CN108358991B - Triterpenoid in chengshuanghua, and preparation method and application thereof - Google Patents

Abstract

The invention relates to a triterpenoid compound in the flower of Chengguang, a preparation method and application thereof, and belongs to the technical field of phytochemistry. The compound of the present invention is named 3-(4'-methoxy-5',6'-dihydroxybenzaldehyde)-3-β-oleananol-14-ene, and its molecular formula is C ₃₇ H ₅₄ O ₄ , As shown in formula (I):

, formula (I). The compound of the present invention is isolated from the branches and leaves of Chengguang flower in Xishuangbanna, Yunnan for the first time. It is determined to be a triterpenoid by nuclear magnetic resonance and mass spectrometry, and its specific structure is characterized. The compounds of the present invention all show moderate inhibitory activity against Sclerotinia sclerotiorum, Strawberry black spot and Potato Verticillium, and the minimum inhibitory concentration is from 25-100ug/mL. At the same time, these two compounds also showed weak antibacterial activity against some bacteria, and the minimum inhibitory concentration ranged from 50-100ug/mL, which had good application prospects.

Description

A kind of triterpenoid in Chengguang flower and its preparation method and application

technical field

The invention belongs to the technical field of phytochemicals, and in particular relates to a triterpenoid compound in Flos Radix, a preparation method and application thereof.

Background technique

Orophea yunnanensis is a plant of the genus Orophea in the family Annuaceae, which is produced in central Yunnan. Chengguang flowers are rich in active triterpenoids. Triterpenoids are a class of terpenoids with a basic core consisting of 30 carbon atoms, which exist in free form or in the form of glycosides or esters combined with sugars. Plants have various biochemical activities. In the present invention, triterpenoid active components with novel structure are obtained by systematically separating the triterpenoid active components of the plant. All have certain activity performance, which provides basic data for the application of such active ingredients. At present, there are no related reports on this compound and its application.

SUMMARY OF THE INVENTION

The first object of the present invention is to provide a triterpenoid compound in Flos Radix; the second object is to provide a preparation method of the compound; the third object is to provide the application of the compound in antifungal and bacterial activity .

For achieving the above object, the technical scheme adopted in the present invention is as follows:

Unless otherwise specified, the percentages used in the present invention are all mass percentages.

A kind of triterpenoid in Chengguang flower, structural formula is shown in formula (I):

英文名为3-(4'-Methox y-5',6'-dihydroxybenzoyl)-3-β-oleanol-14-ene，中文名为3-(4'-甲氧基-5',6'-二 羟基苯甲醛)-3-β-齐墩果醇-14-烯，分子式为C₃₇H₅₄O₄。

English name is 3-(4'-Methox y-5',6'-dihydroxybenzoyl)-3-β-oleanol-14-ene, Chinese name is 3-(4'-methoxy-5',6'- Dihydroxybenzaldehyde)-3-β-oleananol-14-ene, the molecular formula is C ₃₇ H ₅₄ O ₄ .

The present invention simultaneously provides the preparation method of the triterpenoid in the above-mentioned Chengguang flower, comprising the steps:

Step (1), extract extraction: pulverize the dried Chengguang flowers to 30-50 mesh, extract 2-4 times at room temperature with a volume percentage concentration of 70-95% ethanol aqueous solution, each time 6-10h, combine The extract is filtered to obtain a filtrate, which is concentrated under reduced pressure to obtain an extract;

Step (2), silica gel column chromatography: the extract obtained in step (1) is subjected to silica gel column chromatography, and the column silica gel is 200-300 mesh; gradient elution is carried out with petroleum ether and acetone mixed organic solvent, and the volume ratio is sequentially For 15:1, 7:1, 3:1 and 1:1, the gradient eluents of each gradient were collected and concentrated, monitored by TLC, the same fractions were combined and concentrated to obtain four fractions A-D;

Step (3), high-pressure liquid chromatography separation and separation: the B part obtained in step (2) is then subjected to gradient elution with a mixed solvent of chloroform and acetone in a volume ratio of 15:1-2:1 to obtain six parts B1-B6 ; Part B2 is separated and purified by high pressure liquid chromatography to obtain the triterpenoids in the Chengguang flower shown in formula (I).

