Preparation method and quality evaluation method of wine-processed rhizoma polygonati

Abstract

The invention discloses a preparation method and a quality evaluation method of wine-processed rhizoma polygonati, and belongs to the technical field of rhizoma polygonati processing and quality control. The preparation method comprises the following steps: washing and draining raw rhizoma polygonati, adding yellow wine and water according to a ratio, moistening for more than 24 hours, steaming twice in a waterproof manner, drying for three times, slicing, and mixing with rhizoma polygonati juice to finish final drying. The fructose content of the product is greater than or equal to 29.00%. A HPLC-ELSD (High Performance Liquid Chromatography-Evaporative Light Scattering Detector) method is adopted for quality evaluation, after a sample is dried and crushed, 50% methanol is used for ultrasonic extraction, and a specific chromatographic column and acetonitrile-water are used as a mobile phase for detection. Processing parameters are accurately controlled, objective and quantitative quality indexes are provided, and the method can be used for wine rhizoma polygonati quality control and quality identification.

Description

Technical Field

[0001] This invention belongs to the field of Polygonatum processing and quality control technology, and particularly relates to a method for preparing wine-processed Polygonatum and a method for evaluating its quality. Background Technology

[0002] Polygonatum is the dried rhizome of *Polygonatum kingianum* Coll. et Hemsl., *Polygonatum sibiricum* Red., or *Polygonatum cyrtonema* Hua., all belonging to the Liliaceae family. According to the 2020 edition of the *Chinese Pharmacopoeia*, processed Polygonatum products are mainly divided into two categories: raw Polygonatum and wine-processed Polygonatum. Raw Polygonatum is processed at the place of origin, cut into thick slices, and then dried. Wine-processed Polygonatum is made by taking clean Polygonatum, processing it according to the wine-stewing or wine-steaming methods specified in General Processing Rules 0213 of the *Chinese Pharmacopoeia*, and then cutting it into thick slices and drying it. Polygonatum has the effects of tonifying qi and nourishing yin, strengthening the spleen, moistening the lungs, and benefiting the kidneys. Raw Polygonatum has a numbing sensation on the tongue, while the numbing sensation disappears after processing with wine-processed Polygonatum, and its nourishing effect is significantly enhanced.

[0003] The 2020 edition of the Chinese Pharmacopoeia clearly defines the characteristics of processed and wine-processed Polygonatum sibiricum slices: Processed Polygonatum sibiricum slices are irregularly thick slices with a pale yellow to yellowish-brown outer skin. The cut surface is slightly horny, also pale yellow to yellowish-brown, with numerous pale yellow veins visible. It is slightly hard and tough, with a faint odor, a sweet taste, and a sticky texture when chewed. Wine-processed Polygonatum sibiricum slices are also irregularly thick slices with a brownish-black, glossy surface and a brown to light brown center, showing veins. They are softer, sweet, and have a slight wine aroma. Furthermore, ancient texts describe "nine-steamed and nine-dried" Polygonatum sibiricum as "moist and black, with a sweet taste." Practice shows that after steaming, the cut surface of raw Polygonatum sibiricum gradually changes from yellow to brownish-black, eventually turning black. If over-processed, wine-processed Polygonatum sibiricum will have a distinct sour and bitter taste. Currently, research reports on the steaming time and number of steaming times of Polygonatum odoratum are inconsistent. Therefore, developing a method for processing Polygonatum odoratum into wine that can meet the pharmacopoeia requirements, satisfy the public's demand for appearance, and take into account the cost benefits of pharmaceutical companies is of great practical significance.

[0004] According to literature reports, Polygonatum contains abundant saponins, polysaccharides, and small-molecule sugars. The polysaccharides are characterized by a glucose terminal, linked to fructose via β(1→4) glycosidic bonds, and further linked to fructose via β(1→4) glycosidic bonds to form a complete sugar chain. Furthermore, studies on the hydrolysis of polysaccharides to determine monosaccharide composition have confirmed that Polygonatum and its processed products contain both fructose and glucose.

[0005] High-performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) is an analytical method that combines chromatographic separation with a universal detection technique, making it particularly suitable for the analysis of substances lacking ultraviolet absorption or fluorescence properties. Compared to detection using an ultraviolet detector after methyl derivatization, this method significantly simplifies sample pretreatment and offers advantages such as high versatility, good gradient compatibility, and moderate sensitivity. Its detection limits are typically in the ng to μg range, superior to differential refractive index detectors, and its standard curve exhibits a wide linear range, making it particularly suitable for semi-quantitative analysis.

