CN110003293B - Method for extracting parasitoside from Chinese rose - Google Patents
Method for extracting parasitoside from Chinese rose Download PDFInfo
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- CN110003293B CN110003293B CN201910327851.XA CN201910327851A CN110003293B CN 110003293 B CN110003293 B CN 110003293B CN 201910327851 A CN201910327851 A CN 201910327851A CN 110003293 B CN110003293 B CN 110003293B
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- C07H1/00—Processes for the preparation of sugar derivatives
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Abstract
The invention belongs to the technical field of plant extraction, and particularly relates to a method for extracting parasitoside from China rose, which comprises the following steps: 1) pulverizing dried flos Rosae chinensis, cold soaking in petroleum ether at room temperature for defatting, volatilizing petroleum ether from residue, adding ethanol, cold soaking for extraction, and concentrating extractive solution to obtain total ethanol extract; 2) dissolving the ethanol total extract with methanol, mixing, eluting with water, 20% ethanol, and 40% ethanol sequentially through D101 macroporous resin column, and volatilizing solvent to obtain 40% ethanol eluate; 3) detecting and combining 40% ethanol elution components by normal pressure silica gel column chromatography and silica gel thin layer chromatography to obtain 12 components, and sequentially marking the components as components Fr.4-1 to Fr.4-12 according to the polarity of the obtained components from small to large; 4) purifying the 4 th component Fr.4-4 by gel column chromatography and high performance liquid chromatography to obtain 1 pure component, i.e. the parasitoside. The method can quickly and effectively extract and separate the parasitoside contained in the Chinese rose, and the extraction yield of the parasitoside is high.
Description
Technical Field
The invention belongs to the technical field of plant extraction and separation, and particularly relates to a method for effectively and quickly extracting parasitoside from China rose flowers.
Background
The flos Rosae chinensis is Rosa chinensis of Rosa of RosaceaeRosa chinensisDried flower of jacq. The main components of flos Rosae chinensis include flavonoids such as quercetin and kaempferol; other steroid components such as oleanolic acid,βSitosterol, campesterol, cycloeucalenol and the like andvolatile oil components such as gallic acid. The flos Rosae chinensis has various pharmacological effects including inhibiting platelet aggregation, reducing vascular permeability, resisting fungi and virus, resisting oxidation, and enhancing immunity.
At present, there is no report on the extraction and separation of parasitoside from Chinese rose.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a method for effectively and quickly extracting the parasitoside from the Chinese rose, and the method has high extraction yield of the parasitoside.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for extracting parasitoside from Chinese rose flowers comprises the following steps:
1) pulverizing dried flos Rosae chinensis, cold soaking in petroleum ether at room temperature for defatting, volatilizing petroleum ether from residue, adding ethanol, cold soaking for extraction, and concentrating extractive solution to obtain total ethanol extract;
2) dissolving the ethanol total extract with methanol, mixing, eluting with water, 20% ethanol, and 40% ethanol sequentially through D101 macroporous resin column, and volatilizing solvent to obtain 40% ethanol eluate;
3) gradient eluting 40% ethanol components with dichloromethane-methanol by normal pressure silica gel column chromatography, detecting with silica gel thin layer chromatography, mixing to obtain 12 components, and sequentially labeling components from Fr.4-1 to Fr.4-12 according to polarity of the obtained components from small to large;
4) purifying the 4 th component Fr.4-4 by gel column chromatography and high performance liquid chromatography to obtain 1 pure component, and comparing with standard product by NMR spectrogram, reference data to show that the pure component is herba Taxilli glycoside.
Further, in order to obtain a better impregnation extraction effect, the step 1) is preferably carried out according to the following parameter conditions:
in the step 1), the petroleum ether is used for cold-soaking degreasing for 2-4 times, the cold-soaking degreasing time is 2-4 days each time, and the adding ratio range of the dried Chinese rose to the petroleum ether is 1 kg: 4-7L.
