CN1748725A - Process for preparing semen momordicae seed extract containing triterpene saponin component - Google Patents
Process for preparing semen momordicae seed extract containing triterpene saponin component Download PDFInfo
- Publication number
- CN1748725A CN1748725A CN 200510060434 CN200510060434A CN1748725A CN 1748725 A CN1748725 A CN 1748725A CN 200510060434 CN200510060434 CN 200510060434 CN 200510060434 A CN200510060434 A CN 200510060434A CN 1748725 A CN1748725 A CN 1748725A
- Authority
- CN
- China
- Prior art keywords
- semen momordicae
- extract
- triterpene saponin
- semen
- under reduced
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 210000000582 semen Anatomy 0.000 title claims abstract description 78
- 239000000284 extract Substances 0.000 title claims abstract description 52
- 229930182493 triterpene saponin Natural products 0.000 title claims abstract description 15
- 238000004519 manufacturing process Methods 0.000 title 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 19
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 19
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000002360 preparation method Methods 0.000 claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 10
- 239000000843 powder Substances 0.000 claims abstract description 7
- 239000000126 substance Substances 0.000 claims abstract description 6
- 238000010992 reflux Methods 0.000 claims abstract description 4
- 239000011347 resin Substances 0.000 claims abstract description 4
- 229920005989 resin Polymers 0.000 claims abstract description 4
- 239000007864 aqueous solution Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 9
- 239000000287 crude extract Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 239000003480 eluent Substances 0.000 claims description 5
- 239000012535 impurity Substances 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 5
- 239000000470 constituent Substances 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 2
- 239000002994 raw material Substances 0.000 claims description 2
- -1 soak reflux Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 abstract description 11
- 239000007924 injection Substances 0.000 abstract description 11
- 230000002949 hemolytic effect Effects 0.000 abstract description 4
- 229960005486 vaccine Drugs 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 230000000274 adsorptive effect Effects 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 abstract 1
- 231100000053 low toxicity Toxicity 0.000 abstract 1
- 238000000746 purification Methods 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 9
- 241000699670 Mus sp. Species 0.000 description 7
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 6
- 239000007927 intramuscular injection Substances 0.000 description 6
- 238000010255 intramuscular injection Methods 0.000 description 6
- 206010018910 Haemolysis Diseases 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000008588 hemolysis Effects 0.000 description 5
- 238000010253 intravenous injection Methods 0.000 description 5
- 229930182490 saponin Natural products 0.000 description 5
- 150000007949 saponins Chemical class 0.000 description 5
- 235000017709 saponins Nutrition 0.000 description 5
- 150000008130 triterpenoid saponins Chemical class 0.000 description 5
- 231100000331 toxic Toxicity 0.000 description 4
- 230000002588 toxic effect Effects 0.000 description 4
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 3
- 241000287828 Gallus gallus Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 206010042674 Swelling Diseases 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 3
- 230000008961 swelling Effects 0.000 description 3
- 210000003462 vein Anatomy 0.000 description 3
