CN113861250A - A method for preparing high purity sennoside and sennoside or its derivatives - Google Patents
A method for preparing high purity sennoside and sennoside or its derivatives Download PDFInfo
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- CN113861250A CN113861250A CN202111331239.3A CN202111331239A CN113861250A CN 113861250 A CN113861250 A CN 113861250A CN 202111331239 A CN202111331239 A CN 202111331239A CN 113861250 A CN113861250 A CN 113861250A
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- sennoside
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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 61
- 229940124513 senna glycoside Drugs 0.000 title claims abstract description 49
- 229930186851 sennoside Natural products 0.000 title claims abstract description 43
- 238000000034 method Methods 0.000 title claims description 23
- 239000003480 eluent Substances 0.000 claims abstract description 62
- 239000007788 liquid Substances 0.000 claims abstract description 45
- 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 42
- 239000006228 supernatant Substances 0.000 claims abstract description 37
- -1 B compound Chemical class 0.000 claims abstract description 30
- 238000011068 loading method Methods 0.000 claims abstract description 26
- 238000002360 preparation method Methods 0.000 claims abstract description 18
- 238000001556 precipitation Methods 0.000 claims abstract description 15
- 238000004440 column chromatography Methods 0.000 claims abstract description 13
- 239000000284 extract Substances 0.000 claims abstract description 13
- 238000000926 separation method Methods 0.000 claims abstract description 13
- 238000001179 sorption measurement Methods 0.000 claims abstract description 13
- 239000000463 material Substances 0.000 claims abstract description 12
- 238000005342 ion exchange Methods 0.000 claims abstract description 11
- 238000004255 ion exchange chromatography Methods 0.000 claims abstract description 7
- 239000000243 solution Substances 0.000 claims description 76
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 63
- 238000005406 washing Methods 0.000 claims description 40
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 34
- 239000007853 buffer solution Substances 0.000 claims description 31
- 239000011780 sodium chloride Substances 0.000 claims description 31
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 26
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 24
- 239000002244 precipitate Substances 0.000 claims description 17
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 17
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 17
- 239000008055 phosphate buffer solution Substances 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 15
- 239000004471 Glycine Substances 0.000 claims description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 238000001035 drying Methods 0.000 claims description 11
- 238000001914 filtration Methods 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- 238000005377 adsorption chromatography Methods 0.000 claims description 9
- 239000002253 acid Substances 0.000 claims description 8
- 235000019441 ethanol Nutrition 0.000 claims description 7
- 229920005989 resin Polymers 0.000 claims description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 238000012856 packing Methods 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- IVLXQGJVBGMLRR-UHFFFAOYSA-N 2-aminoacetic acid;hydron;chloride Chemical compound Cl.NCC(O)=O IVLXQGJVBGMLRR-UHFFFAOYSA-N 0.000 claims description 4
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 claims description 4
- 229910001424 calcium ion Inorganic materials 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 claims description 4
- 238000011010 flushing procedure Methods 0.000 claims description 4
- 239000011347 resin Substances 0.000 claims description 4
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 claims description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 239000004793 Polystyrene Substances 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
- 239000000945 filler Substances 0.000 claims description 2
- 239000003446 ligand Substances 0.000 claims description 2
- 229920002223 polystyrene Polymers 0.000 claims description 2
- 125000001453 quaternary ammonium group Chemical group 0.000 claims description 2
- 238000002791 soaking Methods 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
- 238000005349 anion exchange Methods 0.000 claims 1
- 125000003011 styrenyl group Chemical class [H]\C(*)=C(/[H])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 claims 1
- 230000007613 environmental effect Effects 0.000 abstract 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 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 238000005070 sampling Methods 0.000 description 9
- 235000006693 Cassia laevigata Nutrition 0.000 description 7
- 241000522641 Senna Species 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- YSCJAYPKBYRXEZ-HZPINHDXSA-N (2s,3s,4s,5r,6r)-6-[[(3s,4ar,6ar,6bs,8as,12as,14ar,14br)-4,4,6a,6b,11,11,14b-heptamethyl-8a-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxycarbonyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-3-hydroxy-4-[(2s,3r,4s, Chemical compound O([C@H]1[C@H](O)[C@H](O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@]1(CCC(C[C@H]14)(C)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C(O)=O)[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O YSCJAYPKBYRXEZ-HZPINHDXSA-N 0.000 description 5
- 229960000583 acetic acid Drugs 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 229940004991 sennoside b Drugs 0.000 description 5