In the step 1), the volume fraction of the added ethanol is 60-80%, and the adding ratio of the residue to the ethanol is 1 kg: 4-7L, cold soaking and extracting for 3 times, wherein the cold soaking and extracting time for each time is 6-8 days, 2-4 days and 2-4 days.
Further, in order to obtain a better purification and separation effect, the steps 3) and 4) are preferably performed according to the following parameter conditions:
in the step 3), dichloromethane-methanol with volume ratio of 10:1, 8:1, 5:1, 3:1 and 2:1 is used for gradient elution.
In step 4), eluting the gel column chromatography with methanol;
the chromatographic conditions of the high performance liquid chromatography are as follows: an RP-18 endclamped chromatographic column (4.6 mm. times.250 mm, 5 μm) is used; the mobile phase is methanol (A) -water (B); gradient elution: 0-30 min, 40-90% of A and 60-10% of B; flow rate: 0.8 mL/min; detection wavelength: 254 nm; sample introduction volume: 20 μ L.
The method quickly and effectively extracts and separates the parasitoside contained in the Chinese rose through the extraction and separation of the Chinese rose ethanol total extract and separation and purification means such as macroporous resin column chromatography, silica gel column chromatography, high performance liquid chromatography and the like, and the extraction yield of the parasitoside is high. Compared with the prior art, the method has the beneficial effects that:
1) at present, no report of extracting and obtaining the parasitoside from the plant China rose exists, and the invention provides a method for extracting and separating the parasitoside from the China rose, which is beneficial to better developing and utilizing the China rose which is a medicinal plant;
2) the compound of the parasitoside extracted by the invention has wide bioactivity, and the invention provides an extraction method of the parasitoside, which is convenient for deep research on the parasitoside;
3) the separation materials used in the invention are macroporous resin, silica gel and gel column chromatography, are insoluble in water and any solvent, are nontoxic and tasteless, have stable chemical properties, have good selectivity to organic matters, are not influenced by inorganic salts and strong ion low molecular compounds, are easy to elute and regenerate, and are beneficial to multiple utilization. The separation of preparative high-performance liquid phase used in the purification process is the most popular novel separation method at present, time and labor are saved, and the yield of samples can be greatly improved.
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FIG. 1 shows the product obtained by extraction in example 11H-NMR spectrum;
FIG. 2 shows the product obtained by extraction in example 113C-NMR spectrum.
Detailed Description
The technical solution of the present invention is further described in detail with reference to the following examples, but the scope of the present invention is not limited thereto.
Example 1
A method for extracting parasitoside from Chinese rose flowers comprises the following steps:
1) taking 2kg of dried Chinese rose, crushing, carrying out cold leaching and degreasing for 3 times by using petroleum ether at room temperature, wherein the degreasing time of each cold leaching is 3 days, volatilizing the petroleum ether from residues, adding ethanol with the volume fraction of 70%, carrying out cold leaching and extraction for 3 times, wherein the cold leaching and extraction time of each time is 7 days, 3 days and 3 days respectively, combining extracting solutions and concentrating to obtain 600 g of total ethanol extract; wherein the adding ratio of the dried Chinese rose flowers to the petroleum ether is 1 kg: 5L, the adding ratio of the residue to the ethanol is 1 kg: 5L;
2) dissolving the total ethanol extract (600 g) with appropriate amount of methanol, mixing with D101 macroporous resin column (600 g), subjecting to D101 macroporous resin column chromatography, sequentially eluting with water, 20% ethanol, and 40% ethanol, and volatilizing solvent to obtain 40% ethanol eluate;
3) gradient eluting 40% ethanol eluate with dichloromethane-methanol at volume ratio of 10:1, 8:1, 5:1, 3:1, 2:1 by normal pressure silica gel column chromatography, detecting with silica gel thin layer chromatography, mixing to obtain 12 fractions, and sequentially labeling the fractions from small to large as fraction Fr.4-1 to Fr.4-12;
4) the 4 th component Fr.4-4 (1 g) is purified by gel column chromatography and high performance liquid chromatography to obtain 1 pure component (150 mg), yellow powder, the purity is 99.388% by high performance liquid chromatography detection, and the pure component is known to be the parasitoside by nuclear magnetic resonance spectrogram, reference data and standard control.
