CN110354205A - A kind of Pale Ephemerantha active component and its preparation method and application - Google Patents
A kind of Pale Ephemerantha active component and its preparation method and application Download PDFInfo
- Publication number
- CN110354205A CN110354205A CN201810250687.2A CN201810250687A CN110354205A CN 110354205 A CN110354205 A CN 110354205A CN 201810250687 A CN201810250687 A CN 201810250687A CN 110354205 A CN110354205 A CN 110354205A
- Authority
- CN
- China
- Prior art keywords
- ephemerantha
- pale
- active component
- gross mass
- extract
- Prior art date
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- Granted
Links
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Classifications
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The present invention relates to technical field of traditional Chinese medicines, a kind of Pale Ephemerantha active component and its preparation method and application is specifically disclosed.Its major secondary metabolic compounds type of the Pale Ephemerantha active component is glucoside compound.The Pale Ephemerantha active component main sugar glycoside composition is pimarane type monoglycosides, flickinflimoside B, bioside fliflidioside A, (+)-syringaresinol-O- β-D- glucopyranoside etc..External activity is studies have shown that the DPPH free radical and OH free radical scavenging capacity that Pale Ephemerantha active component of the invention has had, alpha-glucosaccharase enzyme inhibition activity, and inhibits the damagine activity of the HT22 cell of glutamate induction.The Pale Ephemerantha active component can be developed into treatment Alzheimer's disease, liver protection effect, the ancillary drug or health care product of the diseases such as type II diabetes.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicines, and in particular to a kind of Pale Ephemerantha (Flickingeria fimbriata) have
Imitate position and its preparation method and application.
Background technique
Pale Ephemerantha, that is, tasselled HERBA DENDROBII (Flickingeria fimbriata) be orchid family (Orchidaceae) HERBA DENDROBII
Belong to (Flickingeria) plant, main product is in the ground such as Guangdong (Meizhou region), Guangxi, Hainan, Yunnan, Guizhou.Chinese medicine has melon stone
Dry measure used in former times be Pale Ephemerantha (Flickingeria fimbriata) the drying herb with pseudobulb, alias band melon dendrobium nobile.Chinese medicine has melon
Dendrobium nobile is recorded in " Guangdong Province's Chinese medicine standard ", sweet in flavor, slightly cold, returns stomach, lung, kidney channel, has reinforcing stomach reg fluid, nourishing Yin and clearing heat function
Effect.For impairment of yin body fluid deficiency, dry polydipsia, deficiency of food is retched, and abnormal heat, object are secretly unknown after being ill.Pale Ephemerantha and the " People's Republic of China (PRC)
Pharmacopeia " dendrobium nobile that records effect, cure mainly, in terms of it is identical, Pale Ephemerantha is often made pharmacopeia certified products by In Guangdong Province
Dendrobium nobile substitution uses.Domestic and foreign scholars have carried out research extensively and profoundly to the chemical component of Pale Ephemerantha, find Pale Ephemerantha
Contain the compounds such as bibenzyl, luxuriant and rich with fragrance class, lipid, Diterpenes, sterols, lignanoids, flavonoids, polysaccharide.Pharmacological research is aobvious
Show that Pale Ephemerantha alcohol extract and water extract have the rat hepatocytes activity for repairing peroxide injury.
Currently, with China human mortality aging progress faster, Alzheimer's disease, hepatic injury caused by oxidative stress,
Type II diabetes etc. has become the disease for seriously threatening human health.Chinese medicine has highly-safe, and toxic side effect is small, cheap and easy to get
Advantage, and it is often multicomponent, multiple target point integrated use, therefore to develop activity good for screening from Chinese medicine, has and controls
Alzheimer's disease is treated, hepatic injury caused by oxidative stress, the adjuvant therapy medicaments or health care product of the functions such as type II diabetes,
Have wide practical use.
