Quality detection method and device for steamed polygonatum sibiricum standard decoction based on characteristics of polysaccharide molecules

Abstract

This invention relates to the field of decoction quality testing technology, specifically to a method and apparatus for quality testing of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics. The method separates the steamed Polygonatum sibiricum standard decoction into two components: a polysaccharide precipitate and a small molecule supernatant, through alcohol precipitation fractionation. The absolute weight-average molecular weight and dispersion coefficient of the Polygonatum sibiricum polysaccharides are determined using gel permeation chromatography-laser light scattering (GPS-LS ...

Description

Technical Field

[0001] This invention relates to the field of decoction quality testing technology, and in particular to a method and apparatus for quality testing of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics. Background Technology

[0002] Currently, most existing quality testing methods for steamed Polygonatum sibiricum standard decoctions rely on the content determination of a single or a few indicators. Common methods include using the phenol-sulfuric acid method to determine the total polysaccharide content, with anhydrous glucose as a reference, to quantitatively analyze the polysaccharide components in steamed Polygonatum sibiricum; or using high-performance liquid chromatography (HPLC) to determine the 5-hydroxymethylfurfural content, using it as a marker of the Maillard reaction degree; and methods that evaluate polysaccharide conversion during steaming by measuring the content of reducing sugars such as fructose and glucose. These existing methods are relatively simple to operate and can, to a certain extent, reflect the content level of a certain type of component or the progress of a certain reaction process in the steamed Polygonatum sibiricum standard decoction, providing preliminary analytical means and evaluation basis for the quality control of steamed Polygonatum sibiricum. The current processing of steamed Polygonatum involves multiple interconnected chemical reactions, including polysaccharide degradation, monosaccharide conversion, and Maillard reaction. This means that changes in the content of a single indicator cannot accurately reflect the overall processing level of the steamed Polygonatum. For example, a decrease in total polysaccharide content may be due to insufficient extraction of polysaccharides caused by insufficient steaming, or it may be due to excessive degradation of polysaccharides caused by excessive steaming. An increase in 5-hydroxymethylfurfural content may be a normal result of a full Maillard reaction, or it may be due to abnormal accumulation of byproducts caused by excessive heating. However, existing methods cannot effectively distinguish between the two abnormal quality states of insufficient steaming and excessive steaming, and lack the ability to accurately control the processing endpoint in both directions. As a result, the quality consistency of standard decoctions of steamed Polygonatum from different batches cannot be reliably guaranteed. Summary of the Invention

[0003] The purpose of this invention is to provide a method and apparatus for quality testing of standard decoction of steamed Polygonatum sibiricum based on the molecular characteristics of polysaccharides. By establishing a multidimensional synergistic determination system of absolute molecular weight of polysaccharides, sugar conversion index and Maillard reaction index, it is possible to achieve bidirectional and precise control of the endpoint of steaming Polygonatum sibiricum, and effectively distinguish between under-steaming and over-steaming.

[0004] To achieve the above objectives, this invention provides a method and apparatus for quality testing of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics, comprising the following steps: A standard decoction sample of steamed Polygonatum was obtained, and its total polysaccharide concentration was determined by colorimetry. The sample was then subjected to alcohol precipitation fractionation to separate the polysaccharide precipitate and the small molecule supernatant. The polysaccharide precipitate was reconstituted, and the absolute weight-average molecular weight (Mw) and dispersive index of Polygonatum polysaccharide were determined by gel permeation chromatography-laser light scattering. ; The components of the small molecule supernatant were concentrated and solvent replaced, and the concentrations of characteristic small molecule sugars and 5-hydroxymethylfurfural were determined by hydrophilic interaction chromatography and reversed-phase chromatography, respectively. The characteristic small molecule sugars included fructose, glucose, sucrose, raffinose and stachyose. Based on the total polysaccharide concentration, absolute weight-average molecular weight Mw, and dispersion index By analyzing the characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration, and performing synergistic calculation and threshold determination of multidimensional indicators, a conclusion on the quality qualification of the steamed Polygonatum standard decoction is generated.

[0005] The preparation method of the steamed Polygonatum standard decoction sample is as follows: take steamed Polygonatum slices, add 8 to 12 times the amount of water and soak for 30 to 60 minutes, heat and boil twice for 30 to 60 minutes each time, combine the decoctions, filter while hot, and concentrate the filtrate under reduced pressure to 1 mL equivalent to 0.5 g of steamed Polygonatum slices, thus obtaining the steamed Polygonatum standard decoction sample.

[0006] The specific operation of the alcohol precipitation fractionation process is as follows: anhydrous ethanol is added to the sample under stirring to make the volume percentage of ethanol in the mixture reach 80%, and the mixture is allowed to stand for 2 to 4 hours to allow the polysaccharide to fully precipitate. The mixture is then centrifuged at 4000 to 6000 rpm for 10 to 20 minutes, and the polysaccharide precipitate and the small molecule supernatant are collected separately.

[0007] The gel permeation chromatography-laser light scattering (GPS-LSS) method employs a high-performance gel permeation chromatography-differential refractive index-multi-angle laser light scattering (MLLS) system. The chromatographic columns are a Shodex OHpak SB-806MHQ column and an SB-804HQ column connected in series. The mobile phase is a 0.1 mol / L sodium nitrate aqueous solution containing 0.02% sodium azide, with a flow rate of 0.5 mL / min and a column temperature of 35 °C. The polysaccharide precipitate is reconstituted with hot water at 60–80 °C and diluted to a polysaccharide concentration of 1–5 mg / mL before determination.

[0008] The colorimetric method is the phenol-sulfuric acid method, with anhydrous glucose as the reference standard. The total polysaccharide concentration in the steamed Polygonatum standard decoction is measured to be 10-30 mg / mL.

[0009] The hydrophilic interaction chromatography method uses an amide-based hydrophilic interaction column with dimensions of 4.6 mm × 250 mm and a diameter of 3.5 μm. Mobile phase A is acetonitrile, and mobile phase B is an aqueous solution containing 0.1% triethylamine. The gradient elution program is as follows: from 0 to 15 minutes, the volume percentage of mobile phase B increases linearly from 15% to 35%; from 15 to 25 minutes, the volume percentage of mobile phase B increases linearly from 35% to 45%; and from 25 to 30 minutes, the volume percentage of mobile phase B remains constant at 45%. The detection was performed using an evaporative light scattering detector, with a drift tube temperature of 55°C and a carrier gas flow rate of 1.8 L / min.