In step 4), eluting the gel column chromatography with pure methanol; the chromatographic conditions of the high performance liquid chromatography are as follows: adopting an RP-18 endclamped chromatographic column; the mobile phase is methanol (A) -water (B); gradient elution: 0-30 min, 40-90% of A and 60-10% of B; flow rate: 0.8 mL/min; detection wavelength: 254 nm; sample introduction volume: 20 mu L of the solution; t is tR=14.455 min。
The compound extracted in the above example 1 is identified by structure, and the structural formula of the compound is as follows:
instrument materials: ultraviolet measurement is carried out on a UV-210A ultraviolet spectrum determinator;1H-NMR，13the C-NMR spectrum was determined by Bruker am-400 MHz NMR spectrometer. Measurement conditions and results: yellow powder (DMSO), molecular formula: c20H18O11, ESI-MS m/z: 457 [M＋Na]+. 1H-NMR (DMSO-d 6, 400 MHz) δ: 12.64 (1H, s, 5-OH) ,7.48 (1H, d, J = 1.5 Hz, H-2′), 7.56 (1H, dd, J = 8.5, 1.5 Hz, H-6′), 6.85 (1H, d, J = 8.5 Hz, H-5′), 6.41 (1H, d, J = 2.0 Hz, H-8), 6.20 (1H, d, J = 2.0 Hz, H-6), 5.58 (1H, d, J = 1.2 Hz, H-1″)；13C-NMR (100 MHz, DMSO) δ177.7 (C-4), 164.2 (C-7), 161.2 (C-5), 157.0 (C-9), 156.4 (C-2), 148.5 (C-4 '), 145.1 (C-3 '), 133.4 (C-3), 121.7 (C-6 '), 120.9 (C-1 '), 115.6 (C-5 '), 115.5 (C-2 '), 107.8 (C-1 '), 104.0 (C-10), 98.7 (C-6), 93.6 (C-8), 85.8 (C-4 '), 82.1 (C-2 '), 76.9(C-3 '), 60.6 (C-5 '); specific maps are shown in FIGS. 1 and 2.
Example 2
A method for extracting parasitoside from Chinese rose flowers comprises the following steps:
1) taking 2kg of dried Chinese rose, crushing, carrying out cold leaching and degreasing for 4 times by using petroleum ether at room temperature, wherein the degreasing time of each cold leaching is 4 days, volatilizing the petroleum ether from residues, adding 75% ethanol by volume fraction, carrying out cold leaching and extraction for 2 times, wherein the cold leaching and extraction time of each time is 8 days and 3 days respectively, combining extracting solutions and concentrating to obtain 510 g of total ethanol extract; wherein the adding ratio of the dried Chinese rose flowers to the petroleum ether is 1 kg: 6L, the adding ratio of the residue to the ethanol is 1 kg: 6L;
2) dissolving the ethanol total extract (510 g) with appropriate amount of methanol, mixing with D101 macroporous resin column (510 g), subjecting to D101 macroporous resin column chromatography, sequentially eluting with water, 20% ethanol, and 40% ethanol, and volatilizing solvent to obtain 40% ethanol eluate;
3) gradient eluting 40% ethanol eluate with dichloromethane-methanol at volume ratio of 10:1, 8:1, 5:1, 3:1, 2:1 by normal pressure silica gel column chromatography, detecting with silica gel thin layer chromatography, mixing to obtain 12 fractions, and sequentially labeling the fractions from small to large as fraction Fr.4-1 to Fr.4-12;
4) purifying the 4 th component Fr.4-4 (500 mg) by gel column chromatography and high performance liquid chromatography to obtain 1 pure component (68 mg) and yellow powder with purity of 97.899% by high performance liquid chromatography, and obtaining the herba Taxilli glycoside.