[0006] In the production of Polygonatum sibiricum, the degree of processing is usually judged based on indicators such as skin color, cut surface color, texture, taste, and odor. For example, Chinese invention patent CN117860841A, "A Polygonatum sibiricum slice and its nine-steaming and nine-drying processing method and application," proposes to reduce the steaming time of darker-colored Polygonatum sibiricum and increase the steaming time of lighter-colored Polygonatum sibiricum to avoid over- or under-processing, thereby improving the quality of the finished product and facilitating market sales. However, this method of judging the degree of processing based on appearance is highly dependent on the experience of the operators and is quite subjective. In contrast, objective content determination can more directly and accurately evaluate the quality of processed Polygonatum sibiricum products. Summary of the Invention

[0007] To address the aforementioned technical problems, this invention proposes a method for preparing and evaluating the quality of processed Polygonatum sibiricum. This method simultaneously solves the problems of inaccurate processing time and the lack of objective evaluation indicators for the finished product of processed Polygonatum sibiricum in existing technologies.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] One objective of this invention is to provide a method for preparing Polygonatum sibiricum, comprising the following steps:

[0010] S1. Take raw Polygonatum sibiricum, rinse to remove surface dust, and drain.

[0011] S2. Add rice wine and water to the processed raw Polygonatum medicinal material, mix well, and let it soak for more than 24 hours, turning it over once every 2 hours during this period, until most of the liquid is absorbed and the raw Polygonatum becomes soft.

[0012] S3. Place the soaked raw Polygonatum and the remaining wine into a porcelain plate, place it in a wooden steamer and steam it over water. After the steam rises, steam it at 800W for 6 hours. After steaming, let it sit overnight using the residual heat to obtain Polygonatum and Polygonatum juice.

[0013] S4. Place the Polygonatum in a hot air drying oven and dry it at 55℃ for 24 hours to obtain Polygonatum that is 70% dry.

[0014] S5. Put the 70% dried Polygonatum back into the Polygonatum juice obtained in step S3, put it into a porcelain plate again, and steam it over water. After the steam rises, steam it at 800W power for 2-8 hours (such as 2 hours, 4 hours, 6 hours or 8 hours). After steaming, let it sit overnight using the residual heat to obtain Polygonatum and Polygonatum juice again.

[0015] S6. Place the Polygonatum obtained in step S5 in a hot air drying oven and dry at 55°C for 24 hours to obtain 70% dry Polygonatum. Slice the sliced ​​Polygonatum and then put the sliced ​​Polygonatum back into the Polygonatum juice obtained in step S5. Scrape and wash the Polygonatum juice adhering to the porcelain plate with water 2-3 times, 5 mL each time, and mix it into the Polygonatum. Place the resulting mixture in a hot air drying oven and dry at 55°C for 72 hours.

[0016] S7. Remove from the air, let cool, seal in a self-sealing bag, and store in a cool, dry place.

[0017] Further, the mass ratio of the raw Polygonatum sibiricum to the rice wine and water is 1000:200:(300-600). When the raw Polygonatum sibiricum is Polygonatum yunnanense, the mass ratio of Polygonatum yunnanense to the rice wine and water is 1000:200:600; when the raw Polygonatum sibiricum is Polygonatum multiflorum / Polygonatum sibiricum, the mass ratio of Polygonatum multiflorum / Polygonatum sibiricum to the rice wine and water is 1000:200:300. Here, " / " indicates "or," meaning that the raw Polygonatum sibiricum can be either Polygonatum multiflorum or Polygonatum sibiricum.

[0018] The second objective of this invention is to provide a wine-processed Polygonatum, prepared by the above-mentioned method for preparing wine-processed Polygonatum, with a fructose content ≥29.00%.

[0019] The third objective of this invention is to provide a quality evaluation method for the aforementioned Polygonatum sibiricum product, which uses high performance liquid chromatography-evaporative light scattering to detect the content of small molecule sugars in Polygonatum sibiricum, and the quality standard requires that the fructose content is not less than 29.00%.