[0010] The reversed-phase chromatography method uses a C18 reversed-phase column with dimensions of 4.6 mm × 150 mm and 5 μm. The mobile phase is a mixture of methanol and water at a volume ratio of 15:85. The flow rate is 1.0 mL / min, the column temperature is 30 °C, and the 5-hydroxymethylfurfural is detected at a wavelength of 284 nm using an ultraviolet detector.

[0011] The specific operation of concentration and solvent replacement is as follows: after the ethanol is removed by vacuum concentration of the small molecule supernatant component, a mixed solvent of acetonitrile-water in a volume ratio of 1:1 is added to dissolve and dilute to a final volume to obtain a small molecule assay solution for chromatographic analysis.

[0012] The collaborative calculation and threshold determination of the multidimensional indicators include: The Maillard response index (MRI) is calculated as: MRI = C5 - HMF / (Cfructose + Cglucose) × 100%. Calculate the sugar conversion index (SCI) as follows: (Cfructose + Cglucose) / Ctotal polysaccharides × 100%; Wherein C5-HMF, Cfructose, and Cglucose are the concentrations of 5-hydroxymethylfurfural, fructose, and glucose, respectively, and Ctotal polysaccharide is the total polysaccharide concentration measured by the colorimetric method. All the above concentrations are in mg / mL.

[0013] The generation of a quality compliance conclusion must simultaneously meet all of the following threshold conditions: The absolute weight-average molecular weight (Mw) is 3500 Da to 7500 Da; Dispersion coefficient ; Sucrose concentration C (sucrose ≤ 0.20 mg / mL); Maillard response index on MRI ranged from 0.20% to 0.70%. The sugar conversion index (SCI) is 8%–18%. Furthermore, the molar ratio of fructose to glucose is 0.9:1 to 1.1:1.

[0014] The polysaccharide molecular characteristics-based standard decoction quality detection system for steamed Polygonatum sibiricum, used to implement the method for quality detection of the standard decoction based on polysaccharide molecular characteristics, includes a sample processing module, a polysaccharide molecular weight detection module, a small molecule detection module, and a data processing and judgment module. The polysaccharide precipitation output terminal of the sample processing module is connected to the input terminal of the polysaccharide molecular weight detection module, the supernatant output terminal of the sample processing module is connected to the input terminal of the small molecule detection module, and the input terminal of the data processing and judgment module is connected to the total polysaccharide concentration output terminal of the sample processing module, the output terminal of the polysaccharide molecular weight detection module, and the output terminal of the small molecule detection module, respectively. The sample processing module is used to obtain a standard decoction sample of steamed Polygonatum sibiricum, determine the total polysaccharide concentration, and perform alcohol precipitation fractionation on another sample to separate the polysaccharide precipitate component and the small molecule supernatant component. The polysaccharide molecular weight detection module is used to reconstitute the polysaccharide precipitate and determine the absolute weight-average molecular weight (Mw) and dispersion coefficient of Polygonatum polysaccharide using gel permeation chromatography-laser light scattering. ; The small molecule detection module is used to concentrate and solvent-replace the components of the small molecule supernatant, and to determine the characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration using hydrophilic interaction chromatography and reversed-phase chromatography, respectively. The data processing and judgment module is used to obtain the total polysaccharide concentration, Mw, The characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration were analyzed. The Maillard reaction index (MRI) and sugar conversion index (SCI) were calculated. Multidimensional index threshold determination was implemented to generate a quality qualification conclusion for the steamed Polygonatum sibiricum standard decoction.

[0015] The present invention relates to a method and apparatus for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics. First, two parallel samples are taken from the steamed Polygonatum sibiricum standard decoction sample. One sample is subjected to phenol-sulfuric acid colorimetric method to determine the total polysaccharide concentration, while the other sample undergoes alcohol precipitation fractionation to separate the polysaccharide precipitate and the small molecule supernatant. Subsequently, the polysaccharide precipitate is reconstituted with hot water, and the absolute weight-average molecular weight (Mw) and dispersive index of Polygonatum sibiricum polysaccharides are determined using a high-performance gel permeation chromatography-differential refractive index-multi-angle laser light scattering system. This method accurately reflects the degradation degree and molecular weight distribution uniformity of polysaccharide macromolecules during the steaming process. Simultaneously, after removing ethanol by vacuum concentration of the small molecule supernatant and replacing it with an acetonitrile-water mixed solvent, the content spectra of five characteristic small molecule sugars—fructose, glucose, sucrose, raffinose, and stachyose—were simultaneously determined using amide hydrophilic interaction chromatography-evaporative light scattering detection. The content of the Maillard reaction marker 5-hydroxymethylfurfural was determined using C18 reversed-phase chromatography-UV detection. Finally, based on the above results, two derived indices—the Maillard reaction index MRI (the ratio of 5-hydroxymethylfurfural to the sum of fructose and glucose) and the sugar conversion index SCI (the ratio of the sum of fructose and glucose to the total polysaccharides)—were calculated. The absolute weight-average molecular weight (Mw) and dispersion coefficient were also considered. The method uses a comprehensive threshold determination based on six parameters: sucrose concentration, MRI, SCI, and the fructose to glucose molar ratio. Only when all conditions are met simultaneously is the product deemed to be of acceptable quality. This method, through a complete technical route of "gradual separation - multidimensional characterization - collaborative calculation - comprehensive determination", has for the first time achieved simultaneous and accurate evaluation of the integrity of polysaccharide macromolecules, the degree of polysaccharide to monosaccharide conversion, and the level of Maillard reaction byproducts in steamed Polygonatum standard decoction. It effectively overcomes the fundamental defect of traditional single-index detection that cannot distinguish between under-steaming and over-steaming. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0017] Figure 1 This is a flowchart of the quality testing method for steamed Solomon's Seal standard decoction based on polysaccharide molecular characteristics according to the present invention.