- YDZWHGJRWMQCDP-NKILCQAGSA-N (2s,3s,4s,5r,6r)-6-[[(3s,4ar,6ar,6bs,8as,12as,14ar,14br)-8a-carboxy-4,4,6a,6b,11,11,14b-heptamethyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-yl]oxy]-3-hydroxy-4-[(2s,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-5-[(2s,3r,4 Chemical compound O([C@H]1[C@H](O)[C@H](O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2C1(C)C)C)(C)CC[C@]1(CCC(C[C@H]14)(C)C)C(O)=O)C(O)=O)[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O YDZWHGJRWMQCDP-NKILCQAGSA-N 0.000 description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 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
- 238000004587 chromatography analysis Methods 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 3
- 239000012141 concentrate Substances 0.000 description 3
- 238000005755 formation reaction Methods 0.000 description 3
- 239000012362 glacial acetic acid Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000008213 purified water Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 2
- 239000000920 calcium hydroxide Substances 0.000 description 2
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 2
- 159000000007 calcium salts Chemical class 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000011067 equilibration Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 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
- 238000000746 purification Methods 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 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 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
- 206010010774 Constipation Diseases 0.000 description 1
- 241000196324 Embryophyta Species 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
- 239000008351 acetate buffer Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001154 acute effect Effects 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
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000000872 buffer Substances 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
- 239000000799 cathartic agent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 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 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001543 purgative effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000012487 rinsing solution Substances 0.000 description 1
- 229940076742 senna leaves Drugs 0.000 description 1
- 239000009814 sennoside A&B Substances 0.000 description 1
- 230000001235 sensitizing effect Effects 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 229940031000 streptococcus pneumoniae Drugs 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
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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
-
- 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
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- 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)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a preparation method of high-purity sennoside and sennoside or a derivative prepared by the same. The preparation method comprises the following steps: (1) extracting medicinal materials; (2) ion exchange column chromatography: balancing the ion exchange chromatography column by using 3-5 times of column volume balancing liquid A; loading the acid-adjusted supernatant obtained in the step (1) to a chromatographic column, rebalancing with a 3-5 times column volume balancing solution A, and eluting with a 3-5 times column volume eluent B to obtain an eluent; (3) macroporous adsorption column chromatography: adjusting the pH of the eluent obtained in the step (2) to 3.5-8.0, loading the eluent into a macroporous adsorption chromatographic column, and eluting with eluent E with the volume 3-5 times of the column volume to obtain eluent; (4) and (4) carrying out low-temperature precipitation and solid-liquid separation on the eluent obtained in the step (3) to obtain the extract of the sennoside A/B compound. The sennoside is purified by using a macroporous adsorption chromatographic column and an ion exchange column, and the obtained sennoside extract has high purity, low cost and environmental protection.
Description
Technical Field
The invention relates to the technical field of plant extraction, and particularly relates to a preparation method of high-purity sennoside and sennoside or a derivative prepared by the same.
Background
The main active ingredient of Folium sennae (Folium sennae) is anthraquinone derivatives. The cathartic glycoside A, B, C, D of dianthrone derivative in anthraquinone derivative has stronger purgative effect and irritation than other cathartics containing anthraquinone, and has low oral administration toxicity, and is commonly used for treating acute constipation. 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. The sennoside is purified by macroporous resin, the sennoside content prepared by the method reaches 50 percent, and the total sennoside content reaches more than 50 percent, but the method has complex operation procedure and is easy to degrade by boiling water extraction, and the sennoside is easy to decompose into monoanthrone components by heating and light because of poor stability, and the components are difficult to be efficiently extracted by the conventional methods such as extraction, macroporous adsorption chromatography and the like. Zhang Yang, etc. uses sennoside A content as index, and researches the process condition of extracting dianthranone in senna leaf to obtain the product with 47.2% of total sennoside content, the yield is 2.93%, but the method uses organic solvent n-butyl alcohol and ethanol, the cost is higher and the method is not environment-friendly. Patent CN105254689A discloses a sennoside A.B salt compound and a preparation method and application thereof, wherein senna leaves are used as raw materials to obtain the sennoside A.B salt compound with high-purity effective components, the purity of the sennoside A.B salt compound reaches 80-90%, the method has the same complicated steps, and organic solvents with strong toxicity, such as acetone, methanol and the like, are used in the purification process of the last step.
