CN108558809A - Compound Oleracone F and its extraction separation method in purslane - Google Patents
Compound Oleracone F and its extraction separation method in purslane Download PDFInfo
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- NXGQOPCTFSYSRJ-UHFFFAOYSA-N COc(cc1OC=C2Cc(cccc3)c3[O]=C)cc(OC)c1C2=O Chemical compound COc(cc1OC=C2Cc(cccc3)c3[O]=C)cc(OC)c1C2=O NXGQOPCTFSYSRJ-UHFFFAOYSA-N 0.000 description 1
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
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Abstract
The present invention relates to traditional Chinese medicine extraction, separation fields, more particularly to extracting and developing and the noval chemical compound and its extraction separation method that identify from purslane medicinal material, compound Oleracone F and its extraction separation method in specially a kind of purslane.The present invention provides noval chemical compound, and molecular formula is respectively C18H16O5, it is named as oleracone F.It has been investigated that novel compound of present invention has anti-inflammatory and oxidation resistant effect, while providing a kind of extraction separation method high for the easy, quick, environmentally friendly of noval chemical compound of the present invention, purity.
Description
Technical field
The present invention relates to traditional Chinese medicine extraction, separation field, more particularly to extracting and developing and is identified from purslane medicinal material
Noval chemical compound and its extraction separation method, compound Oleracone F and its extraction separation method in specially a kind of purslane.
Background technology
Purslane (Portulaca oleracea L.) also known as long life dish, horse three-coloured amaranth are portulacaceous plant.Purslane
It is drought-enduring waterlogging, and fast light shade tolerant, widely distributed, resourceful, the wild plant as medicine-food two-purpose is concerned, 2015 editions
《Pharmacopoeia of People's Republic of China》In record the dry aerial parts of purslane and be used as medicine, there is clearing heat and detoxicating, cooling blood and hemostasis, stop dysentery
And other effects, for toxic-heat and blood stasis, carbuncle swells furunculosis, eczema, erysipelas, snakebite and bugbite, have blood in stool, hemorrhoid blood, metrostaxis etc..
Purslane modern pharmacology research shows it with anti-inflammatory analgesic, anti-bacteria and anti-virus, lowering blood pressure and blood fat, antioxygen
Change, anticancer, relaxation skeletal muscle and smooth muscle adjust the effects that immune function.Research shows that the numerous chemical compositions of purslane are it
Various pharmacological action provides material base, and purslane main chemical compositions include flavonoids, cumarin, terpene, steroid, life
Alkaloids, amino acid, various pigments and minerals class etc..Wherein alkaloid is a kind of main chemical composition, mesh in purslane
The preceding composition of alkaloids reported have norepinephrine, dopamine, a small amount of DOPA, adenosine, uracil, adenine, N, N-
Dicyclohexylurea (DCU), allantoin, N- be trans--asafoetide acyl group tyrasamine；Also Cyclic dipeptides alkaloid and amide alkaloid：Purslane acyl
Amine A-I, K, L, N-S.
Most of chemical composition isolated from purslane at present is known, and structure novel is relatively low, therefore, right
The exploitation and separation of noval chemical compound urgently need in purslane.
Invention content
In view of the above-mentioned problems, compound Oleracone F and its extraction separation method in a kind of purslane of present invention offer,
The noval chemical compound specially extracted from purslane, it has been investigated that novel compound of present invention has anti-inflammatory and oxidation resistant work
With, while a kind of extraction separation method high for the easy, quick, environmentally friendly of noval chemical compound of the present invention, purity being provided.
Above-mentioned purpose to realize the present invention, the present invention provide noval chemical compound, molecular formula C18H16O5, it is named as
oleracone F.Chemical structural formula is：
Above-mentioned purpose to realize the present invention, the present invention also provides purslane in compound oleracone F extraction
Separation method, the specific steps are.
Step 1 takes purslane to dry medicinal material, uses alcohol extracting 8~16 times of medicinal material (ethanol consumption for) twice, alcohol extract
Filtration, merging filtrate directly heat concentration, cool to room temperature, it is spare to obtain liquid.
