Tea flower saponin compound as well as preparation method and application thereof

Abstract

The invention discloses a tea flower saponin compound and a preparation method and application thereof.Two saponin monomer new compounds are co-separated and purified from tea flowers, and the inhibitory activity of the two tea flower saponin compounds on candida glabrata, candida albicans and candida tropicalis is measured; the two tea flower saponin compounds have obvious inhibition effects on three kinds of candida; the minimum inhibitory concentrations of the compound 1 to candida glabrata, candida albicans and candida tropicalis are respectively 13.02 mu g / mL, 10.42 mu g / mL and 31.52 mu g / mL; the minimum inhibitory concentrations of the compound 2 to candida glabrata, candida albicans and candida tropicalis are 15.63 [mu] g / mL, 6.51 [mu] g / mL and 26.04 [mu] g / mL respectively. The tea flower saponin compound provided by the invention can be used for preparing natural antifungal drugs, and has great significance in comprehensive utilization of tea flowers.

Description

Technical Field

[0001] The invention relates to the field of saponin compounds, and in particular to a tea tree flower saponin compound and a preparation method and application thereof. Background Art

[0002] Tea flower is a natural resource, and its active ingredients are similar to tea leaves. Every year, a large amount of tea flower resources are not fully utilized. Finding a suitable development path for tea flower can not only prevent tea flower from competing with buds and leaves for nutrients, but also turn waste into treasure, improve the economic benefits of tea farmers, and have great significance for the development of the entire tea industry.

[0003] The dry weight content of tea flower saponins in tea flowers is about 10.35-78.84 mg / g. When the petals account for more than 50% of the total flower weight, saponins accumulate in large quantities. Studies have shown that tea flower saponins have multiple biological activities such as hypoglycemic, anti-allergic, anti-cancer, anti-inflammatory, antibacterial, and whitening. Research on the biological activity functions of tea flower saponins is an important part of the development of tea flower resources. At present, most of the research focuses at home and abroad are on the level of tea flower saponin mixtures, and there is a lack of research on tea flower monomer compounds and structure-activity relationships, which to a certain extent limits the development and utilization of tea flower resources. Summary of the invention

[0004] The main purpose of the present invention is to provide a new tea tree flower saponin compound having a good inhibitory effect on Candida albicans and a preparation method and application thereof.

[0005] To achieve the above object, the present invention provides a tea tree flower saponin compound, the general structural formula of which is shown in Formula 1:

[0006]

[0007] Among them, R 1 is an organic acid or has no substituent, R 2 is an organic acid or a hydroxyl group, R 3 is a hydroxyl group or an organic acid, R 4 is hydroxyl or unsubstituted, R 5 It is a trisaccharide or tetrasaccharide oligosaccharide.

[0008] Furthermore, the structural formula of the tea tree flower saponin compound is shown in Formula 2 or Formula 3:

[0009]

[0010] The present invention also provides a method for preparing the tea tree flower saponin compound, comprising the following steps:

[0011] (1) Raw material crushing

[0012] Taking tea tree flowers, and crushing them to obtain tea tree flower powder;

[0013] (2) Extraction

[0014] Ultrasonic extraction of tea tree flower powder is performed with ethanol aqueous solution, and the extract is filtered to obtain an extract, which is then concentrated under reduced pressure to obtain an extract, and the extract is extracted with n-butanol, and the n-butanol extract is filtered to obtain a n-butanol extract, and finally concentrated under reduced pressure to obtain a paste extract of the n-butanol part;

[0015] (3) Separation and purification

[0016] The paste extract of the n-butanol part is separated and purified to obtain the tea tree flower saponin compound.

[0017] Furthermore, the tea tree flowers are selected from Baiye No. 1 tea tree flowers.

[0018] Furthermore, the separation and purification treatment includes dissolving the paste extract of the n-butanol part and eluting it in sequence through normal phase silica gel column chromatography, reverse phase C-18 column chromatography and high performance liquid chromatography column.

