Fingerprint of millet sprout or roasted millet sprout and preparation method thereof

Abstract

This invention relates to the field of traditional Chinese medicine detection technology, specifically providing characteristic chromatograms of germinated rice or roasted germinated rice and their preparations, and their construction methods. Using octadecylsilane-bonded silica gel as the packing material, the mobile phase includes methanol and formic acid aqueous solution, gradient elution, and a specific elution program obtained through extensive experimental screening, yielding more than 13 common characteristic peaks. Effective separation of characteristic peaks including uridine, adenosine, guanosine, 5-hydroxymethylfurfural, 4-coumaric acid, and ferulic acid is achieved, with good peak shape, stable baseline, complete separation of the measured components, and short detection time. This provides a basis for the quality detection and control of germinated rice or roasted germinated rice and their preparations, fully reflecting the integrity and characteristics of germinated rice or roasted germinated rice and their preparations. The method is simple to operate and applicable to the quality control of different types of germinated rice or roasted germinated rice preparations.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine detection technology, specifically to the characteristic spectra of germinated rice or roasted germinated rice and their preparations and their construction methods. Background Technology

[0002] Germinated millet is a processed product obtained by germinating and drying the mature fruit of *Setaria italica* (L.) Beauv. Its main medicinal effects are aiding digestion, regulating the stomach, and strengthening the spleen and stomach. Fried millet sprouts are a type of processed millet sprouts, prepared by stir-frying the sprouts in a pan over low heat until they turn a deep yellow color, then removing and cooling. Fried millet sprouts are slightly warm in nature and are particularly effective in strengthening the spleen and aiding digestion, often used for those with spleen deficiency and poor appetite. Fried millet sprout granules are produced through the extraction, concentration, drying, and preparation of fried millet sprout slices.

[0003] The 2020 edition of the Chinese Pharmacopoeia discloses quality control methods for germinated rice and germinated rice slices, including appearance, identification, and inspection. However, it does not disclose qualitative and quantitative analysis methods for the main components in germinated rice and germinated rice slices. Existing technologies disclose methods for determining or identifying the content of single components in germinated rice or roasted germinated rice. While these methods can determine single components and thus achieve quality control, they are time-consuming and labor-intensive, making them difficult to widely apply in production practice. Chinese Patent CN117092244A discloses a method for constructing characteristic chromatograms of roasted germinated rice and its preparations, including the preparation of a test solution: weighing the roasted germinated rice and its preparations to be tested, adding 70% ethanol solution, ultrasonically extracting, filtering, and collecting the filtrate to obtain the test solution; using 4-coumaric acid, 5-hydroxymethylfurfural, and ferulic acid as reference standards, and using ultra-high performance liquid chromatography to obtain the characteristic chromatograms of the roasted germinated rice and its preparations. The method for constructing the characteristic spectrum of roasted germinated barley and its preparations disclosed in this patent yields characteristic peaks with moderate peak areas, clear peak shapes, good peak separation, strong repeatability, and better sensitivity. However, the characteristic spectrum constructed in this patent contains only six characteristic peaks, which is insufficient to fully reflect the chemical composition of the medicinal material, leading to inaccurate quality detection and analysis. Summary of the Invention

[0004] The purpose of this invention is to provide characteristic spectra of germinated rice or roasted germinated rice and its preparations, and a method for constructing them. The characteristic spectra constructed using the method of this invention have more than 13 characteristic peaks with good peak separation, which can more comprehensively reflect the chemical composition of the medicinal material and its preparations. Using the characteristic spectra for quality detection can improve the accuracy of the analytical results.

[0005] Therefore, this invention discloses a method for constructing a characteristic spectrum of germinated rice or roasted germinated rice and its preparations, comprising the following steps:

[0006] (1) Preparation of the test solution;

[0007] (2) The test solution was analyzed by ultra-high performance liquid chromatography. The packing material was octadecylsilane bonded silica gel. The mobile phase consisted of methanol and formic acid aqueous solution. Gradient elution was performed. The gradient elution program was: 0 → 8 minutes → 38 minutes → 53 minutes. The volume percentage of methanol in the mobile phase was 0% → 3% → 32% → 32%.