Step 2, upper silicagel column after being evaporated step 1 herb liquid, are eluted with ethyl acetate, and ethyl acetate is recovered under reduced pressure to leaching
Cream obtains ethyl acetate extract.
Step 3 detaches ethyl acetate extract in step 2 through polyamide column, using alcohol-water gradient elution, 50%
Ethyl alcohol upper silicagel column after being evaporated, obtains several elution positions with acetate-methanol gradient elution successively, is carried out through thin-layer chromatography
Detection, colour developing, merges the elution position of colour developing, spare by the elution position after merging through being concentrated to dryness.
Step 4, by gains in step 3 pretreated ODS columns (Octadecylsilyl, octadecylsilane key again
Close silica filler) chromatography obtains several elution positions, is detected through thin-layer chromatography with methanol-water gradient elution, it shows
The elution position of each colour developing is concentrated to dryness, it is spare to obtain concentrate by color respectively.
Gained noval chemical compound in step 4 is detached preparation by step 5 by HPLC (efficient liquid phase), with -0.1% first of methanol
Acid is that mobile phase carries out isocratic elution, finally obtains compound oleracone F.
The preprocessing process of the ODS is that methanol impregnated 24 hours, upper prop, is washed till in instillation water without muddiness with methanol,
It is balanced each other again with initial flow.
Compared with prior art, beneficial effects of the present invention.
The separation of heretofore described purslane noval chemical compound and pharmacology activity research are not reported by existing paper periodical；
The present invention provides noval chemical compound and a kind of extraction separation method for noval chemical compound of the present invention from purslane, using alcohol
Compression leg and HPLC are isolated and purified and are prepared in extraction, silica gel column chromatography, polyamide column, ODS, and successfully extraction is isolated new
Compound, this method operating procedure are only five steps, and operating method is easy and quickly, extraction separation process mainly use alcohol extracting and
Ethyl acetate elutes, process environmental protection, and is all higher than 90% through the isolated compound purity of this method is higher, passes through in addition
Research shows that the above compound has anti-inflammatory and antioxidation.Therefore, noval chemical compound and its salt and derivative of the present invention can be with
As other compound synthesis primers and the raw material of new drug development and pharmacology activity research, also can be used for preparing it is anti-inflammatory and
Oxidation resistant drug.
Description of the drawings
Fig. 1 is the ultraviolet spectrogram of noval chemical compound oleracone F of the present invention.
Fig. 2 is the infrared spectrogram of noval chemical compound oleracone F of the present invention.
The high resolution mass spectrum figure of Fig. 3 noval chemical compound oleracone F of the present invention.
Fig. 4 noval chemical compound oleracone F of the present invention1H-NMR spectrograms.
Fig. 5 invention noval chemical compound oleracone F13C-NMR spectrograms.
Carbon-13 nmr spectra (DEPT) spectrogram of Fig. 6 noval chemical compound oleracone F of the present invention.
The nuclear magnetic resonance of Fig. 7 noval chemical compound oleracone F of the present invention1H-1HCOSY spectrograms.
The nuclear magnetic resonance HMBC spectrograms of Fig. 8 noval chemical compound oleracone F of the present invention.
The nuclear magnetic resonance HSQC spectrograms of Fig. 9 noval chemical compound oleracone F of the present invention.
Specific implementation mode
With reference to specific embodiment, the present invention is described further.
Embodiment 1.
The present invention provides noval chemical compound, molecular formula C18H16O5, being named as oleracone F chemical structural formulas is：
The noval chemical compound is named as oleracone F according to structure, and table 1 is the nuclear magnetic data of the noval chemical compound：1H-
NMR with13C-NMR is in DMSO.
Table 1：The nuclear magnetic data of noval chemical compound oleracone F of the present invention.
Structural Identification is referring to Fig. 1-9.