[0019] Furthermore, the specific process of separation and purification is as follows:

[0020] The paste extract of the n-butanol part was dissolved in methanol and subjected to normal phase silica gel column chromatography. The normal phase silica gel column chromatography was gradient eluted with an eluent system of ethyl acetate: methanol, and the concentration gradient was EtOAc-MeOH volume ratio of 10:0 to 0:10 as the gradient elution. The obtained eluate was developed and colored, and the component with an Rf value of 0.21 was taken;

[0021] The component with an Rf value of 0.21 obtained by normal phase silica gel column chromatography was dissolved in methanol and subjected to reverse phase C-18 column chromatography. The reverse phase C-18 column chromatography was gradient eluted with methanol: water = 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, 100:0, and the elution volume of each gradient was 4 BV. The collected eluate was developed and colored, and the component with an Rf value of 0.23 was taken;

[0022] The component with an Rf value of 0.23 obtained by reverse phase C-18 column chromatography was dissolved in methanol reagent, then filtered with a 0.22 μM microporous organic filter membrane, and then prepared and separated using a semi-preparative liquid chromatograph. The liquid chromatography conditions are as follows: a YMC column is used; a flow rate of 3.0 mL / min; a detection wavelength of 210 nm, 254 nm; an injection volume of 100 μL; a column temperature of 30°C; a mobile phase A of water (containing 0.2% formic acid) and a mobile phase B of acetonitrile; an elution gradient of 0-45 min, 42%-50% B; 45-46 min, 50%-100% B; 46-52 min, 100% B; 52-53 min, 100% to 42% B; 53-60 min, 42% B, and the eluent is collected according to the chromatographic peak signal, and finally concentrated under reduced pressure and dried to obtain the tea tree flower saponin compounds shown in Formulas 2 and 3.

[0023] The present invention also provides application of the tea tree flower saponin compound in preparing anti-Candida drugs.

[0024] The present invention also provides an anti-Candida drug, comprising the tea tree flower saponin compound.

[0025] The beneficial effects of the present invention are embodied in:

[0026] The present invention separates and purifies two new saponin monomer compounds from tea tree flowers, and determines the inhibitory activity of the two tea tree flower saponin compounds on Candida glabrata, Candida albicans, and Candida tropicalis. The two tea tree flower saponin compounds show obvious inhibitory effects on the three Candida; the minimum inhibitory concentrations of compound 1 on Candida glabrata, Candida albicans, and Candida tropicalis are 13.02 μg / mL, 10.42 μg / mL, and 31.52 μg / mL, respectively; the minimum inhibitory concentrations of compound 2 on Candida glabrata, Candida albicans, and Candida tropicalis are 15.63 μg / mL, 6.51 μg / mL, and 26.04 μg / mL, respectively. The tea tree flower saponin compounds of the present invention can be used to prepare natural antifungal drugs, which are of great significance for the comprehensive utilization of tea tree flowers. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a TLC analysis chart of the n-butanol extract of tea tree flowers separated by normal phase silica gel column;

[0028] Figure 2 This is the TLC analysis chart of the Fr.7 component separated by reverse phase C-18 column chromatography;

[0029] Figure 3 This is the antibacterial zone diagram of the graded extracts of tea tree flowers against Candida albicans, where ① to ③ are PE extract, EA extract, and n-Bu extract, respectively;

[0030] Figure 4This is the inhibition zone diagram of each fraction of tea tree flower normal phase column chromatography against Candida albicans, where ① to ⑦ are components Fr.1 to Fr.7 respectively;

[0031] Figure 5 The antibacterial zone diagram of each fraction of tea tree flower reverse phase column chromatography on Candida albicans, ① to They are Fr.7.1 to Fr.7.11 components;

[0032] 6A to 6H is the NMR, mass spectrum and IR spectrum of compound 1, Fig. 6A for 1 H NMR spectrum, Figure 6B for 13 CNMR spectrum, Figure 6C is the HSQC spectrum, Fig.6D is the HMBC spectrum, Fig. 6E for 1 H- 1 H COSYspectrum, Fig. 6F is the NOESY spectrum, Figure 6G This is a high-resolution electrospray mass spectrometer in negative ion mode. Figure 6H is the IR spectrum;

[0033] 7A to 7H is the NMR, mass spectrum and IR spectrum of compound 2, Fig. 7A for 1 H NMR spectrum, Figure 7B for 13 CNMR spectrum, Figure 7C is the HSQC spectrum, Fig.7D is the HMBC spectrum, 7E is 1 H- 1 H COSY spectrum, Figure 7F is the NOESY spectrum, Figure 7G This is a high-resolution electrospray mass spectrometer in negative ion mode. Figure 7H It is the IR spectrum. DETAILED DESCRIPTION

[0034] In order to make the technical scheme of the present invention more clearly understood by those skilled in the art, the following embodiments are listed for illustration. It should be pointed out that the following embodiments do not limit the protection scope of the present invention.

