Pseudo-ginseng stem and leaf rare ginsenoside as well as preparation method and application thereof

Abstract

The invention discloses radix notoginseng stem and leaf rare ginsenoside as well as a preparation method and application thereof, and belongs to the technical field of ginsenoside. The radix notoginseng stem and leaf rare ginsenoside comprises Rk3, Rh4, S-Rg3, R-Rg3, Rk1 and Rg5. The radix notoginseng stem and leaf rare ginsenoside can be used for preparing radix notoginseng stem and leaf rare ginsenoside. The preparation method comprises the following steps: mixing the pseudo-ginseng stem and leaf extract with an acetic acid aqueous solution, reacting at 105-120 DEG C for 10-40 minutes, and drying; in the preparation process, macroporous resin purification is not carried out. According to the scheme, part of chemical substances in the panax notoginseng stem and leaf extract can be rapidly converted at a low temperature, new active substances Rh4, S-Rg3, R-Rg3, Rk1 and Rg5 with high purity are obtained at the same time, and the obtained panax notoginseng stem and leaf rare ginsenoside can effectively inhibit propionibacterium acnes, burkholderia cepacia and malassezia furfur.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of ginsenoside, in particular to a rare ginsenoside in stems and leaves of Panax notoginseng and a preparation method and application thereof. BACKGROUND

[0002] The stems and leaves of Panax notoginseng, a perennial herb of the family Araliaceae, are known as "the forgotten treasure". Panax notoginseng The stems and leaves of Panax notoginseng have a small toxic and side effect and are safe, and can be used as a medicine. In the traditional use of Chinese medicinal materials, the main medicinal parts of Panax notoginseng are roots and rhizomes. Generally, the roots of Panax notoginseng are harvested once every 2-3 years, while the stems and leaves of Panax notoginseng can be harvested once a year. Although the stems and leaves of Panax notoginseng are easier to obtain than the roots, the stems and leaves are often regarded as an accessory, and most of the stems and leaves are used for feed or discarded in the local environment, which causes a waste of a large amount of resources. The stems and leaves of Panax notoginseng contain rich dammarane-type saponins, and the content thereof accounts for about 8%. The stems and leaves of Panax notoginseng, like the roots of Panax notoginseng, have saponins as the main active ingredient to play a pharmacological effect, and have pharmacological effects of anti-inflammatory, anticancer, anti-anxiety, anti-depression, and neuroprotection.

[0003] In recent years, rare ginsenosides have been widely studied due to their superior pharmacological activities and diversified health benefits, and show great development prospects. However, the content of rare ginsenosides in natural plants is extremely low, and Panax notoginseng is difficult to scale up due to the harsh requirements for growth environment and the factors of the genuineness of medicinal materials. The stems and leaves of Panax notoginseng have a high content of ginsenosides and a low price, and are an ideal raw material for producing rare saponins. Therefore, the development and utilization of the stems and leaves of Panax notoginseng is one of the effective ways to solve the sustainable utilization of Panax notoginseng resources.

[0004] Therefore, the present application is provided. SUMMARY

[0005] The present application aims to provide a rare ginsenoside in stems and leaves of Panax notoginseng and a preparation method and application thereof to solve or improve the above technical problems.

[0006] The present application can be achieved as follows: In a first aspect, the present application provides a rare ginsenoside in stems and leaves of Panax notoginseng, which comprises Rk3, Rh4, S-Rg3, R-Rg3, Rk1 and Rg5. The preparation of the rare ginsenoside from the Panax notoginseng stem and leaf includes: mixing the Panax notoginseng stem and leaf extract with an acetic acid aqueous solution, reacting at 105-120℃ for 10-40 minutes, and after centrifugation, the supernatant is concentrated and dried; wherein the Panax notoginseng stem and leaf extract includes S-Rg2, Rb1, Fc, Rb3, Rd and Rk3, and does not contain Rh4, S-Rg3, R-Rg3, Rk1 and Rg5; and the preparation of the rare ginsenoside from the Panax notoginseng stem and leaf does not include macroporous resin purification.

[0007] In an optional embodiment, the rare ginsenoside from the Panax notoginseng stem and leaf includes 1-2wt% of Rk3, 1.5-2.8wt% of Rh4, 7.5-13.5wt% of S-Rg3, 7-13wt% of R-Rg3, 5.5-10.5wt% of Rk1 and 6-12.5wt% of Rg5, in terms of mass percentage.

[0008] In an optional embodiment, the rare ginsenoside from the Panax notoginseng stem and leaf includes 1.3-1.8wt% of Rk3, 1.5-2.2wt% of Rh4, 7.8-10.6wt% of S-Rg3, 7.1-10.2wt% of R-Rg3, 5.6-8.8wt% of Rk1 and 7.2-9.6wt% of Rg5.

[0009] In an optional embodiment, the rare ginsenoside from the Panax notoginseng stem and leaf includes 1.3-1.65wt% of Rk3, 2.05-2.2wt% of Rh4, 9.2-10.6wt% of S-Rg3, 9.85-10.2wt% of R-Rg3, 7.35-8.8wt% of Rk1 and 8.4-9.6wt% of Rg5.

[0010] In an optional embodiment, the Panax notoginseng stem and leaf extract includes 3.1-3.25wt% of S-Rg2, 5.5-6.5wt% of Rb1, 10-11wt% of Fc, 29-32wt% of Rb3, 1-1.5wt% of Rd and 1-2wt% of Rk3, in terms of mass percentage.

[0011] In an optional embodiment, the reaction temperature is 110-120℃.

[0012] In an optional embodiment, the reaction time is 30-40 minutes.

[0013] In an optional embodiment, the concentration of acetic acid in the acetic acid aqueous solution is 0.05-0.2%, in terms of volume percentage.

[0014] In an alternative embodiment, the concentration of acetic acid in the aqueous acetic acid solution is 0.1% to 0.2% by volume.

[0015] In an alternative embodiment, the solid-liquid ratio of the Panax notoginseng stem and leaf extract to the aqueous acetic acid solution is 4 mg: 1 mL to 16 mg: 1 mL.

[0016] In an alternative embodiment, the solid-liquid ratio of the Panax notoginseng stem and leaf extract to the aqueous acetic acid solution is 4 mg: 1 mL to 8 mg: 1 mL.

[0017] In a second aspect, the present application provides a use of the Panax notoginseng stem and leaf rare ginsenoside according to any one of the preceding embodiments in the preparation of a product for inhibiting at least one of Propionibacterium acnes, Burkholderia cenocepacia and Malassezia furfur.

