CN114031654A - Sennoside extraction method - Google Patents
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- CN114031654A CN114031654A CN202111331250.XA CN202111331250A CN114031654A CN 114031654 A CN114031654 A CN 114031654A CN 202111331250 A CN202111331250 A CN 202111331250A CN 114031654 A CN114031654 A CN 114031654A
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- IPQVTOJGNYVQEO-KGFNBKMBSA-N sennoside A Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC2=C1C(=O)C1=C(O)C=C(C(O)=O)C=C1[C@@H]2[C@H]1C2=CC(C(O)=O)=CC(O)=C2C(=O)C2=C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C=CC=C21 IPQVTOJGNYVQEO-KGFNBKMBSA-N 0.000 title claims abstract description 55
- 229940124513 senna glycoside Drugs 0.000 title claims abstract description 42
- 229930186851 sennoside Natural products 0.000 title claims abstract description 36
- 238000000605 extraction Methods 0.000 title claims description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 46
- IPQVTOJGNYVQEO-CXZNLNCXSA-N sennoside A Natural products O=C(O)c1cc(O)c2C(=O)c3c(O[C@H]4[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O4)cccc3[C@@H]([C@H]3c4c(c(O)cc(C(=O)O)c4)C(=O)c4c(O[C@H]5[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O5)cccc34)c2c1 IPQVTOJGNYVQEO-CXZNLNCXSA-N 0.000 claims abstract description 39
- 238000011068 loading method Methods 0.000 claims abstract description 32
- 239000003480 eluent Substances 0.000 claims abstract description 26
- 239000011780 sodium chloride Substances 0.000 claims abstract description 23
- 239000007788 liquid Substances 0.000 claims abstract description 18
- 239000006228 supernatant Substances 0.000 claims abstract description 18
- 238000005406 washing Methods 0.000 claims abstract description 18
- 238000001914 filtration Methods 0.000 claims abstract description 15
- 239000000284 extract Substances 0.000 claims abstract description 14
- -1 B compound Chemical class 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 12
- 238000001035 drying Methods 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims abstract description 10
- 238000010828 elution Methods 0.000 claims abstract description 9
- 238000002791 soaking Methods 0.000 claims abstract description 6
- 238000005342 ion exchange Methods 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 44
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 26
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 13
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 13
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 239000008055 phosphate buffer solution Substances 0.000 claims description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- 239000004471 Glycine Substances 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 4
- 238000011067 equilibration Methods 0.000 claims description 4
- 238000012856 packing Methods 0.000 claims description 4
- 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 claims description 2
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 2
- 238000005349 anion exchange Methods 0.000 claims description 2
- 239000003456 ion exchange resin Substances 0.000 claims description 2
- 229920003303 ion-exchange polymer Polymers 0.000 claims description 2
- 239000003446 ligand Substances 0.000 claims description 2
- 125000001453 quaternary ammonium group Chemical group 0.000 claims description 2
- 150000003440 styrenes Chemical class 0.000 claims description 2
- MZVQCMJNVPIDEA-UHFFFAOYSA-N [CH2]CN(CC)CC Chemical group [CH2]CN(CC)CC MZVQCMJNVPIDEA-UHFFFAOYSA-N 0.000 claims 1
- 125000000217 alkyl group Chemical group 0.000 claims 1
- 239000008363 phosphate buffer Substances 0.000 claims 1
- IPQVTOJGNYVQEO-AIFLABODSA-N sennoside B Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC2=C1C(=O)C1=C(O)C=C(C(O)=O)C=C1[C@H]2[C@H]1C2=CC(C(O)=O)=CC(O)=C2C(=O)C2=C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C=CC=C21 IPQVTOJGNYVQEO-AIFLABODSA-N 0.000 abstract description 11
- 229940004991 sennoside b Drugs 0.000 abstract description 8
- 238000004255 ion exchange chromatography Methods 0.000 abstract description 5
- 229940070486 senna leaf extract Drugs 0.000 abstract description 3
- 238000011084 recovery Methods 0.000 abstract description 2
- 238000001514 detection method Methods 0.000 description 14
- 238000005070 sampling Methods 0.000 description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- 229920005989 resin Polymers 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 206010010774 Constipation Diseases 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- 235000006693 Cassia laevigata Nutrition 0.000 description 7
- 241000522641 Senna Species 0.000 description 7
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- 239000003960 organic solvent Substances 0.000 description 5
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- 238000005571 anion exchange chromatography Methods 0.000 description 4
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Natural products CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 description 4