Further, preferably, the volume percent concentration of the ethanol aqueous solution in the step (1) is 70%.

Further, it is preferred that in the described step (1), the volume of the ethanolic aqueous solution and the mass ratio of Chengguanghua are 8-12L:1Kg during each extraction.

Further, it is preferred that, in step (2), the extract obtained in step (1) is dissolved in 95% methanol aqueous solution with a volume concentration of 1.5-4 times the weight of the extract before being subjected to silica gel column chromatography, and then using The 100-mesh silica gel with 2-4 times the quality of the extract is stirred until it is evenly mixed, and then the sample is loaded;

Further, preferably, in step (2), the weight of the silica gel used for packing the column is 3-5 times the weight of the extract.

Further, preferably, the gradient elution procedure described in the step (2) is as follows: during elution, the flow rate of the eluent is one drop per second, and every 0.5-1.0L of the eluent is concentrated under reduced pressure. If the quality of the extract is less than 1g, the gradient should be replaced;

In the described step (3), during gradient elution, the volume ratio of chloroform and acetone is 15:1, 15:2, 15:3, 15:4, 15:5, 15:7.5 in sequence; gradient elution procedure For: during elution, the flow rate of the eluent is one drop per second, and each 0.5-1.0L eluent is concentrated under reduced pressure. When the mass of the concentrated extract is less than 1g, the gradient is replaced.

Further, preferably, the high-pressure liquid chromatography separation and purification described in step (3) adopts a C ₁₈ chromatographic column of 20 mm×250 mm and 5 μm, with methanol-water as the mobile phase, and the volume ratio of methanol to water is 4 : 6-6:4, the flow rate is 10-14mL/min, the detection wavelength of the UV detector is 254nm, each injection is 180-220μL, the chromatographic peaks of the 10-12min sample are collected, and they are evaporated to dryness after accumulating multiple times to obtain evaporated dry matter. .

Further, preferably, the evaporated-dried product obtained after separation and purification by high performance liquid chromatography is dissolved in methanol, and methanol is used as the mobile phase for separation and purification by gel column chromatography. The eluate is then concentrated to obtain the pure product of the compound represented by formula (I); wherein, a SephadexLH-20 gel column is used for separation and purification by gel chromatography.

The triterpenoid structure in the Chengguang flower prepared by the above-mentioned method is measured by the following method:

3-(4'-Methoxy-5',6'-dihydroxybenzaldehyde)-3-β-oleananol-14-ene (Ⅰ): white powder, [α]20D=+65(c =0.2, CHCl ₃ ), UV(MeOH)λmax(logε) 705(3.54), 383(3.59), 277(0.34), 213(4.07) nm; IR(KBr)ν _max : 3424, 2935, 2852, 1708 , 1613, 1464, 1363, 1227, 1088, 998,776 cm ^-1 . The hydrogen and carbon spectra are shown in Table 1, Figure 1 and Figure 2. Positive HR-ESI-MS: 592.4124 ([M+H] ⁺ , C ₃₈ H ₅₆ O ₅ , cald592.4128).

Table 1 Hydrogen and carbon spectra of compound (I)

In addition, the present invention also provides the application of the triterpenoids in the flower as an antibacterial agent. The specific antifungal and antibacterial results are shown in Tables 2 and 3.