[0018] In an alternative embodiment, the Propionibacterium acnes includes at least one of ATCC11827 Propionibacterium acnes, ATCC12930 Propionibacterium acnes and ATCC6919 Propionibacterium acnes.

[0019] In a third aspect, the present application provides a cosmetic product containing the Panax notoginseng stem and leaf rare ginsenoside according to any one of the preceding embodiments.

[0020] In an alternative embodiment, the amount of the Panax notoginseng stem and leaf rare ginsenoside in the cosmetic product is 0.00625% to 0.55% by mass.

[0021] In an alternative embodiment, the amount of the total amount of Rh4, S-Rg3, R-Rg3, Rk1 and Rg5 in the Panax notoginseng stem and leaf rare ginsenoside in the cosmetic product is 0.00313% to 0.275% by mass.

[0022] In a fourth aspect, the present application provides a pharmaceutical product containing the Panax notoginseng stem and leaf rare ginsenoside according to any one of the preceding embodiments.

[0023] In a fifth aspect, the present application provides a food product containing the Panax notoginseng stem and leaf rare ginsenoside according to any one of the preceding embodiments.

[0024] The beneficial effects of the present application include: The present application creatively proposes mixing the Panax notoginseng stem and leaf extract with the aqueous acetic acid solution and performing a reaction, which can rapidly convert part of the chemical substances in the Panax notoginseng stem and leaf extract at a lower temperature to simultaneously obtain the new active substances Rh4, S-Rg3, R-Rg3, Rk1 and Rg5 with higher purity, and the obtained Panax notoginseng stem and leaf rare ginsenoside can effectively inhibit Propionibacterium acnes, Burkholderia cenocepacia and Malassezia furfur. BRIEF DESCRIPTION OF DRAWINGS

[0025] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required to be used in the embodiments will be briefly introduced as follows. It should be understood that the following drawings only show some of the embodiments of the present application, and therefore should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can also be obtained without creative labor on the basis of these drawings.

[0026] Figure 1 HPLC chromatogram of the mixed standard solution in Test Example 1; Figure 2 HPLC chromatogram of the Panax notoginseng stem and leaf extract in Test Example 1; Figure 3 HPLC chromatogram of the rare ginsenoside of Panax notoginseng stem and leaf in Test Example 1; Figure 4 The rare ginsenoside of Panax notoginseng stem and leaf obtained in Example 1 in Application Example 1, and the Panax notoginseng stem and leaf extract and erythromycin have the bacteriostatic effect on ATCC6919 Propionibacterium acnes. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical solutions and advantages of the embodiments of the present application more clear, the technical solutions in the embodiments of the present application will be clearly and completely described as follows. The specific conditions not mentioned in the embodiments are carried out according to the conventional conditions or the conditions suggested by the manufacturers. The reagents or instruments not mentioned by the manufacturers are all conventional products that can be obtained by market purchase.

[0028] The rare ginsenoside of Panax notoginseng stem and leaf provided by the present application and the preparation method and application thereof will be specifically described as follows.

[0029] The present application provides a rare ginsenoside of Panax notoginseng stem and leaf, which simultaneously comprises Rk3, Rh4, S-Rg3, R-Rg3, Rk1 and Rg5.

[0030] It should be noted that the inventors have found that the above-mentioned Rk3, Rh4, S-Rg3, R-Rg3, Rk1 and Rg5 can cooperate with each other to have a better inhibitory effect on Propionibacterium acnes, Burkholderia cepacia and Malassezia furfur.

[0031] In some optional embodiments, the rare ginsenoside of Panax notoginseng stem and leaf comprises 1wt%-2wt% of Rk3, 1.5wt%-2.8wt% of Rh4, 7.5wt%-13.5wt% of S-Rg3, 7wt%-13wt% of R-Rg3, 5.5wt%-10.5wt% of Rk1 and 6wt%-12.5wt% of Rg5 in terms of mass percentage.

[0032] It should be noted that the five substances described above cannot be effective in inhibiting P. acnes, B. cepacia and M. furfur at the same time. The content of the five substances described above will also affect the inhibition of P. acnes, B. cepacia and M. furfur by the rare ginsenoside in the stems and leaves of Panax notoginseng. If the content of some components is too high or too low, P. acnes, B. cepacia and M. furfur may not be effectively inhibited.

[0033] Exemplarily, the content of Rk3 in the rare ginsenoside in the stems and leaves of Panax notoginseng can be 1wt%, 1.2wt%, 1.4wt%, 1.6wt%, 1.8wt% or 2wt%, etc., and can also be other values within the range of 1wt%~2wt%.

[0034] The content of Rh4 in the rare ginsenoside in the stems and leaves of Panax notoginseng can be 1.5wt%, 1.8wt%, 2wt%, 2.2wt%, 2.5wt% or 2.8wt%, etc., and can also be other values within the range of 1.5wt%~2.8wt%.

[0035] The content of S-Rg3 in the rare ginsenoside in the stems and leaves of Panax notoginseng can be 7.5wt%, 8wt%, 8.5wt%, 9wt%, 9.5wt%, 10wt%, 10.5wt%, 11wt%, 11.5wt%, 12wt%, 12.5wt%, 13wt% or 13.5wt%, etc., and can also be other values within the range of 7.5wt%~13.5wt%.

[0036] The content of R-Rg3 in the rare ginsenoside in the stems and leaves of Panax notoginseng can be 7wt%, 8wt%, 9wt%, 10wt%, 11wt%, 12wt% or 13wt%, etc., and can also be other values within the range of 7wt%~13wt%.

[0037] The content of Rk1 in the rare ginsenoside in the stems and leaves of Panax notoginseng can be 5.5wt%, 6wt%, 6.5wt%, 7wt%, 7.5wt%, 8wt%, 8.5wt%, 9wt%, 9.5wt%, 10wt% or 10.5wt%, etc., and can also be other values within the range of 5.5wt%~10.5wt%.

[0038] The content of Rg5 in the rare ginsenoside in the stems and leaves of Panax notoginseng can be 6wt%, 7wt%, 8wt%, 9wt%, 10wt%, 11wt%, 12wt% or 12.5wt%, etc., and can also be other values within the range of 6wt%~12.5wt%.

[0039] In some preferred embodiments, the rare ginsenosides in Panax notoginseng stems and leaves, by mass percentage, include 1.3wt%~1.8wt% of Rk3, 1.5wt%~2.2wt% of Rh4, 7.8wt%~10.6wt% of S-Rg3, 7.1wt%~10.2wt% of R-Rg3, 5.6wt%~8.8wt% of Rk1 and 7.2wt%~9.6wt% of Rg5. In some preferred embodiments, the rare ginsenosides in Panax notoginseng stems and leaves, by mass percentage, include 1.3wt%~1.65wt% of Rk3, 2.05wt%~2.2wt% of Rh4, 9.2wt%~10.6wt% of S-Rg3, 9.85wt%~10.2wt% of R-Rg3, 7.35wt%~8.8wt% of Rk1 and 8.4wt%~9.6wt% of Rg5.