[0008] Further, step (1) includes:

[0009] 1) Take the analyte, add solvent to extract, and obtain the extract;

[0010] 2) Separate the solid and liquid components of the extract, and take the liquid, which is the test solution;

[0011] Preferably, step (1) also satisfies any one or more of the following AF:

[0012] A. In step 1), the solvent is selected from at least one of water, methanol, and ethanol;

[0013] B. In step 1), the mass ratio of the analyte to the volume of the solvent is 0.2-0.7g:10-40mL;

[0014] C. In step 1), the extraction method is reflux extraction or ultrasonic extraction, and the extraction time is 10 min-1 h;

[0015] D. In step 2), the solid-liquid separation is performed by centrifugation or filtration;

[0016] E. In step 2), after taking the liquid, the steps of drying, dissolving, extracting and collecting the extract and drying and dissolving the liquid are also included; or, the liquid is passed through a separation column, an eluent is added for elution, the eluent is collected and dried and dissolved.

[0017] F. In step 1), the test substance is selected from one or more of the following: germinated rice medicinal materials, germinated rice or roasted germinated rice slices, germinated rice or roasted germinated rice granules, and germinated rice or roasted germinated rice powder.

[0018] Further, the solvent for dissolving is selected from at least one of water, methanol, and ethanol; and / or, the solvent for extraction is ethyl acetate; and / or, the eluent is a methanol solution containing 0.5-2% formic acid by volume; and / or, the separation column is an acidic alumina column; and / or, the height of the separation column is 5cm-12cm (preferably 5cm-6cm); and / or, the drying is evaporation at 55-100°C (preferably evaporation at 55-100°C).

[0019] Furthermore, the chromatographic conditions for ultra-high performance liquid chromatography in step (2) also include: a detection wavelength of 250-330 nm, a flow rate of 0.23-0.27 ml / min, a column temperature of 23-30 °C, and an injection volume of 1-2 μl; preferably, the detection wavelength is 270 nm from 0 to 23 minutes and 310 nm after 23 minutes; preferably, the column temperature is 24-26 °C; preferably, the injection volume is 1-1.5 μl.

[0020] Further, the column length is 100-150 mm, the inner diameter is 2.1 mm, and the particle size is 1.6-1.8 μm; and / or, the volume percentage of formic acid in the formic acid aqueous solution in the mobile phase is 0.08-0.12%.

[0021] Furthermore, the gradient elution program further includes 53 minutes → 54 minutes, with the volume percentage of methanol in the mobile phase changing from 32% to 0%; or, the gradient elution program further includes 53 minutes → 54 minutes → 60 minutes, with the volume percentage of methanol in the mobile phase changing from 32% to 0% to 0%.

[0022] Furthermore, the construction method further includes the step of preparing a reference solution using at least one of uridine, adenosine, guanosine, 5-hydroxymethylfurfural, 4-coumaric acid, and ferulic acid, and the step of obtaining a reference reference spectrum by detecting the reference solution using ultra-high performance liquid chromatography in the above construction method.

[0023] Furthermore, the characteristic spectra of germinated rice or roasted germinated rice and their preparations obtained by the construction method are selected from any one of the following (1)-(2):

[0024] (1) The characteristic chromatogram of the roasted germinated barley and its preparation has 15 characteristic peaks. Peak 4 is used as reference peak 1, and peak 12 as reference peak 2. The relative retention times of peaks 1-6 with reference peak 1 and the relative retention times of peaks 7-15 with reference peak 2 are within ±10% of the specified values. The specified values ​​are: 0.63 (peak 1), 0.72 (peak 2), 0.85 (peak 3), 1.12 (peak 5), 1.15 (peak 6), 0.70 (peak 7), 0.72 (peak 8), 0.83 (peak 9), 0.86 (peak 10), 0.91 (peak 11), 1.03 (peak 13), 1.09 (peak 14), 1.2... 9 (peak 15); or, the characteristic spectrum of the germinated rice and its preparation has 13 characteristic peaks, with peak 4 as reference peak 1 and peak 12 as reference peak 2, and the relative retention times of peaks 1, 4-6 and reference peak 1 and the relative retention times of peaks 7-15 and reference peak 2 are within ±10% of the specified values; the specified values ​​are: 0.63 (peak 1), 1.12 (peak 5), 1.15 (peak 6), 0.70 (peak 7), 0.72 (peak 8), 0.83 (peak 9), 0.86 (peak 10), 0.91 (peak 11), 1.03 (peak 13), 1.09 (peak 14), 1.29 (peak 15);

