CN110354205B - Dendrobium candidum effective part and preparation method and application thereof - Google Patents
Dendrobium candidum effective part and preparation method and application thereof Download PDFInfo
- Publication number
- CN110354205B CN110354205B CN201810250687.2A CN201810250687A CN110354205B CN 110354205 B CN110354205 B CN 110354205B CN 201810250687 A CN201810250687 A CN 201810250687A CN 110354205 B CN110354205 B CN 110354205B
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- dendrobium candidum
- effective part
- water
- dendrobium
- extract
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Abstract
The invention relates to the technical field of traditional Chinese medicines, and particularly discloses a dendrobium candidum effective part and a preparation method and application thereof. The main secondary metabolic compound type of the effective part of the dendrobium candidum is a glucoside compound. The main glycoside components of the dendrobium huoshanense effective part are pimarantene type monoglycoside, flickingflimoside B, difrosidioside A, (+) -syringaresinol-O-beta-D-glucopyranoside and the like. In vitro activity research shows that the dendrobium candidum effective part has good DPPH free radical and OH free radical scavenging activity, alpha-glucosidase inhibition activity and glutamic acid induced HT22 cell damage inhibition activity. The effective part of the dendrobium candidum can be developed into an auxiliary medicine or a health-care product for treating Alzheimer's disease, liver protection, type II diabetes and other diseases.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, and particularly relates to dendrobium candidum (dendrobium officinale) (A. candidum)Flickingeria fimbriata) Effective part and its preparation method and application.
Background
Dendrobium huoshanense, i.e. Dendrobium fimbriatum (A. Merrill. ex Fr.) BenthFlickingeria fimbriata) Is Orchidaceae (Orchidaceae) Dendrobium (C. A. B.) genusFlickingeria) The plants are mainly produced in Guangdong (Meizhou region), Guangxi, Hainan, Yunnan, Guizhou and other places. The traditional Chinese medicine herba Dendrobii is herba Dendrobii (herba Dendrobii) with melonFlickingeria fimbriata) Dried whole plant with pseudobulb is named as Dendrobium loddigesii Rolfe. The traditional Chinese medicine, Guapian dendrobium, is recorded in the Chinese medicinal material Standard of Guangdong province, is sweet in taste and slightly cold, enters the channels of the stomach, lung and kidney, and has the effects of tonifying stomach, promoting fluid production, nourishing yin and clearing heat. Can be used for treating yin deficiency and fluid deficiency, dry mouth with polydipsia, anorexia, retching, and asthenic fever after disease with dim and unclear material. The dendrobium candidum is the same as dendrobium collected in pharmacopoeia of people's republic of China in the aspects of efficacy, main treatment, clinical application and the like, and the dendrobium candidum is often used as a certified dendrobium of the pharmacopoeia in Guangdong. The scholars at home and abroad extensively and intensively research the chemical components of the dendrobium candidum, and the dendrobium candidum is found to contain compounds such as bibenzyls, phenanthrenes, lipids, diterpenes, sterols, lignans, flavonoids, polysaccharides and the like. Pharmacological research shows that the dendrobium candidum alcohol extract and the dendrobium candidum water extract have the activity of repairing rat liver cells with peroxidation damage.
At present, with the accelerating aging process of the population of China, Alzheimer disease, liver damage caused by oxidative stress, type II diabetes and the like become diseases seriously threatening the health of human beings. The traditional Chinese medicine has the advantages of high safety, small toxic and side effects, low price and easy obtainment, and is usually multi-component and multi-target comprehensive application, so that the auxiliary treatment medicine or health care product which has good activity and has the functions of treating Alzheimer's disease, liver injury caused by oxidative stress, type II diabetes and the like is screened and developed from the traditional Chinese medicine, and the application prospect is wide.