Oleracone F：Yellow powder is soluble in methanol, is slightly soluble in chloroform.Point sample sprays trichlorine after silica gel thin-layer plate
It is in cyan to change iron test solution spot.UV(MeOH)λmax:281,255,205nm。IR vmax 2919,2850,1647,1600,1456,
1378,1294,1209,1169,1081,834,756cm-1.HRESI (+) TOFMS provides m/z:313.1066[M+H]+Standard point
Daughter ion peak, molecular weight 312.3210.In conjunction with1H-NMR,13C-NMR and DEPT data, thus it is speculated that possible point of the compound
Minor is C18H16O5, degree of unsaturation 11.13C-NMR is composed and DEPT spectrums show 18 carbon signals, respectively 2 CH3(δ：56.0；
55.9), 1 CH2(δ：25.1), 7 CH (δ：92.9；96.0；115.2；118.8；127.2；130.0；151.1), 8 quaternary carbons
(carbonyl carbon, δ：174.9；The double key carbon of four company O, δ：163.5；160.4；159.5；155.1；Three double key carbons, δ：
125.3；123.4；108.3).
1H-NMR spectrums show an active H signal δ 9.49 (1H, bs), show that there may be a hydroxyl groups.2 first
Base signal is δ 3.84 (3H, s), δ 3.80 (3H, s)；1 methylene signals is δ 3.51 (2H, s)；7 methine signals point
Not Wei δ 6.47 (1H, d, J=2.4), δ 6.60 (1H, d, J=2.3), δ 6.68 (1H, brtd, J=7.4,1.1), δ 6.78 (1H,
Dd, J=8.1,1.1), δ 7.00 (1H, brtd, J=7.6,1.6), δ 7.06 (1H, dd, J=7.5,1.6), δ 7.81 (1H, s).
According to H-H COSY spectrums it is found that the H δ 3.51 and methine δ 7.86 in methylene is coupled；Two methine δ 6.47 and δ 6.60
It is coupled；Methine δ 6.68, δ 6.78, δ 7.00, δ 7.03 intercouples, and illustrates the presence for having phenyl ring.It is composed according to HMBC related
Peak shows H-6, and H-8 is coupled with C-7, C-10 respectively, and H-6, H-8 intercouple, and illustrates and C-7, C-10 are associated；Methoxy
H δ 3.84 and C-7 in base is coupled, and C-7 (δ 163.5) is located at low field area, prompts to be connected with O, illustrates methoxyl group and phenyl ring
On C-7 be connected；H δ 3.80 and C-5 in methoxyl group is coupled, and C-5 (δ 160.4) is located at low field area, prompts to be connected with O,
Illustrate that methoxyl group is connected with the C-5 on phenyl ring；H-8 is coupled with C-9, and H-6 is coupled with C-5, and C-9 (δ 157.73) is located at low
Place prompts to be connected with O；H-2, H-11 intercouple, and H-2, H-11 are coupled with C-3, illustrate related to C-3；C-2(δ
151.1) it is located at low field area, prompts to be connected with O, meanwhile, H-2 is coupled with C-9, illustrates C-2, is connected with O among C-9；H-2,
The carbonyl C of H-11 and C-4 is coupled, and illustrates associated with C-4.H-3 ＇, H-4 ＇, H-5 ＇ and H-6 ＇ intercouple, wherein H-3 ＇,
H-5 ＇ and C-1 ＇ are coupled, and H-3 ＇, H-4 ＇, H-6 ＇ and C-2 ＇ are coupled, and prompt the presence for having phenyl ring and C-1 ＇ and the ortho positions C-2 ＇
Substitution；Wherein C-2 ＇ (δ 155.1) are in low field area, prompt to be connected with O, and illustrating C-2 ＇, even there are one hydroxyl groups；H-11 and C-
1 ＇, C-2 ＇, C-6 ＇ are coupled, and illustrate and C-1 ＇, C-2 ＇, C-6 ＇ are related, to sum up illustrate that C-11 is connected with C-1 ＇.According to the above letter
Breath, it may be determined that this noval chemical compound is above structure.
The extraction separation method of above-mentioned noval chemical compound, the specific steps are.
Step 1：Purslane drying medicinal material 80kg is weighed, is extracted using 50% alcohol reflux, 50% ethanol consumption (v/v) is
10 times of medicinal material, twice, ethyl alcohol is recovered under reduced pressure in each 2h to refluxing extraction, cools to room temperature, it is spare to obtain liquid.
Step 2：It detaches through silica gel column chromatography after gained liquid in step 1 is evaporated, is washed with ethyl acetate (120L) is isocratic
De-, wherein silica gel is 100-200 mesh, and 40 DEG C or less are recovered under reduced pressure ethyl acetate to medicinal extract, obtain ethyl acetate extract.
