CN105106317B - The preparation and application of Southeast Asia herbal combination extract - Google Patents
The preparation and application of Southeast Asia herbal combination extract Download PDFInfo
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- CN105106317B CN105106317B CN201510562784.1A CN201510562784A CN105106317B CN 105106317 B CN105106317 B CN 105106317B CN 201510562784 A CN201510562784 A CN 201510562784A CN 105106317 B CN105106317 B CN 105106317B
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Abstract
The invention discloses a kind of preparation of Southeast Asia herbal combination (ear leaf cassia leaf, Jin Zhuoye and cat's whisker blade of grass) extract and the applications of inhibition alpha-amylase activity and antioxidant activity.The active constituent is by ear leaf cassia leaf, Jin Zhuoye and cat's whisker leaf extract carrying out that alpha-amylase activity and antioxidant activity isoreactivity is inhibited to screen obtained moderately polar active component ingredient.Preparation process is green safe, step is simply original, and extract obtained stability is good, efficacy lasting.It can be used for assisting in the treatment of diabetes and anti-inflammatory, anti-aging application.
Description
1. technical field
The invention belongs to pharmaceutical fields, are related to a kind of Southeast Asia herbal-composition, further relate to the preparation method of the composition
And application.
2. background technique
2.1 ear leaf cassia leafs (Cassia auriculata, CA)
Ear leaf cassia leaf is derived from the leaf of Rosales leguminosae cassia ear leaf Cassia, and dissection uses after drying.
Ear leaf Cassia is referred to as Avarai in light Mir's language, widely distributed in states such as India, Sri Lanka.It is passed in India
In medicine of uniting, ear leaf cassia leaf be used to treat diabetes, rheumatoid, asthma and skin disease.It is main in ear leaf Semen Cassiae extract
Comprising flavonoids and phenolic compound, and it is reported and can be used for Protection hepatic lesion.
2.2 Jin Zhuoye (also known as money banyan, fan banyan;Ficus deltoidea, FD or Mas Cotek)
Jin Zhuoye is derived from the leaf of Urticales Moraceae Ficus Jin Zhuoye, and dissection uses after drying.
Jin Zhuoye is long ago just widely used by the aboriginal on the Malay Peninsula and Borneo, as their tradition
Health body-building, the natural herbal of disease preventing and treating.The authoritative research aircraft of Malaysian agricultural research development office and forest research office
Structure chemical examination test proves: the Jin Zhuoye grown in the original tropical rain forest reserved area of Malaysia contains abundant and good natural
Composition, such as: flavonoids, tannic acid/tannic acid, triterpenoid, phenol etc..Assisting in treating hypertension is used for by locals, is downgraded
Blood glucose is high, alleviates diabetic symptom.
2.3 cat's whisker blades of grass (Orthosiphon aristatus, OA or Misai Kucing)
Cat's whisker blade of grass is derived from the leaf of Lamiaceae plant Mao Xu Cao.Whole year can adopt, dissection, use after drying.
Cat's whisker blade of grass is the medicinal herbs of Malaysian many years institute happiness, and effective component mainly has following three classes: 1, flavonoids
Close object;2, liposoluble ingredient: (caffeic acid, Rosmarinic acid etc.);3, other compositions: Nan Huangsu, amount ramie acid, mono- amyrin of a, paddy
Sterol, Daucosterol, ursolic acid, despinning Inositol etc..Cat's whisker blade of grass has the refrigerant anti-inflammatory effect of diuresis, civil to be mainly used for treating
Acute and chronic nephritis, cystitis, lithangiuria and rheumatic arthritis diabetes patients Mao Xu Cao kidney tea can prevent chronic renal
Functional failure.
It is still insufficient for the research of three kinds of herbal medicine at present although 2.4 have very long applicating history and receive good efficacy,
There is no report on anti-oxidant, anti-inflammatory and hypoglycemic effect caused by three kinds of group subassemblys.So studying and developing excellent
Change anti-oxidant, anti-inflammatory and hypoglycemic effect herbal medicine prescription, it is very necessary.
3. summary of the invention
The object of the invention is intended to provide the preparation method of Southeast Asia herbal combination extract and anti-oxidant, hypoglycemic effect
Using, and it is used for the ancillary drug of exploitation prevention and treatment related disease.