[0020] Furthermore, the quality evaluation method for the aforementioned Polygonatum sibiricum product includes the following steps:

[0021] (1) Place the sample of Polygonatum odoratum in a hot air drying oven and dry it at 60℃ for 24 hours. Then, pulverize it through a No. 4 sieve, put it into a vial, and store it in a desiccator to obtain the sample powder.

[0022] (2) Accurately weigh 0.20 g of powder into a stoppered conical flask, add 10.0 mL of 50% methanol, extract by ultrasonication for 40 min (power 400 W, frequency 50 kHz), cool and then add 50% methanol to make up the weight, shake well, centrifuge at 12000 rpm for 20 min to obtain the test solution.

[0023] (3) The test solution was analyzed by high performance liquid chromatography. The chromatographic column was Nacalai TesqueCOSMOSIL Sugar-D, the mobile phase was acetonitrile-water, isocratic elution was performed, the flow rate was 0.8 mL / min, the column temperature was 35℃, the injection volume was 10 μL, and the elution time was 30 min.

[0024] Furthermore, the volume ratio of acetonitrile to water is 85:15.

[0025] Furthermore, the parameters of the high-performance liquid chromatography method are as follows: drift tube temperature is 100°C, carrier gas pressure is 40 psi, nebulizer temperature is 45°C, and gain is 1.

[0026] The fourth objective of this invention is to provide a quality evaluation method for the aforementioned wine-processed Polygonatum product, which can be applied to the quality control or authenticity identification of wine-processed Polygonatum.

[0027] Compared with the prior art, the present invention has the following advantages and technical effects:

[0028] 1. Based on numerous different processing techniques, this invention precisely specifies the processing time for Polygonatum odoratum to facilitate digital control;

[0029] 2. It provides an objective and quantitative indicator for the quality control of Polygonatum sibiricum, facilitating market supervision. Attached Figure Description

[0030] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings:

[0031] Figure 1 The high-performance liquid chromatograms of the Polygonatum odoratum reference standard and the test sample provided in Example 1;

[0032] Figure 2 This is a graph showing the change in fructose content of Polygonatum rhizome in the first batch of homemade wine in Example 2 over time.

[0033] Figure 3 This is a graph showing the change in fructose content of Polygonatum rhizome in the second batch of homemade wine in Example 2 over time.

[0034] Figure 4 This is a graph showing the change in fructose content of Polygonatum rhizome in the third batch of homemade wine in Example 2 over time.

[0035] Figure 5 This is a graph showing the change in fructose content of Polygonatum rhizome in the fourth batch of homemade wine in Example 2 over time.

[0036] Figure 6 This is a graph showing the change in fructose content of Polygonatum rhizome in the fifth batch of homemade wine in Example 2 over time.

[0037] Figure 7 This is a graph showing the change in fructose content of Polygonatum rhizome in the sixth batch of homemade wine in Example 2 over time.

[0038] Figure 8 This is a graph showing the change in fructose content of Polygonatum rhizome in the seventh batch of homemade wine in Example 2 over time.

[0039] Figure 9 This is a graph showing the change in fructose content of Polygonatum rhizome in the eighth batch of homemade wine in Example 2 over time.

[0040] Figure 10 This is a graph showing the change in fructose content of Polygonatum rhizome in the ninth batch of homemade wine in Example 2 over time.

[0041] Figure 11 This is a graph showing the change in fructose content of Polygonatum rhizome in the tenth batch of homemade wine in Example 2 over time.

[0042] Figure 12 This is a graph showing the change in fructose content of Polygonatum sibiricum in the eleventh batch of homemade wine in Example 2 over time. Detailed Implementation

[0043] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0044] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0045] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0046] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0047] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0048] This invention provides a method for preparing and evaluating the quality of processed Polygonatum sibiricum, relating to the field of Polygonatum sibiricum processing. The preparation method includes the following steps: taking clean Polygonatum sibiricum, adding different amounts of excipients depending on the original material to thoroughly moisten it, steaming it under atmospheric pressure for 6 hours, then letting it sit overnight after the heat is turned off, drying it with hot air at 55°C for 24 hours, mixing it with Polygonatum sibiricum juice, and steaming it again for 6 hours, for a total of 12 hours, then letting it sit overnight after the heat is turned off, and drying it with hot air at 55°C for 24 hours; then slicing it, mixing it with Polygonatum sibiricum juice, and drying it with hot air at 55°C for 72 hours, removing it, cooling it, and sealing it to obtain the final product. The steaming time is examined here; Polygonatum sibiricum steamed for 12 hours has a higher fructose content than that steamed for 8 hours or 14 hours. The finished product of Polygonatum sibiricum contains no less than 29.00% fructose.