Although dendrobium candidum extracts with liver protection effects are reported in the prior art, the extracts cannot define effective parts and components. The dendrobium candidum is used as a raw material, the effective part of the dendrobium candidum is determined, the dendrobium candidum health care product or the auxiliary treatment medicine which has the functions of treating Alzheimer's disease, liver injury caused by oxidative stress, type II diabetes and the like and has high efficiency and low toxicity is developed, and the dendrobium candidum has wide application prospect.
Disclosure of Invention
The invention aims to solve the technical problem of providing the dendrobium candidum effective part and the preparation method and application thereof in order to overcome the problems in the prior art.
The invention provides a dendrobium candidum effective part, which contains glycoside compounds accounting for more than 10-15% of the total mass of the effective part, wherein the glycoside compounds comprise flickingflinoside B, (+) -syringomycins-O-beta-D-glucopyranoside and compounds in a formula (I)
In some embodiments, the glycoside compound is present in an amount of more than 10-20%, for example 15-20% by weight of the total weight of the active site.
In some embodiments, the content of flickiflomoside B accounts for 6-8% of the total mass of the effective parts of the dendrobium officinale, the content of the compound of the formula (I) accounts for 4-6% of the total mass of the effective parts of the dendrobium officinale, and the content of (+) -syringaresinol-O-beta-D-glucopyranoside accounts for 0.5-1% of the total mass of the effective parts of the dendrobium officinale.
In some embodiments, the content of flickiflomoside B accounts for 6-8% of the total mass of the effective parts of the dendrobium officinale, the content of the compound of the formula (I) accounts for 4-6% of the total mass of the effective parts of the dendrobium officinale, and the content of (+) -syringaresinol-O-beta-D-glucopyranoside accounts for 1-2.5% of the total mass of the effective parts of the dendrobium officinale.
In some embodiments, the dendrobium candidum (dendrobium officinale)Flickingeria fimbriata) Produced in Yunnan province.
The invention also provides a preparation method of the dendrobium candidum effective part, which comprises the following steps:
s1, cold soaking and extracting the dendrobium candidum by using a hydrophilic organic solvent-water mixed solution as an extracting solution, filtering, and concentrating the filtrate to obtain the dendrobium candidum extract;
s2, adding a proper amount of water into the dendrobium candidum extract obtained in the step S1 for suspension, performing liquid-liquid extraction by using ethyl acetate, collecting a water phase, and performing spin drying to obtain the dendrobium candidum effective part.
The invention also provides a preparation method of the dendrobium candidum effective part, which comprises the following steps:
s1, cold soaking and extracting the dendrobium candidum by using a hydrophilic organic solvent-water mixed solution as an extracting solution, filtering, and concentrating the filtrate to obtain the dendrobium candidum extract;
and S2, adding a proper amount of methanol into the dendrobium candidum extract obtained in the step S1 for dissolution, loading the dendrobium candidum extract on a solid-phase extraction column for enrichment, eluting the dendrobium candidum extract by water for removing impurities after loading, eluting the dendrobium candidum extract by using an organic solvent-water mixed solution, collecting the organic solvent-water mixed eluent, concentrating and drying to obtain the dendrobium candidum effective part.
In some embodiments, the hydrophilic organic solvent used in step S1 is methanol, acetone, ethanol, or the like. In some embodiments, the volume ratio of the hydrophilic organic solvent (o) to the water (w) in the extract is from 7 to 8: 3-2 (V)O/Vw）。
In some embodiments, the ratio of the amount of the extracting solution used in the step S1 to the amount of the dendrobium candidum serving as the raw material is 8-10L: 1 kg; ultrasonic-assisted extraction is adopted during cold soaking at room temperature, and the extraction times are 2-3.
In some embodiments, the organic solvent used in the liquid-liquid extraction technique of step S2 is ethyl acetate, and the ratio of ethyl acetate to water is 1: 1 (v/v) and the extraction times are 2-3.
In some embodiments, the solid phase extraction column of step S2, wherein the stationary phase is C-18 reverse phase silica gel, non-polar macroporous adsorption resin, or non-polar small pore adsorption resin; the using amount of the stationary phase is that the ratio of the dry extract weight of the dendrobium huoshanense extract obtained in the step S1 to the volume of the stationary phase bed is 0.3 kg: 1L of the compound.
