted regulates pH to 10, and 1:1 adds extraction agent trichloromethane by volume, shakes up, hold over night, take off a layer chloroform phase, HCl regulates pH to 2, suction filtration, obtains lycoremine extraction concentrated solution;
2. cationic exchange resin adsorption: by step 1. in extraction concentrated solution regulate pH to 1 ~ 6 (preferred pH is 2) D-001 type Zeo-karb through HCl, with loading speed 1-3BV/h (preferred 3BV/h) loading to saturated, wash-out is carried out with the elution rate of 3BV/h with 40%-80% (preferably 70%) ethanolic soln containing 0.5-2.5mol/L (preferred 1.5mol/L) ammoniacal liquor, drying, obtains dry thing I;
3. C
18reversed-phase silica gel column chromatography: C
18filler pre-treatment, by 20 times of quality in step 2. in the C of dry thing I
18filler is loaded in chromatography column, then with moving phase (the chromatography thing that moving phase: V (acetonitrile) ︰ V (phosphoric acid salt) is 16 ︰ 84) dissolving layer separates out, loading, adopt methanol-water solution gradation wash-out, collect elutriant, concentrated, vacuum lyophilization, obtains dry thing II;
4. purification on normal-phase silica gel column chromatography: by pretreated 20 times of quality in the column chromatography silica gel (200-300 order) of the step 3. dry thing II of gained with column diameter-post high wet method dress post, the ratio that described column diameter-post is high is 1:10 ~ 30 (preferred 1:10), the step 3. dry thing II of gained mixes with silica gel, add eluent to dissolve, described eluent is two kinds of solvent in sherwood oil, ethyl acetate, chloroform, acetone, methyl alcohol; Dry method loading after evaporating solvent, wash-out, collects elutriant, and concentrated, vacuum lyophilization, obtains dry thing III;
5. recrystallization: 4. the dry thing III of gained is through dissolve with ethanol by step, and recrystallize obtains lycoremine crude product, filters, obtains lycoremine 50 ~ 60 DEG C of vacuum-dryings.
Further, described step 4. in, eluent is sherwood oil and ethyl acetate, and the volume ratio of described ethyl acetate and sherwood oil is 8.4:1.6.
Further, described lycoris plants is Lycoris aurea, Lycoris radiata, rose short-tube lycoris, straw short-tube lycoris or Lycoris (preferred Lycoris aurea).
Further, described short-tube lycoris powder preferred Lycoris aurea bulb powder.
Compared with prior art, the present invention is simple to operate, and extraction time is short, extraction yield is high, extraction time is few, solvent load is few, cost is lower, and adopt ultrasonic synergistic Enzymatic Extraction, lycoremine yield is 0.02840%; Purifying process environmental protection, and purity is high, can reach 99.2%.
Accompanying drawing explanation
Fig. 1 is that enzyme dosage of the present invention affects schematic diagram to lycoremine yield;
Fig. 2 is that enzymolysis time of the present invention affects schematic diagram to lycoremine yield;
Fig. 3 is that ultrasonic time of the present invention affects schematic diagram to lycoremine yield.
Embodiment
Below in conjunction with drawings and Examples, the present invention will be further described.
Embodiment 1
The present embodiment comprises the following steps: accurately take treated short-tube lycoris powder 2.0g, with solid-liquid ratio 1:10, unit is g/mL, adding pH is in the water of 4.5, adds the cellulase of raw materials quality 1.0%-3.0% and the mixed enzyme of polygalacturonase respectively, enzymolysis 20-100min at 50 DEG C, then ultrasonication 5-25min, the centrifugal 20min of 5000r/min, gets supernatant liquor, obtains lycoremine extracting solution;
(2) purifying
1. solvent extraction: lycoremine extracting solution NaOH step (1) extracted regulates pH to 10, and 1:1 adds extraction agent trichloromethane by volume, shakes up, hold over night, take off a layer chloroform phase, HCl regulates pH to 2, suction filtration, obtains lycoremine extraction concentrated solution;
2. cationic exchange resin adsorption: by step 1. in extraction concentrated solution through HCl regulate pH be 2, with D-001 type Zeo-karb, with loading speed 3BV/h loading to saturated, wash-out is carried out with the elution rate of 3BV/h with 70% ethanolic soln containing 1.5mol/L ammoniacal liquor, drying, obtains dry thing I;
3. C
18reversed-phase silica gel column chromatography: C
18filler pre-treatment, by 20 times of quality in step 2. in the C of dry thing I
18filler is loaded in chromatography column, then with moving phase (the chromatography thing that moving phase: V (acetonitrile) ︰ V (phosphoric acid salt) is 16 ︰ 84) dissolving layer separates out, loading, adopt methanol-water solution gradation wash-out, collect elutriant, concentrated, vacuum lyophilization, obtains dry thing II;
4. purification on normal-phase silica gel column chromatography: by pretreated 20 times of quality in the column chromatography silica gel (200-300 order) of the step 3. dry thing II of gained with column diameter-post high wet method dress post, the ratio that described column diameter-post is high is 1:10, the step 3. dry thing II of gained mixes with silica gel, add eluent to dissolve, described eluent is two kinds of solvent in sherwood oil, ethyl acetate, chloroform, acetone, methyl alcohol; Dry method loading after evaporating solvent, wash-out, collects elutriant, and concentrated, vacuum lyophilization, obtains dry thing III;
5. recrystallization: 4. the dry thing III of gained is through dissolve with ethanol by step, and recrystallize obtains lycoremine crude product, filters, obtains 50 ~ 60 DEG C of vacuum-dryings the lycoremine that purity is 99.2%.