[0040] The above-mentioned preferred or better content scheme can have a more significant inhibitory effect on Propionibacterium acnes, Burkholderia cepacia, and Malassezia furfur.

[0041] In this invention, the preparation of rare ginsenosides from Panax notoginseng stems and leaves includes: mixing Panax notoginseng stem and leaf extract with an aqueous acetic acid solution, reacting at 105℃~120℃ for 10min~40min, and drying; and no macroporous resin purification is performed during the preparation of rare ginsenosides from Panax notoginseng stems and leaves.

[0042] In the above method, the inventors creatively proposed that mixing Panax notoginseng stem and leaf extract with an aqueous acetic acid solution and reacting the mixture enables the rapid conversion of some chemical substances in the Panax notoginseng stem and leaf extract at a relatively low temperature (below 130°C) (in less than 1 hour). This is because acidic conditions provide protons (H+). + This method induces the desugaring of glycosidic bonds in ginsenosides, followed by dehydration, double bond rearrangement, and stereoconfiguration, ultimately generating rare ginsenosides with lower polarity and simpler structures. Through this rapid transformation, high-purity new active substances Rh4, S-Rg3, R-Rg3, Rk1, and Rg5 were obtained, which subsequently exhibited good antibacterial effects against *Propionibacterium acnes*, *Burkholderia cepacia*, and *Malassezia furfur*. This method avoids the macroporous resin purification step required in conventional operations, significantly shortening preparation time and process, and reducing costs.

[0043] In this invention, the Panax notoginseng stem and leaf extract includes S-Rg2, Rb1, Fc, Rb3, Rd and Rk3, but does not contain Rh4, S-Rg3, R-Rg3, Rk1 and Rg5.

[0044] In some alternative embodiments, the Panax notoginseng leaf extract comprises 3.1 wt% to 3.25 wt% of S-Rg2, 5.5 wt% to 6.5 wt% of Rb1, 10 wt% to 11 wt% of Fc, 29 wt% to 32 wt% of Rb3, 1 wt% to 1.5 wt% of Rd, and 1 wt% to 2 wt% of Rk3, by mass percentage.

[0045] In some alternative embodiments, the reaction temperature of the Panax notoginseng leaf extract and the acetic acid aqueous solution can be 105°C, 108°C, 110°C, 112°C, 115°C, 118°C, or 120°C, or other values within the range of 105°C to 120°C. In some preferred embodiments, the reaction temperature of the Panax notoginseng leaf extract and the acetic acid aqueous solution is 110°C to 120°C, and a higher content of the newly generated rare ginsenoside can be obtained.

[0046] In some alternative embodiments, the reaction time of the Panax notoginseng leaf extract and the acetic acid aqueous solution can be 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, or 40 min, or other values within the range of 10 min to 40 min. In some preferred embodiments, the reaction time of the Panax notoginseng leaf extract and the acetic acid aqueous solution is 30 min to 40 min, and a higher content of the newly generated rare ginsenoside can be obtained.

[0047] In some alternative embodiments, the concentration of acetic acid in the acetic acid aqueous solution can be 0.05% to 0.2%, such as 0.05%, 0.1%, 0.15%, or 0.2%, or other values within the range of 0.05% to 0.2%, by volume percentage. In some preferred embodiments, the concentration of acetic acid in the acetic acid aqueous solution is 0.1% to 0.2%, by volume percentage, and a higher content of the newly generated rare ginsenoside can be obtained.

[0048] In some alternative embodiments, the solid-liquid ratio of the Panax notoginseng leaf extract and the acetic acid aqueous solution can be 4 mg: 1 mL to 16 mg: 1 mL, such as 4 mg: 1 mL, 6 mg: 1 mL, 8 mg: 1 mL, 10 mg: 1 mL, 12 mg: 1 mL, 14 mg: 1 mL, or 16 mg: 1 mL, or other values within the range of 4 mg: 1 mL to 16 mg: 1 mL. In some preferred embodiments, the solid-liquid ratio of the Panax notoginseng leaf extract and the acetic acid aqueous solution is 4 mg: 1 mL to 8 mg: 1 mL, and a higher content of the newly generated rare ginsenoside can be obtained.

[0049] In addition, the present application also provides the use of the rare ginsenoside in Panax notoginseng stem and leaf, for example, the rare ginsenoside in Panax notoginseng stem and leaf can be used for preparing a product for inhibiting at least one of Propionibacterium acnes, Burkholderia cenocepacia and Malassezia furfur.

[0050] In some optional embodiments, the Propionibacterium acnes can include at least one of Propionibacterium acnes ATCC11827, Propionibacterium acnes ATCC12930 and Propionibacterium acnes ATCC6919.

[0051] In addition, the present application also provides a cosmetic product containing the rare ginsenoside in Panax notoginseng stem and leaf.

[0052] In some optional embodiments, the rare ginsenoside in Panax notoginseng stem and leaf can be used in the cosmetic product in an amount of 0.00625% to 0.55% by mass, such as 0.00625%, 0.0125%, 0.025%, 0.05%, 0.1%, 0.2%, 0.4% or 0.55%, etc., or other values within the range of 0.00625% to 0.55%.

[0053] In some optional embodiments, the total amount of Rh4, S-Rg3, R-Rg3, Rk1 and Rg5 in the rare ginsenoside in Panax notoginseng stem and leaf can be used in the cosmetic product in an amount of 0.00313% to 0.275% by mass, such as 0.00313%, 0.00626%, 0.0125%, 0.025%, 0.05%, 0.1%, 0.2% or 0.275%, etc., or other values within the range of 0.00313% to 0.275%.

[0054] In addition, the present application also provides a pharmaceutical product containing the rare ginsenoside in Panax notoginseng stem and leaf.

[0055] In addition, the present application also provides a food product containing the rare ginsenoside in Panax notoginseng stem and leaf.

[0056] The features and performances of the present application are further described in detail below in combination with embodiments.

[0057] Example 1 The rare ginsenoside in Panax notoginseng stem and leaf was prepared as follows: 160 mg of Panax notoginseng stem and leaf extract (purchased from Jinqi Pharmaceutical Co., Ltd.) was placed in a 250 mL conical flask, 20 mL of 0.1% acetic acid aqueous solution was added, the weight was fixed, the opening was sealed with cowhide paper, and then the reaction was carried out at 110℃ for 40 min and freeze-drying (pre-freezing temperature: -50℃, time: 3 days, vacuum degree: 1 MPa).