In some embodiments, the amount of water used for impurity removal and elution is 4-5 times of the bed volume, and in elution with an organic solvent-water mixed solution, the organic solvent is methanol or ethanol; the volume fraction of methanol or ethanol contained in the eluent is 20-50% (v/v), and the dosage of the organic solvent-water mixed eluent is 5-6 times of the bed volume.
On the other hand, the invention provides the application of the dendrobium candidum effective part in preparing medicines and health products for treating neurodegenerative diseases, liver protection and diabetes.
In some embodiments, the neurodegenerative disease is senile dementia or alzheimer's disease.
In some embodiments, the diabetes is type ii diabetes.
In some embodiments, the liver protecting effect is free radical scavenging activity.
The invention provides a dendrobium candidum effective part with a brand-new composition, and the effective part defines effective components, thereby being beneficial to the quality control and the medication safety of medicaments; experimental results show that the effective part of the dendrobium candidum has stronger oxidation resistance, stronger alpha-glucosidase inhibition activity and stronger glutamic acid induced HT22 cell damage inhibition, and can be used for treatment or adjuvant treatment of neurodegenerative diseases, liver protection and diabetes.
Drawings
FIG. 1 shows the protective effect of the effective fraction of Dendrobium officinale on the induction of HT22 cell death by L-glutamic acid. Positive control NAC: mM, glutamic acid Glu: mM, and the effective part series concentration of dendrobium officinale: μ g/mL.
Detailed Description
The present invention is further explained below with reference to specific examples, which are not intended to limit the present invention in any way.
In the invention, the structural formulas of the monoglycosides flickiflomside B and (+) -syringomycins-O-beta-D-glucopyranoside are as follows:
flickinflimoside B
(+) -syringaresinol-O-beta-D-glucopyranoside
The protons marked by a and c in the figure are the target proton positions selected for the nuclear magnetic resonance quantitative analysis.
The monoglycosides FLICkinflimoside B and (+) -syringaresinol-O-beta-D-glucopyranoside are known in the art and can be prepared by methods described in 1. Chen, J.L., ZHao, Z.M., Xue, X., Tang, G.H., Zhu, L.P., Yang, D.P., Jiang, L.Bioactive noditierpenoids fromFlickingeria fimbriata. RSC Adv. 2014, 4, 14447–14456. DOI: 10.1039/C4RA00835A; 2. Wang, C.Z.; Yu, D.Q. Lignan and acetylenic glycosides from Aster auriculatus. Phytochemistry1998, 48, 711–717. https://doi.org/10.1016/S0031-9422(98) 00019-3。
The compound of formula (I) is a novel compound isolated by the present inventors and having the following structural formula, which is also referred to herein as the diglycoside flifridioside a:
the proton marked by b in the figure is the target proton position selected for nuclear magnetic resonance quantitative analysis.
Example 1 preparation of effective fractions of Dendrobium officinale
S1, cold soaking 1kg of dendrobium candidum in 80% acetone aqueous solution at room temperature, wherein the material-liquid ratio is 8: 1, ultrasonic-assisted extraction, standing for 48 hours at a speed of 80 min/d, filtering, and concentrating the filtrate to obtain a dendrobium candidum extract; the extraction was repeated 3 times.
S2, suspending the dendrobium huoshanense extract obtained in the step S1 in 800mL of water, adding equal volume of ethyl acetate for liquid-liquid extraction, collecting a water phase, and spin-drying to obtain the dendrobium huoshanense effective part. The ethyl acetate extraction was repeated 3 times.