Table 2. Antibacterial activity of compounds (MIC, μg/mL)

Compounds of the present invention Ciprofloxacin Micrococcus lysodeikticus 50 0.78 Bacillus subtilis 100 0.78 Bacillus cereus 50 0.78 Micrococcus luteus 100 0.78 Staphyloccocus aureus 50 3.13 Bacillus megaterium 50 0.78 Bacterium paratyphosum B 100 0.78 Proteusbacillm vulgaris 100 0.78 Salmonella typhi 100 0.78 Psmdomonas aeruginosa 50 0.78 Escherichia coli 100 0.78 Enterobacter aerogenes. 100 0.78

Table 3 Antifungal activity of compounds (MIC, μg/mL)

Compared with the prior art, the present invention has the following beneficial effects:

The compound of the present invention was isolated from the branches and leaves of Chengguang flower in Xishuangbanna, Yunnan for the first time, and was determined to be a triterpenoid by nuclear magnetic resonance and mass spectrometry, and its specific structure was characterized. Compound (I) showed moderate inhibitory activity against Sclerotinia sclerotiorum, Strawberry black spot and Verticillium potato, and the minimum inhibitory concentration was from 25-50ug/mL. At the same time, these two compounds also showed weak antibacterial activity against some bacteria, and the minimum inhibitory concentration ranged from 50-100ug/mL, which has a good application prospect.

Description of drawings

Fig. 1 is the carbon nuclear magnetic resonance spectrum ( ¹³ C NMR) of the compound (I) of the present invention;

Fig. 2 is the hydrogen nuclear magnetic resonance spectrum ( ¹ H NMR) of the compound (I) of the present invention;

Fig. 3 is the HMBC and COSY diagram of the compound of the present invention;

Figure 4 is a ROESY graph of the compounds of the present invention.

Detailed ways

The present invention will be further described in detail below in conjunction with the embodiments.

Those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. If the specific technology or condition is not indicated in the embodiment, it is carried out according to the technology or condition described in the literature in this field or according to the product specification. The materials or equipment used without the manufacturer's indication are conventional products that can be purchased.

The Chengguang flower used in the present invention is a common commercially available dry product, and there is no specific limitation on the moisture content.

Unless otherwise specified in the present invention, the percentages are mass percentages, and the ratios are mass ratios.

When the present invention adopts silica gel column chromatography for separation, TLC thin-layer chromatography spot plate tracking can also be adopted at the same time.

Example 1

A preparation method of triterpenoids in the flower of Chengguang, comprising the steps:

Step (1), extract extraction: pulverize the dried Chengguang flowers to 30-40 mesh, extract 2 times at room temperature with a volume percentage concentration of 70% ethanol aqueous solution, 6h each time, combine the extracts, and filter to obtain The filtrate is concentrated under reduced pressure to obtain extract; the volume of ethanolic aqueous solution and the mass ratio of Chengguanghua are 8L: 1Kg during each extraction;

Step (2), silica gel column chromatography: the extract obtained in step (1) is first dissolved in a 95% methanol aqueous solution with a volume concentration of 1.5 times the weight of the extract, and then stirred with 100 mesh silica gel with 2 times the quality of the extract To mix evenly, then load the sample and carry out silica gel column chromatography, the silica gel for column packing is 200-250 mesh, and the weight of silica gel used for column packing is 3 times the weight of the extract; gradient elution is carried out with petroleum ether and acetone mixed organic solvent, and the volume ratio is successively For 15:1, 7:1, 3:1 and 1:1, the gradient eluents of each gradient were collected and concentrated, monitored by TLC, the same fractions were combined and concentrated to give four fractions A-D;

Wherein, the gradient elution program is: during elution, the flow rate of the eluent is one drop per second, and every 0.5L of the eluent is concentrated under reduced pressure, and when the concentrated extract quality is less than 1g, the gradient is replaced;

Step (3), separation and separation by high pressure liquid chromatography: the volume ratio of part B obtained in step (2) is 15:1, 15:2, 15:3, 15:4, 15:5, 15:7.5 in turn. The mixed solvent of chloroform and acetone is subjected to gradient elution. During elution, the flow rate of the eluent is one drop per second, and each 0.5L of eluent is concentrated under reduced pressure. When the mass of the concentrated extract is less than 1 g, the gradient is replaced to obtain B1 -B6 six parts; the B2 part is separated and purified by high pressure liquid chromatography to obtain the triterpenoid compound in Chengguang flower represented by formula (I).