[0025] (2) The characteristic chromatogram of the roasted germinated barley and its preparation has 15 characteristic peaks. The peak corresponding to the adenosine reference standard peak is designated as reference peak 1, and the peak corresponding to the 4-coumaric acid reference standard peak is designated as reference peak 2. The relative retention times of peaks 1-6 with reference peak 1 and the relative retention times of peaks 7-15 with reference peak 2 are within ±10% of the specified values. The specified values ​​are: 0.63 (peak 1), 0.72 (peak 2), 0.85 (peak 3), 1.12 (peak 5), 1.15 (peak 6), 0.70 (peak 7), 0.72 (peak 8), 0.83 (peak 9), 0.86 (peak 10), 0.91 (peak 11), 1.03 (peak 13), 1.09 (peak 14), 1.2 9 (peak 15); or, the characteristic chromatogram of the germinated rice and its preparation has 13 characteristic peaks, with the peak corresponding to the adenosine reference standard peak as reference peak 1, and the peak corresponding to the 4-coumaric acid reference standard peak as reference peak 2. The relative retention times of characteristic peaks 1 and 4-6 with reference peak 1 and the relative retention times of peaks 7-15 with reference peak 2 are within ±10% of the specified values, which are: 0.63 (peak 1), 1.12 (peak 5), 1.15 (peak 6), 0.70 (peak 7), 0.72 (peak 8), 0.83 (peak 9), 0.86 (peak 10), 0.91 (peak 11), 1.03 (peak 13), 1.09 (peak 14), and 1.29 (peak 15).

[0026] The present invention also provides a quality control method for germinated rice or roasted germinated rice and its preparations, comprising: using the product to be tested as a test sample to construct a characteristic spectrum of the product to be tested according to the above construction method.

[0027] This invention also provides a method for distinguishing between germinated rice and its preparations and roasted germinated rice and its preparations, comprising:

[0028] Using the product to be identified as the test sample, a feature spectrum of the product to be identified is constructed according to the above construction method; the feature spectrum of the product to be identified is compared with the feature spectrum constructed by the above construction method, and identification is performed based on the comparison results.

[0029] Further, when the characteristic spectrum meets the following conditions, the analyte corresponding to the characteristic spectrum is roasted germinated barley and its preparations; the characteristic spectrum has 15 characteristic peaks, with peak 4 as reference peak 1 and peak 12 as reference peak 2, and the relative retention times of peaks 1-6 with reference peak 1 and the relative retention times of peaks 7-15 with reference peak 2 are within ±10% of the specified values; the specified values ​​are: 0.63 (peak 1), 0.72 (peak 2), 0.85 (peak 3), 1.12 (peak 5), 1.15 (peak 6), 0.70 (peak 7), 0.72 (peak 8), 0.83 (peak 9), 0.86 (peak 10), 0.91 (peak 11), 1.03 (peak 13), 1.09 (peak 14), and 1.29 (peak 15).

[0030] Further, when the characteristic spectrum meets the following conditions, the analyte corresponding to the characteristic spectrum is germinated barley and its preparations; the characteristic spectrum has 13 characteristic peaks, with peak 4 as reference peak 1 and peak 12 as reference peak 2, and the relative retention times of peaks 1, 4-6 and reference peak 1, and the relative retention times of peaks 7-15 and reference peak 2 are within ±10% of the specified values; the specified values ​​are: 0.63 (peak 1), 1.12 (peak 5), 1.15 (peak 6), 0.70 (peak 7), 0.72 (peak 8), 0.83 (peak 9), 0.86 (peak 10), 0.91 (peak 11), 1.03 (peak 13), 1.09 (peak 14), and 1.29 (peak 15).

[0031] Furthermore, the germinated rice or roasted germinated rice powder is a freeze-dried powder. The preparation method of the freeze-dried powder includes: mixing germinated rice or roasted germinated rice with water and extracting, separating the solid and liquid, concentrating the liquid, and freeze-drying to obtain the freeze-dried powder of roasted germinated rice decoction; preferably, it also satisfies one or more of the following A and D:

[0032] A. The extraction method is selected from heating reflux extraction or decoction extraction;

[0033] B. The number of extractions is 1-3 times, and the extraction time is 15-60 minutes;

[0034] C. The mass of water added for each extraction should be 6-8 times the mass of the roasted germinated barley;

[0035] D. The relative density of the concentrated extract at 58-62℃ is 1.05-1.10 g / mL.