[0049] The quality evaluation method for the wine-processed Polygonatum product provided in this embodiment of the invention includes:

[0050] Instruments: Waters 2695 high performance liquid chromatograph (Waters Corporation, USA), Waters 2424 evaporative light scattering detector (Waters Corporation, USA), Peak Infinxe 5010 nitrogen generator (Peak Corporation, UK).

[0051] Reagents: Xylose (batch number: J051B219141) and mannose (batch number: F10IB206756) were purchased from Shanghai Yuanye Biotechnology Co., Ltd.; fructose (batch number: JV261220) and sucrose (batch number: J2228685) were purchased from Aladdin Biochemical Technology Co., Ltd.; glucose (batch number: C15122587) was purchased from Shanghai Maclean Biochemical Technology Co., Ltd. All the above reference standards had an HPLC accuracy >98%. Acetonitrile was purchased from Fisher Scientific, USA, and was of chromatographic grade. Purified water was purchased from Hangzhou Wahaha Group Co., Ltd.

[0052] Chromatographic conditions: Nacalai Tesque COSMOSIL Sugar-D (4.6*250mm, 5μm) column; acetonitrile-water (85:15 v / v) mobile phase for isocratic elution; flow rate 0.8 mL / min. -1 The column temperature was 35℃; the injection volume was 10μL; the elution time was 30min; the drift tube temperature was 100℃; the carrier gas pressure was 40psi; the nebulizer temperature was 45℃; the nebulizer output power was 75%; and the gain was 1.

[0053] All raw materials used in this invention were purchased from the market.

[0054] The technical solution of the present invention will be further illustrated by the following embodiments.

[0055] Example 1

[0056] In this embodiment, 30 batches of wine-processed Polygonatum from different origins, manufacturers, and batches were collected. The content of small molecule sugars in the wine-processed Polygonatum was tested by high performance liquid chromatography. The sample information is shown in Table 1.

[0057] Table 130 batches of wine-processed Polygonatum from different origins, manufacturers, and batches.

[0058]

[0059]

[0060] Note: HJJZ indicates that the wine is made from Polygonatum odoratum.

[0061] A quality evaluation method for the above-mentioned Polygonatum odoratum wine products

[0062] 1. Preparation of the test solution: Place the Polygonatum odoratum sample in a hot air drying oven and dry at 60℃ for 24h. Grind it through a No. 4 sieve, put it into a vial, and store it in a desiccator to obtain the test powder. Accurately weigh 0.20g of powder into a stoppered conical flask, add 10.0mL of 50% methanol, and extract by ultrasonication for 40min (power 400W, frequency 50kHz). After cooling, make up the weight with 50% methanol, shake well, and centrifuge at 12000rpm for 20min to obtain the test solution.

[0063] 2. Instruments: Waters 2695 high performance liquid chromatograph (Waters Corporation, USA), Waters 2424 evaporative light scattering detector (Waters Corporation, USA), Peak Infinxe 5010 nitrogen generator (Peak Corporation, UK).

[0064] 3. Reagents: Xylose (batch number: J051B219141) and mannose (batch number: F10IB206756) were purchased from Shanghai Yuanye Biotechnology Co., Ltd.; fructose (batch number: JV261220) and sucrose (batch number: J2228685) were purchased from Aladdin Biochemical Technology Co., Ltd.; glucose (batch number: C15122587) was purchased from Shanghai Maclean Biochemical Technology Co., Ltd. All the above reference standards had an HPLC accuracy >98%. Acetonitrile was purchased from Fisher Scientific, USA, and was of chromatographic grade. Purified water was purchased from Hangzhou Wahaha Group Co., Ltd.

[0065] 4. Chromatographic conditions: A Nacalai Tesque COSMOSIL Sugar-D column (4.6*250mm, 5μm) was used as the chromatographic column; acetonitrile-water (85:15 v / v) was used as the mobile phase for isocratic elution; the flow rate was 0.8 mL / min. -1Column temperature 35℃; injection volume 10μL; elution time 30min. ELSD conditions: drift tube temperature: 100℃, carrier gas pressure: 40psi, nebulizer temperature: 45℃, nebulizer output power: 75%, gain: 1.