[0058] Example 2 differs from Example 1 in that the concentration of the aqueous acetic acid solution is 0.05%.

[0059] Example 3 differs from Example 1 in that the concentration of the aqueous acetic acid solution is 0.2%.

[0060] Example 4 differs from Example 1 in that the amount of the Notoginseng stem and leaf extract is 80 mg.

[0061] Example 5 differs from Example 1 in that the amount of the Notoginseng stem and leaf extract is 320 mg.

[0062] Example 6 differs from Example 1 in that the reaction temperature is 105°C.

[0063] Example 7 differs from Example 1 in that the reaction temperature is 120°C.

[0064] Example 8 differs from Example 1 in that the reaction time is 10 min.

[0065] Example 9 differs from Example 1 in that the reaction time is 20 min.

[0066] Comparative Example 1 differs from Example 1 in that the aqueous acetic acid solution is replaced by pure water.

[0067] Comparative Example 2 differs from Example 1 in that the acetic acid in the aqueous acetic acid solution is replaced by an equal concentration of ethanol.

[0068] Comparative Example 3 differs from Example 1 in that the Notoginseng stem and leaf is replaced by American ginseng stem and leaf.

[0069] Comparative Example 4 uses the corresponding saponin standard to configure the rare ginsenoside of the Notoginseng stem and leaf of Example 1, and differs from Example 1 in that the content of S-Rg3 is reduced to 5 wt%.

[0070] Comparative Example 5 uses the corresponding saponin standard to configure the rare ginsenoside of the Notoginseng stem and leaf of Example 1, and differs from Example 1 in that the content of Rk1 is reduced to 2 wt%.

[0071] Comparative Example 6 uses the corresponding saponin standard to configure the rare ginsenoside of the Notoginseng stem and leaf of Example 1, and differs from Example 1 in that the content of Rg5 is reduced to 5 wt%.

[0072] Test Example 1 The HPLC method is used for analyzing the rare ginsenoside in the Panax notoginseng stem and leaf and the Panax notoginseng stem and leaf extract obtained in Example 1, and the mixed standard solution containing Panax notoginseng saponin R1, ginsenoside Rg1, ginsenoside Re, ginsenoside S-Rg2, ginsenoside Rb1, ginsenoside Rb3, ginsenoside Rd, ginsenoside Rk3, ginsenoside Rh4, ginsenoside S-Rg3, ginsenoside Rk1, ginsenoside Rg5, ginsenoside R-Rg2, ginsenoside R-Rg3 and Panax notoginseng saponin Fc is used as a control. The ginsenosides not prepared by the present application are purchased from Chengdu Keluoma Biological Technology Co., Ltd.

[0073] The chromatographic conditions are as follows: Chromatographic column: Agilent Zorbax SB-C18 column (250 mm x 4.6 mm, 5 μm); Mobile phase: acetonitrile (A) - water (B); Gradient elution: 0-15 min, 20% A; 15-40 min, 20%-45% A; 40-54 min, 45%-75% A; 54-60 min, 75% A; 60-60.1 min, 75%-20% A; 60.1-65 min, 20% A; Flow rate: 1.0 mL / min; Column temperature: 35°C; Detection wavelength: 203 nm; Injection volume: 20 μL.

[0074] The results are shown in Figures 1 to 3 and Tables 1-2.

[0075] Table 1 Chromatographic peaks

[0076] Table 2 Contents of each ginsenoside in the Panax notoginseng stem and leaf extract and the freeze-dried powder after conversion

[0077] As can be seen from Table 2, the method provided in the present application enables the complete conversion of S-Rg2, Rb1, Fc, Rb3 and Rd in the rare ginsenoside in the Panax notoginseng stem and leaf compared with the Panax notoginseng stem and leaf extract, and obtains new saponin substances Rh4, S-Rg3, R-Rg3, Rk1 and Rg5.

[0078] Test Example 3 The rare ginsenoside in the Panax notoginseng stem and leaf obtained in Examples 2-3 is subjected to saponin component and content testing according to the method of Test Example 1, and the results are shown in Table 3.

[0079] Table 3 Contents of each ginsenoside in the freeze-dried powder

[0080] As can be seen from Table 2 and Table 3, with the increase of acetic acid concentration, the total content of ginsenosides shows a growth trend. However, from the rare ginsenoside components, except Rk3, Rh4, S-Rg3, R-Rg3, Rk1, Rg5 all show an increase in content with the increase of acetic acid concentration, indicating that the increase of acetic acid concentration is conducive to the generation of rare ginsenosides. At the same time, Fc, Rb3, Rd and Rk3, which are the same as the stem and leaf extract, appear at the acetic acid concentration of 0.05%, indicating that it is difficult to obtain newly generated rare ginsenosides with higher content at this concentration.

[0081] Test Example 4 The rare ginsenosides obtained from Panax notoginseng stem and leaf in Examples 4-5 were tested for ginsenoside components and content according to the method of Test Example 1, and the results are shown in Table 4.

[0082] Table 4 Ginsenoside content in freeze-dried powder

[0083] As can be seen from Table 2 and Table 4, with the increase of sample concentration, the total content of ginsenosides shows a trend of first increasing and then decreasing, with a peak at 8 mg / ml. At a sample concentration of 8 mg / ml, the contents of low-polarity ginsenoside components Rk3, S-Rg3, R-Rg3, Rk1, Rg5 generally reach a peak, especially R-Rg3 and S-Rg3, whose contents are significantly higher than those of other concentration groups. At the same time, Fc, Rb3, Rd and Rk3, which are the same as the stem and leaf extract, appear at the sample concentration of 16 mg / ml, indicating that it is difficult to obtain newly generated rare ginsenosides with higher content at this concentration.

[0084] Test Example 5 The rare ginsenosides obtained from Panax notoginseng stem and leaf in Examples 6-7 were tested for ginsenoside components and content according to the method of Test Example 1, and the results are shown in Table 5.