In step 4), eluting the gel column chromatography with methanol; the chromatographic conditions of the high performance liquid chromatography are as follows: adopting an RP-18 endclamped chromatographic column; the mobile phase is methanol (A) -water (B); gradient elution: 0-30 min, 40-90% of A and 60-10% of B; flow rate: 0.8 mL/min; detection wavelength: 254 nm; sample introduction volume: 20 mu L of the solution; t is tR=14.455 min。
Claims (4)
1. A method for extracting parasitoside from Chinese rose is characterized by comprising the following steps:
1) pulverizing dried flos Rosae chinensis, cold soaking in petroleum ether at room temperature for defatting, volatilizing petroleum ether from residue, adding ethanol, cold soaking for extraction, and concentrating extractive solution to obtain total ethanol extract;
2) dissolving the ethanol total extract with methanol, mixing, eluting with water, 20% ethanol, and 40% ethanol sequentially through D101 macroporous resin column, and volatilizing solvent to obtain 40% ethanol eluate;
3) gradient eluting 40% ethanol components with dichloromethane-methanol by normal pressure silica gel column chromatography, detecting with silica gel thin layer chromatography, mixing to obtain 12 components, and sequentially labeling components from Fr.4-1 to Fr.4-12 according to polarity of the obtained components from small to large;
4) purifying the 4 th component Fr.4-4 by gel column chromatography and high performance liquid chromatography to obtain 1 pure component, i.e. the parasitoside;
in step 4), eluting the gel column chromatography with methanol;
the chromatographic conditions of the high performance liquid chromatography are as follows: adopting an RP-18 endclamped chromatographic column; the mobile phase is methanol (A) -water (B); gradient elution: 0-30 min, 40-90% of A and 60-10% of B; flow rate: 0.8 mL/min; detection wavelength: 254 nm; sample introduction volume: 20μL。
2. The method for extracting parasitosides from rosa chinensis, according to claim 1, wherein in the step 1), petroleum ether is used for cold-dip degreasing for 2-4 times, the cold-dip degreasing time is 2-4 days, and the adding ratio of the dried rosa chinensis to the petroleum ether is in the range of 1 kg: 4-7L.
3. The method for extracting parasitoside from rosa chinensis, according to claim 1, wherein in the step 1), the volume fraction of ethanol added is 60-80%, and the adding ratio of the residue to the ethanol is in the range of 1 kg: 4-7L, cold soaking and extracting for 3 times, wherein the cold soaking and extracting time for each time is 6-8 days, 2-4 days and 2-4 days.
4. The method for extracting parasitoside from flos Rosae chinensis as claimed in claim 1, wherein step 3) is performed by gradient elution with dichloromethane-methanol at volume ratio of 10:1, 8:1, 5:1, 3:1, 2: 1.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1511583A (en) * | 2002-12-28 | 2004-07-14 | 石家庄制药集团制药技术开发有限公司 | Chinese rose extract and its preparing method and use |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1511583A (en) * | 2002-12-28 | 2004-07-14 | 石家庄制药集团制药技术开发有限公司 | Chinese rose extract and its preparing method and use |
Non-Patent Citations (3)
| Title |
|---|
| Isolation and Identification of Radical Scavenging and Tyrosinase Inhibition of Polyphenols from Tibouchina semidecandra L.;HASNAH M. SIRAT et al.;《J. Agric. Food Chem.》;20101231;第58卷;第10404-10409页 * |
| 月季花的化学成分研究;张宏武;《万方学术期刊数据库》;20100830;第15-16页 * |
| 月季花的化学成分研究;张沛 等;《中草药》;20101031;第41卷(第10期);第1616-1618页 * |
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