[0066] 5. Analyze the test solution using high performance liquid chromatography.

[0067] 6. Preparation of standard solutions: Accurately weigh 51.85 mg xylose, 134.80 mg fructose, 32.95 mg mannose, 52.50 mg glucose, and 25.90 mg sucrose, and dilute to 5 mL with pure water in a volumetric flask. Shake well to obtain a mixed standard solution stock solution. Accurately measure 2 mL of the stock solution, add 1 mL of 50% methanol (v / v), and vortex to mix, obtaining a 2 / 3 dilution. Accurately transfer 2 mL of the 2 / 3 dilution, add 1 mL of 50% methanol (v / v), and vortex to mix, obtaining a 4 / 9 dilution. Repeat this serial dilution process to obtain a total of 11 concentration standard solutions, including the stock solution.

[0068] Inject standard solutions of different concentrations. Plot a standard curve with the natural logarithm of concentration (mg / mL) on the x-axis and the natural logarithm of peak area on the y-axis to obtain the linear regression equation for the pentasaccharide reference standard. Compare the chromatographic peak signal with the baseline signal. The limit of quantitation is when the signal-to-noise ratio is 10:1, and the limit of detection is when the signal-to-noise ratio is 3:1. See Table 2 for details.

[0069] Table 2 Detection Limits of Standard Solutions

[0070]

[0071] Inject the test sample solution under the above chromatographic conditions, record the peak area, calculate the natural logarithm, and then input it into the regression equation to calculate the logarithmic concentration. Calculate the exponent to obtain the concentration of the test sample solution, multiply it by the volume of the extraction solvent, divide it by the sample weight, and multiply by 100% to obtain the content of each small molecule sugar.

[0072] Precision test: Take 2024HJJZ026 Polygonatum odoratum powder, prepare the test solution according to the above method, inject the sample 6 times consecutively, and determine the peak area of ​​xylose, fructose, mannose, glucose and sucrose respectively to examine the instrument precision. The results are shown in Table 3.

[0073] Table 3

[0074] Xylose fructose Mannose glucose sucrose content 1.89％ 27.95％ 2.33％ 12.99％ 5.67％ RSD% 0.06％ 0.69％ 0.08％ 0.38％ 4.17％

[0075] As can be seen from Table 3, the RSD of the Ln peak area for all five sugars is less than 5%, indicating that the instrument has good precision.

[0076] Stability test: Take 2024HJJZ026 Polygonatum odoratum powder, prepare the test solution according to the above method, and inject the sample at 0, 2, 4, 6, 8, 12 and 24 h to determine the peak area of ​​xylose, fructose, mannose, glucose and sucrose, respectively, to investigate the stability of the sample within 24 h. The results are shown in Table 4.

[0077] Table 4

[0078] Xylose fructose Mannose glucose sucrose content 1.92％ 28.46％ 2.33％ 13.33％ 5.13％ RSD% 0.06％ 1.37％ 0.11％ 0.66％ 2.11％

[0079] As can be seen from Table 4, the RSD of the peak area of ​​Ln for all five sugars is less than 5%, indicating that the test solution has good stability within 24 hours.

[0080] Repeatability test: Take 2024HJJZ026 Polygonatum odoratum powder, prepare the test solution according to the above method, and inject it in sequence to determine the peak area of ​​xylose, fructose, mannose, glucose and sucrose respectively to examine the repeatability of the method (see Table 5 for details).

[0081] Table 5

[0082] Xylose fructose Mannose glucose sucrose content 1.89％ 27.98％ 2.28％ 13.16％ 5.93％ RSD% 0.09％ 0.59％ 0.09％ 0.16％ 3.79％

[0083] As can be seen from Table 5, the RSD of the Ln peak area for all five sugars is less than 5%, indicating that the method has good repeatability.