In the prior art, the senna leaf traditional Chinese medicine is usually taken in the form of decoction or granules, and decoction of the decoction is troublesome, laggard and large in dosage, and cannot play a role in first aid. The current sennoside purification method cannot meet the existing requirements.
Disclosure of Invention
The purpose of the invention is as follows: in order to solve the problems of the prior art, the invention provides a preparation method of high-purity sennoside and the sennoside or derivatives prepared by the preparation method. The preparation method is based on macroporous adsorption chromatography column and ion exchange column chromatography to purify sennoside, and the purity and yield of the obtained sennoside extract are superior to those of the prior art, and the preparation method is low in cost and more environment-friendly.
The technical scheme is as follows: the preparation method of high-purity sennoside provided by the invention comprises the following steps:
(1) extracting medicinal materials: adding folium sennae medicinal material into sodium bicarbonate solution, stirring, soaking for 60-120min, filtering to obtain extractive solution, adding acid to adjust pH to 2.5-5.5, filtering to obtain supernatant, and adjusting pH and conductivity of the supernatant again to obtain acid-adjusted supernatant;
(2) ion exchange column chromatography: balancing the ion exchange chromatographic column by using 3-5 times of column volume balancing liquid A, and re-balancing by using 3-5 times of column volume balancing liquid A; loading the acid-adjusted supernatant obtained in the step (1) to a chromatographic column, and eluting by using eluent B with the volume of 3-5 times of that of the column to obtain eluent;
the equilibrium solution A is 0.02-0.2mol/L acetate buffer solution, 0.02-0.2mol/L glycine buffer solution, 0.02-0.2mol/L sodium bicarbonate buffer solution or 0.02-0.2mol/L phosphate buffer solution, the equilibrium solution A also contains 0.0001-0.3mol/L NaCl solution, and the pH value of the equilibrium solution A is 3.5-10.0;
the eluent B is 0.02-0.2mol/L acetate solution, 0.02-0.2mol/L glycine buffer solution, 0.02-0.2mol/L sodium bicarbonate buffer solution or 0.02-0.2mol/L phosphate buffer solution, the eluent B also contains 0.3-1.5mol/L NaCl, and the pH value is 3.5-10.0;
(3) macroporous adsorption column chromatography: adjusting the pH of the eluent obtained in the step (2) to 3.5-8.0, loading the eluent into a macroporous adsorption chromatography column, washing with a washing liquid C with the column volume of 3-5 times, washing with a washing liquid D with the column volume of 1-5 times, and eluting with an eluent E with the column volume of 3-5 times to obtain an eluent;
the flushing liquid C is 0.001-0.2mol/L glycine-hydrochloric acid buffer solution, 0.001-0.2mol/L sodium acetate-acetic acid buffer solution or 0.001-0.2mol/L phosphate buffer solution, and the pH value is 3.5-8.0; the flushing liquid D is pure water; the eluent E is 20-80% ethanol solution;
(4) and (4) carrying out low-temperature precipitation and solid-liquid separation on the eluent obtained in the step (3) to obtain the extract of the sennoside A/B compound.
As a preferred embodiment, in step (4), the low-temperature precipitation comprises the following steps: concentrating under reduced pressure, adding acid, standing at low temperature for precipitation to obtain concentrated precipitate; the solid-liquid separation comprises the following steps: and (3) carrying out solid-liquid separation on the obtained concentrated precipitation solution, dehydrating the solid with absolute ethyl alcohol for 2-3 times, discarding the supernatant, and drying the precipitate.
As a preferable embodiment, 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 or phosphoric acid; the pH value of the acid-adjusting supernatant is 3.5-10.0, and the conductivity of the acid-adjusting supernatant is 0.5-29 mS/cm.
In step (1), the pH of the acid-adjusted supernatant is 3.5, and the conductance is adjusted to 20 mS/cm; or the pH value of the acid-adjusted supernatant is 7.0, and the conductivity is 15 mS/cm; or the pH value of the acid-adjusted supernatant is 10.0, and the conductivity is 7 mS/cm.
In a preferred embodiment, in step (2), the ligand of the ion exchange chromatography column packing is an anion exchange chromatography packing of a diethylaminoethyl group or a trimethylaminylalkyl quaternary ammonium group, and the packing main framework of the ion exchange chromatography column is any one of an acrylic system or a styrene system.