[0018] Figure 2 This is a flowchart of the graded separation process for the standard decoction of steamed Polygonatum sibiricum of the present invention.

[0019] Figure 3 This is a flowchart of the characteristic sugar spectrum of small molecule components and the determination of 5-HMF in this invention.

[0020] Figure 4 This is a flowchart of the multi-dimensional index collaborative calculation and threshold determination process of the present invention.

[0021] Figure 5 This is a schematic diagram of the structure of the quality testing device for steamed Solomon's Seal standard decoction based on polysaccharide molecular characteristics according to the present invention.

[0022] In the diagram: 1-Sample processing module, 2-Polysaccharide molecular weight detection module, 3-Small molecule detection module, 4-Data processing and judgment module. Detailed Implementation

[0023] Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain the present invention, and should not be construed as limiting the present invention.

[0024] In the description of this invention, it should be understood that "a plurality of" means two or more, unless otherwise explicitly specified.

[0025] Please see Figures 1 to 4 This invention provides a method and apparatus for quality testing of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics, comprising the following steps: S1: Obtain a standard decoction sample of steamed Polygonatum sibiricum, determine its total polysaccharide concentration by colorimetric method, and perform alcohol precipitation fractionation on the other sample to separate the polysaccharide precipitate component and the small molecule supernatant component. S2: The polysaccharide precipitate was reconstituted, and the absolute weight-average molecular weight Mw and dispersion coefficient of Polygonatum polysaccharide were determined by gel permeation chromatography-laser light scattering. ; S3: The components of the small molecule supernatant are concentrated and solvent replaced, and the concentrations of characteristic small molecule sugars and 5-hydroxymethylfurfural are determined by hydrophilic interaction chromatography and reversed-phase chromatography, respectively. The characteristic small molecule sugars include fructose, glucose, sucrose, raffinose and stachyose. S4: Based on the total polysaccharide concentration, absolute weight-average molecular weight Mw, and dispersion coefficient... By analyzing the characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration, and performing synergistic calculation and threshold determination of multidimensional indicators, a conclusion on the quality qualification of the steamed Polygonatum standard decoction is generated.

[0026] In this embodiment, when a batch of steamed Polygonatum sibiricum standard decoction samples are sent to the laboratory, the testing personnel first accurately measure two parallel samples. One sample is directly used for the phenol-sulfuric acid colorimetric method to determine the total polysaccharide content and obtain the baseline data of the overall polysaccharide level. The other sample is subjected to alcohol precipitation fractionation. The large molecular polysaccharides in the sample are physically separated from small molecular sugars, 5-hydroxymethylfurfural and other components by ethanol precipitation to obtain polysaccharide precipitate components and small molecule supernatant components.

[0027] Subsequently, the polysaccharide precipitate was reconstituted with hot water and then separated and detected using a gel permeation chromatography-differential refractive index-multi-angle laser light scattering system. The absolute weight-average molecular weight and dispersion coefficient of Polygonatum polysaccharide were directly determined, which truly reflects the degree of degradation and uniformity of polysaccharide macromolecules during the processing.

[0028] Meanwhile, the small molecule supernatant components were concentrated under reduced pressure to remove ethanol and replaced with a suitable solvent system for chromatographic analysis. The samples were then injected into a hydrophilic interaction chromatography system and a reversed-phase chromatography system, respectively, to simultaneously obtain the concentration spectra of five characteristic small molecule sugars, namely fructose, glucose, sucrose, raffinose, and stachyose, as well as the concentration of the Maillard reaction marker 5-hydroxymethylfurfural.

[0029] Finally, the system summarizes all the measurement data, calculates two derived indices: Maillard response index (MRI) and glucose conversion index (SCI), and combines the absolute weight-average molecular weight (Mw) and dispersion coefficient. The six parameters, including sucrose concentration, MRI, SCI, and the molar ratio of fructose to glucose, are compared one by one with the preset threshold conditions. Only when all conditions are met simultaneously is the batch of steamed Solomon's Seal standard decoction deemed to be of qualified quality. This achieves a fundamental shift from single-indicator judgment to multi-dimensional synergistic and precise evaluation, effectively overcoming the technical defects of existing methods that cannot distinguish between under-steaming and over-steaming.

[0030] Furthermore, the preparation method of the steamed Polygonatum standard decoction sample is as follows: take steamed Polygonatum slices, add 8-12 times the amount of water and soak for 30-60 minutes, heat and boil twice for 30-60 minutes each time, combine the decoctions, filter while hot, and concentrate the filtrate under reduced pressure to 0.5g of steamed Polygonatum slices per 1mL, thus obtaining the steamed Polygonatum standard decoction sample.

[0031] In this embodiment, when using the medicine, first take the qualified steamed Solomon's Seal slices, weigh them, place them in a decoction container, add 8 to 12 times the weight of the slices of purified water, and soak them at room temperature for 30 to 60 minutes to allow the medicinal materials to fully swell, which facilitates the subsequent dissolution of the effective components.

[0032] After soaking, heat to a simmer and maintain the simmer for 30 to 60 minutes, then pour out the first decoction. Add 8 to 12 times the amount of water to the dregs and decoct again for 30 to 60 minutes using the same method. Combine the two decoctions. Since the decoction may contain a small amount of dregs, it should be filtered while hot through a 200-mesh filter cloth or filter paper to ensure that the filtrate is clear.

[0033] The filtrate was transferred to a rotary evaporator and concentrated under reduced pressure in a 60°C water bath. The concentration endpoint was determined by volume, with each 1 mL of concentrated solution equivalent to 0.5 g of the original steamed Polygonatum sibiricum slices, thus obtaining the standard decoction sample of the steamed Polygonatum sibiricum. The preparation method of this standard decoction strictly simulates the clinical decoction process, ensuring the representativeness and consistency of the tested subjects and providing a standardized sample basis for subsequent quality testing.

[0034] Furthermore, the specific operation of the alcohol precipitation fractionation process is as follows: anhydrous ethanol is added to the other sample under stirring to make the volume percentage of ethanol in the mixture reach 80%, and the mixture is allowed to stand for 2 to 4 hours to allow the polysaccharide to fully precipitate. The mixture is then centrifuged at 4000 to 6000 rpm for 10 to 20 minutes, and the polysaccharide precipitate and the small molecule supernatant are collected separately.