[0035] Unless otherwise specified, the raw materials, reagents or devices used in the following examples can be obtained from conventional commercial sources or by existing known methods.

[0036] Example 1

[0037] Preparation and detection of saponin compounds from tea tree flowers

[0038] 1. Extraction of tea tree flowers

[0039] The flowers of Baiye No. 1 tea tree were collected, dried outdoors, and then crushed into powder with an electric grinder. 1.5 kg of tea tree flower powder was taken and soaked in pure ethanol solution at a solid-liquid ratio of 1:20 for 2 days, ultrasonically extracted for 4 hours, and then filtered. 70% ethanol water (v / v) was added to the residue at the above-mentioned solid-liquid ratio, and ultrasonically extracted for 4 hours. A total of 3 extractions were performed, and the extracts were combined and concentrated under reduced pressure to obtain 550 g of tea tree flower ethanol crude extract.

[0040] 2.Graded extraction of tea tree flower ethanol crude extract

[0041] The obtained tea tree flower ethanol crude extract was placed in 1L of pure water, fully stirred to make it evenly diffuse in the water, and then extracted with n-butanol (n-Bu). After the extract was concentrated under reduced pressure, 197.8g of n-Bu extract was obtained.

[0042] 3. Separation of n-butanol extract components by normal phase silica gel column chromatography

[0043] Take 197g of n-Bu extract and dissolve it in methanol, mix it with 100-200 mesh silica gel of equal mass, dry-load it with 100-200 mesh silica gel, and dry-load it. Use ethyl acetate: methanol eluent system for gradient elution, the concentration gradient is EtOAc-MeOH volume ratio 10:0 to 0:10 as gradient elution, each eluent gradient elution 6 times column volume, the obtained eluate is developed by thin layer chromatography (TLC), the developing solvent is ethyl acetate: methanol: water (4:1:0.8), and sulfuric acid-ethanol color reaction is used for detection, the components with the same Rf value are combined, and a total of seven components from Fr.1 to Fr.7 are obtained, see Figure 1 , among which the Rf value of Fr.7 is 0.21, which is collected for later use.

[0044] 4. Separation of Fr.7 components by C-18 reverse phase silica gel column chromatography

[0045] Weigh 10.0g of the Fr.7 component with an Rf value of 0.21 obtained by normal phase silica gel column chromatography, dissolve it in methanol until it is completely dissolved, mix it with reverse phase silica gel at a ratio of 1:1, dry and crush it. Weigh 100g of reverse phase C18 filler, activate it with pure methanol and load it into the column. Replace the liquid in the column with 2 times the initial volume of mobile phase before loading the sample, fill the mixed sample into the loaded column, keep the upper surface of the filler flat, and add a layer of absorbent cotton as a protective layer above the sample layer. Gradient elution is performed with methanol: water = 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, 100:0, and the elution volume of each gradient is 4BV. The collected eluate was developed by thin layer chromatography (TLC) with a developing solvent of ethyl acetate: methanol: water (4:1:0.8), and detected by sulfuric acid-ethanol colorimetric reaction. The components with the same Rf value were combined with different elution gradients to obtain 11 components from Fr.7.1 to Fr.7.11, see Figure 2 , where the Rf value of Fr.7.9 is 0.23, which is collected for later use.

[0046] 5. Pre-HPLC separation and purification of Fr.7.9 fraction

[0047] The Fr.7.9 component with an Rf value of 0.23 obtained by C-18 reverse phase silica gel column chromatography was divided into a 30 mg / mL solution (the solvent was a chromatographic methanol solution), and then filtered with a 0.22 μM microporous organic filter membrane, and then prepared and separated by a semi-preparative liquid chromatograph. The first preparative liquid chromatography conditions are as follows: using a YMC chromatographic column; a flow rate of 3.0 mL / min; a detection wavelength of 210 nm, 254 nm; an injection volume of 100 μL; a column temperature of 30°C; an absorption wavelength of 210 nm and 254 nm; a mobile phase A of water (containing 0.2% formic acid), and a phase B of acetonitrile; an elution gradient of 0-45 min, 42%-50% B; 45-46 min, 50%-100% B; 46-52 min, 100% B; 52-53 min, 100%-42% B; 53-60 min, 42% B. The eluate was collected according to the chromatographic peak signal. The signal peaks with an elution time of 40 min, an absorbance of 200 mV at an absorption wavelength of 210 nm, and an absorbance of 100 mV at an absorption wavelength of 254 nm were collected, concentrated under reduced pressure, and dried to obtain compound 1. The signal peaks with an elution time of 46 min, an absorbance of 350 mV at an absorption wavelength of 210 nm, and an absorbance of 100 mV at an absorption wavelength of 254 nm were collected, concentrated under reduced pressure, and dried to obtain compound 2.