The result shows that the content of the glucoside compounds of the effective part of the dendrobium candidum accounts for 12 percent of the total mass of the effective part of the dendrobium candidum. The content of flickifimoside B accounts for 7 percent of the total mass of the effective parts of the dendrobium officinale, the content of the compound shown in the formula (I) accounts for 4 percent of the total mass of the effective parts of the dendrobium officinale, and the content of (+) -syringomycins-O-beta-D-glucopyranoside accounts for 1 percent of the total mass of the effective parts of the dendrobium officinale. The content analysis method adopts a quantitative nuclear magnetic resonance hydrogen spectrum technology, and is detailed in example 4. The pharmacological activity is shown in table 1 and figure 1.
A process for the preparation of compound (I): and (3) separating and purifying the effective part of the dendrobium huoshanense obtained in the step (S2) by using a silica gel column, carrying out gradient elution by using dichloromethane-methanol, collecting corresponding fractions, purifying by using a Sephadex LH-20 column, and using methanol as an eluent to obtain a pure product.
Spectral data: [ a ] 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)。
Example 2 preparation of the effective fractions of Dendrobium officinale
S1, cold soaking 1kg of dendrobium candidum in 80% methanol water solution at room temperature, wherein the material-liquid ratio is 10: 1, ultrasonic-assisted extraction, standing for 48 hours at a speed of 80 min/d, filtering, and concentrating the filtrate to obtain a dendrobium candidum extract; the extraction was repeated 3 times.
S2, adding a proper amount of methanol to dissolve the dendrobium candidum extract obtained in the step S1, and performing solid phase extraction on the dendrobium candidum extract by using a C-18 reverse phase silica gel column, wherein the ratio of the weight of a dry extract of the dendrobium candidum extract to the volume of a fixed phase bed of the C-18 reverse phase silica gel column is 0.3 kg: and 1L, after the sample loading is finished, eluting with water with the volume 5 times that of the bed to remove impurities, eluting with a methanol-water solution with the volume 6 times that of the bed, collecting the methanol-water eluent, concentrating and drying to obtain the dendrobium candidum effective part.
The content of the glucoside compounds of the effective part of the dendrobium candidum accounts for more than 14 percent of the total mass of the effective part. Wherein the content of flickiflomoside B accounts for 7 percent of the total mass of the effective parts of the dendrobium officinale, the content of the compound shown in the formula (I) accounts for 5 percent of the total mass of the effective parts of the dendrobium officinale, and the content of (+) -syringomycins-O-beta-D-glucopyranoside accounts for 2 percent of the total mass of the effective parts of the dendrobium officinale. The content analysis method adopts a quantitative nuclear magnetic resonance hydrogen spectrum technology, and is detailed in example 4. The pharmacological activity is shown in table 1 and figure 1.
Example 3 preparation of the effective fractions of Dendrobium officinale
S1, cold soaking 1kg of dendrobium candidum in 80% ethanol water solution at room temperature, wherein the material-liquid ratio is 10: 1, ultrasonic-assisted extraction, standing for 48 hours at a speed of 80 min/d, filtering, and concentrating the filtrate to obtain a dendrobium candidum extract; the extraction was repeated 3 times.
S2, adding a proper amount of water to the dendrobium candidum extract obtained in the step S1 for dissolving, and performing solid phase extraction on the dendrobium candidum extract by using a macroporous adsorption resin column, wherein the ratio of the weight of dry extract of the dendrobium candidum extract to the volume of a fixed phase bed of the macroporous adsorption resin column is 0.3 kg: 1L, eluting with 5 times bed volume of water to remove impurities, eluting with 6 times 40% ethanol-water solution, collecting ethanol-water eluate, concentrating, and drying to obtain herba Dendrobii effective component;
the result shows that the content of the glucoside compounds of the effective part of the dendrobium candidum accounts for more than 15 percent of the total mass of the effective part. Wherein the content of flickiflomoside B accounts for 8 percent of the total mass of the effective parts of the dendrobium officinale, the content of the compound shown in the formula (I) accounts for 5 percent of the total mass of the effective parts of the dendrobium officinale, and the content of (+) -syringomycins-O-beta-D-glucopyranoside accounts for 2 percent of the total mass of the effective parts of the dendrobium officinale. The content analysis method adopts a quantitative nuclear magnetic resonance hydrogen spectrum technology, and is detailed in example 4. The pharmacological activity is shown in table 1 and figure 1.