[0035] In this embodiment, a precise volume of the steamed Polygonatum sibiricum standard decoction sample is placed in a beaker. Anhydrous ethanol is slowly added along the beaker wall while continuously stirring with a magnetic stirrer until the final volume percentage of ethanol in the mixture reaches 80%. The stirring speed should not be too fast to avoid generating a large number of bubbles that could affect the accuracy of the ethanol measurement.

[0036] After adding the ethanol, continue stirring for several minutes to ensure a homogeneous mixture. Then, allow it to stand at 4°C for 2 to 4 hours to allow the high molecular weight Polygonatum polysaccharides to fully dehydrate and precipitate. After standing, transfer the mixture to a centrifuge tube and centrifuge at 4000 to 6000 rpm for 10 to 20 minutes. After centrifugation, a white or off-white polysaccharide precipitate will be visible at the bottom of the tube, while the supernatant will be a light yellow transparent liquid. Carefully pour off the supernatant to obtain the small molecule supernatant fraction. The precipitate, i.e., the polysaccharide precipitate, can be temporarily stored in the refrigerator for later use.

[0037] The ingenious aspect of this fractionation process is that using an 80% ethanol concentration ensures sufficient precipitation of macromolecular polysaccharides while allowing small-molecule sugars and 5-hydroxymethylfurfural to be almost completely retained in the supernatant, achieving complete separation of the two types of components and laying the foundation for their accurate subsequent determination.

[0038] Furthermore, the gel permeation chromatography-laser light scattering (GPS-LSS) method employs a high-performance gel permeation chromatography-differential refractive index-multi-angle laser light scattering (MLLS) system. The chromatographic columns are a Shodex OHpak SB-806MHQ column and an SB-804HQ column connected in series. The mobile phase is a 0.1 mol / L sodium nitrate aqueous solution containing 0.02% sodium azide, the flow rate is 0.5 mL / min, and the column temperature is 35 °C. The polysaccharide precipitate is reconstituted with hot water at 60–80 °C and diluted to a polysaccharide concentration of 1–5 mg / mL before determination.

[0039] In this embodiment, the polysaccharide precipitate obtained by alcohol precipitation is first transferred to a small beaker, and an appropriate amount of hot purified water at 60 to 80°C is added. The mixture is then stirred with a glass rod or sonicated to completely dissolve the polysaccharide. Finally, the mixture is transferred to a volumetric flask and brought to a final volume to ensure that the polysaccharide concentration is controlled within the range of 1 to 5 mg / mL. This concentration range ensures the detection signal intensity while avoiding column overload or intermolecular aggregation due to excessively high concentrations.

[0040] For the determination, a high-performance gel permeation chromatography-differential refractive index-multi-angle laser light scattering system was used. The chromatographic part was equipped with two gel chromatography columns, Shodex OHpak SB-806M HQ and SB-804 HQ, connected in series. The column temperature was controlled at 35℃. The mobile phase was a 0.1mol / L sodium nitrate aqueous solution containing 0.02% sodium azide as an antibacterial agent, and isocratic elution was performed at a flow rate of 0.5mL / min.

[0041] After the sample solution is injected via an autosampler, the large molecular weight polysaccharides are eluted sequentially according to their molecular size, with the high molecular weight fraction eluting first and the low molecular weight fraction eluting later. The eluent sequentially passes through a multi-angle laser light scattering detector and a differential refractive index detector. The former directly calculates the absolute weight-average molecular weight of each elution fraction by measuring the intensity of scattered light at different angles, without relying on a standard calibration curve; the latter provides the concentration signal. Combining both allows for an accurate plotting of the molecular weight distribution curve, from which the absolute weight-average molecular weight (Mw) and dispersive index of Polygonatum polysaccharides can be obtained. .

[0042] Compared to the traditional relative measurement method that only uses a differential refractive index detector and a dextran standard calibration curve, the molecular weight measured by this method is the true absolute value, which can more accurately reflect the actual degradation state of polysaccharide molecules during steaming, and provides more reliable data support for judging the degree of processing.

[0043] Furthermore, the colorimetric method is the phenol-sulfuric acid method, with anhydrous glucose as the reference standard, and the total polysaccharide concentration in the steamed Polygonatum standard decoction is measured to be 10-30 mg / mL.

[0044] In this embodiment, the total polysaccharide content in the steamed Polygonatum sibiricum standard decoction is determined using the classic phenol-sulfuric acid method. First, an appropriate amount of anhydrous glucose reference standard dried to constant weight at 105℃ is accurately weighed, dissolved in purified water, and serially diluted to prepare a series of glucose standard solutions of different concentrations.

[0045] Accurately measure each standard solution and the appropriately diluted sample solution of the steamed Solomon's Seal standard decoction into stoppered test tubes, add 5% phenol solution and concentrated sulfuric acid in sequence, shake well, heat in a boiling water bath to develop color, remove and cool, and measure absorbance at a wavelength of 490 nm.

[0046] A standard curve was plotted with the concentration of glucose standard solution on the x-axis and absorbance on the y-axis. The corresponding glucose concentration was read from the standard curve based on the absorbance of the sample solution, and then multiplied by the dilution factor to obtain the total polysaccharide concentration in the steamed Polygonatum standard decoction, expressed as anhydrous glucose.

[0047] Verification has shown that the total polysaccharide concentration in qualified steamed Polygonatum sibiricum standard decoction typically falls within the range of 10 to 30 mg / mL. This measurement data not only serves as a benchmark reference for the total polysaccharide content in the sample but is also one of the key parameters for subsequent calculation of the sugar conversion index (SCI). Furthermore, its value itself can preliminarily reflect the overall polysaccharide retention level during steaming. However, it should be noted that the total polysaccharide content alone cannot distinguish whether the polysaccharides have undergone appropriate degradation. This is precisely why this invention introduces subsequent molecular weight determination and conversion index calculation.