[0048] The two compounds separated and purified above were identified by modern spectral analysis methods (ultraviolet spectrum UV, infrared spectrum IR, high resolution mass spectrometry HR-ESI-MS and nuclear magnetic resonance NMR, etc.), and the results are as follows:

[0049] The properties of compound 1 are as follows:

[0050] 1) Soluble in methanol and DMSO, white amorphous powder;

[0051] 2) IR(KBr)vmax(cm -1 ):3431,2929,1706,1632,1414,1388,1305,1235,1161,1077,1045,534;

[0052] 3) Quasi-molecular ion peak in negative ion mode (HR-ESI-MS) m / z-1285.6251 [MH] - , its molecular formula can be deduced to be C 67 H 98 O 27 , thus its unsaturation is 19;

[0053] 4) The NMR spectrum data are shown in Table 1.

[0054] The properties of compound 2 are as follows:

[0055] 1) Soluble in methanol and DMSO, white amorphous powder;

[0056] 2) IR(KBr)vmax(cm -1 ):3427,2959,1706,1632,1414,1388,1305,1235,1161,1077,1045,534;

[0057] 3) Quasi-molecular ion peak m / z-1333.6249 [MH] in negative ion mode (HR-ESI-MS) - , its molecular formula can be deduced to be C 63 H 98 O 27 , thus its unsaturation is 15;

[0058] 4) The NMR spectroscopy data are shown in Table 2.

[0059] Table 1 NMR spectrum data of compound 1

[0060]

[0061]

[0062] Table 2 NMR spectrum data of compound 2

[0063]

[0064]

[0065] Note: 1 H NMR at 600 MHz, 13 C NMR was measured at 150 MHz, with δ in ppm, coupling constant J in Hz, and deuterated pyridine as solvent.

[0066] Also, see 6A to 6H , 7A to 7H It can be seen that all new compounds are characterized by ultraviolet spectroscopy UV, infrared spectroscopy IR and 1 H NMR, 13 C NMR, ESI-HR-MS, 1 H- 1 The assignment of two-dimensional nuclear magnetic resonance spectra such as HCOSY, HSQC, HMBC and ROSEY proved that the structure of the obtained compound 1 was as shown in Formula 2, and the structure of compound 2 was as shown in Formula 3.

[0067]

[0068]

[0069] Example 2

[0070] Verification of antibacterial activity of tea tree flower saponin compounds

[0071] 1. Filter paper diffusion method drug sensitivity test

[0072] Take a loop of bacterial liquid from the Candida glycerol tube with an inoculation loop, inoculate it onto a solid culture plate using the four-zone streak method, place it in an incubator and culture it at 37°C for 24 hours, then select a single colony to continue inoculation and subculture, repeat the subculture twice to ensure the purity and activity of the strain. Pick a small number of single colonies with a cotton swab and adjust it to a fungal suspension with a McFarland concentration of 0.5 for standby use. Use a purchased sterile blank drug-sensitive sheet with a diameter of 6 mm as a filter paper, dissolve saponins at different purification stages with sterile water, and configure the concentration of the fractional extraction fraction and the normal phase column chromatography fraction to 50 mg / mL, and the concentration of the reverse phase column chromatography fraction to 10 mg / mL. Soak these filter paper pieces in saponin solutions at different purification stages that have been prepared. After fully absorbing the solution, take it out and air-dry it in a sterile clean bench for subsequent use, that is, prepare a drug-containing filter paper. Use a purchased Sabour agar medium plate, dip a cotton swab in the fungal suspension, and evenly spread it on the surface of the plate culture medium. After the bacterial suspension is fully adsorbed, paste the drug-containing filter paper on the culture medium. Select a sterile blank drug-sensitive tablet and soak it in physiological saline, take it out and paste it on the contaminated plate as a blank control group.

[0073] After the filter paper is attached, seal the edges of the plate with a sealing film and place it upside down to prevent contamination caused by dripping water vapor. Then, place the plate in a constant temperature and humidity incubator and incubate at 37°C for 24 hours. After the incubation, measure the diameter of each inhibition zone using an inhibition zone measuring instrument.