[0036] The technical solution of this invention has the following advantages:

[0037] 1. The method for constructing characteristic chromatograms of germinated rice or roasted germinated rice and its preparations provided by this invention uses octadecylsilane-bonded silica gel as the packing material, and the mobile phase includes methanol and formic acid aqueous solution. Gradient elution is performed, and a specific elution program is obtained through extensive experimental screening. More than 13 common characteristic peaks are obtained, and effective separation of characteristic peaks including uridine, adenosine, guanosine, 5-hydroxymethylfurfural, 4-coumaric acid, and ferulic acid is achieved. The peaks are well-shaped, the baseline is stable, the components can be completely separated, and the detection time is short. This provides a basis for the quality detection and control of germinated rice or roasted germinated rice and its preparations, and can fully reflect the integrity and characteristics of germinated rice or roasted germinated rice and its preparations. The method is simple to operate and can be applied to the quality control of different types of germinated rice or roasted germinated rice preparations.

[0038] 2. The method for constructing the characteristic chromatogram of germinated rice or roasted germinated rice and its preparations provided by this invention, as identified by reference standards, shows that peaks 1, 4, 5, 6, 12, and 14 are, in order, uridine, adenosine, guanosine, 5-hydroxymethylfurfural, 4-coumaric acid, and ferulic acid, respectively. All peaks are well separated and have moderate areas. Using one or more of uridine, adenosine, guanosine, 5-hydroxymethylfurfural, 4-coumaric acid, and ferulic acid as references, the accuracy of quality control of germinated rice or roasted germinated rice can be achieved qualitatively. The identification of more than 13 characteristic peaks in the characteristic chromatogram ensures the accuracy of this method in the quality control of germinated rice or roasted germinated rice.

[0039] 3. The quality detection method for germinated rice or roasted rice preparations provided by the present invention includes the step of comparing the characteristic spectrum of the germinated rice or roasted rice product to be tested with the control characteristic spectrum of germinated rice or roasted rice and its preparations. This method can control the material transfer of indicator components in germinated rice medicinal materials, germinated rice or roasted rice slices, germinated rice or roasted rice granules, and germinated rice or roasted rice powder, and can also control the overall stability of the quality of characteristic components in germinated rice medicinal materials, germinated rice or roasted rice slices, germinated rice or roasted rice granules, and germinated rice or roasted rice powder. Moreover, the method is simple to operate, has high precision, good stability, good repeatability, high accuracy, and fast analysis rate. Attached Figure Description

[0040] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0041] Figure 1 This is a chromatogram of the uridine and adenosine reference solutions from Example 1.

[0042] Figure 2 This is a chromatogram of the guanosine reference solution from Example 1.

[0043] Figure 3 This is a chromatogram of the 5-hydroxymethylfurfural reference solution from Example 1.

[0044] Figure 4 This is the chromatogram of the 4-coumaric acid reference solution in Example 1.

[0045] Figure 5 This is the chromatogram of the ferulic acid reference solution in Example 1.

[0046] Figure 6 This is the chromatogram of the test solution in Example 1.

[0047] Figure 7 This is the comparative feature map in Example 2.

[0048] Figure 8 This is the chromatogram of the test solution in Example 3.

[0049] Figure 9 It is a superimposed graph of the freeze-dried powder of standard germinated rice decoction and the freeze-dried powder of standard roasted germinated rice decoction in Example 4.

[0050] Figure 10 This is the chromatogram of the test solution in Comparative Example 1.

[0051] Figure 11 This is the chromatogram of the test solution in Comparative Example 2.

[0052] Figure 12 This refers to the chromatogram overlay of the test sample solution in Experiment Example 1 - selection of the detection wavelength.

[0053] Figure 13 This refers to the chromatogram overlay of the test sample solution in Experiment Example 1, specifically the selection of column temperature.

[0054] Figure 14 This is the chromatographic overlay of the test sample solution in Experiment Example 1 - selection of the chromatographic column.

[0055] Figure 15 This refers to the chromatogram overlay of the test sample solution in Experiment Example 1 and the selection of the injection volume.

[0056] Figure 16 This refers to the chromatogram overlay of the test sample solution in Experiment Example 2 – the selection of the treatment method.

[0057] Figure 17 This is an examination of the height of the separation column in the chromatographic overlay of the test sample solution in Experiment Example 2.