- 150000004056 anthraquinones Chemical class 0.000 description 4
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- 238000000746 purification Methods 0.000 description 4
- 229940076742 senna leaves Drugs 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000005377 adsorption chromatography Methods 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
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- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 241001249696 Senna alexandrina Species 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- MGRRGKWPEVFJSH-UHFFFAOYSA-N dianthrone Natural products C12=CC=CC=C2C(=O)C2=CC=CC=C2C1=C1C2=CC=CC=C2C(=O)C2=CC=CC=C21 MGRRGKWPEVFJSH-UHFFFAOYSA-N 0.000 description 2
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- 230000002401 inhibitory effect Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000001543 purgative effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- IPQVTOJGNYVQEO-LALZNDFESA-N 9-[(9r)-2-carboxy-4-hydroxy-10-oxo-5-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-9h-anthracen-9-yl]-4-hydroxy-10-oxo-5-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-9h-anthracene-2-carboxylic acid Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC2=C1C(=O)C1=C(O)C=C(C(O)=O)C=C1C2[C@H]1C2=CC(C(O)=O)=CC(O)=C2C(=O)C2=C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C=CC=C21 IPQVTOJGNYVQEO-LALZNDFESA-N 0.000 description 1
- IPQVTOJGNYVQEO-UHFFFAOYSA-N 9-[2-carboxy-4-hydroxy-10-oxo-5-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-9h-anthracen-9-yl]-4-hydroxy-10-oxo-5-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-9h-anthracene-2-carboxylic acid Chemical class OC1C(O)C(O)C(CO)OC1OC1=CC=CC2=C1C(=O)C1=C(O)C=C(C(O)=O)C=C1C2C1C2=CC(C(O)=O)=CC(O)=C2C(=O)C2=C(OC3C(C(O)C(O)C(CO)O3)O)C=CC=C21 IPQVTOJGNYVQEO-UHFFFAOYSA-N 0.000 description 1
- 206010000060 Abdominal distension Diseases 0.000 description 1
- 208000010392 Bone Fractures Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 208000015220 Febrile disease Diseases 0.000 description 1
- 206010017076 Fracture Diseases 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000193998 Streptococcus pneumoniae Species 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 125000004103 aminoalkyl group Chemical group 0.000 description 1
- MWPLVEDNUUSJAV-UHFFFAOYSA-N anthracene Natural products C1=CC=CC2=CC3=CC=CC=C3C=C21 MWPLVEDNUUSJAV-UHFFFAOYSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- JQVYZJIFFAHQKX-ZAULLPPESA-L calcium;3-carboxy-10-[2-carboxy-4-oxido-10-oxo-5-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-9h-anthracen-9-yl]-9-oxo-8-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-10h-anthracen-1-olate Chemical compound [Ca+2].O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC2=C1C(=O)C1=C(O)C=C(C([O-])=O)C=C1C2C1C2=CC(C([O-])=O)=CC(O)=C2C(=O)C2=C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C=CC=C21 JQVYZJIFFAHQKX-ZAULLPPESA-L 0.000 description 1
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- 201000006549 dyspepsia Diseases 0.000 description 1
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- 230000020477 pH reduction Effects 0.000 description 1
- 230000002980 postoperative effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
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- 239000000047 product Substances 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
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- 238000013341 scale-up Methods 0.000 description 1
- 239000009814 sennoside A&B Substances 0.000 description 1
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- 229940031000 streptococcus pneumoniae Drugs 0.000 description 1
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/244—Anthraquinone radicals, e.g. sennosides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention discloses a method for extracting sennoside, which comprises the following steps: the method comprises the following steps: (1) initially extracting medicinal materials: adjusting pH of extractive solution obtained by soaking folium sennae to 2.5-5.5, filtering to obtain supernatant, adding sodium chloride into the supernatant to adjust conductivity, and adjusting pH of the supernatant to 4.0-10.0 to obtain supernatant; the conductivity of the sample liquid is 2-5mS/cm lower than that of the equilibrium liquid; (2) loading: balancing the ion exchange chromatographic column with a balance liquid in advance, and loading the loading liquid in the step (1) to the chromatographic column; (3) and (3) elution: after the sample loading is finished, washing the column by 3-5 times of column volume by using a balance solution, and then washing the column by using eluent with 3-5 times of column volume; (4) and (4) drying the eluent obtained in the step (3) under reduced pressure to obtain a powdery extract of the sennoside A/B compound. The invention adopts ion exchange chromatography to separate and purify sennoside, can separate sennoside A and sennoside B from senna leaf extract, and has high recovery rate.
