CN110038068A - A kind of application of oil tea exocarp in antithrombotic reagent - Google Patents
A kind of application of oil tea exocarp in antithrombotic reagent Download PDFInfo
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- CN110038068A CN110038068A CN201910136572.5A CN201910136572A CN110038068A CN 110038068 A CN110038068 A CN 110038068A CN 201910136572 A CN201910136572 A CN 201910136572A CN 110038068 A CN110038068 A CN 110038068A
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- exocarp
- oil tea
- antithrombotic reagent
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- antithrombotic
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/82—Theaceae (Tea family), e.g. camellia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The present invention relates to a kind of oil tea exocarp antithrombotic reagent application.The exocarp for first acquiring oil tea, is sieved after dry, pulverize, and is extracted 2-4 times with ethyl alcohol, and filtering, merging filtrate obtains brown solid by being dried under reduced pressure.It takes solids to be dissolved in water, crosses MCI resin, then rush column with various concentration methanol, obtain product drying and stored in the case where 4 DEG C of ﹣.Again by the product of measurement opposed polarity eluate to the TT of rat, APTT, the influence of PT and FIB, the platelet aggregation-against effect of these eluates is determined simultaneously, and the main chemical compositions structure of oil tea exocarp antithrombotic acitivity is further analyzed by LC-Q-TOF/MS, to make a kind of antithrombotic reagent of being developed into of oil tea exocarp provide support.
Description
Technical field
The invention belongs to drug development research field, application of specifically a kind of oil tea exocarp in antithrombotic reagent.
Background technique
The intracorporal blood coagulation of people and it is anticoagulant be homeostasis process under normal condition, coagulation factor and blood platelet are in the balance
It plays an important role in the process.Dysfunction of platelet (as aggregation increases) and blood coagulation will lead to blood " height solidifies ", lead to bolt
Fill in heart and brain diseases, such as coronary heart disease, angina pectoris and myocardial infarction.It is risen year by year in the disease incidence of China, thrombotic diseases,
Have become 21 century it is dead the main reason for one of.
Some Camellia Plants have good anticoagulation and anti thrombotic action, such as black tea and green tea, wherein more containing tea
Phenol can reduce blood clotting, inhibit platelet activation and agglutination, promote fibrinolytic.Oil tea [Camellia
OleiferaAbel], also referred to as tea tree is Theaceae (Camellia) Camellia Plants, also contains specific active physiological object
Matter, such as saponin(e, polysaccharide, flavones and tea polyphenols.Oil tea polyphenol is most important in oil tea and the widest secondary metabolites of distribution
One of, not only have a certain impact to the quality of oil tea, color, flavor and resistance, but also have natural antioxidant activity,
The functions such as antibacterial, removing free radical, antitumor, anticoagulation, antithrombotic, hypoglycemic and to fatty acid synthase inhibiting effect, this
Outer tea saponin has thrombolytic effect.It can prevent various cardiovascular and cerebrovascular diseases caused by vascular sclerosis.
Summary of the invention
It is an object of that present invention to provide a kind of oil tea exocarp antithrombotic reagent application.This programme passes through measurement oil tea
Influence of the exocarp opposed polarity extract to TT, APTT, PT and the FIB of rat, while determining the anti-blood of these extracts
Platelet congregational rate, and pass through the main chemical compositions structure that LC-Q-TOF/MS further analyzes exocarp antithrombotic acitivity,
Support is provided for a kind of antithrombotic reagent of being developed into of oil tea exocarp.
In order to achieve the goal above, The technical solution adopted by the invention is as follows:
The first scheme provided by the invention is to study oil tea exocarp in the application of antithrombotic reagent.
A method of antithrombotic reagent being extracted using oil tea exocarp, includes the following steps: the exocarp for acquiring oil tea,
It is sieved after dry, pulverize, is extracted 2-4 times with ethyl alcohol, filtering, merging filtrate obtains brown solid, take solid by being dried under reduced pressure
Object is dissolved in water, crosses MCI resin, and again with methanol rushes column, obtains product drying and stores in the case where 4 DEG C of ﹣.
Preferably, the concentration of the ethyl alcohol is 70-95%.
Preferably, the concentration of the methanol is 50-100%.
Preferably, described be dried under reduced pressure is dry at being greater than 0.09MPa, temperature lower than 55 DEG C for vacuum degree in pressure.