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- LAHSXXNOJMWHBH-WVPBMNGESA-N Gypsoside Natural products O=C(O)[C@H]1[C@H](O[C@H]2[C@@H](O)[C@H](O)[C@H](O[C@@H]3[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O3)[C@H](CO)O2)[C@H](O[C@H]2[C@@H](O)[C@@H](O)[C@@H](O)CO2)[C@@H](O)[C@@H](O[C@@H]2[C@](C=O)(C)[C@@H]3[C@](C)([C@H]4[C@](C)([C@]5(C)C([C@H]6[C@](C(=O)O[C@H]7[C@H](O[C@@H]8[C@H](O[C@@H]9[C@H](O)[C@H](O)[C@H](O)CO9)[C@H](O)[C@H](O)CO8)[C@@H](O)[C@@H](O[C@@H]8[C@@H](O)[C@@H](O[C@H]9[C@@H](O)[C@@H](O)[C@@H](O)CO9)[C@H](O)[C@@H](C)O8)[C@@H](C)O7)(CC5)CCC(C)(C)C6)=CC4)CC3)CC2)O1 LAHSXXNOJMWHBH-WVPBMNGESA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 235000009815 Momordica Nutrition 0.000 description 2
- 241000218984 Momordica Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 230000002939 deleterious effect Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 241000411851 herbal medicine Species 0.000 description 2
- 239000000568 immunological adjuvant Substances 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 208000004396 mastitis Diseases 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000001117 sulphuric acid Substances 0.000 description 2
- 235000011149 sulphuric acid Nutrition 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- IZWBODJDDBCDFA-DXEZAUPJSA-N virgatic acid Chemical compound CC([C@@H]1CC[C@@]23C)(C)[C@@H](O)CC(=O)[C@]1(C)[C@H]3CC=C1[C@@]2(C)CC[C@@]2(C(O)=O)CCC(C)(C)C[C@H]21 IZWBODJDDBCDFA-DXEZAUPJSA-N 0.000 description 2
- MIJYXULNPSFWEK-GTOFXWBISA-N 3beta-hydroxyolean-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MIJYXULNPSFWEK-GTOFXWBISA-N 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 108010001394 Disaccharidases Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- JKLISIRFYWXLQG-UHFFFAOYSA-N Epioleonolsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4CCC3C21C JKLISIRFYWXLQG-UHFFFAOYSA-N 0.000 description 1
- ITTILXNWLDHOGR-UHFFFAOYSA-N Momordicasaponin I Chemical compound OC1C(O)C(O)C(C)OC1OC1C(OC2C(C(O)C(O)C(CO)O2)O)C(OC2C(C3C(C4C(C5(CCC6(CCC(C)(C)CC6C5=CC4)C(=O)OC4C(C(O)C(O)C(C)O4)OC4C(C(OC5C(C(OC6C(C(O)C(O)CO6)O)C(O)C(CO)O5)O)C(OC5C(C(O)C(O)CO5)O)C(C)O4)O)C)(C)CC3)(C)CC2)(C)C=O)OC(C(O)=O)C1O ITTILXNWLDHOGR-UHFFFAOYSA-N 0.000 description 1
- IZWBODJDDBCDFA-UHFFFAOYSA-N Momordinsaeure Natural products CC12CCC3C(C)(C)C(O)CC(=O)C3(C)C2CC=C2C1(C)CCC1(C(O)=O)CCC(C)(C)CC12 IZWBODJDDBCDFA-UHFFFAOYSA-N 0.000 description 1
- YBRJHZPWOMJYKQ-UHFFFAOYSA-N Oleanolic acid Natural products CC1(C)CC2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C1)C(=O)O YBRJHZPWOMJYKQ-UHFFFAOYSA-N 0.000 description 1
- MIJYXULNPSFWEK-UHFFFAOYSA-N Oleanolinsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MIJYXULNPSFWEK-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 241001501970 Prionailurus bengalensis Species 0.000 description 1
- MQUFAARYGOUYEV-UWEXFCAOSA-N Quillaic acid Natural products CC1(C)CC[C@@]2([C@H](O)C[C@]3(C)C(=CC[C@H]4[C@@]5(C)CC[C@H](O)[C@](C)(C=O)[C@H]5CC[C@@]34C)[C@H]2C1)C(=O)O MQUFAARYGOUYEV-UWEXFCAOSA-N 0.000 description 1
- 206010040943 Skin Ulcer Diseases 0.000 description 1
- HZWXJJCSDBQVLF-UHFFFAOYSA-N acetoxysulfonic acid Chemical compound CC(=O)OS(O)(=O)=O HZWXJJCSDBQVLF-UHFFFAOYSA-N 0.000 description 1
- 231100000215 acute (single dose) toxicity testing Toxicity 0.000 description 1
- 238000011047 acute toxicity test Methods 0.000 description 1
- JZVFJDZBLUFKCA-FXIAWGAOSA-N alpha-Spinasterol Chemical compound C1[C@@H](O)CC[C@]2(C)[C@@H](CC[C@@]3([C@@H]([C@H](C)/C=C/[C@@H](CC)C(C)C)CC[C@H]33)C)C3=CC[C@H]21 JZVFJDZBLUFKCA-FXIAWGAOSA-N 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000001571 immunoadjuvant effect Effects 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 201000007227 lymph node tuberculosis Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- MBOSQJXNWZRRFQ-UHFFFAOYSA-N momordicasaponin ii Chemical compound OC1C(O)C(O)C(C)OC1OC1C(OC2C(C(O)C(O)C(CO)O2)O)C(OC2C(C3C(C4C(C5(CC(O)C6(CCC(C)(C)CC6C5=CC4)C(=O)OC4C(C(O)C(O)C(C)O4)OC4C(C(OC5C(C(OC6C(C(O)C(O)CO6)O)C(O)C(CO)O5)O)C(OC5C(C(O)C(O)CO5)O)C(C)O4)O)C)(C)CC3)(C)CC2)(C)C=O)OC(C(O)=O)C1O MBOSQJXNWZRRFQ-UHFFFAOYSA-N 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000002850 nasal mucosa Anatomy 