[0085] Table 5 Ginsenoside content in freeze-dried powder

[0086] As can be seen from Table 2 and Table 5, with the increase of temperature, the total content of ginsenosides shows a trend of first increasing and then decreasing, and a peak appears at 110°C. During the process of increasing from 105°C to 110°C, the content of other low-polarity ginsenosides such as Rh4 increases except Rk3, which shows that appropriate heating is helpful to the transformation of low-polarity ginsenosides in ginseng and improves the transformation efficiency. However, with the increase of temperature to 120°C, the content of some low-polarity ginsenosides decreases, which may be due to the degradation of low-polarity ginsenosides caused by over-high heating temperature, thereby reducing the transformation efficiency. Meanwhile, the ginsenoside components Fc, Rb3, Rd and Rk3 appear at 105°C, which is the same as the stem and leaf extract, indicating that it is difficult to obtain new rare ginsenosides with higher content at this temperature. Among them, the highest content of new rare ginsenosides can be obtained at the heating temperature of 110°C.

[0087] Test Example 6 The rare ginsenosides obtained from Panax notoginseng stem and leaf in Examples 8-9 were subjected to ginsenoside component and content test according to the method of Test Example 1, and the results are shown in Table 6.

[0088] Table 6 Content of each ginsenoside in freeze-dried powder

[0089] As can be seen from Table 2 and Table 6, with the extension of heating time, the total content of ginsenosides shows a trend of first decreasing and then increasing, and a peak appears at 40 min. However, for low-polarity ginsenosides Rk3, Rh4, S-Rg3, R-Rg3, Rk1 and Rg5, the content shows a growth trend with the increase of heating time, and reaches the highest at 40 min. Meanwhile, the ginsenoside components Fc, Rb3, Rd and Rk3 appear at 10 min and 20 min, which is the same as the stem and leaf extract, and the content of these components gradually decreases with the extension of heating time, and disappears completely at 40 min. This shows that with the extension of heating time, these components are gradually transformed into other rare ginsenosides, and the highest content of new rare ginsenosides can be obtained at 40 min.

[0090] Test Example 7 The ginsenosides obtained from Comparative Examples 1-3 were subjected to ginsenoside component and content test according to the method of Test Example 1, and the results are shown in Table 7.

[0091] Table 7 Content of each ginsenoside in freeze-dried powder

[0092] Application Example 1 In vitro antibacterial effect of rare ginsenosides from Panax notoginseng stem and leaf on Propionibacterium acnes (1) Preparation of culture medium 1) Preparation of P. acnes liquid medium Weigh 37.5 g of P. acnes liquid medium (purchased from Shanghai Ruichu Technology Co., Ltd.) into a conical flask, add 1000 mL of deionized water, heat and boil for 1 min to dissolve, seal, place in a high-pressure steam sterilization pot, sterilize at 115°C for 20 min, after sterilization, shake well, and cool in a refrigerator at 4°C for standby.

[0093] 2) Preparation of P. acnes solid medium Weigh 52.5 g of P. acnes solid medium (purchased from Shanghai Ruichu Technology Co., Ltd.) into a conical flask, add 1000 mL of deionized water, heat and boil for 1 min to dissolve, seal, place in a high-pressure steam sterilization pot, sterilize at 115°C for 20 min, after sterilization, shake well, and cool to 50-55°C, pour into a flat plate, seal with a sealing film after solidification, and place in a refrigerator at 4°C for standby.

[0094] (2) Preparation of bacterial suspension Take the frozen bacteria solution (P. acnes ATCC6919, purchased from Guangdong Microbial Culture Collection Center) and add liquid medium for recovery and expansion. Take 1 mL of bacterial solution with a pipette gun and add it to a centrifuge tube containing liquid medium. Place it in an anaerobic incubator at 37°C for 48 h. Dilute with liquid medium to a final bacterial solution concentration of 1×10 8 CFU / mL.

[0095] (3) Preparation of drug solution 1) Preparation of test drug Weigh the Panax notoginseng stem and leaf extract and the freeze-dried powder prepared in each example and comparative example, dissolve with dimethyl sulfoxide (DMSO), and prepare a test drug solution with a concentration of 20.00 mg / mL. Filter through a nylon 0.22 μm filter membrane and reserve for use.

[0096] 2) Preparation of control solution Weigh an appropriate amount of erythromycin, prepare a control solution with a concentration of 50.0 μg / mL using DMSO, filter through a nylon 0.22 μm filter membrane, and reserve for use.

[0097] (4) Measurement of inhibition zone Use the punch method to measure the inhibition zone of P. acnes by the test drug. Dip a sterile cotton swab in the bacterial suspension (concentration of about 1×10 8CFU / mL), evenly spread on solid culture medium plate, punch with puncher (diameter of 6 mm), pick out agar in the hole, add 50 μL of test liquid (20.00 mg / mL), 50 μL of erythromycin solution (6.4 μg / mL), 50 μL of DMSO solution into the hole, all the above operations are operated under sterile conditions, place in an anaerobic incubator at 37°C for 48 h, measure the diameter of the inhibition zone by cross method, parallel determination for 3 times, take the average value. The following Table 8 evaluates the antibacterial efficacy of each group.

[0098] Table 8 Evaluation standard of antibacterial efficacy

[0099] (5) Determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) The effect of rare ginsenoside on Propionibacterium acnes in the logarithmic growth phase was investigated by micro-doubling dilution method.

[0100] The Propionibacterium acnes in the logarithmic growth phase was diluted with the corresponding liquid medium to a concentration of about 1×10 8 CFU / mL, and then diluted by 10 times successively with liquid medium to prepare 10 6Bacterial solution of 1.0 x 10 CFU / mL, standby. Prepare 2 sterile 96-well plates, add 200 μL of blank medium to rows A, H and columns 1, 12 of the 96-well plate, add 180 μL of bacterial liquid medium to the test hole of column 2 and B10 hole, and add 100 μL of bacterial liquid medium to columns 2-9 and test holes respectively. In the first 96-well plate, add 20 μL of corresponding drug to each hole in column 2, so that the initial concentration of notoginseng stem and leaf extract is 1.0 mg / mL, and the initial concentration of erythromycin is 0.050 mg / mL, mix well by blowing with a row gun, and then transfer 100 μL of liquid medium from column 2 to column 3, mix well, and then transfer 100 μL of liquid medium from column 3 to column 4, and so on, to obtain 8 concentration gradients of test drug-containing medium by the method of dilution by ratio, discard 100 μL of mixed solution in column 9, and finally add 100 μL of blank medium to each hole, so that the final volume of each hole is 200 μL. The transformed freeze-dried powder sample, the freeze-dried powder sample of the comparative example, and the negative control group (DMSO + blank medium) are completed on the same plate, the sample addition method of each freeze-dried powder sample and the notoginseng stem and leaf extract is consistent, and the sample addition method of the negative control group is as follows: the initial hole is 180 μL of blank medium plus 20 μL of DMSO solution, the subsequent holes are gradually diluted by two times, mixed well, and finally 100 μL of blank medium is added to each hole, so that the final volume of each hole is 200 μL. At the same time, the 10th column and the test holes of F11 and G11 of each plate are positive controls (bacterial liquid medium + DMSO), 100 μL of bacterial liquid medium is added to columns C10-G10 and holes F1 and G1, 180 μL of bacterial liquid medium is added to hole B10, 20 μL of DMSO is added, the subsequent holes are gradually diluted by two times, mixed well, 100 μL of mixed solution in the last hole is discarded, and finally 100 μL of blank medium is added to each hole, so that the final volume of each hole is 200 μL. 200 μL of blank medium is added to the remaining holes in column 11. Place in an anaerobic incubator at 37°C for 48 hours, and then detect OD 600 The minimum concentration for inhibiting 90% of P. acnes growth is MIC, and the inhibition rate is calculated according to the following formula: .