The high-pressure liquid chromatography separation and purification described in step (3) is to adopt a C ₁₈ chromatographic column of 20 mm × 250 mm and 5 μm, with methanol-water as the mobile phase, the volume ratio of methanol to water is 4:6, and the flow rate is 10 mL/ min, the detection wavelength of the UV detector was 254 nm, 1800 μL was injected each time, the chromatographic peaks of the 10 min samples were collected, and the chromatographic peaks of the samples were collected for several times and evaporated to dryness to obtain the evaporated dry substance.

Example 2

A preparation method of triterpenoids in the flower of Chengguang, comprising the steps:

Step (1), extract extraction: pulverize the dried Chengguang flower to 40-50 mesh, extract 4 times at room temperature with a volume percentage concentration of 95% ethanol aqueous solution, 10h each time, combine the extracts, filter to obtain The filtrate is concentrated under reduced pressure to obtain the extract; the volume of the ethanolic aqueous solution and the mass ratio of Chengguanghua are 1.2L:1Kg during each extraction;

Step (2), silica gel column chromatography: the extract obtained in step (1) is first dissolved in a 95% methanol aqueous solution with a volume concentration of 4 times the weight of the extract, and then stirred with 100-mesh silica gel that is 4 times the weight of the extract To mix evenly, then load the sample and carry out silica gel column chromatography, the silica gel for column packing is 250-300 mesh, and the weight of silica gel used for column packing is 5 times the weight of the extract; gradient elution is carried out with petroleum ether and acetone mixed organic solvent, and the volume ratio is sequentially For 15:1, 7:1, 3:1 and 1:1, the gradient eluents of each gradient were collected and concentrated, monitored by TLC, the same fractions were combined and concentrated to give four fractions A-D;

Wherein, the gradient elution program is: during elution, the flow rate of the eluent is one drop per second, and every 1.0L of the eluent is concentrated under reduced pressure, and when the concentrated extract quality is less than 1g, the gradient is replaced;

Step (3), separation and separation by high pressure liquid chromatography: the volume ratio of part B obtained in step (2) is 15:1, 15:2, 15:3, 15:4, 15:5, 15:7.5 in turn. The mixed solvent of chloroform and acetone is subjected to gradient elution. During elution, the flow rate of the eluent is one drop per second, and each 1.0 L of eluent is concentrated under reduced pressure. When the mass of the concentrated extract is less than 1 g, the gradient is replaced to obtain B1 -B6 six parts; the B2 part is separated and purified by high pressure liquid chromatography to obtain the triterpenoid compound in Chengguang flower represented by formula (I).

The high-pressure liquid chromatography separation and purification described in step (3) is to adopt a C ₁₈ chromatographic column of 20 mm × 250 mm and 5 μm, with methanol-water as the mobile phase, the volume ratio of methanol to water is 6:4, and the flow rate is 14 mL/ min, the detection wavelength of the UV detector was 254 nm, and each injection was 220 μL, and the chromatographic peaks of the 12 min samples were collected, accumulated for many times, and evaporated to dryness to obtain the evaporated dry substance.

The evaporated-dried product obtained after separation and purification by high performance liquid chromatography was dissolved in methanol, and methanol was used as the mobile phase for separation and purification by gel column chromatography. During separation, the eluate of 120-150 mL was received and concentrated according to the volume of the receiving liquid. , that is, the pure product of the compound represented by formula (I) is obtained; wherein, Sephadex LH-20 gel column is used for separation and purification by gel chromatography.

Example 3

A preparation method of triterpenoids in the flower of Chengguang, comprising the steps:

Step (1), extract extraction: pulverize the dried Chengguang flowers to 30-50 mesh, extract 3 times at room temperature with a volume percentage concentration of 90% ethanol aqueous solution, 8h each time, combine the extracts, and filter to obtain The filtrate is concentrated under reduced pressure to obtain the extract; the volume of the ethanolic aqueous solution and the mass ratio of Chengguanghua are 10 L:1Kg during each extraction;