Although reporting the Pale Ephemerantha extract with hepatoprotective effect in the prior art, these extracts all fail clear
Active component and composition.The present invention specifies its active component using Pale Ephemerantha as raw material, develop have high-efficiency low-toxicity treatment Ah
Er Cihai Mo's disease, hepatic injury caused by oxidative stress, the health care product or adjuvant therapy medicaments of the functions such as type II diabetes have
Wide application prospect.
Summary of the invention
The technical problem to be solved by the present invention is in order to overcome problems of the prior art, providing one kind has melon
Dendrobium nobile active component and its preparation method and application.
One aspect of the present invention provides a kind of Pale Ephemerantha active component, contains and accounts for active component gross mass 10-15%
Above glucoside compound, the glucoside compound include flickinflimoside B, (+)-syringaresinol-O-
The compound of β-D- glucopyranoside and formula (I)
In some embodiments, the glucoside compound content accounts for the 10-20% or more of the active component gross mass, such as
15-20%。
In some embodiments, flickinflimoside B content accounts for the 6- of Pale Ephemerantha active component gross mass
8%, the compounds content of formula (I) accounts for the 4-6% of Pale Ephemerantha active component gross mass, (+)-syringaresinol-O- β-D- pyrrole
Glucopyranoside glycosides content accounts for the 0.5-1% of Pale Ephemerantha active component gross mass.
In some embodiments, flickinflimoside B content accounts for the 6- of Pale Ephemerantha active component gross mass
8%, the compounds content of formula (I) accounts for the 4-6% of Pale Ephemerantha active component gross mass, (+)-syringaresinol-O- β-D- pyrrole
Glucopyranoside glycosides content accounts for the 1-2.5% of Pale Ephemerantha active component gross mass.
In some embodiments, the Pale Ephemerantha (Flickingeria fimbriata) originate from Yunnan.
Another aspect of the present invention provides the preparation method of the Pale Ephemerantha active component, and it includes following steps:
S1. hydrophilic organic solvent-water mixed solution is used to carry out cold soaking extraction to Pale Ephemerantha for extracting solution, filtering will filter
Liquid concentration, obtains Pale Ephemerantha extract；
S2. by Pale Ephemerantha extract obtained by S1, add appropriate aqueous suspension, with ethyl acetate liquid-liquid extraction, collect water phase, be spin-dried for,
Up to Pale Ephemerantha active component.
Another aspect of the present invention provides the preparation method of the Pale Ephemerantha active component, and it includes following steps:
S1. hydrophilic organic solvent-water mixed solution is used to carry out cold soaking extraction to Pale Ephemerantha for extracting solution, filtering will filter
Liquid concentration, obtains Pale Ephemerantha extract；
S3. by Pale Ephemerantha extract obtained by S1, add proper amount of methanol dissolution, the enrichment of upper solid-phase extraction column, loading finishes, and uses water
Elution removal of impurities, then eluted with organic solvent-water mixed liquid, organic solvent-water mixing eluent is collected, is had to obtain the final product after concentrate drying
Melon dendrobium nobile active component.
In some embodiments, hydrophilic organic solvent used in step S1 is methanol, acetone, ethyl alcohol etc..Some
In embodiment, the volume ratio of hydrophilic organic solvent (o) and water (w) is to have 7 8:3 2(V in extracting solutionO/Vw).
In some embodiments, the amount ratio of extracting solution dosage used in step S1 and raw material Pale Ephemerantha be 8~
10L:1kg；Ultrasonic wave assisted extraction is used during room temperature cold soaking, extraction time is 2-3 times.
In some embodiments, organic solvent is ethyl acetate, acetic acid second in liquid-liquid extraction techniques used in step S2
The amount ratio of ester and water is 1:1(v/v), extraction times are 2-3 times.
In some embodiments, solid-phase extraction column described in step S3, stationary phase are C-18 reverse phase silica gel, nonpolar macroporous
Adsorb resin, or the small macroporous adsorbent resin of nonpolarity；Stationary phase dosage is the dry extract weight of Pale Ephemerantha extract obtained by step S1
It is 0.3kg: 1L with stationary phase bed volume ratio.