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 embodiment, in the step (2), the equilibrium liquid a is selected from any one of the following: the balance liquid comprises 0.015-0.025mol/L glycine buffer solution and 0.015-0.25mol/L NaCl solution, the balance liquid comprises 0.015-0.025mol/L phosphate buffer solution and 0.010-0.20mol/L NaCl solution, and the balance liquid comprises 0.15-0.25mol/L sodium bicarbonate buffer solution and 0.010-0.15mol/L NaCl solution.
As a specific embodiment, the equilibrium solution A is a mixture of 0.02mol/L glycine buffer solution and 0.2mol/L NaCl solution, the pH is 3.5, and the conductivity is 23 mS/cm.
As a specific example, the equilibrium solution A is a mixture of 0.02mol/L phosphate buffer solution and 0.15mol/L NaCl solution, the pH is 7.0, and the conductivity is 18 mS/cm.
As a specific embodiment, the equilibrium solution A is a mixed solution of 0.2mol/L sodium bicarbonate buffer solution and 0.1mol/L NaCl solution, the pH value is 10.0, and the conductivity is 11 mS/cm.
As a preferred embodiment, the conductivity of the acid-adjusting supernatant in the step (1) is 3-4mS/cm lower than that of the equilibrium liquid A in the step (2).
As a preferred embodiment, in step (2), the eluent B is selected from any one of the following: an eluent containing 0.015-0.025mol/L glycine buffer solution and 0.3-1.2mol/L NaCl solution, an eluent containing 0.015-0.025mol/L phosphate buffer solution and 0.3-1.5mol/L NaCl solution, and an eluent containing 0.15-0.25mol/L sodium bicarbonate buffer solution and 0.3-1.0mol/L NaCl solution.
As a specific embodiment, the eluent B is a mixed solution of 0.02mol/L glycine buffer solution and 1.0mol/L NaCl solution, the pH value is 3.5, and the conductivity is 93 mS/cm.
As a specific embodiment, the eluent B is a mixed solution of 0.02mol/L phosphate buffer solution and 1.2mol/L NaCl solution, the pH value is 7.0, and the conductivity is 108 mS/cm.
As a specific embodiment, the eluent B is a mixed solution of 0.2mol/L sodium bicarbonate buffer solution and 0.8mol/L NaCl solution, the pH value is 10.0, and the conductivity is 70 mS/cm.
As a preferred embodiment, in step (3), the main framework of the macroporous adsorption chromatographic column filler is macroporous crosslinked polystyrene adsorption resin, and the model is HB-16, HB-60, HB-80, D101 or D201.
In a preferred embodiment, in step (3), the eluent E is 30-80% ethanol solution.
In a specific embodiment, the washing solution C is 0.1mol/L sodium acetate-acetic acid buffer solution with pH of 5.5.
In a specific embodiment, the washing solution C is 0.001mol/L glycine-hydrochloric acid buffer solution with pH of 3.5.
In a specific embodiment, the washing solution C is 0.2mol/L phosphate buffer solution with pH of 7.0.
As a preferred embodiment, the acid is added with hydrochloric acid, acetic acid or phosphoric acid, and the pH is adjusted to 2-3; the low-temperature standing temperature is-20-4 ℃.
In a preferred embodiment, in step (3), the rinsing solution D is pure water, and the rinsing volume is 1 to 3 column volumes.
The invention also provides sennoside prepared by the preparation method; the purity of sennoside A/B prepared by the preparation method is between 85 and 90 percent.
The sennoside A/B compound (the sennoside A and the sennoside B are stereoisomers of each other) obtained by the preparation method has the following structure:
the invention also provides a preparation method of the sennoside calcium salt, which comprises the following steps:
and (4) carrying out solid-liquid separation on the concentrated precipitate obtained in the step (4), washing the solid with pure water, removing supernatant, mixing the precipitate with a compound containing calcium ions, carrying out salt forming reaction, and drying under reduced pressure to obtain the sennoside A/B calcium salt compound.
As a preferred embodiment, a proper amount of water is added in the calcium salt reaction; the calcium salt reaction water adding amount is the volume of the acid adding solution in the step (4).
As a preferred embodiment, the compound containing calcium ions preferably comprises one or more of calcium hydroxide, calcium carbonate and calcium chloride.
As a preferred embodiment, the amount of the calcium ion-containing compound added is 0.7-1.2 molar equivalents, preferably 0.9 molar equivalent of the total of sennoside a and sennoside B contained in the senna leaf material in step (1).