[0048] Furthermore, the hydrophilic interaction chromatography method uses an amide-based hydrophilic interaction column with dimensions of 4.6 mm × 250 mm and a diameter of 3.5 μm. Mobile phase A is acetonitrile, and mobile phase B is an aqueous solution containing 0.1% triethylamine. The gradient elution program is as follows: from 0 to 15 minutes, the volume percentage of mobile phase B increases linearly from 15% to 35%; from 15 to 25 minutes, the volume percentage of mobile phase B increases linearly from 35% to 45%; and from 25 to 30 minutes, the volume percentage of mobile phase B remains constant at 45%. The detection was performed using an evaporative light scattering detector, with a drift tube temperature of 55°C and a carrier gas flow rate of 1.8 L / min.

[0049] In this embodiment, for five highly polar sugar compounds that lack ultraviolet chromophores—fructose, glucose, sucrose, raffinose, and stachyose—an amide-based hydrophilic interaction chromatographic column combined with an evaporative light scattering detector is used for analysis.

[0050] The amide-based stationary phase exhibits differentiated retention of different sugars through hydrophilic partitioning. A higher acetonitrile ratio results in weaker elution but stronger retention. At the start of gradient elution, the mobile phase B ratio is 15%, meaning acetonitrile accounts for 85%, at which point all five sugars are well retained at the column head. Within 0 to 15 minutes, the B phase ratio linearly increases from 15% to 35%, and the gradual increase in the aqueous phase ratio causes monosaccharides and oligosaccharides to elute sequentially in order of increasing polarity, with glucose and fructose eluting first, followed by sucrose. Within 15 to 25 minutes, the B phase ratio increases from 35% to 45%, further increasing the aqueous phase ratio, allowing the trisaccharide raffinose and the tetrasaccharide stachyose, which have stronger retention, to elute sequentially. Finally, within 25 to 30 minutes, the B phase ratio is maintained at 45% to ensure complete elution of all residual components, and the column is restored to its initial equilibrium conditions.

[0051] The evaporative light scattering detector was set with a drift tube temperature of 55℃ and a carrier gas flow rate of 1.8 L / min. Under these conditions, non-volatile triethylamine in the mobile phase and sugars co-formed detectable particles, generating a stable scattering signal. This method can simultaneously separate and quantify five characteristic sugars, providing complete glycospectral information for evaluating the degree of polysaccharide degradation and transformation during the processing of Polygonatum sibiricum.

[0052] Furthermore, the reversed-phase chromatography method uses a C18 reversed-phase column with dimensions of 4.6 mm × 150 mm and 5 μm. The mobile phase is a mixture of methanol and water at a volume ratio of 15:85, the flow rate is 1.0 mL / min, the column temperature is 30 °C, and the 5-hydroxymethylfurfural is detected at a wavelength of 284 nm using an ultraviolet detector.

[0053] In this embodiment, for 5-hydroxymethylfurfural, an important product of the Maillard reaction, the ultraviolet absorption characteristics generated by the conjugated aldehyde group and furan ring in its molecular structure are utilized, and a C18 reversed-phase chromatographic column is used in conjunction with an ultraviolet detector for analysis.

[0054] A methanol-water mixture with a volume ratio of 15:85 was used as the mobile phase. This isocratic elution condition is simple and stable, ensuring good separation of 5-hydroxymethylfurfural from other impurity peaks in the sample while also achieving suitable retention time and peak shape. The flow rate was set at 1.0 mL / min, the column temperature was maintained at 30℃, and the detection wavelength was selected at 284 nm, which is the characteristic maximum absorption wavelength of 5-hydroxymethylfurfural. Under these conditions, both detection sensitivity and specificity were optimal.

[0055] After the sample solution is injected through the autosampler, 5-hydroxymethylfurfural (5-HMF) is retained due to hydrophobic interaction with the C18 stationary phase. After being eluted by the mobile phase, a peak is eluted in about 8 to 12 minutes. Using the external standard method, a series of concentration solutions of 5-HMF standard are prepared to plot a standard curve, and the 5-HMF in the sample is quantified.

[0056] This compound is a hallmark byproduct of the Maillard reaction between reducing sugars and amino acids during the steaming process. Its concentration is closely related to the steaming time and temperature. It cannot be used alone to distinguish between normal process reactions and excessive thermal decomposition. It needs to be correlated with the concentration of fructose and glucose to calculate the Maillard reaction index and then MRI before a scientific judgment can be made.

[0057] Furthermore, the specific operation of concentration and solvent replacement is as follows: after the ethanol is removed by vacuum concentration of the small molecule supernatant component, a mixed solvent of acetonitrile-water volume ratio of 1:1 is added to dissolve and dilute to volume to obtain a small molecule assay solution for chromatographic analysis.

[0058] In this embodiment, the small molecule supernatant obtained from alcohol precipitation fractionation is transferred to a round-bottom flask and concentrated under reduced pressure in a rotary evaporator at a water bath temperature of 40 to 50°C. Since ethanol has a low boiling point, it can be rapidly evaporated at a mild temperature.

[0059] When the solution is concentrated to near dryness, add a small amount of acetonitrile-water mixed solvent (1:1 volume ratio), gently rotate and wash the bottle wall to dissolve the residue, concentrate again under reduced pressure to near dryness, repeat this process twice to ensure that the ethanol is completely replaced, and finally add an appropriate amount of acetonitrile-water mixed solvent to fully dissolve the residue and transfer it to a volumetric flask and dilute to the mark to obtain the small molecule assay solution.

[0060] The choice of a 1:1 volume ratio of acetonitrile to water as the final solvent system was based on two main considerations: First, this solvent composition has good compatibility with the initial mobile phase conditions of subsequent hydrophilic interaction chromatography and reversed-phase chromatography, which can avoid peak distortion caused by solvent effects during injection; Second, this ratio has excellent solubility for fructose, glucose, sucrose, raffinose, stachyose, and 5-hydroxymethylfurfural, which can ensure the full dissolution and recovery of each analyte.

[0061] This step plays a crucial role in the entire process, efficiently converting the alcohol precipitation supernatant—an ethanol-water system unsuitable for direct chromatographic analysis—into a solvent environment compatible with the chromatographic system, thus ensuring the accuracy and reproducibility of the results obtained by both chromatographic analysis methods.