Step 3：Ethyl acetate extract in step 2 is detached through polyamide column, using alcohol-water (0/100,30/70,
50/50,70/30,100/0, v/v) gradient elution, 50% ethyl alcohol detach after being evaporated through silica gel column chromatography, wherein silica gel be 200~
300 mesh use acetate-methanol (5 successively:1、2:1、1:2, v:V) gradient elution is obtained 15 positions and (is obtained 15
A bottle, every bottle of 400mL), it is detected, develops the color through thin-layer chromatography, merge 1~3 elution position of colour developing, by 1~3 after merging
40 DEG C of position or less is concentrated to dryness, spare.
Step 4：By gains in step 3 again the separation of pretreated ODS medium pressure column chromatographies (Octadecylsilyl, ten
Eight alkyl silane bonded silica gel fillers), wherein filler particle size is 20~40 μm, with methanol-water (84/16,93/7,97/3,100/
0, v/v) gradient elution (pressurization, it is 1mL/min to make flow velocity, and temperature is room temperature), obtaining 10 positions, (i.e. gradient elution obtains 10
Bottle, every bottle of 200mL), it is detected, develops the color through thin-layer chromatography, the positions 1-5 of colour developing are retained, 50 DEG C or less are concentrated under reduced pressure into
It is dry, it is spare, obtain noval chemical compound.The preprocessing process of the ODS is that methanol impregnated for 24 hours, and upper prop is washed till instillation water with methanol
Middle no muddiness, then balanced each other with initial flow.
Step 5：Gained noval chemical compound in step 4 is detached through HPLC and is prepared, with methanol：0.1% formic acid (50：50, v/v)
As mobile phase, Detection wavelength 210nm, 280nm, noval chemical compound of the present invention is prepared in separation, and it is equal that normalization method measures purity
It is 90~99%.
The anti-inflammatory effect of noval chemical compound of the present invention.
1, main material.
1.1, drug and reagent：Experiment noval chemical compound used is prepared by the above method, and purity is 90~99%, and precision claims
Take, with DMSO be diluted to following each dosage groups needed for solution.DMEM high glucose mediums, fetal calf serum (Hyclone companies of the U.S.)；
Penicillin, streptomysin (Hangzhou Chinese holly company)；LPS (Sigma Co., USA)；IL-6、TNF-α、PGE2ELISA kit
(Cayman companies of the U.S.)；Cell pyrolysis liquid, Griess reagents (green skies Bioisystech Co., Ltd).
1.2 cell strain：RAW264.7 macrophages (U.S.'s ATCC cell banks).
1.3 grouping：It is divided into control group, LPS groups and experimental group, each one group.
2 experimental methods.
The fetal calf serum of l0%, l% antibiotic (100U/mL penicillin is added in 2.1 cell culture, DMEM high glucose mediums
With 100 μ g/mL streptomysins), it is placed in 37.5%, CO2It is cultivated in incubator.
2.2MTT colorimetric method for determining cell viabilities, above-mentioned three groups of difference logarithmic growth phase RAW264.7 macrophages inoculation
In 96 well culture plates, cell density is 1 × 104A/mL, per 100 μ L of hole, 37 DEG C of temperature, 5%CO2Under the conditions of overnight incubation
Afterwards, the noval chemical compound oleracone F (1-50 μM) of the present invention of various concentration are added in experimental group, are incubated after 1h to LPS groups and reality
The LPS that group is separately added into final concentration of 1 μ g/mL is tested, zeroing group (culture solution of the solvent containing DMSO) is separately set, every group sets 3 multiple holes,
Investigate the influence being added after drug to cell.After above-mentioned each group cell culture for 24 hours, 5mg/mL MTT are added in each hole cell
20 μ L, 37 DEG C of temperature, 5%CO2Under the conditions of continue be incubated 4h after, terminate culture, suction abandon liquid in hole, per hole be added 100 μ L bis-
Methyl sulfoxide (DMSO) vibrates 10min, makes to crystallize fully dissolving into the cell, each hole extinction is measured at microplate reader 570nm wavelength
Value.