[0101] Three replicates are set for each plate, and three parallel determinations are performed. Take 20 μL of each of MIC, 2MIC, and 4MIC from the holes and inoculate into solid culture medium, place in an anaerobic incubator at 37°C for 48 hours, and then take the concentration of sterile colony growth as MBC, with three parallel experiments.

[0102] Inhibition zone determination results The results are shown in Figure 4 , Tables 9 to 11.

[0103] Table 9 Diameter of inhibition zone (average) of each drug and efficacy evaluation (ATCC6919)

[0104] It can be seen from the results that the diameters of the inhibition zone of DMSO and the Panax notoginseng stem and leaf extract are both 6.0 mm, that is, no antibacterial effect. The diameter of the inhibition zone of erythromycin is >20 mm, and the antibacterial effect is high. The diameter of the inhibition zone of the freeze-dried powder of the example is >15 mm, indicating that the rare ginsenoside prepared by the example of the application has strong activity in inhibiting Propionibacterium acnes ATCC6919.

[0105] MIC and MBC determination results The MIC and MBC of the Panax notoginseng stem and leaf extract and the freeze-dried powder sample after conversion, the freeze-dried powder sample of the comparative example were investigated by using the micro two-fold dilution method and the colony counting method, and the results are shown in Table 10.

[0106] Table 10 MIC and MBC of various drugs on Propionibacterium acnes (ATCC6919)

[0107] It can be seen from Table 10 that the MIC of the rare ginsenoside prepared by the example 1 of the application is 62.50 μg / mL, and the MBC is 62.5 μg / mL; the MIC of erythromycin is 0.78 μg / mL, and the MBC is 1.56 μg / mL; and the MIC and MBC of the Panax notoginseng stem and leaf extract cannot be measured.

[0108] Further, the application further provides a molecular docking test.

[0109] Specifically, the 3D molecular structures of rare ginsenosides S-Rg2, Rk3, Rh4, S-Rg3, R-Rg3, Rk1 and Rg5 were downloaded from Pubchem (https: / / pubchem.ncbi.nlm.nih.gov / ). For the components without structure on Pubchem, the molecular structure was drawn by ChemDraw 22.0, and the structure energy was optimized by Chem3D 22.0. Autodock vina software was used to process small molecules (add full hydrogen, add Gasteiger charge processing (no error can be omitted when adding full hydrogen), set as ligand, detect torsion bond, select torsion bond) and save the compound "pdbqt" file. Five key pathogenic protein structures were downloaded from RCSB PDB database (https: / / www.rcsb.org / ), including DNase I (sequence number: 4AWN), PPA0542 (sequence number: 7BCJ), CAMP (sequence number: 7SAY), lipase (sequence number: 5H6G) and DsA1 (sequence number: 7EA4). Pymol software was used to process proteins (remove water, add full hydrogen, remove small molecule ligand), and the grid box of the protein was set to full package on the Autodock vina software. Finally, the Autodock vina software was used to dock the processed five key pathogenic protein receptors with rare ginsenoside small molecule ligands. In each docking, 20 docking conformations were generated, and the one with the lowest docking energy was selected. The results were visualized by Pymol software. The binding activity of the two was evaluated by the binding energy: when the docking energy <0 kcal / mol, it indicated that the receptor and the ligand could spontaneously bind; when the docking energy <-5.0 kcal / mol, it indicated that the two formed a stable conformation structure, and the binding activity was high; when the docking energy <-7.0 kcal / mol, it indicated that the ligand had excellent ability to spontaneously bind to the receptor protein, and the binding activity was strong and the affinity was high.

[0110] Seven rare ginsenosides were selected for docking with five key pathogenic proteins, and the results are shown in Table 11.

[0111] Table 11 Docking energy of rare ginsenosides and five key pathogenic proteins

[0112] As can be seen from the above table, the docking energy of each rare ginsenoside with the five pathogenic proteins ranges from-6.7 to-10.5 kcal / mol, except that the binding energy between S-Rg2 and DNase I is-6.7 kcal / mol, and the docking energy of other rare ginsenosides with the five key pathogenic proteins is <-7.0 kcal / mol, which indicates that the rare ginsenosides have excellent ability to spontaneously bind to the five key pathogenic proteins and form a stable structure, and the binding activity and affinity are high.

[0113] The mechanism of the rare ginsenoside provided by the present application to play the anti-P. acnes includes: (1) The hydrophobicity of the rare ginsenoside is enhanced after deglycosylation, that is, the liposolubility is enhanced, and it is easier to penetrate the bacterial cell membrane, so it has good antibacterial activity; (2) There may be structural synergy and structural complementarity, because the lipophilicity of the rare ginsenoside is enhanced, there may be superposition and synergistic enhancement of lipophilicity to the penetration efficiency of the P. acnes bacterial membrane, and the aglycone of different types of rare ginsenosides may form a complex through hydrophobic interaction to enhance the insertion ability to the membrane phospholipid layer; (3) There may be biofilm inhibition, as can be seen from the molecular docking result, the rare ginsenoside Rh4 may play a role by binding to the proteins DNase I and CAMP, thereby reducing the formation of biofilm, and then having antibacterial activity; (4) It may block the nutrient source of P. acnes, and the rare ginsenoside R-Rg3 may bind to lipase to make it unable to hydrolyze skin triacylglycerol, making it difficult to release glycerol and free fatty acids, thereby blocking the growth and reproduction of P. acnes; (5) It may block the colonization of P. acnes, and the rare ginsenoside binds to the main surface glycoprotein DsA1 of P. acnes, so that it cannot specifically bind to dermatan sulfate, thereby blocking the invasion of host cells mediated by dermatan sulfate, and thus blocking the colonization of bacteria.