[0074] 2. Determination of minimum inhibitory concentration (MIC) by microdilution method

[0075] According to the M27-A3 document published by the Clinical Laboratory Standards Institute (CLSI) of the United States, the minimum inhibitory concentration of tea tree flower extract was determined.

[0076] Experimental group: 100 μL of extract solution with different gradient concentrations was added to each well, and then 100 μL of fungal suspension diluted 100 times was added. Negative control group: 200 μL of RPMI 1640 culture medium was added, and no solution and fungal suspension were added. Positive control group: 100 μL of RPMI 1640 culture medium was added, and then 100 μL of fungal suspension diluted 100 times was added.

[0077] Place the 96-well plate needed for the experiment in the biosafety cabinet in advance, turn on the ultraviolet light for 30 minutes, and then add 100 μL of sterile RPMI 1640 culture medium to wells 1 to 11. Add 100 μL of 50 mg / mL saponin solution at different purification stages to well 1, as well as the final monomer compound (diluted and prepared with RPMI 1640 culture medium). Use a pipette to mix the solution in well 1 evenly, then accurately draw 100 μL and add it to well 2, and mix it again with a pipette. Accurately draw 100 μL of the mixed solution from well 2 and add it to well 3, and so on, so that the extract solution is doubled diluted on the 96-well plate. The same operation method is used until it is diluted to well 10, and 100 μL is discarded after mixing. Three replicate wells are set for each concentration. Use a pipette to draw 100 μL of the prepared fungal suspension and add it to wells 1 to 11. Add 100 μL of the prepared fungal suspension to the 11th well as a positive control group to observe and compare the growth of Candida albicans. Add 200 μL of RPMI 1640 culture medium to the 12th well as a negative control group to observe whether the culture medium is contaminated. After the operation, place the 96-well plate in a constant temperature and humidity incubator at 37°C for 24 hours, and take out to observe the growth of each well.

[0078] Experimental results: The process of extracting the crude ethanol extract of tea tree flowers in step 2 of Example 1 was replaced by petroleum ether (PE) and ethyl acetate (EA) for extraction. After the extract was concentrated under reduced pressure, PE extract (4.1 g) and EA extract (50.3 g) were obtained. The n-Bu extract, PE extract and EAC extract were subjected to the antibacterial experiment of Candida albicans. The results are as follows: Figure 3 As shown, it can be seen that the n-Bu extract has the best antibacterial effect, and its specific inhibition zone diameter and MIC value are shown in Table 3.

[0079] Table 3 Inhibition zone diameter and MIC of tea tree flower n-Bu extract against Candida albicans

[0080]

[0081] The n-Bu extract of tea tree flower was subjected to normal phase silica gel column chromatography, and 7 fractions were collected. Figure 4 It was found that fraction No. 7 (Fr.7) had the strongest antibacterial activity, among which the inhibition zone diameters and MIC values ​​of fraction No. 6 (Fr.6) and fraction No. 7 (Fr.7) are shown in Table 4. When the drug concentration was 50 mg / mL, the inhibition zone diameters of fraction No. 7 against Candida glabrata, Candida albicans and Candida tropicalis were 22 mm, 19 mm and 14 mm, and the minimum inhibitory concentrations for Candida glabrata, Candida albicans and Candida tropicalis were 0.10 mg / mL, 0.13 mg / mL and 0.33 mg / mL, respectively.

[0082] Table 4 Inhibition zone diameter and MIC of tea tree flower normal phase column chromatography fractions 6 and 7 against Candida albicans

[0083]

[0084] Fraction Fr.7 was chromatographed on a reverse phase C-18 column. Among the 11 fractions collected, see Figure 5 The No. 9 fraction (Fr.7.9) had the best inhibitory activity, and the MIC values ​​of the No. 6 fraction (Fr.7.6), No. 7 fraction (Fr.7.7) and No. 9 fraction (Fr.7.9) against Candida are shown in Table 5. The minimum inhibitory concentrations of the No. 9 fraction against Candida glabrata, Candida albicans and Candida tropicalis were 9.77 μg / mL, 16.28 μg / mL and 39.06 μg / mL, respectively.