3.1 solutions of the invention
3.1.1. extracting method:
1) powder is made in the Southeast Asia herbal medicine dried under natural conditions (ear leaf cassia leaf is dry, Jin Zhuoye and cat's whisker blade of grass)
End;
2) ear leaf cassia leaf, Jin Zhuoye and cat's whisker blade of grass powder are mixed with ethanol solution respectively by following mass volume ratio
It is put into round-bottomed flask, every Southeast Asia 1g herbal powder uses 30ml~60ml ethanol solution;The quality of ethyl alcohol in ethanol solution
Then concentration heats soak extraction 3h~3.5h twice under the conditions of 25 DEG C~40 DEG C between 50%~60%, merging mentions twice
Liquid is taken, crude extract is obtained;
3) Southeast Asia herbal medicine ethanol crude extract is concentrated under reduced pressure and removes and recycle ethyl alcohol, suspended with hot water and take off the extraction of ethyl alcohol
Object obtains suspension;
4) petroleum ether extraction suspension is used, the petroleum ether 3: 1,5: 2 or 1: 1 mixes by volume with suspension, removes
Chlorophyll and tannin, obtain refining liquid;
5) with the activity extract in ethyl acetate ultrasonic extraction refining liquid, the ethyl acetate and refining liquid are by volume
3: 1,5: 2 or 1: 1 mixes, and activity extract is dried in vacuo into lotion after concentration and recovery ethyl acetate.
3.1.2. Minitab software design Factorial Design is utilized, Southeast Asia herbal combination extract is optimized.Its feature exists
In being made of the herbal medicine dried extract of following weight proportion: ear leaf cassia leaf dried extract: Jin Zhuoye dried extract:
Cat's whisker blade of grass dried extract=1: 1: 1.
3.1.3. its effective ingredient includes that the compounds such as flavonoids, phenols, triterpenes, phenolic acid class can become through encapsulating sterilizing
Oral liquid formulation, can also add auxiliary material or auxiliary material granulation agent, tablet is not added etc..It is characterized by: the product be tablet, granula,
Aqua, creme, finish or emulsion.
3.2 compared with prior art, the invention has the following advantages that
1) it selects application time long, have the local herbal medicine of medicable tradition for raw material and develops natural plant active component,
High value added product is produced, there is very high economic value, and ecological environment can be adjusted to a certain extent.
2) it extracts ethyl alcohol used and has been depressurized concentration removal.
3) preparation takes low temperature to extract, high-temperature digestion, and active constituent has preferable high-temperature stability.
4) optimize prescription, more preferably than simple efficiency.
4. specific embodiment
The present invention chooses from the herbal medicine of Southeast Asia people's prolonged application has good anti-oxidant, hypoglycemic effect ear
Leaf cassia leaf, Jin Zhuoye and cat's whisker blade of grass therefrom extract the active component of middle polarity ingredient and are subject to rational proportion, have
More excellent anti-oxidant, anti-inflammatory and hypoglycemic effect.
4.1 the present invention from the herbal medicine of Southeast Asia extract active constituents the following steps are included:
1) powder is made in the Southeast Asia herbal medicine dried under natural conditions (ear leaf cassia leaf, Jin Zhuoye and cat's whisker blade of grass);
2) ear leaf cassia leaf, Jin Zhuoye and cat's whisker blade of grass powder are mixed with ethanol solution respectively by following mass volume ratio
It is put into round-bottomed flask, every Southeast Asia 1g herbal powder uses 30ml~60ml ethanol solution;The quality of ethyl alcohol in ethanol solution
Then concentration heats soak extraction 3h~3.5h twice under the conditions of 25 DEG C~40 DEG C between 50%~60%, merging mentions twice
Liquid is taken, crude extract is obtained;
3) Southeast Asia herbal medicine ethanol crude extract is concentrated under reduced pressure and removes and recycle ethyl alcohol, suspended with hot water and take off the extraction of ethyl alcohol
Object obtains suspension;
4) petroleum ether extraction suspension is used, the petroleum ether 3: 1,5: 2 or 1: 1 mixes by volume with suspension, removes
Chlorophyll and tannin, obtain refining liquid;
5) with the activity extract in ethyl acetate ultrasonic extraction refining liquid, the ethyl acetate and refining liquid are by volume
3: 1,5: 2 or 1: 1 mixes, and activity extract is dried in vacuo into lotion after concentration and recovery ethyl acetate.
6) Minitab software design Factorial Design is utilized, Southeast Asia herbal combination extract is optimized.It is characterized in that, by
The herbal medicine dried extract of following weight proportion forms: ear leaf cassia leaf dried extract: Jin Zhuoye dried extract: Mao Xu Cao
Leaf dried extract=1: 1: 1
7) its effective ingredient includes that the compounds such as flavonoids, phenols, triterpenes, phenolic acid class can sterilize through encapsulating as oral
Liquor type, can also add auxiliary material or auxiliary material granulation agent, tablet is not added etc..It is characterized by: the product be tablet, granula, aqua,
Creme, finish or emulsion.