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229940100243 oleanolic acid Drugs 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- HZLWUYJLOIAQFC-UHFFFAOYSA-N prosapogenin PS-A Natural products C12CC(C)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OCC(O)C(O)C1O HZLWUYJLOIAQFC-UHFFFAOYSA-N 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- MQUFAARYGOUYEV-UAWZMHPWSA-N quillaic acid Chemical compound C1C[C@H](O)[C@@](C)(C=O)[C@@H]2CC[C@@]3(C)[C@]4(C)C[C@@H](O)[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MQUFAARYGOUYEV-UAWZMHPWSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 231100000019 skin ulcer Toxicity 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000925 very toxic Toxicity 0.000 description 1
- 239000000273 veterinary drug Substances 0.000 description 1
- 238000009333 weeding Methods 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Steroid Compounds (AREA)
Abstract
The preparation process of semen momordicae extract containing triterpene saponin includes crushing semen momordicae, alcohol reflux extraction, ethyl ether extraction to eliminate liposulable component, n-butanol extraction to eliminate n-butanol insoluble component, and purification with adsorptive macroporous resin to obtain yellowish scented semen momordicae extract powder. Physical and chemical analysis and hemolytic experiment show that the semen momordicae extract contains triterpene saponin component. The semen momordicae extract has low toxicity, may be used in orally taking and injection safely and has wide application foreground in vaccine preparing field.
Description
Technical field
The invention belongs to Chinese medicine extracts active ingredients technology, relate to a kind of preparation method of Chinese medicine semen momordicae seed extract containing triterpene saponin component.
Background technology
Semen Momordicae (Semen momordicae) is the dry mature seed of cucurbitaceous plant Semen Momordicae Momordica Cochinchinesis (Lour.) Spreng..Semen Momordicae is perennial liana, mainly is distributed in the southern most of area of country in Southeast Asia, Mongolia and China, as provinces such as Guangxi, Sichuan, Hubei, Hunan, Guizhou, Yunnan, Guangdong, Anhui.The mature fruit of gathering general winter is cut open, shines to half-dried, removes sarcocarp, takes out seed, dries standby.Semen Momordicae is a conventional Chinese medicine, and bitter in the mouth, little sweet, cool in nature has mass dissipating and swelling eliminating, and counteracting toxic substances is treated the function of skin ulcer.Record in the Northern Song Dynasty " Kaibao Bencao " (973 years Christian eras) Semen Momordicae is controlled " married woman's acute mastitis, swelling and pain of anus ".This medicine has recorded in " Chinese pharmacopoeia and " Chinese veterinary drug allusion quotation " now.Be usually used in treating infectious disease such as acute mastitis, diarrhoea, toxic swelling, scrofula on people doctor and the veterinary clinic.
Semen Momordicae has been used more than 1000 year clinically as an infection Chinese medicine.Semen Momordicae can be used by single, also can form prescription with the other drug compatibility and use.
Chemical analysis shows compositions such as Semen Momordicae fatty acids, Saponin, protein and hitodesterol, oleanolic acid, momordic acid.Nearest a kind of water extract of discovering Semen Momordicae has antitumor (Tien et al., 2005; Tsoi et al., 2004; Wonget al., 2004) and antioxidant activity (Tsoi et al., 2005).Iwamnoto etc. (1985) isolate two kinds of Saponins from Semen Momordicae, momordica saponin I and II, and confirm that I is a gypsoside, II is a kind of Saponin that contains quillaic acid.Gypsoside is the triterpenoid saponin that contains the disaccharidase chain.Some Saponins with similar chemical constitution have been proved to be has immunoadjuvant function and vaccine is used the immune effect that can improve vaccine simultaneously.