[0085] Table 5 MIC of tea tree flower reverse phase column chromatography fractions 6, 7, and 9 against Candida albicans

[0086]

[0087] Fr 7.9 was further separated and purified by high performance liquid chromatography and other methods to obtain two new triterpenoid saponin monomers, namely compound 1 shown in formula 2 and compound 2 shown in formula 3. The MICs of the two against Candida are shown in Table 6. The minimum inhibitory concentrations of compound 1 against Candida glabrata, Candida albicans, and Candida tropicalis were 13.02 μg / mL, 10.42 μg / mL, and 31.52 μg / mL, respectively; the minimum inhibitory concentrations of compound 2 against Candida glabrata, Candida albicans, and Candida tropicalis were 15.63 μg / mL, 6.51 μg / mL, and 26.04 μg / mL, respectively.

[0088] Table 6 MIC of tea tree flower saponin compounds 1 and 2 against Candida albicans

[0089]

[0090] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A tea tree flower saponin compound, characterized in that: Its general structure is shown in Formula 1: Among them, R1 is an organic acid or no substituent, R2 is an organic acid or a hydroxyl group, R3 is a hydroxyl group or an organic acid, R4 is a hydroxyl group or no substituent, and R5 is a trisaccharide or tetrasaccharide oligosaccharide.

2. The tea tree flower saponin compound according to claim 1, characterized in that Its structural formula is shown in Formula 2 or Formula 3:

3. The method for preparing tea tree flower saponin compound according to claim 1, characterized in that: The following steps are involved: (1) Raw material crushing Taking tea tree flowers, and crushing them to obtain tea tree flower powder; (2) Extraction Ultrasonic extraction of tea tree flower powder is performed with ethanol aqueous solution, and the extract is filtered to obtain an extract, which is then concentrated under reduced pressure to obtain an extract, and the extract is extracted with n-butanol, and the n-butanol extract is filtered to obtain a n-butanol extract, and finally concentrated under reduced pressure to obtain a paste extract of the n-butanol part; (3) Separation and purification The paste extract of the n-butanol part is separated and purified to obtain the tea tree flower saponin compound.

4. The method for preparing tea tree flower saponin compound according to claim 3, characterized in that: The tea tree flowers are selected from Baiye No. 1 tea tree flowers.

5. The method for preparing tea tree flower saponin compound according to claim 3 or 4, characterized in that: The separation and purification process includes dissolving the paste extract of the n-butanol part and then eluting it in sequence through normal phase silica gel column chromatography, reverse phase C-18 column chromatography and high performance liquid chromatography column.

6. The method for preparing tea tree flower saponin compound according to claim 5, characterized in that: The specific process of separation and purification is as follows: The paste extract of the n-butanol part was dissolved in methanol and subjected to normal phase silica gel column chromatography. The normal phase silica gel column chromatography was gradient eluted with an eluent system of ethyl acetate: methanol, and the concentration gradient was EtOAc-MeOH volume ratio of 10:1 to 0:10 as a gradient elution. The obtained eluate was developed and colored, and the component with an Rf value of 0.21 was taken; The component with an Rf value of 0.21 obtained by normal phase silica gel column chromatography was dissolved in methanol and subjected to reverse phase C-18 column chromatography. The reverse phase C-18 column chromatography was gradient eluted with methanol: water = 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, 100:0, and the elution volume of each gradient was 4 BV. The collected eluate was developed and colored, and the component with an Rf value of 0.23 was taken; The component with an Rf value of 0.23 obtained by reverse phase C-18 column chromatography was dissolved in methanol reagent, then filtered with a 0.22 μM microporous organic filter membrane, and then prepared and separated using a semi-preparative liquid chromatograph. The liquid chromatography conditions are as follows: a YMC column is used; a flow rate of 3.0 mL / min; a detection wavelength of 210 nm, 254 nm; an injection volume of 100 μL; a column temperature of 30°C; a mobile phase A of water (containing 0.2% formic acid) and a mobile phase B of acetonitrile; an elution gradient of 0-45 min, 42%-50% B; 45-46 min, 50%-100% B; 46-52 min, 100% B; 52-53 min, 100%-42% B; 53-60 min, 42% B, and the eluent is collected according to the chromatographic peak signal, and finally concentrated under reduced pressure and dried to obtain the tea tree flower saponin compounds shown in Formulas 2 and 3.

7. Use of the tea tree flower saponin compound as claimed in claim 1 or 2 in the preparation of anti-Candida drugs.

8. An anti-Candida drug, characterized in that: It comprises the tea tree flower saponin compound as described in claim 1 or 2.

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