[0114] Further, the panax notoginseng root is extracted to obtain a panax notoginseng root extract. Then, the panax notoginseng root extract is mixed with water, and extracted at 130℃ for 4h, and then purified by a macroporous resin to obtain an active component (named as G95).

[0115] The saponin components and content of the panax notoginseng root extract and the active component G95 are tested, and the results are shown in Table 12.

[0116] In addition, the MIC and MBC of the panax notoginseng root extract and the active component G95 are investigated by using the micro two-fold dilution method and the colony counting method, and the results are also shown in Table 12.

[0117] Table 12 Content of each ginsenoside in Panax notoginseng root extract and G95 and MIC and MBC results

[0118] Taking Panax notoginseng root as raw material, the ginsenoside components in Panax notoginseng root were obtained by the preparation method of rare ginsenosides in Panax notoginseng stem and leaf provided in the application through different conditions (temperature 105℃~120℃, time 20min~40min, solid-liquid ratio of extract and acetic acid aqueous solution 4mg:1mL~8mg:1mL), and were named as G1 to G9, and the specific composition of G1 to G9 is shown in Table 13. The MIC and MBC of G1 to G9 were investigated by micro double dilution method and colony counting method, and the results are also shown in Table 13.

[0119] Table 13 Content of each ginsenoside in G1 to G9 and MIC and MBC results

[0120] Application Example 2 In vitro antibacterial effect of rare ginsenosides in Panax notoginseng stem and leaf on Burkholderia cepacia Experimental strains: purchased ATCC25416 and CMCC(B)23005; Culture medium: Tryptose Soya Agar (TSA), Beijing Land Bridge Technology Co., Ltd., batch number: 240524; Tryptose Soya Broth (TSB), Guangdong Huan Kai Microbial Science and Technology Co., Ltd., batch number: 250612A10; (all are commercial culture media).

[0121] Experimental steps: Preparation of bacterial solution: prepare bacterial solution with McFarland turbidity of 0.5 (about 10 8 CFU / mL) from freshly cultured ATCC25416 and CMCC(B)23005 plates, and then dilute with TSB by 10 times successively to prepare bacterial solution with 10 6 CFU / mL for standby; Preparation of drug solution: dilute the rare ginsenosides in Panax notoginseng stem and leaf provided in Example 1 with TSB to 500μg / mL, 250μg / mL, 125μg / mL and 62.5μg / mL respectively.

[0122] Antibacterial experiment: Experimental group: add the diluted drug solution to the 96-well plate, 10 wells for each concentration of drug solution, 100μL per well, and add the diluted bacterial solution to the drug solution, 5 wells for each kind of bacteria, 100μL per well.

[0123] Positive control group: in 96-well plates, add 100 μL TSB, a total of 10 holes, 5 holes add 100 μL ATCC25416, and the other 5 holes add 100 μL CMCC(B)23005.

[0124] Place the 96-well plate at 36°C and incubate overnight.

[0125] Determine the bacterial concentration (OD 600 ) The experimental results show that the MIC and MBC values of the panax notoginseng stem and leaf rare ginsenoside provided in Example 1 for ATCC25416 are both 125 μg / mL, and the MIC and MBC values for CMCC(B)23005 are 250 μg / mL.

[0126] In addition, the antibacterial effect of panax notoginseng stem and leaf extract on Burkholderia cepacia was tested according to the above experimental method, and the results showed that the panax notoginseng stem and leaf extract had no inhibitory effect on Burkholderia cepacia.

[0127] Application Example 3 In Vitro Antibacterial Effect of Panax Notoginseng Stem and Leaf Rare Ginsenoside on Malassezia furfur Experimental materials: Malassezia furfur (Shanghai Center for Collection and Authentication of Microorganisms), number ATCC4344, culture temperature 25°C~28°C, aerobic culture. Configuration: freeze-dried powder strain, culture solution; Malt extract powder, oxgall powder, pH 7.0±0.2 (25°C) (Shanghai Ruichu Biological Technology Co., Ltd.), number T2761A; Tween 40 (Shanghai Ruichu Biological Technology Co., Ltd.), number T2761B, 1 g x 20 bags; Glycerol monooleate (Shanghai Ruichu Biological Technology Co., Ltd.), number T2280A, 0.25 g x 20 bags; Agar powder (Biyun Tian Biological Technology Co., Ltd.), number ST004E, 500 g.

[0128] Preparation of liquid culture medium: Weigh 75.0 g of Malassezia furfur liquid culture medium (malt extract powder, oxgall powder) in 1 L of distilled water or deionized water, heat and boil for more than 1 minute to dissolve, dispense, add 1 g of Tween 40 and 0.25 g of glycerol monooleate to each 100 mL of culture medium, mix well, autoclave at 121°C for 15 minutes, cool to about 60°C, shake well, pour flat, solidify, and prepare for use.

[0129] Inoculation (activation): Prepare the following materials for the experiment: lyophilized powder tubes, dissolving solution, sterile water, alcohol lamp, two solid slant culture media (flat solid culture media), pipettes, and several 200ul pipette tips.

[0130] Using a clean workbench, wipe the ampoule clean with cotton wool soaked in 75% alcohol. Heat the tip of the ampoule with a flame, and drip a small amount of sterile water onto the heated tip to break it. Use a file or tweezers to knock off the broken tip of the ampoule. Use a pipette (1000μL tip) to draw 2-3mL of culture medium into the ampoule. After the culture medium and the ampoule are thoroughly mixed, inoculate the bacterial suspension with a pipette (or spread it on a flat surface with a spreader) onto two solid slant agar plates (flat solid agar plates), distributing 200μL to each plate.

[0131] Cultivation (Generation): Seal the solid slant culture medium (flat solid medium) with sealing film and incubate it in a constant temperature incubator at 25℃~28°C for 48 hours until visible colonies form on the surface of the medium. Collect a small number of colonies with an inoculation loop and transfer them to an Erlenmeyer flask containing liquid medium (flame-sterilized mouth) to obtain a homogeneous bacterial suspension. Incubate at 25℃~28°C with shaking at 180 rpm. Subculture again after about 3 to 4 days.

[0132] Measurement and adjustment of microbial concentration: The bacterial concentration was determined using an ELISA reader and a serial dilution method: Take several sterile tubes and add 900 μL of culture medium to each. Add 100 μL of bacterial suspension to the first tube, mix well, then change the pipette tip and add 100 μL to the second tube. Repeat this serial dilution process. Take 10... -4 10 -5 10 -6 10 μL of the dilution solution (3-4 drops per dilution, replicated) is vertically dropped onto a solid culture medium. The medium is then incubated under the appropriate conditions (30°C for *Malassezia furfur*) in an incubator (colonizations are visible in 3-4 days). The colony count is determined to obtain the concentration of the stock solution (CFU / mL). Using a 96-well plate, 200 μL of the stock solution is diluted 0, 2, 3, 4, 5, and 8 times in each well, along with sterile culture medium. The OD value at 600 nm is measured using a microplate reader. The measured OD values ​​are correlated with CFU / mL. The *Malassezia furfur* bacterial suspension is adjusted to a concentration of 1 × 10⁻⁶. 6 CFU / mL available for use.