[0058] Figure 18 This is an investigation of the chromatographic overlay of the test sample solution in Experiment Example 2 and the solvent evaporation method.

[0059] Figure 19 This is the chromatogram of the negative control solution in Experiment Example 3 - specificity test.

[0060] Figure 20 This is the chromatogram-specificity test of the test solution in Experimental Example 3 of this invention.

[0061] Figure labels: Peak 1: uridine; Peak 4 (S1): adenosine; Peak 5: guanosine; Peak 6: 5-hydroxymethylfurfural; Peak 12 (S2): 4-coumaric acid; Peak 14: ferulic acid. Detailed Implementation

[0062] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0063] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0064] The instruments and reagents used in this invention are as follows:

[0065] 1. Chromatograph: (1) Waters ACQUITYUPLC I-Class chromatography system, including a quaternary solvent manager (ACQ-QSM), an autosampler (ACQ-FTN), an original imported column oven (ACQ-CM), a diode array UV detector (ACQ-TUV), and an Empower chromatography management system.

[0066] (2) Thermo Vanquish Flex UHPLC system: Quaternary solvent manager (VF-P20-A), autosampler (VF-A10-A-02), column oven (VH-C10-A-02), DAD detector (VF-D40-A).

[0067] 2. Electronic analytical balances: Mettler Toledo NewClassic MS 0.0001 g balance, Jing TianFA2044A 0.0001 g balance.

[0068] 3. Chromatographic columns: (1) Waters ACQUITYUPLC HSS T3, 150mm×2.1mm, 1.8μm; (2) LunaOmega PS C18100A, 150mm×2.1mm, 1.6μm; (3) Waters CORTECS T3, 150mm×2.1mm, 1.6μm; (4) Agilent sb-sq column, 150mm×2.1mm, 1.8μm.

[0069] 4. Reagents: Methanol (chromatographic grade), water (ultrapure water), formic acid (analytical grade).

[0070] 5. Test reagents: uridine (110887-202305, 99.6%), adenosine (110879-202204, 99.4%), guanosine (111977-202202, 88.6%), 5-hydroxymethylfurfural (111626-202316, 98.4%), 4-coumaric acid (112037-202102, 99.7%), ferulic acid (110773-202316, 99.3%), purchased from the National Institutes for Food and Drug Control.

[0071] The preparation method of the standard decoction freeze-dried powder of roasted barley sprouts is as follows: take roasted barley sprout slices, heat and reflux extract twice. For the first extraction, add water with a weight of 8 times the weight of the slices and soak for 30 minutes, then heat and reflux extract for 30 minutes and filter. For the second extraction, add water with a weight of 6 times the weight of the slices and extract for 25 minutes and filter. Combine the filtrates from the two filtrations and concentrate the filtrate to a relative density of 1.05 g / mL (detected at 60℃). Freeze-dry the concentrate to obtain the standard decoction freeze-dried powder.

[0072] Replace the roasted germinated barley slices with regular barley slices, and prepare the standard barley malt decoction freeze-dried powder according to the "Preparation Method of Roasted Germinated Barley Standard Decoction Freeze-dried Powder".

[0073] Example 1

[0074] This embodiment provides a method for constructing characteristic spectra of roasted germinated barley and its preparations, including the following steps:

[0075] Chromatographic conditions and system suitability tests were performed using octadecylsilane-bonded silica gel as the packing material (Waters ACQUITYUPLC HSS T3, column length 150 mm, column inner diameter 2.1 mm, particle size 1.8 μm); methanol as mobile phase A and 0.1% formic acid aqueous solution (V:V) as mobile phase B, with gradient elution performed according to the gradients specified in Table 1; flow rate 0.25 ml / min; column temperature 25 °C; detection wavelength 270 nm for 0-23 minutes and 310 nm for 23-53 minutes.

[0076] Table 1 Elution gradient

[0077]

[0078] Preparation of reference solutions: Take appropriate amounts of uridine, adenosine, guanosine, 5-hydroxymethylfurfural, 4-coumaric acid, and ferulic acid reference standards, and add 50% methanol aqueous solution to prepare a mixed reference solution containing 30 μg of uridine, 30 μg of adenosine, 30 μg of guanosine, 30 μg of 5-hydroxymethylfurfural, 50 μg of 4-coumaric acid, and 50 μg of ferulic acid per 1 ml.