As a preferred embodiment, the temperature of the reduced pressure drying is 55 ℃ to 80 ℃, preferably 70 ℃.
In the present invention, "%" is a mass% unless otherwise specified.
Has the advantages that: (1) the invention provides a method for purifying sennoside based on ion exchange chromatography and macroporous adsorption chromatography, and repeated experiments prove that the method has stable process, is beneficial to industrialized and large-scale production, can well and characteristically separate and purify sennoside A/B, has the yield of each batch of target products higher than 50 percent and the purity higher than 85 percent, and is easy for enlarged production; (2) the method solves the defects that the sennoside A/B is similar to other anthraquinone impurities in the senna and the physicochemical property is similar, and the sennoside A/B cannot be purified characteristically by using a conventional organic solvent extraction method, elutes the sennoside A/B characteristically by using sodium chloride salt, and solves the defects that the organic solvent has higher cost and is not environment-friendly; (3) the method combining the ion exchange chromatography and the macroporous resin has the advantages of simple operation, low cost and good separation effect, and can efficiently obtain the target product; (4) the method adopts a method of combining ion exchange chromatography and macroporous adsorption chromatography to separate and purify the sennoside, and the obtained extract has high purity and convenient use.
Drawings
FIG. 1 is an HPLC analytical map of the calcium salt compound of example 1 of the present invention.
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, reserving supernate as an extracting solution (sampling detection of the extracting solution), adding 6mol/L HCl to adjust the pH value to 3.0, removing part of impurities, filtering to reserve supernate, adding sodium chloride to adjust the 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) ion exchange column chromatography: 300mL of a macroporous weakly basic anion exchange chromatography column of A451 type (ZG A451, Hangzhou Guanguang resin Co., Ltd.) was previously loaded, 5 column volumes were equilibrated at a linear speed of 60cm/h with an equilibration solution A, which was 0.02mol/L glycine buffer solution containing 0.2mol/L NaCl, having a pH of 3.5 and a conductivity of 23mS/cm, and the loading solution in step (1) was loaded onto the chromatography column at a linear speed of 60 cm/h. After the sample loading is finished, firstly, washing the column by 4 times of column volume by using the equilibrium solution A at a linear speed of 60cm/h, then washing the column by using an eluent B with 5 times of column volume at a linear speed of 40cm/h, wherein the eluent B 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. The eluate (eluent 1 was sampled and detected), the pH was adjusted to 4.0, and the column was prepared for D101.
(3) Macroporous adsorption column chromatography: loading 100mL of D101 (Shanghai Min Yongchun Co., Ltd.) macroporous adsorption chromatographic column in advance, loading the sample loading solution in the step (2) into the chromatographic column at a linear speed of 40cm/h, washing the column by using washing liquid C at a linear speed of 40cm/h for 4 times of the column volume after loading, washing the column by using pure water (washing liquid D) at a linear speed of 40cm/h for 3 times of the column volume, washing the column by using eluent E at a linear speed of 40cm/h for 5 times of the column volume, wherein the washing liquid C is 0.001mol/L glycine-hydrochloric acid buffer solution, the pH is 3.5, and the eluent E is 30% ethanol solution; the eluate was collected (eluent 2 was sampled and detected).
(4) Low-temperature precipitation: and (4) concentrating the eluent in the step (3) under reduced pressure for 1 hour at the temperature of 60 ℃ and the vacuum degree of-0.08 MPa, adjusting the pH of the obtained concentrate to 2.0 by using 6mol/L HCl, and standing overnight at the temperature of 4 ℃.
(5) Solid-liquid separation: and (4) centrifuging the concentrated precipitation solution obtained in the step (4) at 4000rpm for 10min, dehydrating the solid twice by using 10mL of absolute ethyl alcohol, removing the supernatant, and drying the precipitate to obtain the powdery extract of the high-purity sennoside A/B compound.
Preparation of sennoside calcium salt: centrifuging the concentrated precipitate obtained in step (4) of the example at 4000rpm for 10min, washing the obtained solid with 10mL of purified water twice, discarding the supernatant, adding 80mg of calcium chloride into the precipitate, mixing with water equal to 6mol/L HCl in step (4), performing salt formation reaction on the obtained mixture at 55 ℃ and under the vacuum degree of-0.08 MPa, and drying under reduced pressure for 30min to obtain the sennoside A/B calcium salt compound (sampling and detecting the calcium salt compound, and detecting the HPLC chromatogram shown in figure 1). The results are shown in Table 1.