Description
Technical Field
The invention relates to the technical field of plant extraction, and particularly relates to a sennoside extraction method.
Background
Senna (Folium sennae) is one of the commonly used purgative and indigestion-removing drugs, and is the dried leaves of senna angustifolia (Cassia angustifolia Vahl) or senna acutifolia (C. The main active component in senna is anthraquinone derivative. In recent years, research shows that the cathartic sennoside A, B, C, D of dianthrone derivatives in anthraquinone derivatives has stronger purgative effect and irritation than other cathartic containing anthraquinone derivatives, has little toxicity when being taken orally, and is commonly used for acute constipation clinically. The dianthracene ketone compound is formed by mutually connecting two molecules of anthracene ketone C-C, wherein sennosides A and B and sennosides C and D are mutually stereoisomers. Sennoside has inhibitory effect on Streptococcus pneumoniae, Klebsiella pneumoniae and Staphylococcus aureus. In addition, sennoside has insulin sensitizing and alpha-glucoamylase inhibiting effects. Clinically, the medicine containing the sennoside is mainly used for treating constipation and clearing intestines, and comprises various kinds of constipation such as senile constipation, constipation in puerperium, functional constipation of young people, constipation caused by postoperative and febrile diseases, thoracolumbar fracture, abdominal distension and constipation, and the like.
As for the purification technology of sennoside, one of the prior arts adopts macroporous resin to purify sennoside, and the technology is that 20 times of boiling water is added into senna leaves, the senna leaves are soaked for 2 times at 80 ℃ with heat preservation for 45min each time, the filtrate is cooled to room temperature, the pH value is adjusted to 3, and the sennoside is centrifuged. And (3) putting the centrifugate on a DM-301 resin column, washing with pure water until the molish reaction is negative, and eluting with 0.5mol/L sodium bicarbonate aqueous solution and pure water sequentially. Collecting when the color band is about to flow out, neutralizing the collected solution with hydrochloric acid, loading on another DM-301 resin column, washing with water, eluting with 50% ethanol, concentrating the eluate under reduced pressure to obtain fluid extract, drying under reduced pressure at 60 deg.C, and pulverizing to obtain brown powder with sennoside content of 50% and sennoside content of more than 50%. The operation procedure is complicated and boiling water extraction is liable to cause degradation of sennoside. In the prior art, the process conditions for extracting dianthrone in senna leaves are researched by taking the sennoside A content as an index, and the optimal extraction process is as follows: soaking folium sennae powder in 3 times of 50% ethanol for 12 hr, percolating with 6 times of 50% ethanol at 600 ml/hr kg flow rate, and extracting to obtain extractive solution. Concentrating the extractive solution, purifying with X-5 macroporous resin column, washing with 10 times of resin amount of water, washing with 7.5 times of 30% ethanol, collecting eluate, and concentrating to dry to obtain extract. Further acidifying the extract, and extracting with n-butanol to obtain product with total sennoside content of 47.2%, with yield of 2.93%. The organic solvents of n-butanol and ethanol are used, so the cost is higher and the environment is not protected. Patent document CN 105254689A discloses a sennoside a.b salt compound and a preparation method and application thereof, which uses senna leaves as a raw material, and the sennoside a.b salt compound with high purity of effective components is obtained by extraction, filtration, acidification, fine filtration, extraction, refining, salification of the compound, precipitation and purification, and the purity reaches 80-90%. The steps are complicated, and organic solvents with strong toxicity, such as acetone, methanol and the like, are used in the purification process of the last step. In addition, sennoside has poor stability, is easily decomposed and converted into monoanthrone components by heat and light, and is difficult to extract the components with high efficiency by using conventional extraction, macroporous adsorption chromatography and other methods.