Step (2), silica gel column chromatography: the extract obtained in step (1) is first dissolved in a 95% methanol aqueous solution with a volume concentration of 2 times the weight of the extract, and then stirred with 100-mesh silica gel that is 3 times the weight of the extract To mix evenly, then load the sample and carry out silica gel column chromatography, the silica gel for column packing is 220-280 mesh, and the weight of silica gel used for column packing is 4 times the weight of the extract; gradient elution is carried out with petroleum ether and acetone mixed organic solvent, and the volume ratios are sequentially For 15:1, 7:1, 3:1 and 1:1, the gradient eluents of each gradient were collected and concentrated, monitored by TLC, the same fractions were combined and concentrated to give four fractions A-D;

Wherein, the gradient elution program is: during elution, the eluent flow rate is one drop per second, and every 0.8L eluent is concentrated under reduced pressure, and when the concentrated extract quality is less than 1g, the gradient is replaced;

Step (3), separation and separation by high pressure liquid chromatography: the volume ratio of part B obtained in step (2) is 15:1, 15:2, 15:3, 15:4, 15:5, 15:7.5 in turn. The mixed solvent of chloroform and acetone is subjected to gradient elution. During elution, the flow rate of the eluent is one drop per second, and each 0.8L of eluent is concentrated under reduced pressure. When the mass of the concentrated extract is less than 1g, the gradient is replaced to obtain B1 -B6 six parts; the B2 part is separated and purified by high pressure liquid chromatography to obtain the triterpenoid compound in Chengguang flower represented by formula (I).

The high-pressure liquid chromatography separation and purification described in step (3) is to adopt a C ₁₈ chromatographic column of 20 mm × 250 mm and 5 μm, with methanol-water as the mobile phase, the volume ratio of methanol to water is 5:5, and the flow rate is 13 mL/ min, the detection wavelength of the UV detector is 254 nm, and each injection is 200 μL, and the chromatographic peaks of the sample in 11.2 min are collected, accumulated for many times and evaporated to dryness to obtain the evaporated dry substance.

The evaporated-dried product obtained after separation and purification by high performance liquid chromatography was dissolved in methanol, and methanol was used as the mobile phase for separation and purification by gel column chromatography. During separation, the eluate of 120-150 mL was received and concentrated according to the volume of the receiving liquid. , that is, the pure product of the compound represented by formula (I) is obtained; wherein, Sephadex LH-20 gel column is used for separation and purification by gel chromatography.

Example 4

A preparation method of triterpenoids in the flower of Chengguang, comprising the steps:

Step (1), extract extraction: pulverize the dried Chengguang flowers to 30-50 mesh, extract 3 times at room temperature with a volume percentage concentration of 85% ethanol aqueous solution, each 7h, combine the extracts, and filter to obtain The filtrate is concentrated under reduced pressure to obtain the extract; the volume of the ethanolic aqueous solution and the mass ratio of Chengguanghua are 9L: 1Kg during each extraction;

Step (2), silica gel column chromatography: the extract obtained in step (1) is first dissolved in a 95% methanol aqueous solution with a volume concentration of 3 times the weight of the extract, and then stirred with 100 mesh silica gel with 3.5 times the weight of the extract To mix evenly, then load the sample and carry out silica gel column chromatography, the silica gel for column packing is 200-300 mesh, and the weight of silica gel used for column packing is 4.5 times the weight of the extract; gradient elution is carried out with petroleum ether and acetone mixed organic solvent, and the volume ratio is sequentially For 15:1, 7:1, 3:1 and 1:1, the gradient eluents of each gradient were collected and concentrated, monitored by TLC, the same fractions were combined and concentrated to give four fractions A-D;

Wherein, the gradient elution program is: during elution, the eluent flow is one drop per second, and every 0.9L eluent is concentrated under reduced pressure, and when the concentrated extract quality is less than 1g, the gradient is replaced;

Step (3), separation and separation by high pressure liquid chromatography: the volume ratio of part B obtained in step (2) is 15:1, 15:2, 15:3, 15:4, 15:5, 15:7.5 in turn. The mixed solvent of chloroform and acetone is subjected to gradient elution. During elution, the flow rate of the eluent is one drop per second, and each 0.6L of eluent is concentrated under reduced pressure. When the mass of the concentrated extract is less than 1g, the gradient is replaced to obtain B1 -B6 six parts; the B2 part is separated and purified by high pressure liquid chromatography to obtain the triterpenoid compound in Chengguang flower represented by formula (I).