[0084] Take 0.2 g of Polygonatum odoratum powder (2024HJJZ026) with known sugar content, accurately add it to the mixed reference standard, and apply it in three concentration gradients so that the theoretical sugar content is equivalent to 80%, 100%, and 120% of the sugar mass in 0.4 g of Polygonatum odoratum powder, respectively. Each gradient is repeated in triplicate. Prepare the test solution according to the method in section 1, and determine the peak areas of xylose, fructose, mannose, and glucose under the chromatographic conditions in section 4. Calculate the content, recovery rate, and RSD. The results are shown in Table 6.

[0085] Table 6

[0086]

[0087]

[0088]

[0089]

[0090] As shown in Table 6, the average recoveries of the five small molecule sugars in the test solution were 101.05%, 101.87%, 100.67%, 102.09%, and 101.40%, respectively. The RSDs were 1.74%, 1.99%, 4.12%, 2.24%, and 2.59%, respectively. This indicates that the method has good accuracy.

[0091] Table 730 shows the test results (%) of small molecule sugar content of Polygonatum in wine.

[0092]

[0093]

[0094] Note: Blank spaces indicate that no data was detected (the same applies below).

[0095] Table 7 shows that the fructose content is generally high, with an average of 37.04% for the 30 batches of *Polygonatum odoratum*. According to the General Rules of the Pharmacopoeia, Volume IV, a 20% reduction from the average value is applied, stipulating that *Polygonatum odoratum* must contain at least 29.60% fructose as the standard for content determination. If the content limit is set based on more than 80% of batches being qualified, then the standard for fructose content in *Polygonatum odoratum* should be at least 29.8%. Therefore, a comprehensive determination of 29.0% is adopted as the standard for content determination of *Polygonatum odoratum*. These results can provide a reference for the quality control of *Polygonatum odoratum*.

[0096] Figure 1 The high-performance liquid chromatograms of 30 batches of Polygonatum odoratum provided in Example 1.

[0097] Example 2

[0098] Eleven batches of raw Polygonatum sibiricum from different origins and places of origin were collected as raw materials. Wine-processed Polygonatum sibiricum was prepared using the method of the present invention. Samples were taken at 8, 10, 12 and 14 hours (total time) of steaming. The small molecule sugar content in the wine-processed Polygonatum sibiricum was tested according to the quality evaluation method of wine-processed Polygonatum sibiricum provided in Example 1 to determine the optimal steaming time with fructose content as the measurement index. The sample information is shown in Table 8.

[0099] Table 811 shows 11 batches of raw Polygonatum sibiricum from different origins and places of origin.

[0100]

[0101] Note: HJYC - Polygonatum sibiricum medicinal material; DHJYC - Polygonatum sibiricum medicinal material from Yunnan; DHHJYC - Polygonatum sibiricum medicinal material from multiple flowers.

[0102] (I) A method for preparing Polygonatum sibiricum wine, comprising the following steps:

[0103] S1. Take raw Polygonatum sibiricum, rinse to remove surface dust, and drain.

[0104] S2. Add rice wine and water to the processed raw Polygonatum medicinal material, mix well, and let it soak for more than 24 hours, turning it over once every 2 hours during this period, until most of the liquid is absorbed and the raw Polygonatum becomes soft.

[0105] S3. Place the soaked raw Polygonatum and the remaining wine into a porcelain plate, place it in a wooden steamer and steam it over water. After the steam rises, steam it at 800W for 6 hours. After steaming, let it sit overnight using the residual heat to obtain Polygonatum and Polygonatum juice.

[0106] S4. Place the Polygonatum in a hot air drying oven and dry it at 55℃ for 24 hours to obtain Polygonatum that is 70% dry.

[0107] S5. Put the 70% dried Polygonatum back into the Polygonatum juice obtained in step S3, put it into a porcelain plate again, and steam it over water. After the steam rises, steam it at 800W for different times (2h, 4h, 6h and 8h respectively). After steaming, let it sit overnight using the residual heat to obtain Polygonatum and Polygonatum juice again.

[0108] S6. Place the Polygonatum obtained in step S5 in a hot air drying oven and dry at 55℃ for 24 hours to obtain 70% dry Polygonatum. Cut it into thick slices with a thickness of 2-4 mm. Then put the sliced ​​Polygonatum back into the Polygonatum juice obtained in step S5. Scrape and wash the Polygonatum juice adhering to the porcelain plate with water 2-3 times, 5 mL each time, and mix it into the Polygonatum. Place the resulting mixture in a hot air drying oven and dry at 55℃ for 72 hours.