In some embodiments, the dosage of removal of impurities elution water used is 4 ~ 5 times of bed volumes, the organic solvent-of use
In water mixed liquid elution, organic solvent is methanol or ethyl alcohol；Volume fraction containing methanol or ethyl alcohol in eluent is 20 ~ 50%(v/
V), organic solvent-water mixing eluent dosage is 5 ~ 6 times of bed volumes.
Another aspect of the present invention provides the Pale Ephemerantha active component and is preparing neurodegenerative disease, liver protection effect,
The therapeutic agent of diabetes and the application in health care product.
In some embodiments, the neurodegenerative disease is senile dementia or Alzheimer's disease.
In some embodiments, the diabetes are type II diabetes.
In some embodiments, the liver protection effect is free radical scavenging activity.
The present invention provides the Pale Ephemerantha active component that one kind completely newly forms, which specifies effective component,
Be conducive to quality control and the drug safety of drug；The experimental results showed that the Pale Ephemerantha active component has stronger resist
Oxidability, stronger inhibition alpha-glucosidase activity, the damage of the stronger HT22 cell for inhibiting glutamate induction can be used
In neurodegenerative disease, the treatment or adjuvant treatment of liver protection effect, diabetes.
Detailed description of the invention
Fig. 1 shows protective effect of the Pale Ephemerantha active component to Pidolidone induction HT22 cell death.It is positive right
According to NAC:mM, glutamic acid Glu:mM, Pale Ephemerantha active component series of concentrations: μ g/mL.
Specific embodiment
The present invention is explained further below in conjunction with specific embodiment, but embodiment does not do any type of limit to the present invention
It is fixed.
In the present invention, monoglycosides flickinflimoside B, (+)-syringaresinol-O- β-D- glucopyranose
Glycosides structural formula is as follows:
flickinflimoside B
(+)-syringaresinol-O- β-D- glucopyranoside
。
The proton that a, c are marked in figure is target proton position selected by nmr quantitative analysis.
Monoglycosides flickinflimoside B and (+)-syringaresinol-O- β-D- glucopyranoside are abilities
Compound known to domain, preparation method can be found in 1. Chen of document, J.L.; Zhao, Z.M.; Xue, X.; Tang,
G.H.; Zhu, L.P.; Yang, D.P.; Jiang, L. Bioactive norditerpenoids fromFlickingeria fimbriata. RSC Adv. 2014, 4, 14447–14456. DOI: 10.1039/
C4RA00835A; 2. Wang, C.Z.; Yu, D.Q. Lignan and acetylenic glycosides fromAster auriculatus. Phytochemistry1998, 48, 711–717. https://doi.org/10.1016/
S0031-9422(98) 00019-3。
The compound of formula (I) is the new compound that the present inventor isolates, and structural formula is as follows, herein also referred to as
For bioside fliflidioside A:
The proton that b is marked in figure is target proton position selected by nmr quantitative analysis.
The preparation of embodiment 1-Pale Ephemerantha active component
S1. by 80% aqueous acetone solution room temperature cold soaking of 1kg Pale Ephemerantha, solid-liquid ratio 8:1, ultrasonic wave assisted extraction, 80
Min/d, places 48h, and filtering is concentrated filtrate, obtains Pale Ephemerantha extract；It repeats to extract 3 times.
S2. it by Pale Ephemerantha extract obtained by S1, is suspended in 800mL water, isometric acetic acid ethyl fluid liquid extraction is added
It takes, collects water phase, be spin-dried for get Pale Ephemerantha active component.Ethyl acetate is repeated to extract 3 times.
As a result it measures Pale Ephemerantha active component glucoside compound content and accounts for Pale Ephemerantha active component gross mass
12%.Flickinflimoside B content accounts for the 7% of Pale Ephemerantha active component gross mass, and formula (I) compound represented contains
Amount accounts for the 4% of Pale Ephemerantha active component gross mass, and (+)-syringaresinol-O- β-D- glucopyranoside content occupies melon
The 1% of dendrobium nobile active component gross mass.Content analysis method is using quantitative hydrogen nuclear magnetic resonance spectral technology, detailed in Example 4.Pharmacology
Activity is shown in Table 1, Fig. 1.