Table 1 test results of example 1 of the present invention
| Name (R) | Content of glycoside A | Content of glycoside B | Total glycoside A + glycoside B content | Yield of |
| Extract liquid | 11.00％ | 12.41％ | 23.41％ | 88.20％ |
| Acid-regulating sample loading liquid | 10.08％ | 12.57％ | 22.65％ | 88.20 |
| Eluent | ||||
| 1 | 28.93％ | 36.71％ | 65.64％ | 98.93 |
| Eluent | ||||
| 2 | 27.66％ | 38.68％ | 66.34％ | 96.44％ |
| Calcium salt compound | 29.76％ | 55.48％ | 85.24％ | 95.57％ |
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, taking supernatant as an extracting solution (sampling detection of the extracting solution), adding glacial acetic acid to adjust the pH value to 4.5, filtering to obtain supernatant, adding sodium chloride to adjust the conductance to 15mS/cm, adding 5mol/L NaOH to adjust the pH value to 7.0, and obtaining a supernatant (sampling detection of an acid-adjusted supernatant);
(2) ion exchange column chromatography: 300mL of A351 type (ZG A351 type, Hangzhou Guanguang resin Co., Ltd.) macroporous weakly basic anion exchange chromatography column is loaded in advance, 5 times of column volume is balanced by equilibrium liquid A at a linear speed of 60cm/h, the equilibrium liquid A is 0.02mol/L phosphate buffer solution, contains 0.15mol/L NaCl, has a pH value of 7.0 and an electric conductivity of 18mS/cm, and then the sample loading liquid in the step (1) is loaded on the chromatography column at a linear speed of 60 cm/h. After the sample loading is finished, firstly, washing the column by 4 times of column volume by using the equilibrium solution A at a linear speed of 60cm/h, then washing the column by using the eluent B with 5 times of column volume at a linear speed of 40cm/h, wherein the eluent B is 0.02mol/L phosphate buffer solution, contains 1.2mol/L NaCl, has the pH value of 7.0 and has the conductivity of 108 mS/cm. The eluate (eluent 1 was sampled and detected) was collected, and the pH was adjusted to 6.0.
(3) Macroporous adsorption column chromatography: pre-loading 100mL of HB-16 (Shanghai Min Yongchun Co., Ltd.) macroporous adsorption chromatography column, loading the loading solution in the step (2) into the chromatography column at a linear speed of 40cm/h, washing the column by using washing liquid C at a linear speed of 40cm/h for 4 times of column volume after loading is finished, washing the column by using pure water (washing liquid D) at a linear speed of 40cm/h for 3 times of column volume, washing the column by using eluent E at a linear speed of 40cm/h for 5 times of column volume, wherein the washing liquid C is 0.1mol/L sodium acetate-acetic acid buffer solution, the pH is 5.5, and the eluent E is 80% ethanol solution; the eluate was collected (eluent 2 was sampled and detected).
(4) Low-temperature precipitation: concentrating the eluate in step (3) under reduced pressure at 60 deg.C and vacuum degree of-0.08 MPa for 1 hr, adjusting pH of the obtained concentrate to 2.5 with 6mol/L HCl, and standing at-10 deg.C overnight.
(5) Solid-liquid separation: and (4) centrifuging the concentrated precipitation solution obtained in the step (4) at 4000rpm for 10min, dehydrating the solid twice by using 10mL of absolute ethyl alcohol, removing the supernatant, and drying the precipitate to obtain the powdery extract of the high-purity sennoside A/B compound.
Preparation of sennoside calcium salt: centrifuging the concentrated precipitate obtained in the step (4) of the embodiment at 4000rpm for 10min, washing the solid with 10mL of purified water twice, discarding the supernatant, adding 142mg of calcium carbonate into the precipitate, mixing with water equal to 6mol/L HCl in the step (4), performing salt formation reaction on the obtained mixture at the temperature of 80 ℃ and the vacuum degree of-0.08 MPa, and drying under reduced pressure for 40min to obtain the sennoside A/B calcium salt compound (sampling and detecting the calcium salt compound). The results are shown in Table 2.