Preferably, the LC-Q-TOF/MS analysis is 1260Infinity series HPLC, SupelcosilABZ+PLUS column
(150mm × 4.6mm, 3 μm), wherein 0.1% formic acid-water is as mobile phase A, 0.1% formic acid-methanol is as Mobile phase B.Linearly
Gradient is 0-3min, 30%B；3-7min, 30-100%B；7-10min, 100%B, flow velocity 0.3mL/min, Detection wavelength 210
And 254nm, 0.3 μ L of sample volume.Electrospray ionisation source (ESI), anion scanning, scanning range (m/z): 100~1500；Dry gas
Flow velocity 8L/min, nebulizer pressure 20psi, 350 DEG C, sheath gas 11L/min, capillary voltage 3500V of sheath temperature degree.
Preferably, when the concentration of the methanol is 100%, the main component of extracted antithrombotic reagent are as follows:
Ellagic acid and 3,3 ', 4 '-trimethoxy ellagic acids, structural formula are respectively as follows:
Preferably, the detection method of the product is to use thrombin time, activated partial factor I time
Coagulating effectiveness is assessed with thrombin time test, and then determines the anticoagulation of oil tea exocarp.
Preferably, the detection method of the product is that antithrombotic effect is assessed using platelet aggregation rate measurement, in turn
Determine the anti thrombotic action of oil tea exocarp.
The method of this programme can be in product such as Cenxi switch oil tea, Luchuan oil-tea camellia, big fruit oil tea, safflower oil tea, fragrant flower oil teas
Kind uses.
Compared with prior art, advantages of the present invention and the utility model has the advantages that
This programme passes through TT, APTT, PT and FIB of the crude extract to rat of measurement oil tea exocarp opposed polarity extract
Influence, while determining the platelet aggregation-against effect of these extracts, and oil is further analyzed by LC-Q-TOF/MS
A kind of antithrombotic reagent of being developed into of the main chemical compositions structure of tea exocarp antithrombotic acitivity, oil tea exocarp provides
Support, expands the industrial chain of oil tea.
Detailed description of the invention
The coagulating effectiveness test of Fig. 1, Fig. 2 TF, Fr-1, Fr-2 and Fr-3；
Fig. 3 is the total ion figure of Fr-3, and Fig. 4, Fig. 5 are the mass spectrogram of Fr-3.
Specific embodiment
Invention is further described in detail With reference to embodiment.It is emphasized that following the description is only
It is merely exemplary, the range and its application being not intended to be limiting of the invention.
Embodiment 1
A method of antithrombotic reagent being extracted using oil tea exocarp, includes the following steps: to acquire oil in Nanning
Tea exocarp, dry, pulverize, and pass through 80 meshes.Three times with the extraction of 80% ethyl alcohol, filtering, merging filtrate is greater than in vacuum degree
0.09MPa, temperature are lower than drying at 55 DEG C, obtain brown solid (TF).Appropriate solid is taken, water is dissolved in, MCI resin is crossed, is used in combination
Various concentration (0% (water), 50% and methanol 100%) rush column, obtained eluate is respectively Fr-1, Fr-2 and Fr-3, will
Every kind of eluate is dry and stores in the case where 4 DEG C of ﹣.
The coagulating effectiveness of TF, Fr-1, Fr-2 and Fr-3 are tested
Test method: the Wistar rat of selection weight 220-250g is tested.With 1% anaesthetized with pentobarbital Wistar
Then rat collects blood sample into 0.109mol/L sodium citrate vacuum tube from abdominal aorta.With 160 × g at 25 DEG C
It is centrifuged 10min and obtains supernatant (platelet rich plasma, PRP).Then, remaining blood sample is centrifuged 10min with 2000 × g
To obtain platelet poor plasma (PPP).PRP by adding different extracts carries out platelet aggregation survey in clean test cup
It is fixed, wherein being induced with ADP with the final concentration of 5 μm of ol/L.Physiological saline (NS) is as blank control and aspirin as positive
The maximum aggregation rate (MAR) in 6min is observed in control.Calculating aggregation inhibiting rate (AIR, %)=(NCAR ﹣ EAR)/NCAR ×
100%
NCAR: control group maximum aggregate rate
EAR: experimental group maximum aggregate rate
Test result is as shown in Fig. 1:
Compared with the control group, TF, Fr-2 and Fr-3 reduce the maximum aggregation rate of blood platelet in rat plasma.Fr-3 pairs
The inhibitory effect of AIR is best, is 37.61%, is secondly Fr-2, AIR 36.04%, and difference is statistically significant (P < 0.05).
In addition, the AIR of blood platelet is 29.80% in positive group (aspirin), there is significant difference (P < 0.05) with blank group.TF's
AIR is 32.07%, shows the trend (P=0.064) for inhibiting platelet aggregation.The result shows that oil tea exocarp extract energy
Enough inhibit the trend of platelet aggregation, Fr-2 and Fr-3 have significant inhibiting effect to blood platelet, can be used for extracting anti-blood
The active component of bolt formation effect.