The preparation method of compound (I): by the resulting Pale Ephemerantha active component of S2 through silica gel column separating purification, dichloromethane
Alkane-methanol elution gradient collects corresponding fraction, then through Sephadex LH-20 column purification, methanol makees eluant, eluent, obtains pure
Product.
Spectral data: [a]20 D - 47 (c 0.074, MeOH); UV (MeOH) λmax (logε): 203 (1.42)
nm; ECD (MeOH) λmax (Δε): 206 (- 6.03), 300 (+ 0.39); IR (KBr) νmax: 3352,
2936, 1709, 1363, 1074 cm−1; HRESIMS m/z: 643.33388 [M - H]− (calcd for
C32H51O13, 643.33351).；13C NMR (100MHz, CD3OD) δ: 213.7 (C), 143.5 (C), 124.8
(d), 104.2 (d), 101.9 (d), 85.8 (d), 78.2 (d), 78.1 (d), 77.7 (d), 77.6 (d),
75.1 (d), 74.9 (d), 72.4 (t), 71.9 (d), 71.4 (d), 62.9 (t), 62.7 (t), 55.9
(d), 51.9 (d), 48.6 (s), 40.4 (s), 39.4 (s), 37.7 (t), 36.7 (t), 33.6 (t),
29.1 (q), 28.1 (t), 27.5 (q), 23.2 (t), 21.2 (t), 17.3 (q), 15.1 (q); 1H NMR
(400MHz, CD3OD) δ: 5.52 (s), 4.26 (d, 7.6 Hz) , 4.33 (d, 7.6 Hz), 3.38 (m),
3.37-3.27 (m), 3.37-3.27 (m), 3.37-3.27 (m), 3.37-3.27 (m), 3.26-3.18 (m),
3.26-3.18 (m), 4.86 (d, 18.4 Hz), 4.49 (d, 18.4 Hz), 3.29 (m), 3.29 (m), 3.67
(m), 3.86 (m), 3.67 (m), 3.86 (m), 1.15 (m), 1.78 (m), 1.69 (m), 2.04 (m), 2.11
(m), 2.29 (m), 1.14 (m), 1.07 (s), 1.12 (m), 1.14 (s), 1.61 (m), 1.46 (m),
1.63 (m), 1.63 (m), 0.87 (s), 0.73 (s)。
The preparation of embodiment 2-Pale Ephemerantha active component
S1. by 80% methanol aqueous solution room temperature cold soaking of 1kg Pale Ephemerantha, solid-liquid ratio 10:1, ultrasonic wave assisted extraction, 80
Min/d, places 48h, and filtering is concentrated filtrate, obtains Pale Ephemerantha extract；It repeats to extract 3 times.
S2. by Pale Ephemerantha extract obtained by S1, add proper amount of methanol dissolution, upper C-18 reverse phase silica gel column solid phase extraction, have
Melon Herba Dendrobii extract dry extract weight and C-18 reverse phase silica gel column stationary phase bed volume ratio are 0.3kg: 1L, and loading finishes, with 5
The water elution removal of impurities of times bed volume, then with 6 times of 30% methanol-water solution elution, collect methanol-water eluent, be after concentrate drying
Obtain Pale Ephemerantha active component.
The Pale Ephemerantha active component glucoside compound content accounts for 14% or more of the active component gross mass.Its
Middle flickinflimoside B content accounts for the 7% of Pale Ephemerantha active component gross mass, and formula (I) compound represented content accounts for
The 5% of Pale Ephemerantha active component gross mass, (+)-syringaresinol-O- β-D- glucopyranoside content account for Pale Ephemerantha
The 2% of active component gross mass.Content analysis method is using quantitative hydrogen nuclear magnetic resonance spectral technology, detailed in Example 4.Pharmacological activity
It is shown in Table 1, Fig. 1.