Disclosure of Invention
The purpose of the invention is as follows: the invention provides a sennoside extraction method, which adopts ion exchange chromatography to separate and purify sennoside, can separate sennoside A and sennoside B from senna leaf extract, has a recovery rate of more than 90 percent, has obvious impurity removal effect, and provides convenience for the separation and purification of high-purity sennoside in the later period.
The technical scheme is as follows: the invention provides a sennoside extraction method, wherein the sennoside is a sennoside A/B compound (sennoside A and sennoside B are stereoisomers of each other), and has the following structure:
the extraction method of sennoside comprises the following steps: the method comprises the following steps:
(1) initially extracting medicinal materials: adjusting pH of extractive solution obtained by soaking folium sennae to 2.5-5.5, filtering to obtain supernatant, adding sodium chloride into the supernatant to adjust conductivity, and adjusting pH of the supernatant to 4.0-10.0 to obtain supernatant; the conductivity of the sample liquid is 2-5mS/cm lower than that of the equilibrium liquid;
(2) loading: balancing the ion exchange chromatographic column with a balance liquid in advance, and loading the loading liquid in the step (1) to the chromatographic column; the equilibrium solution is 0.02-0.2mol/L acetate, glycine, sodium bicarbonate or phosphate buffer solution, the equilibrium solution contains 0.0001-0.3mol/L NaCl, and the pH value is 3.5-10.0;
(3) and (3) elution: after the sample loading is finished, washing the column by 3-5 times of column volume by using a balance solution, and then washing the column by using eluent with 3-5 times of column volume; the eluent is 0.02-0.2mol/L acetate, glycine, sodium bicarbonate or phosphate buffer solution, the eluent contains 0.3-1.5mol/L NaCl, and the pH value is 3.5-10.0;
(4) and (4) drying the eluent obtained in the step (3) under reduced pressure to obtain a powdery extract of the sennoside A/B compound.
As a preferable example, in the step (1), the pH of the extract is adjusted to 3.0 to 5.5, and acids used for adjusting the pH are hydrochloric acid, acetic acid and phosphoric acid.
As a preferable example, in the step (3), the eluents are 0.02mol/L phosphate buffer solution and 1.2mol/L NaCl solution, and the pH value is 7.0.
As a preferable example, in step (2), the ligand of the ion exchange chromatography column is anion exchange chromatography packing with diethyl aminoethyl group and trimethyl aminoalkyl quaternary ammonium group, and the main framework of the packing is any one of acrylic series or styrene series.
As a preferred embodiment, the type of the ion exchange resin is A313 type, A313FC type, D620 type, D630 type, D640 type, A351 type or A451 type.
As a preferred example, in the step (1), the extract is prepared by the following method: adding 5-10 times of sodium bicarbonate solution into folium sennae, stirring, soaking for 60-120min, and filtering to obtain extractive solution.
As a preferred embodiment, in the step (1), the conductivity of the sample solution is 0.5mS/cm-29 mS/cm; in the step (2), the conductivity of the equilibrium liquid is 1-30 mS/cm.
As a preferred embodiment, in the step (3), the conductivity of the eluent is 30-108 mS/cm.
As a preferable example, in the step (4), the reduced pressure drying is performed under the conditions that the temperature is 55-85 ℃ and the vacuum degree is-0.05-0.1 MPa.