[0062] Furthermore, the collaborative calculation and threshold determination of the multidimensional indicators include: The Maillard response index (MRI) is calculated as: MRI = C5 - HMF / (Cfructose + Cglucose) × 100%. Calculate the sugar conversion index (SCI) as follows: (Cfructose + Cglucose) / Ctotal polysaccharides × 100%; Wherein C5-HMF, Cfructose, and Cglucose are the concentrations of 5-hydroxymethylfurfural, fructose, and glucose, respectively, and Ctotal polysaccharide is the total polysaccharide concentration measured by the colorimetric method. All the above concentrations are in mg / mL.

[0063] In this embodiment, after all instrument measurements are completed, the raw data are summarized, and two key derived indicators are calculated. The Maillard reaction index MRI is calculated by dividing the concentration of 5-hydroxymethylfurfural by the sum of the concentrations of fructose and glucose, and then multiplying by 100%. This formula is based on the basic principle of the Maillard reaction. 5-hydroxymethylfurfural is generated by the reaction of reducing sugars and amino acids. Its generation is affected by both substrate concentration and reaction conditions. Simply measuring the absolute concentration of 5-hydroxymethylfurfural is insufficient to reflect the true progress of the reaction. By correlating it with the instantaneous concentrations of the reaction substrates fructose and glucose, a ratio index reflecting the conversion efficiency of the Maillard reaction is obtained, which is more kinetically significant than a single concentration value.

[0064] The Sugar Conversion Index (SCI) is calculated by dividing the sum of fructose and glucose concentrations by the total polysaccharide concentration and then multiplying by 100%. This index directly quantifies the percentage of conversion from macromolecular polysaccharides to reducing monosaccharides and is a core parameter for measuring the degree of polysaccharide degradation and conversion during the steaming process.

[0065] A low SCI indicates insufficient polysaccharide degradation, suggesting that the steaming of the medicinal slices may not have reached the required level; a high SCI indicates excessive polysaccharide degradation, which may lead to the loss of medicinal components. The calculation of these two indices transforms the original chromatographic data into quality evaluation indicators with clear process implications, enabling the judgment of the endpoint of steamed Polygonatum processing to shift from empirical qualitative description to data-driven precise quantification. This is the core innovation of this method.

[0066] Furthermore, the generation of a quality compliance conclusion must simultaneously meet all of the following threshold conditions: The absolute weight-average molecular weight (Mw) is 3500 Da to 7500 Da; Dispersion coefficient ; Sucrose concentration C (sucrose ≤ 0.20 mg / mL); Maillard response index on MRI ranged from 0.20% to 0.70%. The sugar conversion index (SCI) is 8%–18%. Furthermore, the molar ratio of fructose to glucose is 0.9:1 to 1.1:1.

[0067] In this embodiment, all six parameters measured and calculated in the aforementioned steps are compared one by one with preset threshold conditions to form a final comprehensive quality judgment. The absolute weight-average molecular weight (Mw) is controlled between 3500 Da and 7500 Da. This range corresponds to the appropriate molecular weight range of Polygonatum polysaccharides during the steaming process. Too low a Mw indicates excessive degradation of the polysaccharide and possible decrease in activity, while too high a Mw indicates insufficient steaming and inadequate degradation of the polysaccharide molecular chains. The dispersion coefficient... To ensure uniformity in the molecular weight distribution of polysaccharides, an excessively wide distribution indicates uneven steaming. A sucrose concentration ≤0.20 mg / mL ensures that native sucrose has been fully hydrolyzed and converted during steaming; this is a key negative indicator of whether steaming is adequate. The Maillard reaction index (MRI) should be controlled between 0.20% and 0.70%. Within this range, the Maillard reaction produces the characteristic color and aroma of steamed Polygonatum without generating excessive byproducts. The sugar conversion index (SCI) should be controlled between 8% and 18%. This range reflects the ideal conversion level of polysaccharides to reducing sugars; insufficient steaming results in a low SCI, while excessive steaming results in a high SCI. The fructose to glucose molar ratio should be limited to a narrow range of 0.9:1 to 1.1:1. The stability of this ratio is evidence of the controllability of the polysaccharide degradation process; deviations from this range suggest possible abnormal degradation pathways or abnormal raw materials.

[0068] All six conditions mentioned above must be met simultaneously. Failure to meet any one condition will result in the batch of steamed Polygonatum standard decoction being deemed substandard. This multi-dimensional threshold design enables the quality evaluation to have a two-way control capability, which can identify both insufficient steaming (high sucrose residue, low SCI, high Mw) and excessive steaming (low Mw, high SCI, high MRI), achieving unprecedentedly precise control over the endpoint of steamed Polygonatum processing.

[0069] Please see Figure 5 The quality detection system for steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics, used to implement the quality detection method for steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics, includes a sample processing module 1, a polysaccharide molecular weight detection module 2, a small molecule detection module 3, and a data processing and judgment module 4. The polysaccharide precipitation output terminal of the sample processing module 1 is connected to the input terminal of the polysaccharide molecular weight detection module 2, the supernatant output terminal of the sample processing module 1 is connected to the input terminal of the small molecule detection module 3, and the input terminal of the data processing and judgment module 4 is connected to the total polysaccharide concentration output terminal of the sample processing module 1, the output terminal of the polysaccharide molecular weight detection module 2, and the output terminal of the small molecule detection module 3, respectively. The sample processing module 1 is used to obtain a standard decoction sample of steamed Polygonatum sibiricum, determine the total polysaccharide concentration, and perform alcohol precipitation fractionation on the other sample to separate the polysaccharide precipitate component and the small molecule supernatant component. The polysaccharide molecular weight detection module 2 is used to reconstitute the polysaccharide precipitate and determine the absolute weight-average molecular weight Mw and dispersion coefficient of Polygonatum polysaccharide using gel permeation chromatography-laser light scattering. ; The small molecule detection module 3 is used to concentrate and solvent-replace the components of the small molecule supernatant, and to determine the characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration using hydrophilic interaction chromatography and reversed-phase chromatography, respectively. The data processing and judgment module 4 is used to obtain the total polysaccharide concentration, Mw, The characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration were analyzed. The Maillard reaction index (MRI) and sugar conversion index (SCI) were calculated. Multidimensional index threshold determination was implemented to generate a quality qualification conclusion for the steamed Polygonatum sibiricum standard decoction.