Step 4. in, eluent is sherwood oil and ethyl acetate, and the volume ratio of described ethyl acetate and sherwood oil is 8.4:1.6.
With reference to accompanying drawing 1, enzyme dosage affect situation, accurately take treated short-tube lycoris powder 2.000g, add the water 20mL that pH is 4.5, add the cellulase of raw materials quality 1.0%, 1.5%, 2.0%, 2.5%, 3.0% and the mixed enzyme of polygalacturonase respectively, enzymolysis 1h at 50 DEG C, supersound process 15min, the centrifugal 20min of 5000r/min, gets supernatant liquor, is settled to 50mL.Result shows, and within the scope of 1%-2.5%, lycoremine yield raises with the increase of enzyme dosage, and enzyme dosage is the best results of 2.5%.
With reference to accompanying drawing 2, the impact of enzymolysis time, take short-tube lycoris powder 2.000g, add the water 20mL that pH is 4.5, add the cellulase of raw materials quality 2.0% and the mixed enzyme of polygalacturonase, at 50 DEG C enzymolysis 20,40,60,80,100min, supersound process 15min, the centrifugal 20min of 5000r/min, gets supernatant liquor, is settled to 50mL.Result is known, and enzymolysis time is in 20-60min, and lycoremine yield increases in time and raises, and slightly reduces afterwards.60min best results.
With reference to accompanying drawing 3, the impact of ultrasonic time, takes short-tube lycoris powder 2.000g, adds the water 20mL that pH is 4.5, add the cellulase of raw materials quality 2.0% and the mixed enzyme of polygalacturonase, 50 DEG C of enzymolysis 1h, then supersound process 5,10,15,20, the centrifugal 20min of 25min, 5000r/min, get supernatant liquor, be settled to 50mL, lycoremine yield increases with the growth of sonication treatment time in 5-15min, reaches maximum value during 15min.
With lycoremine yield for index, carry out intuitive analysis and the results of analysis of variance in table 1, table 2.
Table 1 ultrasonic synergistic Enzymatic Extraction lycoremine orthogonal design and result
The variance analysis of table 2 ultrasonic synergistic Enzymatic Extraction lycoremine
| Soruces of variation | Deviation square | Degree of freedom | All square | F value | P value |
| A | 7.152 | 2 | 3.576 | 12.460 | ＜0.1 |
| B | 15.107 | 2 | 7.554 | 26.319 | ＜0.05 |
| C | 13.105 | 2 | 6.552 | 22.831 | ＜0.05 |
| Error | 0.57 | ||||
| Summation | 35.934 |
By table 1 intuitive analysis, known lycoremine optimum extraction process is A
2b
2c
1.According to variance analysis (table 2), enzymolysis time, ultrasonic time are on the impact of lycoremine yield significantly (P < 0.05); The extraction of enzyme dosage on lycoremine has impact (P < 0.1).The each factor primary and secondary order affecting lycoremine yield is enzymolysis time > ultrasonic time > enzyme dosage.The optimal procedure parameters of ultrasonic synergistic Enzymatic Extraction lycoremine is enzymolysis time 1.0h, ultrasonic time 10min, enzyme dosage 2.5%.
Be 0.02840% in this Optimal technique process lycoremine yield.
Comparative example 1.