As a preferable embodiment, the equilibrium solution contains 0.0001-0.3mol/L NaCl; the eluent contains 0.3-1.5mol/L NaCl.
In the present invention, "%" is a mass percentage unless otherwise specified.
Has the advantages that: (1) on the basis of the prior art, the invention provides a simple, high-efficiency and easily-amplified separation method, which adopts ion exchange chromatography to separate and purify sennoside, and separates sennoside A and sennoside B from senna leaf extract, thereby solving the problem of high cost caused by elution of a macroporous adsorption resin organic solvent for the first time, and the sennoside A/B is eluted characteristically by sodium chloride, and the yield is higher than 90%; (2) the invention solves the problems that the sennoside A/B is similar to other anthraquinone impurities in senna in physicochemical properties and cannot be purified out characteristically by using a conventional organic solvent extraction method in the prior art, and simultaneously solves the problems of lower organic solvent elution yield, high cost, low resolution and poorer use effect of macroporous adsorption resin; (3) the extraction method has stable process and is beneficial to industrial and large-scale production. The invention is verified by repeated experiments, the sennoside A/B can be separated and purified with good characteristics by ion exchange chromatography, and the method is easy for scale-up production.
Drawings
FIG. 1 is an HPLC chromatogram of the anion exchange column eluate of example 1;
FIG. 2 is an HPLC chromatogram of the eluate from the macroporous adsorption chromatography column of example 4.
Detailed Description
Example 1: (1) extracting medicinal materials: adding 100g of senna leaf medicinal material (wherein the content of sennoside A and sennoside B is 1.25%) into 8 times (g/mL) of 0.1% sodium bicarbonate solution, stirring and extracting for 60min, filtering to obtain an extracting solution (sampling detection of the extracting solution), adding 6mol/L HCl to adjust the pH value to 3.0, removing impurities, filtering to obtain a supernatant, adding sodium chloride to adjust the electric conductivity to 20mS/cm, adding 5mol/L NaOH to adjust the pH value to 3.5, and obtaining a supernatant (sampling detection of an acid-adjusted supernatant);
(2) loading: 300mL of a macroporous weakly basic anion exchange chromatography column of A451 type (ZG A451, Hangzhou Guanguang resin Co., Ltd.) was previously loaded, equilibrated with 5 column volumes of an equilibration solution at a linear speed of 60cm/h, 0.02mol/L of a glycine buffer solution containing 0.2mol/L of NaCl, having a pH of 3.5 and a conductivity of 23mS/cm, and then the loading solution in step (1) was loaded onto the chromatography column at a linear speed of 60 cm/h.
(3) And (3) elution: after the sample loading is finished, firstly washing 4 times of column volume (upper punch T sampling detection) by using equilibrium liquid at a linear speed of 60cm/h, then washing the column by using eluent with 5 times of column volume at a linear speed of 40cm/h (eluent sampling detection), wherein the eluent is 0.02mol/L glycine buffer solution, contains 1.0mol/L NaCl, has the pH value of 3.5 and has the conductivity of 93 mS/cm. Collecting eluate, drying under reduced pressure at 60 deg.C and vacuum degree of-0.08 MPa to obtain powdered extract of sennoside A/B compound, and detecting by HPLC with chromatogram shown in figure 1.
(4) The samples were sampled and the content was measured as follows:
example 2: (1) extracting medicinal materials: taking 100g of senna leaf medicinal material (wherein the content of sennoside A and sennoside B is 1.23%), adding 10 times (g/mL) of 0.25% sodium bicarbonate solution, stirring and extracting for 90min, filtering to obtain an extracting solution (sampling detection), adding 6mol/L HCl to adjust the pH value to 4.5, filtering to obtain a supernatant, adding sodium chloride to adjust the conductivity to 15mS/cm, adding 5mol/L NaOH to adjust the pH value to 7.0, and obtaining a loading solution (adjusting the acid and sampling detection of the loading solution);
(2) loading: 300mL of a A351 type (ZG A351 type, Hangzhou Guanguang resin Co., Ltd.) macroporous weakly basic anion exchange chromatography column is loaded in advance, the column is equilibrated with 5 times of column volume of an equilibration solution at a linear speed of 60cm/h, the equilibration solution is 0.02mol/L phosphate buffer solution which contains 0.15mol/L NaCl, the pH value is 7.0, the conductivity is 18mS/cm, and then the sample loading solution in the step (1) is loaded on the chromatography column at a linear speed of 60 cm/h.