Example 4 content determination method of Dendrobium huoshanense effective part
The content analysis of the glucoside compounds in the effective part of the dendrobium candidum adopts a hydrogen nuclear magnetic resonance quantitative analysis technology. The 500MHz NMR spectrometer is adopted, and the experimental parameters are set as follows: the method comprises the following steps of pulse sequence zg30, spectrum width SW =14 ppm, center frequency O1P =6 ppm, sampling time AQ =3 s, sampling point number TD =41976, waiting time D1=2 s, scanning frequency NS =64, conversion point number SI =65536, and quantitative peaks such as a-position CH proton peak in a flickinflside B structure, B-position CH proton peak of a compound shown in formula (I) and c-position CH proton peak of (+) -syringaresinol-O-beta-D-glucopyranoside. The proton on the aromatic ring in the salicylic acid molecule is used as an external standard, and the quantitation is carried out by an ERETIC2 method.
Sample operation, accurately weighing 30mg of extract of the effective part of the dendrobium candidum which is dried to constant weight, adding deuterated methanol, performing fixed dissolution to 1mL, sealing a container, dissolving the sample by vortex, performing centrifugal sedimentation on insoluble solids, taking 600 mu L of supernatant, putting the supernatant into a nuclear magnetic tube (5 mm), and immediately performing nuclear magnetic resonance hydrogen spectrum analysis. The operations were performed in parallel for 3 times. The same procedure was carried out for salicylic acid, a standard sample. The content of glycoside compounds in the effective parts of salicylic acid and dendrobium candidum is measured by an ERETIC2 method.
Example 5 determination of the radical scavenging ability (by DPPH radical scavenging method)
Reagent: preparing DPPH solution with 150 μ M concentration by using methanol; the dendrobium candidum effective part solution is prepared by DMSO, the initial concentration is 30mg/mL, and the dendrobium candidum effective part solution is diluted into a series of concentrations in equal proportion when in use. Positive control: 2,6-ditertbutyl-4-methyl-phenol (BHT) solution, DMSO, initial concentration 10 μmol/mL, when using, equal ratio dilution to series concentration.
The method comprises the following steps: taking 96 holesIn the cell culture plate, 180 mu L of DPPH solution, 20 mu L of the solution of the effective part of the dendrobium huoshanense with each gradient concentration and 20 mu L of the solution of the positive control product BHT with each gradient concentration are respectively added into each hole, and meanwhile, a DPPH control group is arranged, and the absorbance value (A) of each hole is measured at 517 nm on an enzyme labeling instrument. The measurement is carried out every 10 minutes for 60 min. The clearance (%) was calculated as [ (a)Control- ASample (I))/AControl]X 100%. The results are shown in table 1.
2+EXAMPLE 6 measurement of free hydroxyl radical elimination ability (by o-diazaphenanthrene-Fe oxidation)
Reagent: an aqueous solution of phenanthroline, at a concentration of 5 mM. FeSO4Aqueous solution, concentration 7.5 mM. PBS buffer, 0.75M, pH 7.4. Determination application liquid: when in use, the o-diazepine solution is prepared by the following steps: PBS buffer: redistilling water: FeS04Solution, as 3: 8: 8: 2 are mixed sequentially. The dendrobium candidum effective part solution is prepared by DMSO, the initial concentration is 30mg/mL, and the dendrobium candidum effective part solution is diluted into a series of concentrations in equal proportion when in use. The positive control, Vit C solution, was prepared in DMSO at an initial concentration of 10 μmol/mL and diluted to serial concentrations at equal ratios for use. 1% hydrogen peroxide.