Table 2 test results of example 2 of the present invention
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, taking supernatant as an extracting solution (sampling detection of the extracting solution), adding 6mol/L HCl to adjust the pH value to 5.5, filtering to obtain 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 supernatant (sampling detection of an acid-adjusted supernatant);
(2) ion exchange column chromatography: the column was previously loaded with 1500mL of a313 type (ZG A313, Hangzhou Chongguang resin Co., Ltd.) macroporous weakly basic anion exchange chromatography column, 5 column volumes were equilibrated at a linear speed of 60cm/h with an equilibration solution A, which was 0.2mol/L sodium bicarbonate buffer containing 0.1mol/L NaCl, having a pH of 10.0 and an electrical conductivity of 11mS/cm, and the loading solution in step (1) was loaded onto the column at a linear speed of 60 cm/h. After the sample loading is finished, firstly, washing the column by 4 times of column volume by using the equilibrium solution A at a linear speed of 60cm/h, then washing the column by using an eluent B with 5 times of column volume at a linear speed of 40cm/h, wherein the eluent B is 0.2mol/L sodium bicarbonate buffer solution, contains 0.8mol/L NaCl, has the pH value of 10.0 and has the conductivity of 70 mS/cm. The eluate (eluent 1 was sampled and detected) was collected, and the pH was adjusted to 8.0.
(3) Macroporous adsorption column chromatography: pre-loading 1000mL of HB-60 (Shanghai Min Yongchun Co., Ltd.) macroporous adsorption chromatography column, loading the sample loading solution in the step (2) into the chromatography column at a linear speed of 40cm/h, washing the column by using washing solution C at a linear speed of 40cm/h for 4 times of column volume after loading, washing the column by using pure water (washing solution D) at a linear speed of 40cm/h for 3 times of column volume, washing the column by using eluent E at a linear speed of 40cm/h for 5 times of column volume, wherein the washing solution C is 0.2mol/L of phosphate buffer solution, and the pH is 7.0; the eluent E is 60% ethanol solution; the eluate was collected (eluent 2 was sampled and detected).
(4) Low-temperature precipitation: concentrating the eluate in step (3) under reduced pressure at 60 deg.C and vacuum degree of-0.08 MPa for 8 hr, adjusting pH of the obtained concentrate to 3.0 with glacial acetic acid, and standing at-20 deg.C overnight.
(5) Solid-liquid separation: and (4) centrifuging the concentrated precipitation solution obtained in the step (4) at 4000rpm for 10min, dehydrating the solid twice by using 50mL of absolute ethyl alcohol, removing the supernatant, and drying the precipitate to obtain the powdery extract of the high-purity sennoside A/B compound.
Preparation of sennoside calcium salt: centrifuging the concentrated precipitate obtained in the step (4) of the embodiment at 4000rpm for 10min, washing the solid with 50mL of purified water twice, discarding the supernatant, adding 1180mg of calcium hydroxide into the precipitate, mixing with water equal in volume to the glacial acetic acid obtained in the step (4), performing salt formation reaction on the obtained mixture at the temperature of 80 ℃ and the vacuum degree of-0.08 MPa, and drying under reduced pressure for 90min to obtain the sennoside A/B calcium salt compound (sampling and detecting the calcium salt compound). The results of the measurements are shown in Table 3 below
Table 3 test results of example 3 of the present invention
| Name (R) | Content of glycoside A | Content of glycoside B | Total glycoside A + glycoside B content | Yield of |
| Extract liquid | 12.09％ | 15.11％ | 27.20％ | 96.65％ |
| Acid-regulating sample loading liquid | 11.01％ | 14.85％ | 25.86％ | 96.01 |
| Eluent | ||||
| 1 | 30.69％ | 36.31％ | 67.00％ | 92.27 |
| Eluent | ||||
| 2 | 30.14％ | 38.02％ | 68.16％ | 93.26％ |
| Calcium salt compound | 40.18％ | 50.49％ | 90.67％ | 90.13％ |
Claims (10)
1. A preparation method of high-purity sennoside is characterized by comprising the following steps:
(1) extracting medicinal materials: adding folium sennae medicinal material into sodium bicarbonate solution, stirring, soaking for 60-120min, filtering to obtain extractive solution, adding acid to adjust pH to 2.5-5.5, filtering to obtain supernatant, and adjusting pH and conductivity of the supernatant again to obtain acid-adjusted supernatant;
(2) ion exchange column chromatography: balancing the ion exchange chromatographic column by using a 3-5 times column volume balancing solution A, loading the acid-adjusted supernatant obtained in the step (1) to the chromatographic column, rebalancing by using the 3-5 times column volume balancing solution A, and eluting by using a 3-5 times column volume eluent B to obtain an eluent;