Lycoremine in ultrasonic-assisted extraction short-tube lycoris: use the H of 0.5%
2sO
4the aqueous solution, solid-liquid ratio 1:10 (g:mL), lixiviate 2.5h then ultrasonic treatment time 30min under 70 DEG C of water-baths, extract 2 times with this understanding, lycoremine yield is 0.02530%.
Compared with embodiment 1, lycoremine yield is low.
Comparative example 2.
Lycoremine in Enzymatic Extraction short-tube lycoris: adding pH with solid-liquid ratio 1:10 (g:mL) is in the water of 4.5, add cellulase and polygalacturonase mixed enzyme enzymolysis 2h under 50 DEG C of water-baths of 4%, extract 2 times with this understanding, lycoremine yield is 0.02940%.
Compared with embodiment 1, though embodiment 1 lycoremine yield is lower than the present embodiment enzymolysis process 3.42%, but the enzyme dosage that embodiment 1 is extracted is 62.5% of simple enzymolysis process, enzymolysis time only has 1/2 of the present embodiment enzymolysis process, therefore consider from the angle of economy, embodiment 1 is carried out lycoremine extraction and is had larger advantage.
Claims (6)
1. from lycoris plants, extract a method for lycoremine, it is characterized in that, comprise the following steps:
(1) extract: take treated short-tube lycoris powder, with solid-liquid ratio 1:10, unit ratio is g/mL, adding pH is in the water of 4.5, adds the cellulase of short-tube lycoris powder quality 1.0%-3.0% and the mixed enzyme of polygalacturonase, and the mass ratio of described cellulase and polygalacturonase is 1:1, enzymolysis 20-100min at 50 DEG C, then the centrifugal 20min of ultrasonication 5-25min, 5000r/min, gets supernatant liquor and namely obtains lycoremine extracting solution;
(2) purifying:
1. solvent extraction: lycoremine extracting solution NaOH step (1) extracted regulates pH to 10, and 1:1 adds extraction agent trichloromethane by volume, shakes up, hold over night, take off a layer chloroform phase, HCl regulates pH to 2, suction filtration, obtains lycoremine extraction concentrated solution;
2. cationic exchange resin adsorption: extract concentrated solution through HCl adjustment pH to 1 ~ 6 (with D-001 type Zeo-karb by 1. middle for step, with loading speed 1-3BV/h loading to saturated, wash-out is carried out with the elution rate of 3BV/h with the 40%-80% ethanolic soln containing 0.5-2.5mol/L ammoniacal liquor, drying, obtains dry thing I;
3. C
18reversed-phase silica gel column chromatography: C
18filler pre-treatment, by 20 times of quality in step 2. in the C of dry thing I
18filler is loaded in chromatography column, then with the chromatography thing that moving phase dissolving layer separates out, and loading, adopt methanol-water solution gradation wash-out, collect elutriant, concentrated, vacuum lyophilization, obtains dry thing II;
4. purification on normal-phase silica gel column chromatography: by pretreated 20 times of quality in the column chromatography silica gel of the step 3. dry thing II of gained with column diameter-post high wet method dress post, the ratio that described column diameter-post is high is 1:10 ~ 30, the step 3. dry thing II of gained mixes with silica gel, add eluent to dissolve, described eluent is two kinds of solvent in sherwood oil, ethyl acetate, chloroform, acetone, methyl alcohol; Dry method loading after evaporating solvent, wash-out, collects elutriant, and concentrated, vacuum lyophilization, obtains dry thing III;
5. recrystallization: 4. the dry thing III of gained is through dissolve with ethanol by step, and recrystallize obtains lycoremine crude product, filters, obtains lycoremine 50 ~ 60 DEG C of vacuum-dryings.