Yet Semen Momordicae is considered to a deleterious Chinese medicine in many medical literatures.Zhang Jingyue is put down in writing in " book on Chinese herbal medicine is just ": " Semen Momordicae, very toxic, its sweet temperature of book on Chinese herbal medicine speech is nontoxic, and Miao is also.The present is seen malicious Canis familiaris L. person, can get killed in momentariness." the Semen Momordicae infusion, animals such as ethanol infusion and alcohol leaching liquid vein or intramuscular injection Canis familiaris L. cat rabbit are injected animal blood pressure and reduced, and are dead within a few days; The mice LD of Semen Momordicae Saponin
50Studies show that intravenous injection is 32.35mg/kg, lumbar injection is 37.34mg/kg.
In order to open up the using value of China's Chinese medicine, have at present semen momordicae extract containing triterpene saponin at the applied research aspect the immunological adjuvant, yet, the approach that adjuvant uses for be included in vaccine in oral or inject, this makes tradition can not directly apply to adjuvant based on the deleterious Semen Momordicae of external, the objective of the invention is to prepare the extract that contains the triterpenoid saponin effective ingredient in the Semen Momordicae, remove its toxic component, make the application safety, reliable of Semen Momordicae.
Summary of the invention
The invention provides a kind of preparation method of Chinese medicine semen momordicae seed extract containing triterpene saponin component.Resulting Semen Momordicae extract is a kind of toxic yellow powder that contains triterpene saponin component of removing.
The preparation method of semen momordicae seed extract containing triterpene saponin component of the present invention may further comprise the steps:
(1) the raw material Semen Momordicae is pulverized the back and in 40～50% ethanol, soak reflux, extract, 2～5 times, concentrating under reduced pressure behind the extracting liquid filtering;
(2) in concentrated solution, add clarifier and remove impurity, get supernatant after leaving standstill, remove liposoluble constituent, keep the aqueous solution of water layer with extracted with diethyl ether;
(3) aqueous solution water saturation n-butanol extraction, the concentrating under reduced pressure butanol extraction liquid gets yellow powder shape Semen Momordicae crude extract to doing;
(4) with 50～80 ℃ of hot water dissolving's crude extracts, regulate pH to 6～7, fully after the dissolving, water-soluble Semen Momordicae crude extract is added the adsorptivity macroporous resin column, leave standstill 15～30 hours after, the impurity of non-active ingredients is removed in washing;
(5) with 40～50% ethanol elutions, the concentrating under reduced pressure eluent with spissated eluent vacuum drying, gets yellow powder shape Semen Momordicae extract.
The Semen Momordicae crude extract or the Semen Momordicae extract product that obtain by above method are product of the present invention, contain the triterpenoid saponin chemical constituent in this product.
The specific embodiment
Embodiment one
30 kilograms of Semen Momordicaes (available from Bozhou, Anhui Province Chinese Medicinal Materials Markets) divide 3 extractions, each 10 kilograms.At first Semen Momordicae is pulverized, in 40～50% ethanol, soaked 24 hours; Then, alcohol reflux 3 times, each 2 hours; Extracting liquid filtering, concentrating under reduced pressure filtrate (R502B rotary evaporator is available from Shensheng Science ﹠ Tech. Co., Ltd., Shanghai); Concentrated solution adds 101 clarifiers (inferior biological engineering company limited is irrigated in Shanghai), and making ultimate density is 5 ‰ (101 clarifiers and concentrated solution w/vs), behind the abundant mixing, heating in water bath to 60 ℃, leaves standstill 24 hours; Get supernatant, use extracted with diethyl ether 3 times, with the weeding of grease soluble components; Water layer adding water-saturated n-butanol extraction 4 times, to remove the composition that is insoluble to n-butyl alcohol, concentrating under reduced pressure n-butyl alcohol collection liquid gets yellow residue to doing; Hot water washes out residue, regulates pH to 6～7 with the 1N sodium hydroxide, and residue is fully dissolved; Solution is added to adsorptivity macroporous resin (D101A, Tianjin sea light chemical industry company limited product) post, left standstill 24 hours, the absorption effective ingredient; Remove impurity, reuse 40～50% ethanol eluting with distillation washing post; The concentrating under reduced pressure eluent; The concentrated solution vacuum drying gets faint yellow Semen Momordicae extract powder, and slightly fragrant has zest to nasal mucosa.Three extractions obtain Semen Momordicae 20.58g, 22.10g and 19.63g respectively, obtain extract 62.31g from the 30kg Semen Momordicae altogether, and average yield is 2.077g/kg.
