Processing method for increasing content of ginsenoside Rb1 in ginseng

Abstract

The invention discloses a processing method capable of effectively increasing the content of ginsenoside Rb1 in ginseng. The processing method comprises the following steps: 1) selecting fresh ginseng which is just harvested, washing and airing; 2) placing the ginseng in a dark environment and irradiating the ginseng with an ultraviolet lamp, and 3) performing high-temperature fixation on the ginseng slices obtained in the step 2), and then drying the ginseng slices to obtain the ginsenoside Rb1. By adopting the method, the content of ginsenoside Rb1 in the ginseng can be effectively increased, and the method has the advantages of being remarkable in effect, simple and easy to implement and convenient to operate.

Description

Technical Field

[0001] The invention relates to the field of traditional Chinese medicine processing, and in particular to a processing method for increasing the content of ginsenoside Rb1 in ginseng. Background Art

[0002] Ginseng ( Panax ginseng CA Meyer) is the dried root of a perennial herbaceous plant of the genus Panax in the Araliaceae family. It is a precious Chinese medicinal material with extremely high medicinal value. Ginseng is rich in ginsenosides and ginseng polysaccharides, and secondly contains active ingredients such as amino acids, alkaloids, enzymes, proteins and volatile oils. Ginsenosides are a class of steroid compounds, also known as triterpenoid saponins, and are the main active natural compounds of ginseng. They have been shown to have a wide range of pharmacological activities, such as antioxidant, anti-inflammatory and anti-tumor effects. Ginsenosides can be divided into dammarane type and oleanane type according to different glycoside skeletons. Among them, the dammarane type is one of the main active ingredients of ginseng, which is mainly divided into two categories: ginsenodiols and ginsenotriols. Ginsenodiols cover the most types of ginsenosides, mainly ginsenosides Rb1, Rb2, Rb3, Rc, Rd, Rg3, Rh2 and glycoside PD. Ginsenosides include ginsenosides Re, Rg1, Rg2, Rh1 and glycoside PT. Ginseng is known as the "king of herbs" and is widely used in health food, medicine and cosmetics industries. The demand for high-quality ginseng in the pharmaceutical market is rising year by year. Ginsenoside Rb1 is one of the main components of ginsenosides. It has anti-inflammatory, antioxidant, anti-apoptotic and autophagy regulating effects. In addition, it has the advantages of high availability, low cost, high efficiency and low risk. Therefore, ginsenoside Rb1 has attracted much attention in recent years. A large number of research results show that diol ginsenoside Rb1 is the active ingredient of ginseng that exerts the main pharmacological effects and has a significant protective effect on the central nervous system; ginsenoside Rb1 is often used as an anti-inflammatory and learning and memory improvement drug, which exerts an anti-depressant effect by inhibiting inflammatory reactions and promoting neurogenesis.

[0003] The saponin content in ginseng currently on the market is easily affected by cultivation methods, planting management techniques, origin, growing years and primary processing methods, resulting in a low saponin content in ginseng. Currently, there are not many reports on increasing the content of ginsenoside Rb1.

[0004] It has been reported that ultraviolet light can stimulate plants to produce secondary metabolites. For example, irradiating blueberries after harvest can significantly increase the content of anthocyanins and polysaccharides. The effects of ultraviolet light of different wavelengths are also different. The wavelength range of ultraviolet light is 100~400 nm, which is divided into two sections. The section with a wavelength of 100~200 nm is called the far ultraviolet region. This section can only be studied in a vacuum and is currently of little use in practice. The section with a wavelength of 200~400 nm is called the near ultraviolet region. The general ultraviolet spectrum refers to the absorption spectrum of this region.

[0005] Currently, the main function of using ultraviolet light to irradiate ginseng is to kill bacteria. No reports have been found that irradiating ultraviolet light can increase the content of ginsenoside Rb1 in ginseng. Summary of the invention

[0006] In order to overcome the shortcomings of the existing ginseng processing methods, the present invention provides a processing method for effectively increasing the content of ginsenoside Rb1 in ginseng, which has the advantages of significant effect, simple method and convenient operation.

[0007] The present invention provides a processing method for increasing the content of ginsenoside Rb1 in ginseng, characterized in that the processing method comprises the step of irradiating the ginseng with ultraviolet light.

[0008] The present invention has determined the specific steps of the processing method for increasing the Rb1 content of ginseng by irradiating ginseng with ultraviolet light through a large number of experiments.

[0009] Specifically, the processing method for increasing the content of ginsenoside Rb1 in ginseng according to the present invention comprises the following steps:

[0010] A processing method for increasing the content of ginsenoside Rb1 in ginseng, characterized by comprising the steps of: 1) selecting freshly harvested ginseng, washing it, and drying it.