[0070] In this embodiment, the standard decoction sample of Polygonatum sibiricum to be tested first enters the sample processing module 1. This module automatically completes the sample separation and quantification: one sample is transferred to the colorimetric analysis unit, and the total polysaccharide concentration is determined by the phenol-sulfuric acid method, expressed as anhydrous glucose; the other sample enters the alcohol precipitation unit, where anhydrous ethanol is automatically added to a final concentration of 80% under stirring conditions according to the set program. After standing at 4°C for 2 to 4 hours, the centrifuge is started and centrifuged at 4000 to 6000 rpm for 10 to 20 minutes. The obtained polysaccharide precipitate is transferred to the polysaccharide molecular weight detection module 2 through the polysaccharide precipitation output terminal, while the small molecule supernatant is transferred to the small molecule detection module 3 through the supernatant output terminal.

[0071] After receiving the polysaccharide precipitate, the polysaccharide molecular weight detection module 2 automatically adds hot water at 60-80℃ for reconstitution and adjusts the volume to a polysaccharide concentration of 1-5 mg / mL. The sample is then injected into a high-performance gel permeation chromatography-differential refractive index-multi-angle laser light scattering (MLLS) system. Separation and detection are performed using a Shodex OHpak SB-806M HQ column and an SB-804 HQ column in series, with a mobile phase of 0.1 mol / L sodium nitrate (containing 0.02% sodium azide), a flow rate of 0.5 mL / min, and a column temperature of 35℃. The absolute molecular weight of each elution fraction is directly determined using a multi-angle laser light scattering detector, while the differential refractive index detector simultaneously provides the concentration signal. The combined results output the absolute weight-average molecular weight (Mw) and dispersion coefficient of the Polygonatum sibiricum polysaccharide. The detection results are then transmitted in real time to the data processing and judgment module 4.

[0072] After receiving the small molecule supernatant components, the small molecule detection module 3 first initiates a reduced-pressure concentration program to remove ethanol, then adds a 1:1 acetonitrile-water mixed solvent to dissolve and dilute to volume, obtaining the small molecule assay solution. Subsequently, this assay solution is automatically split into two streams: one stream enters a hydrophilic interaction chromatography system, using an XBridge Amide column (4.6 mm × 250 mm, 3.5 μm), with acetonitrile as mobile phase A and an aqueous solution containing 0.1% triethylamine as mobile phase B, and performs separation using a gradient elution program (0-15 min B phase 15% → 35%, 15-25 min B phase 35% → 45%, 25-30 min B phase 45%), simultaneously determining the concentrations of fructose, glucose, sucrose, raffinose, and stachyose using an evaporative light scattering detector (drift tube temperature 55℃, carrier gas flow rate 1.8 L / min); the other stream enters a reversed-phase chromatography system, using a C18 column (4.6 mm × 150 mm, 3.5 μm), and performs separation using a C18 column (4.6 mm × 150 mm, 3.5 μm). The concentration of 5-hydroxymethylfurfural was measured at a wavelength of 284 nm on a column with a flow rate of 1.0 mL / min and a column temperature of 30 °C. All the above small molecule detection data were transmitted to the data processing and judgment module 4 through the module output terminal.

[0073] The data processing and judgment module 4 serves as the analysis center of the entire system, receiving in real time the total polysaccharide concentration data from the sample processing module 1 and the Mw and... from the polysaccharide molecular weight detection module 2. The data includes the characteristic sugar spectrum and 5-hydroxymethylfurfural concentration data of the small molecule detection module 3. The module's built-in calculation unit automatically calculates two derived indices based on the acquired data: Maillard reaction index MRI = C5-HMF / (C-fructose + C-glucose) × 100% and sugar conversion index SCI = (C-fructose + C-glucose) / C-total polysaccharides × 100%. Subsequently, the judgment unit compares each of the above parameters with six preset threshold conditions, determining the threshold only if the absolute weight-average molecular weight Mw is between 3500 Da and 7500 Da, and the dispersion coefficient is... When the sucrose concentration (Csucrose) is ≤ 0.20 mg / mL, the MRI value is 0.20% to 0.70%, the SCI value is 8% to 18%, and the fructose to glucose molar ratio is 0.9:1 to 1.1:1, the system outputs a conclusion that the quality is qualified; if any one of these conditions is not met, the system outputs a conclusion that the quality is unqualified, thus completing the automated, multi-dimensional, and precise determination of the quality of the steamed Polygonatum sibiricum standard decoction.

[0074] The above-disclosed embodiments are merely one or more preferred embodiments of this application and should not be construed as limiting the scope of this application. Those skilled in the art can understand that all or part of the processes for implementing the above embodiments and equivalent changes made in accordance with the claims of this application still fall within the scope of this application.

Claims

1. A method for quality detection of standard decoction of steamed Polygonatum sibiricum based on polysaccharide molecular characteristics, characterized in that, Includes the following steps: A standard decoction sample of steamed Polygonatum was obtained, and its total polysaccharide concentration was determined by colorimetry. The sample was then subjected to alcohol precipitation fractionation to separate the polysaccharide precipitate and the small molecule supernatant. The polysaccharide precipitate was reconstituted, and the absolute weight-average molecular weight Mw and dispersion coefficient K of Polygonatum polysaccharide were determined by gel permeation chromatography-laser light scattering. The components of the small molecule supernatant were concentrated and solvent replaced, and the concentrations of characteristic small molecule sugars and 5-hydroxymethylfurfural were determined by hydrophilic interaction chromatography and reversed-phase chromatography, respectively. The characteristic small molecule sugars included fructose, glucose, sucrose, raffinose and stachyose. Based on the total polysaccharide concentration, absolute weight-average molecular weight Mw, dispersion coefficient K, characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration, multidimensional index synergistic calculation and threshold determination are performed to generate a quality qualification conclusion for the steamed Polygonatum standard decoction. The preparation method of the standard decoction sample of steamed Polygonatum is as follows: take steamed Polygonatum slices, add 8 to 12 times the amount of water and soak for 30 to 60 minutes, heat and boil twice for 30 to 60 minutes each time, combine the decoctions, filter while hot, and concentrate the filtrate under reduced pressure to 1 mL equivalent to 0.5 g of steamed Polygonatum slices, thus obtaining the standard decoction sample of steamed Polygonatum.