2.3 measure the content of NO using Ge Lisi (Griess) method, investigate the mouse that noval chemical compound of the present invention induces LPS
The inhibiting effect of the NO yields of macrophage RAW264.7.Containing 10% tire ox blood after mouse macrophage RAW264.7 passages
It is cultivated in the sugared cell culture medium DMEM of clear height, the noval chemical compound oleracone F (1- of the present invention of various concentration are added in experimental group
20 μM), at 37 DEG C, 5%CO2Under the conditions of be incubated 1h after with LPS (final concentration of 1 μ g/mL) induce inflammatory reaction, collect afterwards for 24 hours
Supernatant, every group of processing repeat 3 holes.Griess methods measure the content of NO in cell supernatant, and according to various concentration, the present invention is new
Compound discharges the RAW264.7 cells that LPS is induced the influence of NO, to reflect NO levels.
2.4ELISA methods measure inflammatory factor IL-6, TNF-α and inflammatory mediator PGE2：Exponential phase RAW264.7 is huge
Phagocyte is inoculated in 24 well culture plates, and cell density is 1 × 105A/mL, per hole 1mL, 37 DEG C of temperature, 5%CO2Under the conditions of train
It supports overnight, noval chemical compound oleracone F (1-20 μM) of the present invention are added in experimental group, and after cultivating 1h, LPS is added (eventually in every hole
A concentration of 1 μ g/mL), it is incubated altogether for 24 hours, every group of processing repeats 3 holes.ELISA method measures purslane source noval chemical compound, and treated
IL-6, TNF-α and the PGE of RAW264.7 macrophages secretes2Content.
3 experimental results.
The experimental results showed that special compound of the present invention on the proliferation of the LPS macrophage RAW264.7 induced without influence,
It is safe and non-toxic；And it can effectively inhibit excessive inflammatory cytokine IL-6, TNF- produced by the macrophage RAW264.7 of LPS inductions
α and inflammatory mediator NO, PGE2, and be in concentration dependant.
Comparative survival rate of cells experimental result is as shown in table 2.
Table 2：Influence of the present invention to RAW264.7 macrophage relative survival rates.
Note：*P<0.05 compared with the control group (the significant difference of high concentration group),
The content experimental result that NO is measured using Ge Lisi (Griess) method is shown in Table 3.
Table 3：The present invention discharges the RAW264.7 cells that LPS is induced the influence (mean ± standard deviation, n=3) of NO.
Note：*P<0.05 compared with the control group,#P<0.05 compared with LPS groups.
ELISA method measures inflammatory factor IL-6, TNF-α and inflammatory mediator PGE2The results are shown in Table 4.
Table 4：IL-6, TNF-α and PGE of the present invention to the LPS RAW264.7 cells secretions induced2The influence of content is (
Number ± standard deviation, n=3).
Note：*P<0.05 compared with the control group,#P<0.05 compared with LPS groups.
The antioxidation of noval chemical compound of the present invention.
1 main material.
1.1 drugs and reagent：Experiment noval chemical compound used is prepared by the above method, and purity is 90~99%, and precision weighs,
The solution needed for methanol dilution to following each dosage groups.DPPH (1,1- diphenyl -2- picryl hydrazines free radical) (Sigma-Fluka
Company)；BHA (tert-butyl hydroxyanisole) (Shanghai auspicious sign Science and Technology Ltd.)；Methanol, chromatographically pure (the prosperous limited public affairs of Taixing industry
Department).
1.2 grouping：Control group, experimental group, blank group.
2. experimental method.
Colorimetric method for determining eliminates the ability of DPPH free radicals, and sample sets take 1mL DPPH solution (126.80 μM) to be added to
In 4mL cuvettes, the sample solution (8.01,16.02,32.04,48.06,64.08 μM) of 1mL various concentrations is added；Control
Group takes 1mL methanol solutions to be added in 4mL cuvettes, adds the sample solution of 1mL various concentrations；Blank group takes 1mL DPPH
Solution is added in 4mL cuvettes, adds 1mL methanol solutions.Three groups mix well, and room temperature, which is protected from light, stands 5min, 517nm
Lower measurement light absorption value.Each sample average measurement is averaged three times, and positive control is the BHA solution of various concentration.According to
Lower formula calculates clearance rate of the sample to DPPH free radicals, and further calculates its free radical scavenging activity IC50Value.