The high-pressure liquid chromatography separation and purification described in step (3) adopts a C ₁₈ chromatographic column of 20 mm×250 mm and 5 μm, with methanol-water as the mobile phase, the volume ratio of methanol to water is 4.5:5.5, and the flow rate is 1 mL /min, the detection wavelength of UV detector is 254nm, and each injection is 190 μL, and the chromatographic peaks of the 10.8-min sample are collected, accumulated for many times and evaporated to dryness to obtain evaporated dry substance.

The evaporated-dried product obtained after separation and purification by high performance liquid chromatography was dissolved in methanol, and methanol was used as the mobile phase for separation and purification by gel column chromatography. During separation, the eluate of 120-150 mL was received and concentrated according to the volume of the receiving liquid. , that is, the pure product of the compound represented by formula (I) is obtained; wherein, Sephadex LH-20 gel column is used for separation and purification by gel chromatography.

Example 5

------ Identification of compound structures

The structure of the compound prepared by the method of Example 1 was determined by the following method:

3-(4'-Methoxy-5',6'-dihydroxybenzaldehyde)-3-β-oleananol-14-ene (Ⅰ): white powder, [α]20D=+65(c =0.2, CHCl ₃ ), UV(MeOH)λmax(logε) 705(3.54), 383(3.59), 277(0.34), 213(4.07) nm; IR(KBr)ν _max : 3424, 2935, 2852, 1708 , 1613, 1464, 1363, 1227, 1088, 998,776 cm ^-1 . The hydrogen and carbon spectra are shown in Table 1, Figure 1 and Figure 2. HMBC and COSY plots are shown in Figure 3, and ROESY plots are shown in Figure 3. Positive HR-ESI-MS: 592.4124 ([M+H] ⁺ , C ₃₈ H ₅₆ O ₅ , cald 592.4128). So far, the structure of the compound has been determined.

Example 6

The compounds prepared in Examples 2-4 were taken as white powders. The measurement method is the same as that of Example 5, and it is confirmed that the compound prepared in Example 2-4 is 3-(4'-methoxy-5',6'-dihydroxybenzaldehyde)-3-β-oleananol-14 - ene.

Application example

The present invention also provides antifungal and antibacterial tests of the triterpenoids in Flos Chilli, the results are shown in Table 4 and Table 5.

Table 4. Antibacterial activity of compounds (MIC, μg/mL)

Compounds of the present invention Ciprofloxacin Micrococcus lysodeikticus 50 0.78 Bacillus subtilis 100 0.78 Bacillus cereus 50 0.78 Micrococcus luteus 100 0.78 Staphyloccocus aureus 50 3.13 Bacillus megaterium 50 0.78 Bacterium paratyphosum B 100 0.78 Proteusbacillm vulgaris 100 0.78 Salmonella typhi 100 0.78 Psmdomonas aeruginosa 50 0.78 Escherichia coli 100 0.78 Enterobacter aerogenes. 100 0.78

Table 5 Antifungal activity of compounds (MIC, μg/mL)

Compounds of the present invention Ketoconazole Sclerotinia sclerotiorum 25 6.25 Rhizoctorzia solani 100 12.5 Ceratocystis fimbriata 25 0.78 Verticillium dahliae Kleb 50 0.56 Fusarium oxysporum schlecht 100 12.5

The above results show that the compound of the present invention can be used as an antibacterial agent and has a good application prospect.

The foregoing has shown and described the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above-mentioned embodiments, and the descriptions in the above-mentioned embodiments and the description are only to illustrate the principle of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have Various changes and modifications fall within the scope of the claimed invention. The claimed scope of the present invention is defined by the appended claims and their equivalents.