[0133] Antibacterial activity test MIC determination: similar to that of *Bacillus cereus*.

[0134] Minimum bactericidal concentration (MBC) determination: 4MIC, 2MIC, 1MIC concentration of ginsenoside solution is equal to liquid medium by plate coating method coated in the culture dish. 30℃ incubator inverted culture 48h~60h observation of the growth of malassezia in the medium, if no sterile growth, according to the situation using lower concentration of ginsenoside solution, if no sterile growth and the lowest concentration, that is the minimum bactericidal concentration (MBC).

[0135] The experimental results show that the MIC value of the panax notoginseng stem and leaf rare ginsenoside provided by example 1 on malassezia furfur is 64 μg / mL, and the MBC value is 64 μg / mL.

[0136] In addition, according to the above experimental method, the antibacterial property of panax notoginseng stem and leaf extract on malassezia furfur was tested, and the results showed that the panax notoginseng stem and leaf extract had no inhibitory effect on malassezia furfur.

[0137] In summary, the scheme provided by the present application can rapidly convert part of the chemical substances in the panax notoginseng stem and leaf extract at a lower temperature, so as to obtain new active substances Rh4, S-Rg3, R-Rg3, Rk1 and Rg5 with high purity at the same time, and the obtained panax notoginseng stem and leaf rare ginsenoside can effectively inhibit propionibacterium acnes, burkholderia cepacia and malassezia furfur.

[0138] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A rare ginsenoside of Panax notoginseng stem leaf, characterized in that, The rare ginsenoside in the Panax notoginseng stem and leaf comprises Rk3, Rh4, S-Rg3, R-Rg3, Rk1 and Rg5. The preparation of the rare ginsenoside in the Panax notoginseng stem and leaf comprises: mixing the Panax notoginseng stem and leaf extract with an acetic acid aqueous solution, reacting at 105-120℃ for 10-40 min, and drying; wherein the Panax notoginseng stem and leaf extract comprises S-Rg2, Rb1, Fc, Rb3, Rd and Rk3, and does not contain Rh4, S-Rg3, R-Rg3, Rk1 and Rg5; and the preparation of the rare ginsenoside in the Panax notoginseng stem and leaf does not involve macroporous resin purification.

2. The panax quinquefolium stem and leaf rare ginsenoside of claim 1, characterized in that, The rare ginsenoside in the Panax notoginseng stem and leaf comprises 1-2wt% of Rk3, 1.5-2.8wt% of Rh4, 7.5-13.5wt% of S-Rg3, 7-13wt% of R-Rg3, 5.5-10.5wt% of Rk1 and 6-12.5wt% of Rg5 by mass percentage.

3. The panax quinquefolium stem and leaf rare ginsenoside of claim 2, characterized in that, The rare ginsenoside in the Panax notoginseng stem and leaf comprises 1.3-1.8wt% of Rk3, 1.5-2.2wt% of Rh4, 7.8-10.6wt% of S-Rg3, 7.1-10.2wt% of R-Rg3, 5.6-8.8wt% of Rk1 and 7.2-9.6wt% of Rg5 by mass percentage.

4. The panax quinquefolium stem and leaf rare ginsenoside of claim 3, characterized in that, The rare ginsenoside in the Panax notoginseng stem and leaf comprises 1.3-1.65wt% of Rk3, 2.05-2.2wt% of Rh4, 9.2-10.6wt% of S-Rg3, 9.85-10.2wt% of R-Rg3, 7.35-8.8wt% of Rk1 and 8.4-9.6wt% of Rg5 by mass percentage.

5. The panax quinquefolium stem and leaf rare ginsenoside of claim 1, characterized in that, The Panax notoginseng stem and leaf extract comprises 3.1-3.25wt% of S-Rg2, 5.5-6.5wt% of Rb1, 10-11wt% of Fc, 29-32wt% of Rb3, 1-1.5wt% of Rd and 1-2wt% of Rk3 by mass percentage.

6. The panax quinquefolium stem and leaf rare ginsenoside of any one of claims 1-5, characterized in that, The reaction temperature is 110-120℃.

7. The panax quinquefolium stem and leaf rare ginsenoside of any one of claims 1-5, characterized in that, The reaction time is 30-40 min.

8. The panax quinquefolium stem and leaf rare ginsenoside of any one of claims 1-5, characterized in that, The concentration of acetic acid in the acetic acid aqueous solution is 0.05-0.2% by volume percentage.

9. The panax quinquefolium stem and leaf rare ginsenoside of any one of claims 1-5, characterized in that, The solid-liquid ratio of the Panax notoginseng stem and leaf extract to the acetic acid aqueous solution is 4mg:1mL-16mg:1mL.

10. The use of the rare ginsenoside in the stem and leaf of Panax notoginseng according to any one of claims 1-9, characterized in that, The rare ginsenoside in the Panax notoginseng stem and leaf is used for preparing a product for inhibiting at least one of Propionibacterium acnes, Burkholderia cenocepacia and Malassezia furfur.

11. Use according to claim 10, characterized in that, The Propionibacterium acnes comprises at least one of Propionibacterium acnes ATCC11827, Propionibacterium acnes ATCC12930 and Propionibacterium acnes ATCC6919.

12. A cosmetic product, characterized by, The cosmetic contains the rare ginsenoside in the Panax notoginseng stem and leaf according to any one of claims 1-9.

13. The cosmetic product of claim 12, wherein, The rare ginsenoside in the Panax notoginseng stem and leaf is used in the cosmetic in an amount of 0.00625% to 0.55% by mass.

14. The cosmetic product of claim 13, wherein, The total amount of Rh4, S-Rg3, R-Rg3, Rk1 and Rg5 in the rare ginsenoside in the Panax notoginseng stem and leaf is used in the cosmetic in an amount of 0.00313% to 0.275% by mass.

15. A medicine, characterized in that, The medicine contains the rare ginsenoside in the Panax notoginseng stem and leaf as claimed in any one of claims 1 to 9.

16. A food product, characterized by, The food contains the rare ginsenoside in the Panax notoginseng stem and leaf as claimed in any one of claims 1 to 9.

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