DPPH clearance rates (%)=1- (A1- A2)/A0× 100%, wherein A0For the absorbance value of blank group；
A1For the absorbance value of sample sets；A2For the absorbance value of control group.
3. experimental result.
The experimental results showed that noval chemical compound of the present invention has scavenging effect to DPPH free radicals, and increase with drug concentration,
Clearance rate is also significantly raised.Noval chemical compound of the present invention is to DPPH free radicals IC50Value is shown in Table 5.
Scavenging effect of 5 noval chemical compound of the present invention of table to DPPH free radicals.
| Group | IC50(μM) |
| BHA | 56.96 |
| Oleracone F | 17.78 |
In conclusion the present invention provides special compound and its extraction separation method, carried successively using 50% alcohol reflux
It takes, compression leg isolates and purifies in silica gel column chromatography, polyamide column chromatography, ODS, a kind of successful isolated noval chemical compound, the party
Method is easy, quickly, environmental protection, and it is higher through the isolated compound purity of this method, since gained compound chemical structure is only
Spy extracts from conventional Chinese medicine purslane, with anti-inflammatory and antioxidation, therefore special compound of the present invention and its
Salt and derivative can be used as natural products to develop new Chinese medicine, have broad prospects.
Claims (4)
1. compound Oleracone F in purslane, which is characterized in that the molecular formula of the compound is C18H16O5, it is named as
Oleracone F, chemical structural formula are.
2. compound Oleracone F in purslane as described in claim 1, which is characterized in that the specific steps are：
Step 1 takes purslane to dry medicinal material, uses alcohol extracting 8~16 times of medicinal material (ethanol consumption be) twice, and alcohol extract is filtered
It crosses, merging filtrate directly heats concentration, cools to room temperature, it is spare to obtain liquid；
Step 2, upper silicagel column after being evaporated step 1 herb liquid, are eluted with ethyl acetate, ethyl acetate are recovered under reduced pressure to medicinal extract,
Obtain ethyl acetate extract；
Step 3 detaches ethyl acetate extract in step 2 through polyamide column, using alcohol-water gradient elution, 50% ethyl alcohol
Upper silicagel column, obtains several elution positions with acetate-methanol gradient elution successively, is examined through thin-layer chromatography after being evaporated
It surveys, colour developing merges the elution position of colour developing, spare by the elution position after merging through being concentrated to dryness；
Step 4, by gains in step 3, pretreated ODS column chromatography for separation is obtained several with methanol-water gradient elution again
Position is eluted, is detected through thin-layer chromatography, develops the color, the elution position of each colour developing is concentrated to dryness respectively, is concentrated
Object is spare；
Gained noval chemical compound in step 4 is detached preparation by step 5 by HPLC, is carried out by mobile phase of -0.1% formic acid of methanol
Isocratic elution finally obtains compound oleracone F.
3. the extraction separation method of compound Oleracone F in purslane as described in claim 1, which is characterized in that tool
Body step is：
Step 1：Purslane drying medicinal material 80kg is weighed, is extracted using 50% alcohol reflux, 50% ethanol consumption (v/v) is medicinal material
10 times, twice, ethyl alcohol is recovered under reduced pressure in each 2h to refluxing extraction, cools to room temperature, it is spare to obtain liquid；
Step 2：It is detached through silica gel column chromatography after gained liquid in step 1 is evaporated, with ethyl acetate 120L isocratic elutions, wherein
Silica gel is 100-200 mesh, and 40 DEG C or less are recovered under reduced pressure ethyl acetate to medicinal extract, obtain ethyl acetate extract；
Step 3：Ethyl acetate extract in step 2 is detached through polyamide column, using alcohol-water (0/100,30/70,50/
50,70/30,100/0, v/v) gradient elution, 50% ethyl alcohol detach after being evaporated through silica gel column chromatography, and wherein silica gel is 200~300
Mesh uses acetate-methanol (5 successively:1、2:1、1:2, v:V) gradient elution is detected through thin-layer chromatography, is developed the color, and is merged
40 DEG C of position or less after merging is concentrated to dryness by the elution position of colour developing, spare；
Step 4：By the pretreated ODS medium pressure column chromatographies separation again of gains in step 3, wherein filler particle size is 20~40 μ
M is detected with methanol-water (84/16,93/7,97/3,100/0, v/v) gradient elution through thin-layer chromatography, and colour developing will develop the color
Position retain, 50 DEG C or less are concentrated to dryness, spare, obtain noval chemical compound；The preprocessing process of the ODS soaks for methanol
It steeped for 24 hours, upper prop, and was washed till to instill in water with methanol and be balanced each other without muddiness, then with initial flow；
Step 5：Gained noval chemical compound in step 4 is detached through HPLC and is prepared, with methanol：0.1% formic acid (50：50, v/v) conduct
Compound OleraconeF is prepared in mobile phase, Detection wavelength 210nm, 280nm, separation, and normalization method measures purity and is
90~99%.