The method comprises the following steps: and (3) taking a 96-well cell culture plate, and respectively adding 170 mu L of the determination application liquid, 20 mu L of the effective part solution of the dendrobium huoshanense with each concentration gradient, or 20 mu L of the positive control Vit C solution and 10 mu L of 1% hydrogen peroxide into each well. Setting blank group, control group and sample group, keeping temperature at 37 ℃ for 10 min, and measuring absorbance value (A) of each hole at 510 nm within 10-120 min. Hydroxyl radical scavenging rate (%) - (A)Sample (A)- AInjury of the skin)/ （AWithout damage- AInjury of the skin）]X 100%. The results are shown in table 1.
Example 7-measurement of alpha-glucosidase inhibitory Activity
Reagent: KH (Perkin Elmer)2PO4-K2HPO4Buffer solution at 50 mM, PH = 7. The substrate p-nitrophenol-alpha-glucoside (PNPG) solution is used at the concentration of 1 mM and KH2PO4-K2HPO4BufferAnd (4) solution preparation. The alpha-glucosidase solution is applied by KH2PO4-K2HPO4The buffer solution is formulated to the desired concentration. Positive control: acarbose, DMSO, initial concentration of 10 u mol/mL, use equal ratio dilution series concentration. The effective part DMSO solution of Dendrobium officinale has an initial concentration of 30mg/mL, and is diluted into a series of concentrations at equal ratio when in use.
The method comprises the following steps: the 96-well plate was taken and the corresponding solution was added to the corresponding well according to the following table. The 1min absorbance change was measured at λ =400 nm. The test was performed 3 times in parallel and the average was taken.
Calculating the relative activity of the enzyme inhibitor: relative enzyme inhibitory activity = (A)0-A）/A0*100%。
A0: no enzyme catalytic activity (OD/min) of the sample was added.
A: inhibitory activity of the sample on the enzyme (OD/min). The results are shown in table 1.
Example 8 protective Effect on L-glutamate induced HT22 cell death
Reagent: the dendrobium huoshanense effective part solution is prepared by DMSO, the initial concentration is 30mg/mL, and the dendrobium huoshanense effective part solution is diluted into a series of concentrations in equal proportion when in use. L-glutamate solution in DMSO at a concentration of 100 mM. Positive control N-acetyl cysteine (NAC), in DMSO, at an initial concentration of 10. mu. mol/mL.
The method comprises the following steps: mouse hippocampal neuron cell line HT22 was cultured in DMEM complete medium containing 10% fetal calf serum at 37 deg.C and saturated humidity, and containing 5% CO by volume2The carbon dioxide incubator of (1) is used for conventional culture. Taking cells in logarithmic growth phase, digesting with 0.25% pancreatin, completely suspending the cells in a culture medium, counting the cells by a cell counting plate under a microscope, adjusting the cell concentration to 10 multiplied by 104 cells/ml, inoculating 96-well cell culture plates, culturing at 100 mu L/well overnight, and enabling the cells to adhere to the wall. Sucking the culture medium in a 96-well plate, and adding the solution with the serial concentration of the effective parts of the dendrobium officinale into the 96-well plate at a concentration of 100 mu L/well. Preparation ofAfter incubation for 30min, 2. mu.L of 100mM L-glutamate was added. Model group without sample, 2. mu.L of 100mM L-glutamate was added directly. After 24h incubation, 10. mu.L of 5mg/mL MTT was added to each well, incubation was carried out for 2h, the supernatant was discarded, 100. mu.L DMSO/well was added, the resultant formazan was sufficiently dissolved by shaking, the absorbance value of each well was measured on a microplate reader, and the wavelength was measured at 570 nm. Calculating the survival rate of the cells:
protection against L-glutamate induced HT22 cell death (%) -100% (a)Sample to be tested-ABlank space)/(AModel (model)-ABlank space). The results are shown in FIG. 1.
TABLE 1 biological activity test results of the effective part of Dendrobium huoshanense
/: no test; n = 3.
