CN109045100A - Red kidney bean bark extract, and its preparation method and application - Google Patents

Red kidney bean bark extract, and its preparation method and application Download PDF

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Publication number
CN109045100A
CN109045100A CN201811033012.9A CN201811033012A CN109045100A CN 109045100 A CN109045100 A CN 109045100A CN 201811033012 A CN201811033012 A CN 201811033012A CN 109045100 A CN109045100 A CN 109045100A
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China
Prior art keywords
kidney bean
red kidney
bark extract
skin
red
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CN201811033012.9A
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Chinese (zh)
Inventor
李志勇
黄建湘
武庆荣
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Shanxi Lulihua Biotechnology Development Co Ltd
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Shanxi Lulihua Biotechnology Development Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

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  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Epidemiology (AREA)
  • Medical Informatics (AREA)
  • Botany (AREA)
  • Biotechnology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention discloses red kidney bean bark extract, and its preparation method and application, belong to biomedicine field.A kind of red kidney bean bark extract is that ethanol solution after red kidney bean skin drying and crushing, will be added to carry out ultrasonic extraction acquisition.The preparation method of red kidney bean bark extract, step are as follows: (1) red kidney bean is impregnated, separate skin of beancurd, dry, crushing, is made red kidney bean skin powder by the red kidney bean skin separated；(2) by red kidney bean skin powder alcohol solution dipping, ultrasonic extraction is concentrated under reduced pressure after filtering extracting solution to get red kidney bean bark extract.Ultrasonic extraction red kidney bean skin effective component of the present invention, recovery rate is high, and simple process, extract has very strong inhibiting effect to melanoma cells, disclosing red kidney bean bark extract is the potential drug for treating melanoma, is laid a good foundation for exploitation anti-melanin tumor medicine.