(3) And (3) elution: after the sample loading is finished, firstly, washing the column by 4 times of column volume by using equilibrium liquid at a linear speed of 60cm/h (upper punch T sampling detection), and then washing the column by using eluent with 5 times of column volume at a linear speed of 40cm/h, wherein the eluent is 0.02mol/L phosphate buffer solution, contains 1.2mol/L NaCl, has a pH value of 7.0 and has a conductivity of 108 mS/cm. Collecting eluate, and drying under reduced pressure at 70 deg.C and vacuum degree of-0.08 MPa to obtain powdered extract of sennoside A/B compound.
(4) The samples were sampled and the content was measured as follows:
example 3: (1) extracting medicinal materials: adding 1000g of senna leaf medicinal material (wherein the content of sennoside A and sennoside B is 1.15%) into 10 times (g/mL) of 0.25% sodium bicarbonate solution, stirring and extracting for 120min, filtering to obtain an extracting solution (sampling detection of the extracting solution), adding 6mol/L HCl to adjust the pH value to 5.5, filtering to obtain a supernatant, adding sodium chloride to adjust the conductivity to 7mS/cm, adding 5mol/L NaOH to adjust the pH value to 10.0, and obtaining a loading solution (sampling detection of the acid-adjusted loading solution);
(2) loading: loading 1500ml of A313 (ZG A313, Hangzhou Chongguang resin Co., Ltd.) macroporous weakly basic anion exchange chromatography column in advance, balancing 5 times of column volume with a balance solution at a linear speed of 60cm/h, wherein the balance solution is 0.2mol/L sodium bicarbonate buffer solution containing 0.1mol/L NaCl, the pH value is 10.0, the conductivity is 11mS/cm, and loading the sample loading solution in the step (1) onto the chromatography column at a linear speed of 60 cm/h.
(3) And (3) elution: after the sample loading is finished, firstly, washing the column by 4 times of column volume by using equilibrium liquid at a linear speed of 60cm/h (upper punch T sampling detection), then, washing the column by using eluent with 5 times of column volume at a linear speed of 40cm/h (eluent sampling detection), wherein the eluent is 0.2mol/L sodium bicarbonate buffer solution, contains 0.8mol/L NaCl, has the pH of 10.0 and has the conductivity of 70 mS/cm. Collecting eluate, and drying under reduced pressure at 80 deg.C and vacuum degree of-0.08 MPa to obtain powdered extract of sennoside A/B compound.
(4) The samples were sampled and the content was measured as follows:
example 4: (1) extracting medicinal materials: adding 10 times (g/mL) of 0.25% sodium bicarbonate solution into 100g of folium sennae (sennoside A + sennoside B content is 1.23%), stirring, extracting for 90min, filtering to obtain extractive solution (sampling detection), adding 6mol/L HCl to adjust pH to 3.5, filtering, and collecting supernatant (adjusting acid and sampling detection);
(2) loading: the loading solution in the step (1) was loaded on a D101 type adsorption chromatography column (300 mL) in advance at a linear speed of 60 cm/h.
(3) And (3) elution: after the sample loading is finished, washing the column by 4 times of the column volume by using water at a linear speed of 60cm/h (upward T sampling detection), and then washing the column by using eluent with 5 times of the column volume at a linear speed of 40cm/h, wherein the eluent is 50% ethanol solution. Collecting eluate, drying under reduced pressure at 55 deg.C and vacuum degree of-0.08 MPa to obtain powdered extract of sennoside A/B compound, and detecting the obtained extract as shown in FIG. 2.