Claims (8)
1. The dendrobium candidum effective part contains 10-20% of glucoside compounds in the total mass of the effective part, wherein the glucoside compounds comprise flickingflinoside B, (+) -syringomycins-O-beta-D-glucopyranoside and compounds of formula (I)
the dendrobium huoshanense effective part is prepared by the following steps:
s1, cold soaking and extracting the dendrobium candidum by using a mixed solution of a hydrophilic organic solvent and water as an extracting solution, filtering, and concentrating the filtrate to obtain the dendrobium candidum extract;
s2, adding a proper amount of water into the dendrobium candidum extract obtained in the step S1 for suspension, performing liquid-liquid extraction by using ethyl acetate, collecting a water phase, and performing spin drying to obtain the dendrobium candidum effective part.
2. The dendrobium candidum effective part contains 10-20% of glucoside compounds in the total mass of the effective part, wherein the glucoside compounds comprise flickingflinoside B, (+) -syringomycins-O-beta-D-glucopyranoside and compounds of formula (I)
the dendrobium huoshanense effective part is prepared by the following steps:
s1, cold soaking and extracting the dendrobium candidum by using a mixed solution of a hydrophilic organic solvent and water as an extracting solution, filtering, and concentrating the filtrate to obtain the dendrobium candidum extract;
s2, adding a proper amount of methanol to dissolve the dendrobium candidum extract obtained in the step S1, subjecting the dendrobium candidum extract to C-18 reverse phase silica gel column or macroporous adsorption resin column solid phase extraction and enrichment, eluting the extract with water to remove impurities after sample loading is finished, eluting the extract with a water-containing organic solution, collecting the water-containing organic eluent, concentrating and drying to obtain the dendrobium candidum effective part.
3. The dendrobium candidum effective part according to claim 1, wherein the content of flickiflomioside B accounts for 6-8% of the total mass of the dendrobium candidum effective part, the content of the compound shown in the formula (I) accounts for 4-6% of the total mass of the dendrobium candidum effective part, and the content of (+) -syringaresinol-O-beta-D-glucopyranoside accounts for 0.5-1% of the total mass of the dendrobium candidum effective part.
4. The dendrobium candidum effective part according to claim 2, wherein the content of flickiflomioside B accounts for 6-8% of the total mass of the dendrobium candidum effective part, the content of the compound shown in the formula (I) accounts for 4-6% of the total mass of the dendrobium candidum effective part, and the content of (+) -syringaresinol-O-beta-D-glucopyranoside accounts for 1-2.5% of the total mass of the dendrobium candidum effective part.
5. Use of the dendrobium candidum effective part of any one of claims 1 to 4 in the preparation of a medicament for treating diabetes.
6. The use according to claim 5, wherein the diabetes is type II diabetes.
7. A method for preparing the effective component of dendrobium candidum of claim 1 or 3, which comprises the following steps:
s1, cold soaking and extracting the dendrobium candidum by using a mixed solution of a hydrophilic organic solvent and water as an extracting solution, filtering, and concentrating the filtrate to obtain the dendrobium candidum extract;
s2, adding a proper amount of water into the dendrobium candidum extract obtained in the step S1 for suspension, performing liquid-liquid extraction by using ethyl acetate, collecting a water phase, and performing spin drying to obtain the dendrobium candidum effective part.
8. A method for preparing the effective component of dendrobium candidum of claim 2 or 4, which comprises the following steps:
s1, cold soaking and extracting the dendrobium candidum by using a mixed solution of a hydrophilic organic solvent and water as an extracting solution, filtering, and concentrating the filtrate to obtain the dendrobium candidum extract;
s2, adding a proper amount of methanol to dissolve the dendrobium candidum extract obtained in the step S1, subjecting the dendrobium candidum extract to C-18 reverse phase silica gel column or macroporous adsorption resin column solid phase extraction and enrichment, eluting the extract with water to remove impurities after sample loading is finished, eluting the extract with a water-containing organic solution, collecting the water-containing organic eluent, concentrating and drying to obtain the dendrobium candidum effective part.
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