Using thrombin time (PT), activated partial factor I time (APTT) and thrombin time (TT) are surveyed
Fixed to assess influence of the oil tea exocarp to the coagulation function of rat, test results are shown in figure 2.The fresh blood of rat aorta
Liquid is mixed with 9 ︰, 1 ratio with anticoagulant (0.109mol/L sodium citrate), and 3000rpm is centrifuged 10min to obtain blood plasma.With sky
White control group (physiological saline group) is compared, and TF, Fr-2 and the Fr-3 in oil tea exocarp can significantly shorten the TT of rat plasma, tool
There is statistical difference (P < 0.05), shows that these extract/eluates have anticoagulative substance, rat TT and anti-hemostasis can be shortened
Liquid condensation.PT and APTT no significant difference (P > 0.05) between two groups prompts the effect and blank group of each group PT and APTT
Without significant difference.
The total ion figure of the Fr-3 that the present invention extracts is as shown in Fig. 3, and high resolution mass spectrum figure as shown in figures 4 and 5, is tied
Structure formula is respectively as follows:
Therefrom learn that the two kinds of ellagic acid ingredients of main component for the Fr-3 that this programme extracts are not disclosed in documents.
The above content is combine it is specific/further detailed description of the invention for preferred embodiment, cannot
Assert that specific implementation of the invention is only limited to these instructions.General technical staff of the technical field of the invention is come
It says, without departing from the inventive concept of the premise, some replacements or modifications can also be made to the embodiment that these have been described,
And these substitutions or variant are regarded as belonging to the scope of protection of the present invention.
Claims (7)
1. a kind of application of oil tea exocarp, it is characterised in that: application of the oil tea exocarp in antithrombotic reagent.
2. a kind of method for extracting antithrombotic reagent using oil tea exocarp, it is characterised in that: described method includes following steps:
The exocarp for acquiring oil tea, is sieved after dry, pulverize, and is extracted 2-4 times with ethyl alcohol, filtering, merging filtrate, by being dried under reduced pressure,
Brown solid is obtained, solids is taken to be dissolved in water, crosses MCI resin, again with methanol rushes column, obtains product drying and stores in the case where 4 DEG C of ﹣.
3. the method for extracting antithrombotic reagent using oil tea exocarp according to claim 2, it is characterised in that: the ethyl alcohol
Concentration be 70-95%.
4. the method for extracting antithrombotic reagent using oil tea exocarp according to claim 2, it is characterised in that: the methanol
Concentration be 50-100%.
5. the method for extracting antithrombotic reagent using oil tea exocarp according to claim 2, it is characterised in that: the decompression
It is drying at vacuum degree is greater than 0.09MPa, temperature for lower than 55 DEG C that drying, which is in pressure,.
6. the method for extracting antithrombotic reagent using oil tea exocarp according to claim 4, it is characterised in that: the methanol
Concentration be 100%.
7. the method for extracting antithrombotic reagent using oil tea exocarp as claimed in claim 6, it is characterised in that: described anti-
The main component of thrombus drug is ellagic acid and 3, and 3 ', 4 '-trimethoxy ellagic acids, structural formula is respectively as follows:
。
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| CN201910136572.5A CN110038068A (en) | 2019-02-25 | 2019-02-25 | A kind of application of oil tea exocarp in antithrombotic reagent |
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| CN201910136572.5A CN110038068A (en) | 2019-02-25 | 2019-02-25 | A kind of application of oil tea exocarp in antithrombotic reagent |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114605428A (en) * | 2022-03-31 | 2022-06-10 | 广西壮族自治区林业科学研究院 | Method for extracting and purifying ellagic acid from camellia oleifera shells |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108578461A (en) * | 2018-01-22 | 2018-09-28 | 河南大学 | Cercis extract and its extracting method and the application in terms of preparing antithrombotic reagent |
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2019
- 2019-02-25 CN CN201910136572.5A patent/CN110038068A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108578461A (en) * | 2018-01-22 | 2018-09-28 | 河南大学 | Cercis extract and its extracting method and the application in terms of preparing antithrombotic reagent |
Non-Patent Citations (3)
| Title |
|---|
| YI CHANG ET AL.: "Novel Bioactivity of Ellagic Acid in Inhibiting Human Platelet Activation", 《HINDAWI》 * |
| 沈建福等: "大孔树脂分离纯化油茶蒲鞣质工艺研究", 《中国食品学报》 * |
| 田瑛等: "中药地锦草酚性成分", 《中国中药杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114605428A (en) * | 2022-03-31 | 2022-06-10 | 广西壮族自治区林业科学研究院 | Method for extracting and purifying ellagic acid from camellia oleifera shells |
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