The preparation of implementation column 3-Pale Ephemerantha active component
S1. by 80% ethanol water room temperature cold soaking of 1kg Pale Ephemerantha, solid-liquid ratio 10:1, ultrasonic wave assisted extraction, 80
Min/d, places 48h, and filtrate is concentrated, obtains Pale Ephemerantha extract by filtering；It repeats to extract 3 times.
S2. by Pale Ephemerantha extract obtained by S1, add suitable quantity of water dissolution, upper large pore resin absorption column Solid Phase Extraction, there is melon
The dry extract weight and macroporous resin adsorption resin column stationary phase bed volume ratio of Herba Dendrobii extract are 0.3kg: 1L, and loading finishes,
It is cleaned with the water elution of 5 times of bed volumes, then is eluted with 6 times of 40% ethanol-water solution, collect alcohol-water eluent, be concentrated and dried
Afterwards up to Pale Ephemerantha active component；
As a result it measures Pale Ephemerantha active component glucoside compound content and accounts for 15% or more of the active component gross mass.Wherein
Flickinflimoside B content accounts for the 8% of Pale Ephemerantha active component gross mass, and formula (I) compound represented content is occupied
The 5% of melon dendrobium nobile active component gross mass, (+)-syringaresinol-O- β-D- glucopyranoside content, which accounts for Pale Ephemerantha, to be had
Imitate the 2% of position gross mass.Content analysis method is using quantitative hydrogen nuclear magnetic resonance spectral technology, detailed in Example 4.Pharmacological activity is shown in
Table 1, Fig. 1.
The content assaying method of implementation column 4-Pale Ephemerantha active component
Pale Ephemerantha active component glucoside compound content analysis uses hydrogen nuclear magnetic resonance quantitative analysis tech.Using 500MHz
Nuclear Magnetic Resonance, experiment parameter setting are as follows: pulse train zg30, spectrum width SW=14 ppm, centre frequency O1P=6 ppm are adopted
Sample time AQ=3 s, sampling number TD=41976, the s of waiting time D1=2, scanning times NS=64, transformation points SI=
65536, the position a CH proton peak in flickinflimoside B structure, the position b of compound shown in formula (I) are chosen in quantitative peak
CH proton peak, the position the c CH proton peak of (+)-syringaresinol-O- β-D- glucopyranoside.With fragrant in bigcatkin willow acid molecule
Proton is external standard ,-ERETIC2 standard measure on ring.
Sample operation accurately weighs the Pale Ephemerantha active component medicinal extract 30mg done to constant weight, and deuterated methanol is added, fixed molten
To 1mL, closed container is vortexed sample dissolution, and centrifugal sedimentation insoluble solids take 600 μ L of supernatant to enter nuclear magnetic tube (5mm), immediately
Carry out hydrogen nuclear magnetic resonance spectrum analysis.Parallel 3 operations.Standard sample salicylic acid is equally operated.It is measured with ERETIC2 method
Salicylic acid and Pale Ephemerantha active component glucoside compound content.
5-free radical of embodiment eliminates the measurement (using DPPH free radical null method) of ability
Reagent: DPPH solution, methanol are prepared, and concentration is 150 μM；Pale Ephemerantha active component solution, DMSO are prepared, initial dense
30mg/mL is spent, proportional diluted is at series of concentrations when use.Positive reference substance: 2,6-ditertbutyl-4-methyl-phenol
(BHT) solution, DMSO are prepared, and 10 μm of ol/mL of initial concentration, proportional diluted is at series of concentrations when use.