Embodiment two
The kind Renhe hard kind skin of Semen Momordicae is separated, get kernel 0.597kg and hard kind skin 0.401kg.Extract kind of a Renhe hard kind skin as stated above respectively.Obtain 0.45g from 0.401kg hard episperm, yield is 1.12g/kg; Obtain 2.02g from the 0.597kg kernel, yield is 3.38g/kg.
The analysis of Semen Momordicae extract character: be the character of confirmation the inventive method gained Semen Momordicae extract, and whether contain triterpenoid saponin, the spy carries out following test.
1. acetic anhydride-strong sulfuric acid response (Liebermann-Burchard reaction): get the 5mg Semen Momordicae extract and be dissolved in the 1ml acetic anhydride, add concentrated sulphuric acid-acetic anhydride (1: 20) 5ml, begin to appear as yellowish-brown, become dark brown then.
2. chloroform-strong sulfuric acid response (Salkowski reaction): get the 5mg Semen Momordicae extract and be dissolved in the 1ml chloroform, the back adds the 1ml concentrated sulphuric acid, and the concentrated sulphuric acid layer of bottom begins to be yellowish-brown, becomes dark brown then.
3. surface activity: the aqueous solution 2ml that gets Semen Momordicae extract is in tool plug test tube, and firmly jolting can produce the persistency foam, and not disappear because of heating.
4. ultraviolet spectral analysis: preparation 0.1mg/ml Semen Momordicae extract aqueous solution, with ultraviolet spectrophotometer (Ultrospec3300pro, Biochrom Ltd company) scanning, the uv absorption of observing the Semen Momordicae extract aqueous solution.Under ultraviolet spectrophotometer, in the 190nm-1100nm scope, scan, maximum absorption band is arranged at the 200nm place.
Above results suggest Semen Momordicae extract contains triterpenoid saponin.
5, the haemolysis of Semen Momordicae extract
Get sodium citrate anticoagulant Sanguis Gallus domesticus, 1500rpm10 minute is centrifugal, supernatant discarded, and it is inferior to give a baby a bath on the third day after its birth with normal saline, is made into 2% chicken erythrocyte suspension.The Semen Momordicae extract aqueous solution of preparation 5mg/ml adds the 50ul normal saline in 1～12 hole of V-type 96 hole blood-coagulation-boards, add 50ul Semen Momordicae extract solution in the 1st hole, the two-fold dilution, add the 50ul chicken erythrocyte suspension in each hole then, cultivated observation haemolysis situation 1 hour in 37 ℃.According to formula V/PS calculate hemolytic index (hemolytic index, HI), V in the formula: the cumulative volume (ml) of every hole liquid; P: the percent concentration that makes the minimum Semen Momordicae extract of erythrocyte hemolysis; S: the volume (ml) that adds Semen Momordicae extract solution in the hand-hole.The least concentration Semen Momordicae extract solution that occurs haemolysis on 96 orifice plates is positioned at the 7th hole, and this hole Semen Momordicae extract concentration is 0.0039%, according to the hemolytic index computing formula, and HI=V/PS=100 μ l/ (0.0039% * 50 μ l)=51200.
Haemolysis prompting Semen Momordicae extract contains triterpene saponin component.
6, the acute toxicity test of Semen Momordicae extract
(1) Semen Momordicae extract intravenous injection LD
50Mensuration
Get body weight and be 80 of the ICR mices of 19～21g, male and female half and half are divided into 4 groups at random, and 20 every group, male and female half and half.Tail vein injection Semen Momordicae extract aqueous solution, each is organized injected dose and is respectively 115.50mg/kg, 109.73mg/kg, 104.24mg/kg and 99.03mg/kg.With the sterilization of 75% alcohol swab afterbody, slowly injection after the injection, is observed the reaction of mice immediately, carries out record before the injection.The LD of intravenous injection Semen Momordicae extract
50Be 108.05mg/kg, 95% fiducial limit scope is 105.78～110.36mg/kg;
(2) Semen Momordicae extract intramuscular injection LD
50Mensuration
Get body weight and be 100 of the ICR mices of 19～21g, male and female half and half are divided into 5 groups at random, and 20 every group, male and female half and half.Leg muscle injection Semen Momordicae extract aqueous solution, each is organized injected dose and is respectively 240.00mg/kg, 192.00mg/kg, 153.60mg/kg, 122.90mg/kg and 98.30mg/kg, sterilize with 75% alcohol swab shank before the injection, slowly injection, after the injection, observe the reaction of mice immediately, carry out record.The LD of intramuscular injection Semen Momordicae extract
50Be 178.09mg/kg, 95% fiducial limit scope is 160.10～198.09mg/kg.