Description

Red kidney bean bark extract, and its preparation method and application
Technical field
The invention belongs to biomedicine fields, are related to red kidney bean, and in particular to a kind of red kidney bean bark extract and its preparation Methods and applications.
Background technique
Red kidney bean (Phaseolus vulgaris Linn. sp.) red Kidney bean of scientific name is Papillionoideae (Faboideae) Phaseolus (Phaseolus L.) plant.The main component of red kidney bean is protein and crude fibre, also contains ammonia Base acid, vitamin and calcium, iron wait until various trace elements, have very high edible and medical value, human body itself can be improved Immunity, enhancing disease resistance.At present about the further technological processing way of red kidney bean, drink mainly is prepared by raw material of red kidney bean, Or supercritical carbon dioxide extracting its effective component, and red kidney bean skin is then used as byproduct to be used to prepare feed etc..
Red kidney bean skin as the byproduct in red kidney bean process, equally rich in protein, cellulose, vitamin and Calcium, iron wait until various trace elements, meanwhile, rich in there is selenium in the red kidney bean skin of Shanxi area plantation, selenium is in animal and human body The important component of some antioxidases (glutathione peroxidase) and selenium-P albumen, plays balance oxidation in vivo The effect of atmosphere is restored, research, which has shown that have, improves animal immunizing power effect, and selenium is for immunity influence and cancer in the world The research of prevention is the hot issue in the field.
The purpose of the present invention is carrying out waste utilization to red kidney bean skin, its effective component is extracted, red kidney bean deep processing is widened Approach improves its application value.
Summary of the invention
The object of the present invention is to provide red kidney bean bark extract, and its preparation method and application.
The present invention is achieved by the following technical solutions:
A kind of red kidney bean bark extract is that ethanol solution after red kidney bean skin drying and crushing, will be added to carry out ultrasonic extraction acquisition.
Further, the invention discloses the preparation method of red kidney bean bark extract, steps are as follows:
(1) red kidney bean is impregnated, separates skin of beancurd, dry, crushing, is made red kidney bean skin powder by the red kidney bean skin separated；
(2) by red kidney bean skin powder alcohol solution dipping, ultrasonic extraction is concentrated under reduced pressure after filtering extracting solution to get red rue Skin of beancurd extract.
Wherein, the liquid-solid ratio of the mass concentration 40-90% of the ethyl alcohol, ethanol solution and red kidney bean skin powder is 3-20ml/ g。
When the described ultrasonic extraction, ultrasonic extraction 1-3 times, each 10-45 min.
Further, the application the invention also discloses red kidney bean bark extract in preparation anti-melanin tumor medicine.
Red kidney bean skin waits until various trace elements rich in protein, cellulose, vitamin and calcium, iron, selenium, crushed ... After mesh, under normal temperature conditions plus ethanol solution carry out ultrasonic extraction, can be effectively prevented heat-sensitive ingredients and chemically unstable at The oxidation and decomposition divided, to achieve the purpose that sufficiently to extract the effective component in red kidney bean.
Melanoma is clinically one of relatively conventional, the fastest-rising malignant tumour of disease incidence, and annual growth is 3%—5%.China's melanoma is also in the trend that is doubled and redoubled in recent years, about 20,000 people of annual new cases.Since melanoma is one The skin neoplasin for the high malignancy that kind easily shifts, the death rate is also malignant tumour of skin first, and survival rate is less than 5% within 5 years. The malignant mela noma phenomenon that there are misdiagnosis rates is higher, treatment rate is low and prognosis is poor at present.Therefore, melanoma has become sternly Jeopardize one of the disease of the health of our people again.
In order to verify red kidney bean bark extract to the resistance of melanoma, external melanoma activity test is carried out.
(1) cell culture
DMEM culture medium is added after the B16-F10 mouse skin melanoma cells recovery frozen, is placed in 37 DEG C, 5% titanium dioxide It is cultivated in carbon incubator；Cell in culture bottle carries out cell passage when growing into 85%~95%；When passage, old culture is discarded PBS repeated flushing 2 times, 0.25% pancreatin digestive juice, 500 μ L is added in liquid, and after attached cell falls off, 2 mL culture is added Liquid terminates digestion, and 1500 r/min are centrifuged 5 min, abandons supernatant, is transferred in new sterile culture flask and is cultivated with 1:3 ratio.
(2) medicine ordinance
Red kidney bean bark extract powder is accurately weighed, is made into 22 mg/mL solution, -20 DEG C of preservations with dimethyl sulfoxide (DMSO) It is spare, face the initial concentration that the used time is diluted to 0.1 mg/mL with culture medium, then be diluted to a series of gradient concentrations i.e. by 2 times 0.05、0.025、0.0125、0.00625、0.0030、0.0015 mg/mL。
(3) Activity determination
The B16-F10 mouse skin melanoma cells of logarithmic growth phase, with 5 × 104 The density of a/mL is inoculated in 96 holes In plate, every 100 μ L of hole；After being incubated for 24 h in 37 DEG C, the insulating box of 5%CO2, supernatant is abandoned, gained is added by 100 holes μ L/ Red kidney bean bark extract, is incubated for 24 h, and every hole is added 10 μ L of MTT reagent, is placed in 37 DEG C of incubators and is incubated for 4 h, microplate reader Absorbance (A) value that each hole is detected at 490 nm calculates red kidney bean skin and takes object to B16-F10 mouse skin melanoma cells Inhibiting rate and IC50Value.
Inhibiting rate (%)=(OD control-OD experiment)/OD control × 100%.
Experiment is set to be repeated three times, is as a result averaged.Red kidney bean bark extract is thin to B16-F10 mouse skin melanoma The inhibiting rate of born of the same parents, experimental result are shown in Table 1.
Calculation of half inhibitory concentration IC50, experimental result is shown in Table 1.
Inhibiting rate and half-inhibitory concentration of the 1 red kidney bean bark extract of table to B16-F10 mouse skin melanoma cells (IC50, μ g/mL)
The experimental results showed that red kidney bean bark extract has very strong inhibiting effect to B16-F10 mouse skin melanoma cells, And IC50Value is 35.42 μ g/mL.
Ultrasonic extraction red kidney bean skin effective component of the present invention, recovery rate is high, and simple process, extract is to melanoma cells There is very strong inhibiting effect, disclosing red kidney bean bark extract is the potential drug for treating melanoma, to develop anti-melanin Tumor medicine is laid a good foundation.
Specific embodiment
The invention will be further described with reference to embodiments.
Embodiment 1
A kind of preparation method of red kidney bean bark extract, step are as follows:
(1) 500 g red kidney beans are taken, soaked overnight separates skin of beancurd, dry, pulverize, and crosses 400 meshes, obtains red kidney bean skin powder；
(2) ethanol solution for being 14 ml/g by red kidney bean skin powder addition liquid-solid ratio, concentration of alcohol 60%, ultrasonic extraction 3 times, often Secondary 30 min, gained extracting solution merge, filtering, and filtrate is concentrated under reduced pressure and obtains 2.2 g red kidney bean bark extracts.
Embodiment 2
A kind of preparation method of red kidney bean bark extract, step are as follows:
(1) 500 g red kidney beans are taken, soaked overnight separates skin of beancurd, dry, pulverize；
(2) ethanol solution that the red kidney bean skin powder addition liquid-solid ratio obtained step (1) is 10 ml/g, 70 % of concentration of alcohol, Ultrasonic extraction 2 times, 45 min, gained extracting solution merge every time, filtering, and filtrate is concentrated under reduced pressure and obtains 2.1 g red kidney bean bark extracts.
The preparation of 3 red kidney bean bark extract of embodiment
(1) 500 g red kidney beans are taken, soaked overnight separates skin of beancurd, dry, pulverize；
(2) ethanol solution that the red kidney bean skin powder addition liquid-solid ratio obtained step (1) is 3 ml/g, 40 % of concentration of alcohol, Ultrasonic extraction 3 times, 20 min, gained extracting solution merge every time, filtering, and filtrate is concentrated under reduced pressure and obtains 2.2 g red kidney bean bark extracts.
The preparation of 4 red kidney bean bark extract of embodiment
(1) 500 g red kidney beans are taken, soaked overnight separates skin of beancurd, dry, pulverize；
(2) ethanol solution that the red kidney bean skin powder addition liquid-solid ratio obtained step (1) is 20ml/g, 90 % of concentration of alcohol, Ultrasonic extraction 3 times, 10 min, gained extracting solution merge every time, filtering, and filtrate is concentrated under reduced pressure and obtains 2.2 g red kidney bean bark extracts.