[0011] 2) Place the ginseng in a dark environment and irradiate it with ultraviolet light.

[0012] 3) Slice the ginseng obtained in step 2) and sterilize at high temperature, then dry to obtain the ginseng.

[0013] Wherein, the wavelength of the ultraviolet lamp in step 2) is light in a specific wavelength band of 287-307 nm.

[0014] Preferably, in step 2), the energy of the wavelength of the ultraviolet light is 0.161-0.214 W / m 2 .

[0015] Preferably, in step 2), the energy of the wavelength of the ultraviolet lamp is 0.161 W / m 2 .

[0016] Preferably, the lamp irradiation time in step 2) is 10 min.

[0017] Preferably, in step 2), the sample is irradiated with light for 10 min and then kept in darkness for 24 h.

[0018] Preferably, in step 2), the sample is irradiated with light for 10 min and then kept in darkness for 24 h, and this step is repeated 1-5 times.

[0019] Preferably, the fixing temperature in step 3) is 105° C. and the fixing time is 15 min.

[0020] Preferably, the drying temperature in step 3) is 70°C.

[0021] The present invention also provides a ginseng product obtained by the processing method as described above.

[0022] The ginseng sample obtained in step 3) was crushed and passed through a No. 4 sieve to prepare the test solution: about 1 g of the powder of the product was accurately weighed and placed in a Soxhlet extractor, dichloromethane was added and heated to reflux for 3 h, the dichloromethane solution was discarded, the solvent of the residue was evaporated, and the residue was transferred into a 100 mL conical flask together with the filter paper tube, 50 mL of water-saturated n-butanol was accurately added, the flask was sealed, and it was left overnight. Ultrasonic treatment (power 250 W, frequency 50 kHz) was performed for 30 min, and the initial filtrate was discarded. 25 mL of the subsequent filtrate was accurately measured and placed in an evaporating dish to evaporate to dryness. The residue was dissolved in methanol and transferred to a 5 mL volumetric flask, and methanol was added to dilute to the scale, shaken well, filtered, and the subsequent filtrate was taken. The obtained solution was used for qualitative and quantitative analysis of ginsenoside Rb1 by high performance liquid chromatography.

[0023] The processing method for increasing the content of ginsenoside Rb1 in ginseng provided by the present invention can significantly increase the content of ginsenoside Rb1 in ginseng, and the method is simple and easy to operate and easy to promote and popularize. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 is the HPLC chromatogram of the standard sample ginsenoside Rb1; Figure 2 The high performance liquid chromatograms of the products obtained in Examples 1-3 and Comparative Examples 1-3 are shown. DETAILED DESCRIPTION

[0025] The principles and features of the present invention are described below in conjunction with examples. The examples are only used to explain the present invention and are not used to limit the scope of the present invention.

[0026] Embodiment 1: 1) Select freshly harvested ginseng, remove the remaining soil to avoid affecting the accuracy of the experimental results, and place it in a cool and ventilated place to dry 2) Use the ginseng from step 1) and place it in a dark environment. Use a wavelength of 287-307 nm and an energy of 0.214 W / m 2 The cells were irradiated with light for 10 min and then kept in darkness for 24 h after the light was turned off.

[0027] 3) Slice the ginseng obtained in step 2), and inactivate it at 105°C for 15 min, and then dry it at 70°C to obtain the ginseng.

[0028] Weigh the ginseng sample obtained in step 3), crush and sieve (No. 4 sieve), and prepare the test solution: take about 1 g of the powder of this product, weigh it accurately, put it in a Soxhlet extractor, add dichloromethane and heat it under reflux for 3 h, discard the dichloromethane liquid, evaporate the solvent from the residue, move it into a 100 mL conical flask together with the filter paper tube, accurately add 50 mL of saturated n-butanol with water, plug it tightly, leave it overnight, ultrasonically treat it (power 250 W, frequency 50 kHz) for 30 min, filter it, discard the initial filtrate, accurately measure 25 mL of the subsequent filtrate, evaporate it to dryness in an evaporating dish, dissolve the residue in methanol and transfer it to a 5 mL volumetric flask, dilute it to the mark with methanol, shake it well, filter it, and take the subsequent filtrate. The obtained solution was used for qualitative and quantitative analysis of ginsenoside Rb1 using high performance liquid chromatography.

[0029] Embodiment 2: The difference between this embodiment and embodiment 1 is that the energy of the lamp in step 2) is 0.161 W / m 2 , the number of light irradiation + dark treatment was 3 times.

[0030] Embodiment 3: The difference between this embodiment and embodiment 2 is that the number of times of light irradiation + dark treatment in step 2) is 5 times.