Method: taking 96 porocyte culture plates, and each hole is separately added into 180 μ L of DPPH solution, the Pale Ephemerantha of each gradient concentration
20 μ L of active component solution, the 20 μ L of positive reference substance BHT solution of each gradient concentration, while DPPH control group is set, in enzyme mark
In 517 nm on instrument, each hole absorbance value (A) is measured.It was surveyed once at interval of 10 minutes, continuously surveys 60min.Calculate clearance rate
(%)=[(AControl- ASample)/AControl] × 100%.As a result it is displayed in Table 1.
The measurement that 6-hydroxyl free of embodiment eliminates ability (uses Phen-Fe2+Oxidizing process)
Reagent: Phen aqueous solution, concentration are 5 mM.FeSO4Aqueous solution, concentration are 7.5 mM.PBS buffer solution, concentration are
0.75 M, pH7.4.Liquid is applied in measurement: by Phen solution when use: PBS buffer solution: redistilled water: FeS04Solution, by 3:
8:8:2 sequence mixes.Pale Ephemerantha active component solution, DMSO are prepared, initial concentration 30mg/mL, when use proportional diluted at
Series of concentrations.Positive reference substance Vit C solution, DMSO are prepared, and 10 μm of ol/mL of initial concentration, proportional diluted is at series when use
Concentration.1% hydrogen peroxide.
Method: taking 96 porocyte culture plates, and each hole is separately added into measurement using 170 μ L of liquid, and each concentration gradient has melon stone
Positive reference substance Vit the C solution 20 μ L, 1% hydrogen peroxide, 10 μ L of 20 μ L of dry measure used in former times active component solution or each concentration gradient.Point
If blank group, control group, sample sets, 37 DEG C of 10 min of heat preservation, respectively in 10~120 min time ranges, in 510 nm survey
Fixed each hole absorbance value (A).Scavenging action to hydroxyl free radical (%)=[(ASample- ADamage)/(AIt does not damage- ADamage)] × 100%.As a result in table
It is shown in 1.
The measurement of embodiment 7-alpha-glucosaccharase enzyme inhibition activity
Reagent: KH2PO4-K2HPO4Buffer solution, concentration are 50 mM, PH=7.Substrate p-nitrophenol-alpha-glucosaccharase
(PNPG) solution, concentration are 1 mM, and when use uses KH2PO4-K2HPO4Buffer preparation.Alpha-glucosaccharase enzyme solution, when use
Use KH2PO4-K2HPO4Buffer preparation is at required concentration.Positive control: acarbose, DMSO are prepared, initial concentration
For 10 μm of ol/mL, proportional diluted is at series of concentrations when use.Pale Ephemerantha active component DMSO solution, initial concentration 30mg/
ML, proportional diluted is at series of concentrations when use.
Method: taking 96 orifice plates, and corresponding solution is added according to the form below in corresponding aperture.1min absorbance is surveyed at λ=400nm to become
Change.It parallel testing 3 times, is averaged.
Press down enzyme relative activity to calculate: suppression enzyme relative activity=(A0- A)/A0*100%。
A0: the enzymatic activity (OD/min) of sample is not added.
A: inhibitory activity (OD/min) of the sample to enzyme.As a result it is displayed in Table 1.
Protective effect of the embodiment 8-to L-glutamate induction HT22 cell death
Reagent: Pale Ephemerantha active component solution, DMSO are prepared, and initial concentration 30mg/mL, proportional diluted is at series when use
Concentration.L-glutamate solution, DMSO are prepared, and concentration is 100 mM.Positive control N-acetyl cysteine (NAC),
DMSO is prepared, and initial concentration is 10 μm of ol/mL.