The LD of intravenous injection Semen Momordicae extract
50Be 108.05mg/kg, the LD that the intramuscular injection Semen Momordicae extract is
50Be 178.09mg/kg, vein and lumbar injection are all fool proof.
Above result shows, mainly contains triterpene saponin component by the resulting Semen Momordicae extract of preparation method of the present invention, has removed toxicity, and intravenous injection and intramuscular injection are all fool proof to mice.Intramuscular injection mice, milch cow and pig do not see the local excitation reaction.Therefore, the preparation method place to go that the present invention adopts the toxic component in the Semen Momordicae, make the application of semen momordicae seed extract containing triterpene saponin component safer.
Claims (2)
1, Semen Momordicae contains the preparation method of triterpene saponin component extract, it is characterized in that following steps:
(1) the raw material Semen Momordicae is pulverized the back and in 40～50% ethanol, soak reflux, extract, 2～5 times, concentrating under reduced pressure behind the extracting liquid filtering;
(2) in concentrated solution, add clarifier and remove impurity, get supernatant after leaving standstill, remove liposoluble constituent, keep the aqueous solution of water layer with extracted with diethyl ether;
(3) aqueous solution water saturation n-butanol extraction, the concentrating under reduced pressure butanol extraction liquid gets yellow powder shape Semen Momordicae crude extract to doing;
(4) with 55～65 ℃ of hot water dissolving's crude extracts, regulate pH to 6～7, fully after the dissolving, water-soluble Semen Momordicae crude extract is added the adsorptivity macroporous resin column, leave standstill 15～30 hours after, the impurity of non-active ingredients is removed in washing;
(5) with 40～50% ethanol elutions, the concentrating under reduced pressure eluent with spissated eluent vacuum drying, gets yellow powder shape Semen Momordicae extract.
2, contain the preparation method of triterpene saponin component extract by the described Semen Momordicae of claim 1, it is characterized in that: a kind of Semen Momordicae crude extract or Semen Momordicae extract product that obtains by above method contains the triterpene saponin chemical constituent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100604341A CN100335073C (en) | 2005-08-19 | 2005-08-19 | Process for preparing semen momordicae seed extract containing triterpene saponin component |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100604341A CN100335073C (en) | 2005-08-19 | 2005-08-19 | Process for preparing semen momordicae seed extract containing triterpene saponin component |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1748725A true CN1748725A (en) | 2006-03-22 |
| CN100335073C CN100335073C (en) | 2007-09-05 |
Family
ID=36604502
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100604341A Expired - Fee Related CN100335073C (en) | 2005-08-19 | 2005-08-19 | Process for preparing semen momordicae seed extract containing triterpene saponin component |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100335073C (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101224231B (en) * | 2008-01-25 | 2010-06-09 | 浙江大学 | Method of extracting immunologic adjuvant component from cochinchina momordica seed |
| EP2207559A2 (en) * | 2007-10-22 | 2010-07-21 | SK Chemicals, Co., Ltd. | Wound-healing agent containing momordicae semen extract |
| CN104710487A (en) * | 2008-04-08 | 2015-06-17 | 索隆-基特林癌症研究协会 | Triterpene saponins, methods of synthesis, and uses thereof |
| CN106243163A (en) * | 2016-08-05 | 2016-12-21 | 上海交通大学 | The method preparing high-purity chemical reference substance momordin I |
| CN116298054A (en) * | 2023-03-30 | 2023-06-23 | 北京康仁堂药业有限公司 | Thin layer identification method for Momordica cochinchinensis products |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1108308C (en) * | 1999-04-05 | 2003-05-14 | 桂林集琦药业股份有限公司 | Gymnema sylvestre extract, its preparation method and application |
-
2005
- 2005-08-19 CN CNB2005100604341A patent/CN100335073C/en not_active Expired - Fee Related