4.2 activity extracts obtained using the above scheme have good high-temperature stability, list of the inventor to production
Only extract and merging component carry out anti-oxidant, free radical resisting and the detection of anti-alpha-amylase activity, find living in preparation process
Property ingredient is retained very well, the merging component activity with higher of acquisition.
4.2.1. anti-oxidant, free radical resisting detection method: 1,1- diphenyl -2- hardship diazanyl free radical (DPPH) method
1) precision weighs DPPH standard items, and dehydrated alcohol is added, and configures the liquid storage of 0.05mM concentration in 10mL volumetric flask.
2) 2.00mL is taken to configure 6.5X 10 with dehydrated alcohol-5Herb extracts are separately dissolved in anhydrous by mol/L DPPH solution
Ethyl alcohol is configured to series of concentrations solution as sample liquid.Accurate absorption 2.5mL DPPH solution and 0.5mL sample liquid shake up, and room temperature is kept away
Optical path 1cm cuvette is poured into after light placing response 10min, measures light absorption value under 517nm.It replication 3 times, is as a result averaged
Value.Then DPPH free radical scavenging activity is calculated as follows.
3) clearance rate (%)=[I- (Ai-Aj)/AO] X 100%:AO is 2.5mL DPPH solution+0.5mL dehydrated alcohol;
Ai is 2.5mL DPPH solution+0.5mL sample liquid;Aj is 2.5mL dehydrated alcohol+0.5mL sample liquid.
4) extract antioxidant activity IC50See attached drawing 1 and table one
Table one: extract antioxidant activity IC50
Extract | IC50(μg/ml) |
Ear leaf cassia leaf | 3.17±1.08 |
Jin Zhuoye | 15.08±1.13 |
Cat's whisker blade of grass | 18.85±1.14 |
Ear leaf cassia leaf: Jin Zhuoye: cat's whisker blade of grass (1: 1: 1) | 4.11±1.32 |
Three kinds of ingredient optimum quality proportions are 1: 1: 1.Use the component of the proportion in concentration for the anti-oxidant work of 10ug/ml
Property be (65.15 ± 5.84) %, 95% credibility interval (60.26,71.98) %.The IC of the combination50It is only slightly higher than ear leaf cassia leaf
Extract is lower than Jin Zhuoye and cat's whisker leaf extract.
4.2.2. alpha-amylase activity inhibits test
1) amylase solution will be mixed with from pig pancreatic juice alpha-amylase and 0.02M sodium phosphate buffer (pH6.5).With
8% soluble starch and the sodium phosphate buffer (pH6.5) of 0.08M prepare substrate solution.Enzyme, substrate solution and test is molten
37 DEG C of agent composition are reacted 7 minutes, are control with the mixture of enzyme, substrate solution and water.It is heated to 100 DEG C of stopped reactions.With
HPLC method detects the maltose to be formed, using arabinose as internal reference.Inhibitor activity is calculated with following formula.
2) the anti-alpha-amylase activity %=(1- (malt formed in the germinatus sugar amount/control sample formed in test sample
Sugar amount)) × 100
3) the anti-alpha-amylase activity IC of extract50See attached drawing 2 and table two
Table two: the anti-alpha-amylase activity IC of extract50
Extract | IC50(mg/ml) |
Ear leaf cassia leaf | 4.57±1.21 |
Jin Zhuoye | 6.82±1.05 |
Cat's whisker blade of grass | 9.15±1.33 |
Ear leaf cassia leaf: Jin Zhuoye: cat's whisker blade of grass (1: 1: 1) | 3.34±1.06 |
Three kinds of ingredient optimum quality proportions are 1: 1: 1.Use anti alpha-shallow lake of the component of the proportion when concentration is 10mg/ml
Powder enzymatic activity is (66.12 ± 2.92) %, 95% credibility interval (41.72,88.57) %.The IC of the combination50It is single lower than all
Component, including ear leaf Cassia leaf extract, Jin Zhuoye and cat's whisker leaf extract.
4.2.3. Factorial Design is carried out with optimization of C/C composites using Minitab
Using Minitab software design Factorial Design, by ear leaf cassia leaf, Jin Zhuoye and cat's whisker leaf extract are pressed respectively
According to the high component of quality (10), middle component (5), low component (1) mixing match, every kind of combination is in triplicate.Observation composition exists
Anti- alpha-amylase activity when optimal antioxidant activity and concentration under the concentration of 10ug/ml are 10mg/ml.