Method: mouse hippocampal neuron cell strain HT22, with contain 10% fetal calf serum DMEM complete medium, 37
DEG C, saturated humidity is 5%CO containing volume fraction2Carbon dioxide incubator in routine culture.Logarithmic growth phase cell, with
After the digestion of 0.25% pancreatin, complete medium is resuspended, and it is 10 × 104 that cell counting board, which counts and adjusts cell concentration, under microscope
A/ml, is inoculated with 96 porocyte culture plates, 100 holes μ L/, and overnight incubation keeps cell adherent.Culture medium in 96 orifice plates is siphoned away, is had
Melon dendrobium nobile active component series of concentrations solution is added in 96 orifice plates, 100 holes μ L/.After preincubate 30min, 2 μ L 100mM are added
L-glutamate.Model group is not added to sample, is directly added into 2 μ L 100mML-glutamate.After being incubated for for 24 hours, every hole is added
10 μ L 5mg/mL MTT are incubated for 2h, discard supernatant, and add 100 hole μ L/ DMSO, and oscillation dissolves product formazan sufficiently,
Each hole absorbance value is measured in microplate reader, measures wavelength 570nm.Calculate cell survival rate:
To protective effect (%)=100%* (A of L-glutamate induction HT22 cell deathSample to be tested-ABlank)/(AModel-ABlank)。
As a result it is shown in Fig. 1.
1 Pale Ephemerantha active component biological activity test result of table
/: do not test；n=3.
Claims (10)
1. a kind of Pale Ephemerantha active component contains the glycoside chemical combination for accounting for the 10-15% or more of the active component gross mass
Object, the glucoside compound include flickinflimoside B, (+)-syringaresinol-O- β-D- glucopyranoside
With the compound of formula (I)
。
2. Pale Ephemerantha active component according to claim 1, wherein the glucoside compound content accounts for effective portion
The 10-20% of position gross mass.
3. Pale Ephemerantha active component according to claim 1, wherein flickinflimoside B content occupies melon stone
The 6-8% of dry measure used in former times active component gross mass, formula (I) compound represented content account for the 4-6% of Pale Ephemerantha active component gross mass,
(+)-syringaresinol-O- β-D- glucopyranoside content accounts for the 0.5-1% of Pale Ephemerantha active component gross mass.
4. Pale Ephemerantha active component according to claim 1, wherein lickinflimoside B content accounts for Pale Ephemerantha
The 6-8% of active component gross mass, formula (I) compound represented content account for the 4-6% of Pale Ephemerantha active component gross mass, (
+)-syringaresinol-O- β-D- glucopyranoside content accounts for the 1-2.5% of Pale Ephemerantha active component gross mass.
5. the described in any item Pale Ephemerantha active components of claim 1-4 are in preparation neurodegenerative disease, liver protection effect, sugar
Urinate the application in the therapeutic agent and health care product of disease.
6. application according to claim 5, wherein the neurodegenerative disease is senile dementia or Alzheimer
Family name's disease.
7. application according to claim 5, wherein the diabetes are type II diabetes.
8. application according to claim 5, wherein the liver protection effect is free radical scavenging activity.
9. a kind of method for preparing the described in any item Pale Ephemerantha active components of claim 1-4, comprises the following steps:
S1. it uses hydrophilic organic solvent-water mixed solution to carry out cold soaking extraction to Pale Ephemerantha for extracting solution, filters, it will
Filtrate concentration, obtains Pale Ephemerantha extract；
S2. by Pale Ephemerantha extract obtained by S1, add appropriate aqueous suspension, with ethyl acetate liquid-liquid extraction, collect water phase, be spin-dried for,
Up to Pale Ephemerantha active component.
10. a kind of method for preparing the described in any item Pale Ephemerantha active components of claim 1-4, comprises the following steps:
S1. it uses hydrophilic organic solvent-water mixed solution to carry out cold soaking extraction to Pale Ephemerantha for extracting solution, filters, it will
Filtrate concentration, obtains Pale Ephemerantha extract；
S3. by Pale Ephemerantha extract obtained by S1, add proper amount of methanol dissolution, the enrichment of upper solid-phase extraction column, loading finishes, and uses water
Elution removal of impurities, then eluted with aqueous organic solution, aqueous organic eluent is collected, up to the effective portion of Pale Ephemerantha after concentrate drying
Position.
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| CN106822599A (en) * | 2017-01-09 | 2017-06-13 | 中山大学 | A kind of dendrobium loddigesii Rolfe extract and its anti-diabetes use |
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