Claims (10)
1. The sennoside extraction method is characterized by comprising the following steps:
(1) initially extracting medicinal materials: adjusting pH of extractive solution obtained by soaking folium sennae to 2.5-5.5, filtering to obtain supernatant, adding sodium chloride into the supernatant to adjust conductivity, and adjusting pH of the supernatant to 4.0-10.0 to obtain supernatant; the conductivity of the sample liquid is 2-5mS/cm lower than that of the equilibrium liquid;
(2) loading: balancing the ion exchange chromatographic column with a balance liquid in advance, and loading the loading liquid in the step (1) to the chromatographic column; the equilibrium solution is 0.02-0.2mol/L acetate, glycine, sodium bicarbonate or phosphate buffer solution, the equilibrium solution contains 0.0001-0.3mol/L NaCl, and the pH value is 3.5-10.0;
(3) and (3) elution: after the sample loading is finished, firstly washing the column by using 3-5 times of column volume balance liquid, and then washing the column by using 3-5 times of column volume eluent; the eluent is 0.02-0.2mol/L acetate, glycine, sodium bicarbonate or phosphate buffer solution, the eluent contains 0.3-1.5mol/L NaCl, and the pH value is 3.5-10.0;
(4) and (4) drying the eluent obtained in the step (3) under reduced pressure to obtain a powdery extract of the sennoside A/B compound.
2. The method for extracting sennoside as claimed in claim 1, wherein in the step (1), the pH of the extract is adjusted to 3.0-5.5, and the acid used for adjusting the pH is hydrochloric acid, acetic acid and phosphoric acid.
3. The sennoside extraction method according to claim 1, wherein in step (3), the eluents are 0.02mol/L phosphate buffer and 1.2mol/L NaCl solution, and the pH value is 7.0.
4. The sennoside extraction method according to claim 1, wherein in the step (2), the ligand of the ion exchange chromatographic column is anion exchange chromatographic packing with diethyl aminoethyl and trimethyl alkyl quaternary ammonium groups, and the main framework of the packing is any one of acrylic series or styrene series.
5. The sennoside extraction method of claim 4, wherein the ion exchange resin is of type A313, A313FC, D620, D630, D640, A351 or A451.
6. The sennoside extraction method according to claim 1, characterised in that in step (1), the extraction solution is prepared by: adding 5-10 times of sodium bicarbonate solution into folium sennae, stirring, soaking for 60-120min, and filtering to obtain extractive solution.
7. The sennoside extraction process of claim 1, wherein in step (1), the conductivity of the loading solution is between 0.5mS/cm and 29 mS/cm; in the step (2), the conductivity of the equilibrium liquid is 1-30 mS/cm.
8. The sennoside extraction process of claim 1, wherein in step (3), the conductivity of the eluent is 30-108 mS/cm.
9. The method for extracting sennoside as claimed in claim 1, wherein in the step (4), the reduced pressure drying is performed under the conditions of a temperature of 55-85 ℃ and a vacuum degree of-0.05-0.1 MPa.
10. The sennoside extraction method of claim 1, wherein the equilibration solution contains 0.0001-0.3mol/L NaCl; the eluent contains 0.3-1.5mol/L NaCl.
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| CN102040637A (en) * | 2010-09-27 | 2011-05-04 | 南京泽朗医药科技有限公司 | Method for extracting sennoside |
| CN105254689A (en) * | 2015-11-04 | 2016-01-20 | 安徽九方药物研究院有限公司 | Sennoside A.B salt compound as well as preparation method and application thereof |
| CN109966337A (en) * | 2019-03-28 | 2019-07-05 | 湖南华诚生物资源股份有限公司 | A method of extracting sennoside from folium sennae |
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| CN102040637A (en) * | 2010-09-27 | 2011-05-04 | 南京泽朗医药科技有限公司 | Method for extracting sennoside |
| CN105254689A (en) * | 2015-11-04 | 2016-01-20 | 安徽九方药物研究院有限公司 | Sennoside A.B salt compound as well as preparation method and application thereof |
| CN109966337A (en) * | 2019-03-28 | 2019-07-05 | 湖南华诚生物资源股份有限公司 | A method of extracting sennoside from folium sennae |
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