[0109] S7. Remove from the air, let cool, seal in a self-sealing bag, and store in a cool, dry place.

[0110] For the samples in Table 8, when the raw Polygonatum medicinal material is Yunnan Polygonatum medicinal material, the mass ratio of Yunnan Polygonatum medicinal material to rice wine and water is 1000:200:600; when the raw Polygonatum medicinal material is Polygonatum multiflorum / Polygonatum medicinal material, the mass ratio of Polygonatum multiflorum / Polygonatum medicinal material to rice wine and water is 1000:200:300.

[0111] (II) A quality evaluation method for the above-mentioned Polygonatum odoratum wine products

[0112] 1. Preparation of the test solution: Place the Polygonatum odoratum sample in a hot air drying oven and dry at 60℃ for 24h. Grind it through a No. 4 sieve, put it into a vial, and store it in a desiccator to obtain the test powder. Accurately weigh 0.20g of powder into a stoppered conical flask, add 10.0mL of 50% methanol, and extract by ultrasonication for 40min (power 400W, frequency 50kHz). After cooling, make up the weight with 50% methanol, shake well, and centrifuge at 12000rpm for 20min to obtain the test solution.

[0113] 2. Instruments: Waters 2695 high performance liquid chromatograph (Waters Corporation, USA), Waters 2424 evaporative light scattering detector (Waters Corporation, USA), Peak Infinxe 5010 nitrogen generator (Peak Corporation, UK).

[0114] 3. Reagents: Xylose (batch number: J051B219141) and mannose (batch number: F10IB206756) were purchased from Shanghai Yuanye Biotechnology Co., Ltd.; fructose (batch number: JV261220) and sucrose (batch number: J2228685) were purchased from Aladdin Biochemical Technology Co., Ltd.; glucose (batch number: C15122587) was purchased from Shanghai Maclean Biochemical Technology Co., Ltd. All the above reference standards had an HPLC accuracy >98%. Acetonitrile was purchased from Fisher Scientific, USA, and was of chromatographic grade. Purified water was purchased from Hangzhou Wahaha Group Co., Ltd.

[0115] 4. Chromatographic conditions: A Nacalai Tesque COSMOSIL Sugar-D column (4.6*250mm, 5μm) was used as the chromatographic column; acetonitrile-water (85:15 v / v) was used as the mobile phase for isocratic elution; the flow rate was 0.8 mL / min. -1 Column temperature 35℃; injection volume 10μL; elution time 30min. ELSD conditions: drift tube temperature: 100℃, carrier gas pressure: 40psi, nebulizer temperature: 45℃, nebulizer output power: 75%, gain: 1.

[0116] 5. Analyze the test solution using high performance liquid chromatography.

[0117] 6. Preparation of standard solutions: Accurately weigh 51.85 mg xylose, 134.80 mg fructose, 32.95 mg mannose, 52.50 mg glucose, and 25.90 mg sucrose, and dilute to 5 mL with pure water in a volumetric flask. Shake well to obtain a mixed standard solution stock solution. Accurately measure 2 mL of the stock solution, add 1 mL of 50% methanol (v / v), and vortex to mix, obtaining a 2 / 3 dilution. Accurately transfer 2 mL of the 2 / 3 dilution, add 1 mL of 50% methanol (v / v), and vortex to mix, obtaining a 4 / 9 dilution. Repeat this serial dilution process to obtain a total of 11 concentration standard solutions, including the stock solution.

[0118] Inject standard solutions of different concentrations. Plot a standard curve with the natural logarithm of concentration (mg / mL) on the x-axis and the natural logarithm of peak area on the y-axis to obtain the linear regression equations for the five sugar reference standards.

[0119] Inject the above-mentioned test solution under the above chromatographic conditions, record the peak area, calculate the natural logarithm and then enter it into the regression equation to calculate the logarithmic concentration. Calculate the exponent to obtain the concentration of the test solution, multiply it by the volume of the extraction solvent, divide it by the sample weight, and multiply by 100% to obtain the content of each small molecule sugar.