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2207559A2 (en) * | 2007-10-22 | 2010-07-21 | SK Chemicals, Co., Ltd. | Wound-healing agent containing momordicae semen extract |
| JP2011510912A (en) * | 2007-10-22 | 2011-04-07 | エスケー ケミカルズ カンパニー リミテッド | Lacanka extract-containing wound healing agent |
| EP2207559A4 (en) * | 2007-10-22 | 2011-10-26 | Sk Chemicals Co Ltd | Wound-healing agent containing momordicae semen extract |
| CN101224231B (en) * | 2008-01-25 | 2010-06-09 | 浙江大学 | Method of extracting immunologic adjuvant component from cochinchina momordica seed |
| CN104710487A (en) * | 2008-04-08 | 2015-06-17 | 索隆-基特林癌症研究协会 | Triterpene saponins, methods of synthesis, and uses thereof |
| US9718850B2 (en) | 2008-04-08 | 2017-08-01 | Memorial Sloan-Kettering Institute For Cancer Research | Triterpene saponins, methods of synthesis and uses thereof |
| US11447515B2 (en) | 2008-04-08 | 2022-09-20 | Sloan Kettering Institute For Cancer Research | Triterpene saponins, methods of synthesis and uses thereof |
| CN106243163A (en) * | 2016-08-05 | 2016-12-21 | 上海交通大学 | The method preparing high-purity chemical reference substance momordin I |
| CN116298054A (en) * | 2023-03-30 | 2023-06-23 | 北京康仁堂药业有限公司 | Thin layer identification method for Momordica cochinchinensis products |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100335073C (en) | 2007-09-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US7087252B2 (en) | Medicinal preparation containing phenylethanoid glycosides extracted from herbaceous plant, Cistanche tubulosa (Schrenk.) Wight, process of making the same, and uses of the same | |
| US4511559A (en) | Biologically active polysaccharide concentrates and process for production of preparates containing such substances | |
| KR20040029072A (en) | Composition comprising wenguanguo extracts, methods for preparing same and uses thereof | |
| CN1468859A (en) | Longbract cattail general flavone extractive and its prepn and use | |
| CN100335073C (en) | Process for preparing semen momordicae seed extract containing triterpene saponin component | |
| JP2009542634A (en) | Pharmaceutical composition for the prevention and treatment of inflammatory diseases comprising extract | |
| CN112190593B (en) | Polysaccharide composition for inhibiting TRPV1 pathway and preparation method and application thereof | |
| CN113413392A (en) | Eucommia ulmoides extract composition and preparation method and application thereof | |
| CN105998103A (en) | Chestnut flower activated extract and preparation method and application thereof | |
| CN1846752A (en) | Belladonna extract preparing process | |
| JP4448034B2 (en) | Application of ramyokan extract in drug preparation | |
| WO1997004791A1 (en) | Antiviral agent and process for preparing the same | |
| CN112791137A (en) | Three schisandra fruit extracts, preparation process and application thereof | |
| CN1225998C (en) | Cigarette additive extracted from natural plant, and extracting method and use in cigarette thereof | |
| CN105943572A (en) | Chinese chestnut flower ethyl acetate extract and preparation method and application thereof | |
| CN112250655B (en) | Two novel cyclic diphenylheptanes compounds, preparation method and application thereof | |
| EP2793917B1 (en) | Extract of rhus copallina extract for use as a medicament | |
| CN113082085A (en) | Cistanche tubulosa extract and preparation method thereof | |
| EP1498131B1 (en) | Medicinal preparation containing phenylethanoid glycosides extracted from Cistanche tubulosa | |
| TWI618540B (en) | Composition for preventing renal toxicity caused by drug toxicity, preparation method thereof and Its use | |
| KR100672036B1 (en) | Asthma Therapeutics using Lonicera japonica extracts | |
| CN103083376B (en) | Can be used for the Rhizoma Ligustici extract for the treatment of rhinitis | |
| CN1628754A (en) | Local externally applied itch stopping garlic paint | |
| Aldhafiri et al. | Hepatoprotective studies of methanolic leaf extract of Barringtonia acutangula (L) against carbon tetrachloride induced liver injury in wistar rats | |
| AU2008300612C1 (en) | Production of leaf extracts of Pittosporum phillyraeoides and the use thereof in medicine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070905 Termination date: 20110819 |