4. compound Oleracone F are used to prepare anti-inflammatory and oxidation resistant drug in the purslane as described in claim 1-3.
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| CN109824685A (en) * | 2019-04-03 | 2019-05-31 | 辽宁中医药大学 | Compound oleracone G and its extraction separation method and application in purslane |
| CN110194755A (en) * | 2019-04-03 | 2019-09-03 | 辽宁中医药大学 | Compound Oleracone H in purslane, extraction and separation method and application thereof |
| CN110294733A (en) * | 2019-04-03 | 2019-10-01 | 辽宁中医药大学 | One kind Oleracone I of key compound containing peroxide and its extraction separation method and application in purslane |
| CN110305094A (en) * | 2019-07-16 | 2019-10-08 | 辽宁中医药大学 | Two kinds of flavone compounds and its extraction separation method and purposes in purslane |
| CN111217773A (en) * | 2020-03-12 | 2020-06-02 | 辽宁中医药大学 | Furan ring compound in purslane and extraction and separation method and application thereof |
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| CN109824685A (en) * | 2019-04-03 | 2019-05-31 | 辽宁中医药大学 | Compound oleracone G and its extraction separation method and application in purslane |
| CN110194755A (en) * | 2019-04-03 | 2019-09-03 | 辽宁中医药大学 | Compound Oleracone H in purslane, extraction and separation method and application thereof |
| CN110294733A (en) * | 2019-04-03 | 2019-10-01 | 辽宁中医药大学 | One kind Oleracone I of key compound containing peroxide and its extraction separation method and application in purslane |
| CN109824685B (en) * | 2019-04-03 | 2021-03-23 | 辽宁中医药大学 | Compound oleracene G in purslane, extraction and separation method and application thereof |
| CN110294733B (en) * | 2019-04-03 | 2021-09-07 | 辽宁中医药大学 | Peroxide bond-containing compound Oleracone I in purslane, and extraction separation method and application thereof |
| CN110194755B (en) * | 2019-04-03 | 2022-05-13 | 辽宁中医药大学 | Compound Oleracone H in purslane, extraction and separation method and application thereof |
| CN110305094A (en) * | 2019-07-16 | 2019-10-08 | 辽宁中医药大学 | Two kinds of flavone compounds and its extraction separation method and purposes in purslane |
| CN110305094B (en) * | 2019-07-16 | 2022-06-17 | 辽宁中医药大学 | Two flavonoid compounds in purslane and extraction and separation method and application thereof |
| CN111217773A (en) * | 2020-03-12 | 2020-06-02 | 辽宁中医药大学 | Furan ring compound in purslane and extraction and separation method and application thereof |
| CN111217773B (en) * | 2020-03-12 | 2022-05-13 | 辽宁中医药大学 | Furan ring compound in purslane and extraction and separation method and application thereof |
| CN115385884A (en) * | 2022-08-23 | 2022-11-25 | 辽宁中医药大学 | Extraction and separation method of new chromone alcohols in purslane and application thereof |
| CN115385884B (en) * | 2022-08-23 | 2023-04-25 | 辽宁中医药大学 | Extraction and separation method of neochronol in purslane and application thereof |
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