2. The method for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics as described in claim 1, characterized in that, The specific operation of the alcohol precipitation fractionation process is as follows: anhydrous ethanol is added to the other sample under stirring to make the volume percentage of ethanol in the mixture reach 80%, and the mixture is allowed to stand for 2 to 4 hours to allow the polysaccharide to fully precipitate. The mixture is then centrifuged at 4000 to 6000 rpm for 10 to 20 minutes, and the polysaccharide precipitate and the small molecule supernatant are collected separately.

3. The method for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics as described in claim 1, characterized in that, The gel permeation chromatography-laser light scattering (GPS-LSS) method employs a high-performance gel permeation chromatography-differential refractive index-multi-angle laser light scattering (LSIS-LSS) system. The chromatographic columns are a Shodex OHpak SB-806MHQ column and an SB-804HQ column connected in series. The mobile phase is a 0.1 mol / L sodium nitrate aqueous solution containing 0.02% sodium azide, with a flow rate of 0.5 mL / min and a column temperature of 35 °C. The polysaccharide precipitate is reconstituted with hot water at 60–80 °C and diluted to a polysaccharide concentration of 1–5 mg / mL before determination.

4. The method for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics as described in claim 1, characterized in that, The colorimetric method is the phenol-sulfuric acid method, with anhydrous glucose as the reference standard. The total polysaccharide concentration in the steamed Polygonatum standard decoction was measured to be 10-30 mg / mL.

5. The method for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics as described in claim 1, characterized in that, The hydrophilic interaction chromatography method uses an amide-based hydrophilic interaction column with dimensions of 4.6 mm × 250 mm and a diameter of 3.5 μm. Mobile phase A is acetonitrile, and mobile phase B is an aqueous solution containing 0.1% triethylamine. The gradient elution program is as follows: from 0 to 15 minutes, the volume percentage of mobile phase B increases linearly from 15% to 35%; from 15 to 25 minutes, the volume percentage of mobile phase B increases linearly from 35% to 45%; and from 25 to 30 minutes, the volume percentage of mobile phase B remains constant at 45%. The detection was performed using an evaporative light scattering detector, with a drift tube temperature of 55°C and a carrier gas flow rate of 1.8 L / min.

6. The method for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics as described in claim 1, characterized in that, The reversed-phase chromatography method used a C18 reversed-phase column with dimensions of 4.6 mm × 150 mm and 5 μm. The mobile phase was a mixture of methanol and water at a volume ratio of 15:

85. The flow rate was 1.0 mL / min, the column temperature was 30 °C, and the 5-hydroxymethylfurfural was detected at a wavelength of 284 nm using an ultraviolet detector.

7. The method for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics as described in claim 1, characterized in that, The specific operation of concentration and solvent replacement is as follows: after the ethanol is removed by vacuum concentration of the small molecule supernatant component, a mixed solvent of acetonitrile-water volume ratio of 1:1 is added to dissolve and dilute to volume to obtain a small molecule assay solution for chromatographic analysis.

8. The method for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics as described in claim 1, characterized in that, The collaborative calculation and threshold determination of the multidimensional indicators include: The Maillard response index (MRI) is calculated as: MRI = C5 - HMF / (Cfructose + Cglucose) × 100%. Calculate the sugar conversion index (SCI) as follows: (Cfructose + Cglucose) / Ctotal polysaccharides × 100%; Wherein C5-HMF, Cfructose, and Cglucose are the concentrations of 5-hydroxymethylfurfural, fructose, and glucose, respectively, and Ctotal polysaccharide is the total polysaccharide concentration measured by the colorimetric method. All the above concentrations are in mg / mL.

9. The method for quality detection of steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics as described in claim 8, characterized in that, The generation of a quality compliance conclusion must simultaneously meet all of the following threshold conditions: The absolute weight-average molecular weight (Mw) is 3500 Da to 7500 Da; The dispersion coefficient Đ ≤ 1.6; Sucrose concentration C (sucrose ≤ 0.20 mg / mL); Maillard response index on MRI ranged from 0.20% to 0.70%. The sugar conversion index (SCI) is 8%–18%. Furthermore, the molar ratio of fructose to glucose is 0.9:1 to 1.1:

1.

10. A quality detection system for steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics, used to implement the quality detection method for steamed Polygonatum sibiricum standard decoction based on polysaccharide molecular characteristics according to any one of claims 1-9, characterized in that, It includes a sample processing module, a polysaccharide molecular weight detection module, a small molecule detection module, and a data processing and judgment module; The polysaccharide precipitation output terminal of the sample processing module is connected to the input terminal of the polysaccharide molecular weight detection module, the supernatant output terminal of the sample processing module is connected to the input terminal of the small molecule detection module, and the input terminal of the data processing and judgment module is connected to the total polysaccharide concentration output terminal of the sample processing module, the output terminal of the polysaccharide molecular weight detection module, and the output terminal of the small molecule detection module, respectively. The sample processing module is used to obtain a standard decoction sample of steamed Polygonatum sibiricum, determine the total polysaccharide concentration, and perform alcohol precipitation fractionation on another sample to separate the polysaccharide precipitate component and the small molecule supernatant component. The polysaccharide molecular weight detection module is used to re-dissolve the polysaccharide precipitate and determine the absolute weight-average molecular weight Mw and dispersion coefficient K of Polygonatum polysaccharide using gel permeation chromatography-laser light scattering coupled method. The small molecule detection module is used to concentrate and solvent-replace the components of the small molecule supernatant, and to determine the characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration using hydrophilic interaction chromatography and reversed-phase chromatography, respectively. The data processing and judgment module is used to obtain the total polysaccharide concentration, Mw, K, characteristic small molecule sugar spectrum and 5-hydroxymethylfurfural concentration, calculate Maillard reaction index MRI and sugar conversion index SCI, perform multidimensional index threshold judgment, and generate a quality qualification conclusion for the steamed Polygonatum standard decoction.

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