Three kinds of ingredient optimum quality proportions are 1: 1: 1.Use the component of the proportion in concentration for the anti-oxidant work of 10ug/ml
Property be (65.15 ± 5.84) %, 95% credibility interval (60.26,71.98) %;Anti- alpha-amylase when concentration is 10mg/ml
Activity is (66.12 ± 2.92) %, 95% credibility interval (41.72,88.57) %.(see attached drawing 3)
4.2.4 pass through the detection discovery of the above external activity: herbal combination extract in Southeast Asia prepared by the present invention has good
The effect of good inhibition alpha-amylase activity and antioxidant activity.And the combined extracts have good thermal stability and light
Stability.Its effective ingredient includes that the compounds such as flavonoids, phenols, triterpenes, phenolic acid class can sterilize through encapsulating as oral solution
Dosage form, can also add auxiliary material or auxiliary material granulation agent, tablet is not added etc..It is characterized by: the product is tablet, granula, aqua, frost
Agent, finish or emulsion.
5. Detailed description of the invention
1) Fig. 1 extract antioxidant activity IC50, including ear leaf Cassia leaf extract (Figure 1A), Jin Zhuoye leaf extract
(Figure 1B), cat's whisker leaf extract (Fig. 1 C), ear leaf cassia leaf: Jin Zhuoye: Mao Xu Cao extract combination (1: 1: 1, mass ratio)
(Fig. 1 D).(see attached drawing 1)
2) Fig. 2 extract anti alpha amylase activity IC50, including ear leaf Cassia leaf extract (Fig. 2A), Jin Zhuoyeye extraction
Object (Fig. 2 B), cat's whisker leaf extract (Fig. 2 C), ear leaf cassia leaf: Jin Zhuoye: Mao Xu Cao extract combination (1: 1: 1, quality
Than) (Fig. 2 D).(see attached drawing 2)
3) Fig. 3 is using Minitab software to ear leaf cassia leaf: Jin Zhuoye: Mao Xu Cao extract combination carries out Factorial Design
It was found that: three kinds of ingredient optimum quality proportions are 1: 1: 1.Use the component of the proportion in concentration for the antioxidant activity of 10ug/ml
For (65.15 ± 5.84) %, 95% credibility interval (60.26,71.98) %;Anti alpha-diastatic activity when concentration is 10mg/ml
Property be (66.12 ± 2.92) %, 95% credibility interval (41.72,88.57) %.
Claims (2)
1. Southeast Asia herbal combination extract inhibits the application in alpha-amylase activity drug in preparation, it is characterised in that by following
The herbal medicine dried extract of weight proportion forms: ear leaf cassia leaf dried extract: Jin Zhuoye dried extract: cat's whisker blade of grass is dry
Dry extract=1: 1: 1;And it is characterized in that herbal medicine dried extract is made according to following extracting method:
1) average particle size is made in 100 mesh powders in the ear leaf cassia leaf that will be dried under natural conditions, Jin Zhuoye and cat's whisker blade of grass;
2) ear leaf cassia leaf, Jin Zhuoye and cat's whisker blade of grass powder are mixed with ethanol solution be put into respectively by following mass volume ratio
In round-bottomed flask, every Southeast Asia 1g herbal powder uses 30ml~60ml ethanol solution;The mass concentration of ethyl alcohol in ethanol solution
Between 50%~60%, soak extraction 3h~3.5h then is heated twice under the conditions of 25 DEG C~40 DEG C, merges extracting solution twice,
Obtain crude extract;
3) above-mentioned ethanol crude extract is concentrated under reduced pressure and removes and recycle ethyl alcohol, suspended with hot water and take off the extract of ethyl alcohol, hanged
Supernatant liquid;
4) petroleum ether extraction suspension is used, the petroleum ether 3: 1,5: 2 or 1: 1 mixes by volume with suspension, and removing leaf is green
Element and tannin, obtain refining liquid;
5) with the activity extract in ethyl acetate ultrasonic extraction refining liquid, the ethyl acetate and refining liquid by volume 3: 1,
5: 2 or 1: 1 mixes, and activity extract is dried in vacuo into lotion after concentration and recovery ethyl acetate.
2. purposes according to claim 1, it is characterised in that: tablet, granula, aqua, creme, finish can further be made
Or emulsion.
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CN101690705A (en) * | 2009-08-21 | 2010-04-07 | 珈侬生化科技(中国)有限公司 | Preparation method of alpine pine needle active extract and whitening anti-aging active cosmetic |
CN101999999A (en) * | 2009-09-01 | 2011-04-06 | 珈侬生化科技(中国)有限公司 | Method for preparing active extract of Juniperus sibirica and whitening and anti-ageing active cosmetic |
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