Claims (5)

1. a kind of red kidney bean bark extract, which is characterized in that be that ethanol solution after red kidney bean skin drying and crushing, will be added to carry out ultrasound Extract acquisition.
2. the preparation method of red kidney bean bark extract described in claim 1, which is characterized in that step are as follows:
(1) red kidney bean is impregnated, separates skin of beancurd, dry, crushing, is made red kidney bean skin powder by the red kidney bean skin separated；
(2) by red kidney bean skin powder alcohol solution dipping, ultrasonic extraction is concentrated under reduced pressure after filtering extracting solution to get red rue Skin of beancurd extract.
3. the preparation method of red kidney bean bark extract according to claim 2, which is characterized in that the quality of the ethyl alcohol The liquid-solid ratio of concentration 40-90%, ethanol solution and red kidney bean skin powder is 3-20ml/g.
4. the preparation method of red kidney bean bark extract according to claim 2, which is characterized in that the ultrasonic extraction When, ultrasonic extraction 1-3 times, each 10-45 min.
5. application of the red kidney bean bark extract described in claim 1 in preparation anti-melanin tumor medicine.
CN201811033012.9A 2018-09-05 2018-09-05 Red kidney bean bark extract, and its preparation method and application Pending CN109045100A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109528811A (en) * 2018-12-24 2019-03-29 山西大学 Red kidney bean bark extract is preparing the application in medicament for resisting cervical cancer
CN112970909A (en) * 2021-04-07 2021-06-18 山西农业大学 Method for extracting effective components of green kernel and black bean skin, composition, preparation method of medicinal tea and application

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102453014A (en) * 2010-10-22 2012-05-16 王明军 Method for extracting proanthocyanidin in broad bean skin
CN106038618A (en) * 2016-05-11 2016-10-26 南京泽朗医药科技有限公司 A method of preparing a Chinese torreya extract product and an antitumor function of the extract product

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102453014A (en) * 2010-10-22 2012-05-16 王明军 Method for extracting proanthocyanidin in broad bean skin
CN106038618A (en) * 2016-05-11 2016-10-26 南京泽朗医药科技有限公司 A method of preparing a Chinese torreya extract product and an antitumor function of the extract product

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ERLEI WANG ET AL.: ""Antiproliferative and proapoptotic activities of anthocyanin and anthocyanidin extracts from blueberry fruits on B16-F10 melanoma cells"", 《FOOD&NUTRITION RESEARCH》 *
郝瑞林等: ""大孔树脂纯化红芸豆种皮花色苷"", 《安徽农业科学》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109528811A (en) * 2018-12-24 2019-03-29 山西大学 Red kidney bean bark extract is preparing the application in medicament for resisting cervical cancer
CN112970909A (en) * 2021-04-07 2021-06-18 山西农业大学 Method for extracting effective components of green kernel and black bean skin, composition, preparation method of medicinal tea and application
CN112970909B (en) * 2021-04-07 2023-03-21 山西农业大学 Method for extracting effective components of green kernel and black bean skin, composition, preparation method of medicinal tea and application

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