[0120] Table 9. Small molecule sugar content (%) of the 11 test solutions

[0121]

[0122]

[0123]

[0124] As can be seen from Table 9, among the 11 batches of samples, 9 batches reached the peak fructose content after 12 hours of steaming, and 2 batches reached the peak fructose content after 10 hours of steaming. Extending the steaming time reduced the fructose content. Therefore, 12 hours of steaming was selected as the optimal steaming time for wine-processed Polygonatum sibiricum. Moreover, the fructose content of wine-processed Polygonatum sibiricum prepared according to this process was higher than the average value of commercially available wine-processed Polygonatum sibiricum.

[0125] The above are merely preferred embodiments of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A method for preparing Polygonatum sibiricum, characterized in that, Includes the following steps: S1. Take raw Polygonatum sibiricum, rinse to remove surface dust, and drain. S2. Add rice wine and water to the processed raw Polygonatum medicinal material, mix well, let it soak for more than 24 hours, and turn it over regularly until most of the liquid is absorbed and the raw Polygonatum medicinal material becomes soft. S3. Place the soaked raw Polygonatum medicinal material and the remaining wine into a porcelain plate, place it in a wooden steamer and steam it over water. After the steam rises, steam it at 800W power for 6 hours. After steaming, let it sit overnight to obtain Polygonatum and Polygonatum juice. S4. Dry the Polygonatum sibiricum to obtain Polygonatum sibiricum that is 70% dry; S5. Put the 70% dry Polygonatum back into the Polygonatum juice obtained in step S3, put it into a porcelain plate again, and steam it over water. After the steam rises, steam it at 800W power for 2-8 hours. After steaming, let it sit overnight to get Polygonatum and Polygonatum juice again. S6. Dry the Polygonatum obtained in step S5 a second time to obtain Polygonatum that is 70% dry, slice it, and then put the sliced ​​Polygonatum back into the Polygonatum juice obtained in step S5. The Polygonatum juice adhering to the porcelain plate is scraped and washed with water 2-3 times and mixed into the Polygonatum. The resulting mixture is then dried a third time. S7. Let cool, seal in a self-sealing bag, and store in a cool, dry place.

2. The method for preparing Polygonatum sibiricum according to claim 1, characterized in that, The mass ratio of raw Polygonatum sibiricum medicinal material to rice wine and water is 1000:200:(300-600).

3. The method for preparing Polygonatum sibiricum according to claim 1, characterized in that, The raw Polygonatum medicinal materials include Polygonatum yunnanense, Polygonatum multiflorum, and Polygonatum sibiricum.

4. A type of wine-processed Polygonatum, characterized in that, The product is prepared by the preparation method according to any one of claims 1-3, and its fructose content is ≥29.00%.

5. A quality evaluation method for the Polygonatum sibiricum product as described in claim 4, characterized in that, The content of small-molecule sugars in Polygonatum sibiricum was determined by high performance liquid chromatography-evaporative light scattering, and the quality standard required that the fructose content be no less than 29.00%.

6. The quality evaluation method for the Polygonatum sibiricum product according to claim 5, characterized in that, Includes the following steps: (1) Dry the sample of Polygonatum odoratum at 60℃ for 24h, pulverize it and pass it through a No. 4 sieve to obtain the sample powder; (2) Accurately weigh 0.20g of powder, add 10.0mL of methanol with a volume concentration of 50%, extract by ultrasonication for 40min, cool and make up the weight with methanol, centrifuge to obtain the supernatant as the test solution; (3) The test solution was analyzed by high performance liquid chromatography. The chromatographic column was Nacalai Tesque COSMOSIL Sugar-D, the mobile phase was acetonitrile-water, isocratic elution was performed, the flow rate was 0.8 mL / min, the column temperature was 35℃, the injection volume was 10 μL, and the elution time was 30 min.

7. The quality evaluation method for the Polygonatum sibiricum product according to claim 6, characterized in that, The ultrasonic extraction conditions are: power 400W, frequency 50kHz.

8. The quality evaluation method for the Polygonatum sibiricum product according to claim 6, characterized in that, The volume ratio of acetonitrile to water is 85:

15.

9. The quality evaluation method for the Polygonatum sibiricum product according to claim 6, characterized in that, The parameters for the high-performance liquid chromatography method are as follows: drift tube temperature 100℃, carrier gas pressure 40psi, nebulizer temperature 45℃, and gain 1.

10. The application of a quality evaluation method as described in any one of claims 6-9 in the quality control or quality identification of Polygonatum odoratum in wine.

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