2. the method extracting lycoremine from lycoris plants according to claim 1, is characterized in that, comprise the following steps:
(1) extract: take treated short-tube lycoris powder, with solid-liquid ratio 1:10, unit ratio is g/mL, adding pH is in the water of 4.5, adds the cellulase of short-tube lycoris powder quality 2.0%-2.8% and the mixed enzyme of polygalacturonase, and the mass ratio of described cellulase and polygalacturonase is 1:1, enzymolysis 40-80min at 50 DEG C, then the centrifugal 20min of ultrasonication 5-25min, 5000r/min, gets supernatant liquor and namely obtains lycoremine extracting solution;
(2) purifying
1. solvent extraction: lycoremine extracting solution NaOH step (1) extracted regulates pH to 10, and 1:1 adds extraction agent trichloromethane by volume, shakes up, hold over night, take off a layer chloroform phase, HCl regulates pH to 2, suction filtration, obtains lycoremine extraction concentrated solution;
2. cationic exchange resin adsorption: by step 1. in extraction concentrated solution through HCl regulate pH to 2, with D-001 type Zeo-karb, with loading speed 3BV/h loading to saturated, wash-out is carried out with the elution rate of 3BV/h with 70% ethanolic soln containing 1.5mol/L ammoniacal liquor, drying, obtains dry thing I;
3. C
18reversed-phase silica gel column chromatography: C
18filler pre-treatment, by 20 times of quality in step 2. in the C of dry thing I
18filler is loaded in chromatography column, then with the chromatography thing that moving phase dissolving layer separates out, and loading, adopt methanol-water solution gradation wash-out, collect elutriant, concentrated, vacuum lyophilization, obtains dry thing II;
4. purification on normal-phase silica gel column chromatography: by pretreated 20 times of quality in the column chromatography silica gel of the step 3. dry thing II of gained with column diameter-post high wet method dress post, the ratio that described column diameter-post is high is 1:10, the step 3. dry thing II of gained mixes with silica gel, add eluent to dissolve, described eluent is two kinds of solvent in sherwood oil, ethyl acetate, chloroform, acetone, methyl alcohol; Dry method loading after evaporating solvent, wash-out, collects elutriant, and concentrated, vacuum lyophilization, obtains dry thing III;
5. recrystallization: 4. the dry thing III of gained is through dissolve with ethanol by step, and recrystallize obtains lycoremine crude product, filters, obtains lycoremine 50 ~ 60 DEG C of vacuum-dryings.
3. the method extracting lycoremine from lycoris plants according to claim 1 and 2, is characterized in that, described step 4. in, eluent is sherwood oil and ethyl acetate, and the volume ratio of described ethyl acetate and sherwood oil is 8.4:1.6.
4. the method extracting lycoremine from lycoris plants according to claim 1 and 2, is characterized in that, described lycoris plants is Lycoris aurea, Lycoris radiata, rose short-tube lycoris, straw short-tube lycoris or Lycoris.
5. the method extracting lycoremine from lycoris plants according to claim 4, is characterized in that, described lycoris plants is Lycoris aurea.
6. the method extracting lycoremine from lycoris plants according to any one of claim 1 or 2, is characterized in that, described lycoris plants powder is Lycoris aurea bulb powder.
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| CN201510066820.5A CN104628736B (en) | 2015-02-07 | 2015-02-07 | A kind of method extracting galantamine from lycoris plants |
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| CN201510066820.5A CN104628736B (en) | 2015-02-07 | 2015-02-07 | A kind of method extracting galantamine from lycoris plants |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106967080A (en) * | 2017-04-10 | 2017-07-21 | 吉首大学 | A kind of method that galanthamine is extracted from short-tube lycoris |
| CN107462656A (en) * | 2017-07-04 | 2017-12-12 | 上海市农业科学院 | A kind of method of galanthamine content in quick detection amrallid |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101704822A (en) * | 2009-10-20 | 2010-05-12 | 南京荣世医药科技有限公司 | Method for preparing galanthamine |
| CN101781305A (en) * | 2010-03-23 | 2010-07-21 | 泰州市今朝伟业精细化工有限公司 | Method for artificially synthesizing galanthamine |
| CN101157947B (en) * | 2007-09-13 | 2011-04-20 | 华中科技大学 | Method for extracting active alkaloid from lycoris herb |
| CN103772403A (en) * | 2014-02-27 | 2014-05-07 | 湖南中医药大学 | Method for extracting galanthamine by using lycoris aurea |
-
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- 2015-02-07 CN CN201510066820.5A patent/CN104628736B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101157947B (en) * | 2007-09-13 | 2011-04-20 | 华中科技大学 | Method for extracting active alkaloid from lycoris herb |
| CN101704822A (en) * | 2009-10-20 | 2010-05-12 | 南京荣世医药科技有限公司 | Method for preparing galanthamine |
| CN101781305A (en) * | 2010-03-23 | 2010-07-21 | 泰州市今朝伟业精细化工有限公司 | Method for artificially synthesizing galanthamine |
| CN103772403A (en) * | 2014-02-27 | 2014-05-07 | 湖南中医药大学 | Method for extracting galanthamine by using lycoris aurea |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106967080A (en) * | 2017-04-10 | 2017-07-21 | 吉首大学 | A kind of method that galanthamine is extracted from short-tube lycoris |
| CN107462656A (en) * | 2017-07-04 | 2017-12-12 | 上海市农业科学院 | A kind of method of galanthamine content in quick detection amrallid |
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