Claims (9)

1. A triterpenoid in the chenguang flower is characterized in that the structural formula is shown as the formula (I):

formula (I).

2. The method for preparing triterpenes from the chenopodium album linn of claim 1, which comprises the following steps:

step (1), extract extraction: pulverizing dried Chengni flos into 30-50 mesh, extracting with 70-95% ethanol water solution at room temperature for 6-10 hr for 2-4 times, mixing extractive solutions, filtering to obtain filtrate, and concentrating under reduced pressure to obtain extract;

step (2), silica gel column chromatography: performing silica gel column chromatography on the extract obtained in the step (1), wherein silica gel filled in the column is 200-300 meshes; gradient elution is carried out by using a mixed organic solvent of petroleum ether and acetone according to the volume ratio of 15:1, 7:1, 3:1 and 1:1 in sequence, gradient eluents of all gradients are collected and concentrated, monitoring by TLC is carried out, the same parts are combined and concentrated, and four parts A-D are obtained;

and (3) separating and separating by high pressure liquid chromatography: performing gradient elution on the part B obtained in the step (2) by using a mixed solvent of chloroform and acetone in a volume ratio of 15:1-2:1 to obtain six parts B1-B6; separating and purifying part B2 by high pressure liquid chromatography to obtain triterpenes of the chenguang flower shown in formula (I);

the high pressure liquid chromatography separation and purification of the step (3) adopts the method of adopting 20mm multiplied by 250mm, 5μC of m₁₈A chromatographic column, wherein methanol-water is taken as a mobile phase, the volume ratio of methanol to water is 4:6-6:4, the flow rate is 10-14mL/min, the detection wavelength of an ultraviolet detector is 254nm, and 180-220 materials are injected each timeμAnd L, collecting chromatographic peaks of the sample for 10-12min, accumulating for multiple times, and evaporating to dryness to obtain an evaporated dry matter.

3. The method for preparing triterpenes from wide flowers as claimed in claim 2, wherein the ethanol water solution in step (1) has a concentration of 70% by volume.

4. The method for preparing triterpenes from the chenopodium album linn of claim 2, wherein the mass ratio of the volume of the ethanol aqueous solution to the chenopodium album linn is 8-12L:1Kg in each extraction in the step (1).

5. The method for preparing triterpenes in the chenopodium album (linn) Pat. according to claim 2, wherein in the step (2), the extract obtained in the step (1) is dissolved by a 95% methanol aqueous solution with volume concentration 1.5-4 times of the weight of the extract before silica gel column chromatography, and then is stirred by 100 mesh silica gel with mass 2-4 times of the extract until being uniformly mixed, and then is loaded.

6. The method for preparing triterpenes from the chenopodium album linn of claim 2, wherein the weight of the silica gel used for loading the column in the step (2) is 3-5 times of the weight of the extract.

7. The method for preparing triterpenes from the chenopodium album linn of claim 2, wherein the gradient elution procedure of the step (2) is as follows: during elution, the flow of the eluent is one drop per second, and the reduced pressure concentration is carried out on 0.5-1.0L of eluent, when the mass of the concentrated extract is less than 1g, the gradient is changed;

in the step (3), the volume ratio of chloroform to acetone is 15:1, 15:2, 15:3, 15:4, 15:5 and 15:7.5 in sequence during gradient elution; the gradient elution procedure was: during elution, the flow of the eluent is one drop per second, and the pressure reduction concentration is carried out on 0.5-1.0L of eluent, when the mass of the concentrated extract is less than 1g, the gradient is changed.

8. The method for preparing triterpenoids in wide orange flowers as claimed in claim 2, wherein the dried substances obtained after separation and purification by HPLC are dissolved in methanol, and then separated and purified by gel column chromatography with methanol as mobile phase, and the 120 th and 150mL eluents are received and then concentrated according to the volume of the received solution during separation to obtain the pure compounds of formula (I); wherein, the gel chromatographic separation and purification uses a Sephadex LH-20 gel column.

9. Use of the triterpenes from the chenopodium album of claim 1 as antibacterial agents.

Images