the equilibrium solution A is 0.02-0.2mol/L acetate solution, 0.02-0.2mol/L glycine buffer solution, 0.02-0.2mol/L sodium bicarbonate buffer solution or 0.02-0.2mol/L phosphate buffer solution, the equilibrium solution A also contains 0.0001-0.3mol/L NaCl solution, and the pH value of the equilibrium solution A is 3.5-10.0;
the eluent B is 0.02-0.2mol/L acetate solution, 0.02-0.2mol/L glycine buffer solution, 0.02-0.2mol/L sodium bicarbonate buffer solution or 0.02-0.2mol/L phosphate buffer solution, the eluent B also contains 0.3-1.5mol/L NaCl, and the pH value is 3.5-10.0;
(3) macroporous adsorption column chromatography: adjusting the pH of the eluent obtained in the step (2) to 3.5-8.0, loading the eluent into a macroporous adsorption chromatography column, washing with a washing liquid C with the column volume of 3-5 times, washing with a washing liquid D with the column volume of 1-5 times, and eluting with an eluent E with the column volume of 3-5 times to obtain an eluent;
the flushing liquid C is 0.001-0.2mol/L glycine-hydrochloric acid buffer solution, 0.001-0.2mol/L sodium acetate-acetic acid buffer solution or 0.001-0.2mol/L phosphate buffer solution, and the pH value is 3.5-8.0; the flushing liquid D is pure water; the eluent E is 20-80% ethanol solution;
(4) and (4) carrying out low-temperature precipitation and solid-liquid separation on the eluent obtained in the step (3) to obtain the extract of the sennoside A/B compound.
2. The method for preparing sennoside according to claim 1, wherein in step (4), the low-temperature precipitation comprises the following steps: concentrating under reduced pressure, adding acid, standing at low temperature for precipitation to obtain concentrated precipitate; the solid-liquid separation comprises the following steps: and (3) carrying out solid-liquid separation on the obtained concentrated precipitation solution, dehydrating the solid with absolute ethyl alcohol for 2-3 times, discarding the supernatant, and drying the precipitate.
3. The method for producing sennoside according to 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 or phosphoric acid; the pH value of the acid-adjusting supernatant is 3.5-10.0, and the conductivity of the acid-adjusting supernatant is 0.5-29 mS/cm.
4. The method for preparing sennoside 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 method for producing sennoside according to claim 1, wherein in the step (2), the equilibrium liquid A is selected from any one of the following: the balance liquid comprises 0.015-0.025mol/L glycine buffer solution and 0.015-0.25mol/L NaCl solution, the balance liquid comprises 0.015-0.025mol/L phosphate buffer solution and 0.010-0.20mol/L NaCl solution, and the balance liquid comprises 0.15-0.25mol/L sodium bicarbonate buffer solution and 0.010-0.15mol/L NaCl solution.
6. The method for producing sennoside according to claim 1, wherein in the step (2), the eluent B is selected from any one of the following: an eluent containing 0.015-0.025mol/L glycine buffer solution and 0.3-1.2mol/L NaCl solution, an eluent containing 0.015-0.025mol/L phosphate buffer solution and 0.3-1.5mol/L NaCl solution, and an eluent containing 0.15-0.25mol/L sodium bicarbonate buffer solution and 0.3-1.0mol/L NaCl solution.
7. The method for preparing sennoside according to claim 1, wherein in the step (3), the main framework of the macroporous adsorption chromatography column filler is macroporous crosslinked polystyrene adsorption resin.
8. The method for producing sennoside according to claim 1, wherein in the step (2), the type of the ion exchange chromatography column is A313 type, A313FC type, D620 type, D630 type, D640 type, A351 type or A451 type; in the step (3), the model of the macroporous adsorption chromatographic column is HB-16 type, HB-60 type, HB-80 type, D101 type or D201 type.
9. A sennoside or derivative prepared by the method of any one of claims 1 to 8; the derivative is a sennoside calcium salt.
10. A process for the preparation of the sennoside calcium salt according to claim 9, characterised in that it comprises the following steps: subjecting the concentrated precipitate obtained in step (4) of claim 1 to solid-liquid separation, washing the solid with pure water, removing the supernatant, mixing the precipitate with a calcium ion-containing compound, performing salt-forming reaction, and drying under reduced pressure to obtain a sennoside A/B calcium salt compound.
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