[0031] Comparative Example 1: Select freshly harvested ginseng, remove the residual soil to avoid affecting the accuracy of the experimental results, and place it in a cool and ventilated place to dry; place the ginseng in a light-proof environment and darken it for 24 h; slice the obtained ginseng, fix it at 105°C for 15 min, and then dry it at 70°C. Weigh the obtained ginseng sample, crush it and sieve it (No. 4 sieve) to prepare the test solution: take about 1 g of the powder of this product, accurately weigh it, put it in a Soxhlet extractor, add dichloromethane and heat it to reflux for 3 h, discard the dichloromethane liquid, evaporate the solvent from the residue, move it into a 100 mL conical flask together with the filter paper tube, accurately add 50 mL of saturated n-butanol with water, plug it, place it overnight, treat it with ultrasound (power 250 W, frequency 50 kHz) for 30 min, filter it, discard the initial filtrate, accurately measure 25 mL of the subsequent filtrate, evaporate it to dryness in an evaporating dish, dissolve the residue in methanol and transfer it to 5 mL volumetric flask, add methanol to dilute to the scale, shake well, filter, and take the filtrate to obtain; use high performance liquid chromatography to perform qualitative and quantitative analysis of ginsenoside Rb1 on the obtained solution.

[0032] Comparative Example 2: The difference between this comparative example and Example 1 is that the wavelength of the lamp in step 2) is 355-375 nm and the energy is 6.500 W / m 2 .

[0033] Comparative Example 3: The difference between this embodiment and embodiment 1 is that the energy of the lamp in step 2) is 0.161 W / m 2 .

[0034] Ginsenoside Rb1 content detection: The product obtained in the example of the present invention was qualitatively and quantitatively analyzed by high performance liquid chromatography, and its specific parameters were as follows: chromatographic column: Poroshell 120 EC-C18 (4.6×100 mm, 2.7 μm); water was used as mobile phase A, acetonitrile was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; flow rate was 5 mL / min; column temperature was 35°C; detection wavelength was 203 nm; injection volume: 10 μL.

[0035] Table 1: Gradient elution Time (minutes) Mobile phase A (%) Mobile phase B (%) 0~35 81 19 35~55 81→71 19→29 55~70 71 29 70~100 71→60 29→40 Preparation of reference solution: Accurately weigh ginsenoside Rb1 reference substance, add methanol to make a mixed solution containing 0.2 mg per 1 mL, shake well, and obtain.

[0036] Preparation of test solution: Take about 1 g of the powder of the product (passed through a No. 4 sieve), accurately weigh it, place it in a Soxhlet extractor, add dichloromethane and heat to reflux for 3 h, discard the dichloromethane solution, evaporate the solvent from the residue, move it into a 100 mL conical flask together with the filter paper tube, accurately add 50 mL of water-saturated n-butanol, plug it tightly, leave it overnight, treat it with ultrasound (power 250 W, frequency 50 kHz) for 30 min, filter, discard the initial filtrate, accurately measure 25 mL of the subsequent filtrate, evaporate it to dryness in an evaporating dish, dissolve the residue in methanol and transfer it to a 5 mL volumetric flask, add methanol to dilute to the scale, shake well, filter, and take the subsequent filtrate.

[0037] After calibration and measurement using ginsenoside Rb1 standard, the peak time of ginsenoside Rb1 was determined to be 72.985 min.

[0038] Data processing and statistical methods: Each experiment was repeated three times, and the results were expressed as mean ± standard deviation. SPSS27.0 was used for variance analysis. P < 0.05 was considered significant.

[0039] The results of Rb1 quantification using high performance liquid chromatography were compared between the products obtained in the embodiment and the comparative example, as shown in Table 2: Table 2: Quantitative data of ginsenoside Rb1 in the products obtained in the examples and comparative examples Ginsenoside Rb1 content (%) Comparative Example 1 0.136±0.008 Comparative Example 2 0.146±0.008 Comparative Example 3 0.181±0.031 Example 1 <![CDATA[0.224±0.024** # ]]> Example 2 <![CDATA[0.230±0.034** # ]]> Example 3 <![CDATA[0.213±0.007* # ]]> *P<0.05, compared with comparative example 1, **P<0.01, compared with comparative example 1; # P<0.05, compared with comparative example 2.

[0040] From the results in Table 2, it can be seen that after being irradiated for different periods of time with lamps of different wavelengths and energies, the content of ginsenoside Rb1 in ginseng will also change. Not all wavelengths of lamps can increase the content of ginsenoside Rb1 in ginseng. For example, when using a wavelength of 287-307 nm and an energy of 0.161 W / m 2 When the ginseng was irradiated with a lamp for three times (Example 2), the content of ginsenoside Rb1 in ginseng reached the highest level, which was 0.230%, which was 1.69 times that of the ginseng without lamp irradiation (Comparative Example 1). 2 When the wavelength is 287-307nm and the energy is 0.214 W / m 2 When the ginseng was irradiated once with a lamp (Example 1), the content of ginsenoside Rb1 in ginseng was higher than that when it was not irradiated with a lamp (Comparative Example 1), the wavelength was 355-375 nm, the energy was 6.500 W / m 2When irradiated with a lamp of wavelength 287-307 nm and energy 0.161 W / m 2 The results show that the use of a lamp with a specific wavelength of 287-307 nm can significantly increase the content of ginsenoside Rb1 in ginseng compared with no irradiation or irradiation with lamps of other wavelengths.

[0041] When the wavelength is 287-307 nm and the energy is 0.161 W / m 2 After irradiation with a lamp for 3 times (Example 2), the content of ginsenoside Rb1 in ginseng was high; after irradiation for 5 times, it became low (compared with Example 1). After irradiation for 5 times (Example 3), the content began to decrease.

[0042] The above experiments show that the wavelength, energy and frequency of irradiation will affect the content of ginsenoside Rb1.

[0043] The embodiments described above are merely embodiments that can simply and clearly embody the technical effects during the research process of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A processing method for increasing the content of ginsenoside Rb1 in ginseng, characterized in that Includes steps: 1) Select freshly harvested ginseng, wash it and dry it; 2) Place the ginseng in a dark environment and irradiate it with ultraviolet light; 3) The ginseng slices obtained in step 2) are sterilized at high temperature and then dried to obtain the ginseng slices. The wavelength of the ultraviolet lamp in step 2) is a light of a specific wavelength band of 287-307 nm.

2. A processing method for increasing the content of ginsenoside Rb1 in ginseng as claimed in claim 1, characterized in that: In step 2), the energy of the wavelength of the ultraviolet lamp is 0.161-0.214 W / m 2 .

3. A processing method for increasing the content of ginsenoside Rb1 in ginseng as claimed in claim 1, characterized in that: In step 2), the energy of the wavelength of the ultraviolet light is 0.161 W / m 2 .

4. A processing method for increasing the content of ginsenoside Rb1 in ginseng as claimed in claim 1, characterized in that: The time of the light irradiation in step 2) is 10 min.

5. A processing method for increasing the content of ginsenoside Rb1 in ginseng as claimed in claim 1, characterized in that: In step 2), the lamp is irradiated for 10 minutes and then kept in darkness for 24 hours, and this step is repeated 1-5 times.

6. A method for increasing the content of ginsenoside Rb1 in ginseng as claimed in claim 1, characterized in that: In step 2), the lamp was irradiated for 10 min and then treated in darkness for 24 h. This treatment step was repeated 3 times.

7. A method for increasing the content of ginsenoside Rb1 in ginseng as claimed in claim 1, characterized in that: The fixing temperature in step 3) is 105°C and the fixing time is 15 min.

8. A processing method for increasing the content of ginsenoside Rb1 in ginseng as claimed in claim 1, characterized in that: The drying temperature in step 3) is 70°C.

9. A ginseng product obtained by the processing method according to any one of claims 1 to 6.

10. The method for detecting ginseng products according to claim 7, characterized in that Includes steps: The obtained ginseng sample was crushed and passed through a No. 4 sieve to prepare the test solution: about 1 g of the powder of this product was taken, accurately weighed, placed in a Soxhlet extractor, added with dichloromethane and heated under reflux for 3 h, the dichloromethane solution was discarded, the solvent of the drug residue was evaporated, and it was transferred into a 100 mL conical flask together with the filter paper tube, and 50 mL of saturated n-butanol with water was accurately added, the bottle was sealed, and left overnight, ultrasonically treated at a power of 250 W and a frequency of 50 kHz for 30 min, filtered, and the initial filtrate was discarded. 25 mL of the subsequent filtrate was accurately measured and placed in an evaporating dish to evaporate to dryness. The residue was dissolved in methanol and transferred to a 5 mL volumetric flask, diluted to the mark with methanol, shaken well, filtered, and the subsequent filtrate was obtained; Preparation of reference solution: Accurately weigh ginsenoside Rb1 reference substance, add methanol to make a mixed solution containing 0.2 mg per 1 mL, shake well, and obtain; High performance liquid chromatography was used for qualitative and quantitative analysis, and its specific parameters were as follows: chromatographic column: Poroshell 120 EC-C18, 4.6×100 mm, 2.7 μm; water was used as mobile phase A, acetonitrile was used as mobile phase B, and gradient elution was performed according to the provisions in the following table; flow rate was 5 mL / min; column temperature was 35°C; detection wavelength was 203 nm; injection volume: 10 μL.

Images