[0079] Prepare reference solutions containing 30 μg of uridine and 30 μg of adenosine per 1 mL by dissolving appropriate amounts of uridine and adenosine in 70% methanol aqueous solution. Prepare individual reference solutions containing 30 μg of guanosine, 30 μg of 5-hydroxymethylfurfural, 50 μg of 4-coumaric acid, and 50 μg of ferulic acid per 1 mL by dissolving appropriate amounts of guanosine, 5-hydroxymethylfurfural, and 4-coumaric acid in 70% methanol aqueous solution, respectively.

[0080] Preparation of the test solution: Take an appropriate amount of freeze-dried powder of roasted barley sprout standard decoction, about 0.5 g, accurately weigh it, place it in a stoppered conical flask, accurately add 10 ml of 70% methanol aqueous solution (V:V), stopper tightly, weigh it, sonicate (power 250W, frequency 40kHz) for 30 minutes, cool it, and make up the weight loss with 70% methanol aqueous solution (V:V), shake well, filter, and pass the filtrate through an acidic alumina column (inner diameter 1.5 cm, packing weight 5 g, column length...). The column is 5 cm long. Before loading the sample, activate the acidic alumina column with about 3 column volumes of 70% methanol aqueous solution. Collect the sample solution after passing through the column, and then elute with methanol solvent (V:V) containing 1% formic acid (V:V) (about 3 column volumes). Continue to collect the eluent, and evaporate the solvent to dryness by rotary evaporation at low temperature (55℃). Dissolve the residue in 50% methanol aqueous solution (V:V) and transfer it to a 5 ml volumetric flask. Add 50% methanol aqueous solution to the mark, shake well, filter, and collect the filtrate.

[0081] The assay involves precisely pipetting 1 μl of both the reference solution and the test solution into a liquid chromatograph and measuring the results.

[0082] The test results are shown in Table 2. Figure 1-6 The chromatogram of the test solution showed 15 characteristic peaks, among which peaks 1, 4, 5, 6, 12, and 14 corresponded to the retention times of the reference peaks of uridine, adenosine, guanosine, 5-hydroxymethylfurfural, 4-coumaric acid, and ferulic acid. The peak corresponding to the adenosine reference standard peak is peak S1. Calculate the relative retention times of peaks 1-6 with peak S1. The peak corresponding to the 4-coumaric acid reference standard peak is peak S2. Calculate the relative retention times of peaks 7-15 with peak S2. The relative retention times are within ±10% of the specified values, which are: 0.63 (peak 1), 0.72 (peak 2), 0.85 (peak 3), 1.12 (peak 5), 1.15 (peak 6), 0.70 (peak 7), 0.72 (peak 8), 0.83 (peak 9), 0.86 (peak 10), 0.91 (peak 11), 1.03 (peak 13), 1.09 (peak 14), and 1.29 (peak 15).

[0083] Table 2 System Adaptability Parameters

[0084]

[0085] Example 2

[0086] This embodiment provides a method for constructing the characteristic spectrum of roasted germinated barley and its preparations, including:

[0087] Preparation of the test solution: The preparation method is the same as in Example 1.

[0088] Preparation of reference solution: Take appropriate amounts of adenosine and 4-coumaric acid reference standards, accurately weigh them, and add 50% methanol aqueous solution (V:V) to prepare a reference solution containing 30 μg per ml.

[0089] The chromatographic conditions were the same as in Example 1.

[0090] The common pattern of the characteristic spectrum of the freeze-dried powder of the standard decoction of roasted barley sprouts was established using the method described above.

[0091] Fifteen batches of freeze-dried powder of roasted barley sprout standard decoction were taken and the test solution was prepared according to the method of this embodiment. The characteristic chromatogram of the freeze-dried powder of roasted barley sprout standard decoction was obtained by ultra-high performance liquid chromatography. The results are shown in Table 3-4.

[0092] Table 3.15: Results of Relative Retention Time Determination of Characteristic Spectra of Standard Decoctions (Freeze-dried Powder) for 15 Batches of Fried Germinated Rice Slices

[0093]

[0094]

[0095] Table 4.15: Results of relative peak area determination of characteristic chromatograms for standard decoctions (freeze-dried powder) of roasted barley sprouts (batch 15).

[0096]

[0097]

[0098]

[0099] Using the fingerprint chromatogram similarity evaluation software "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System 2012 Edition" compiled by the Pharmacopoeia Commission, a reference characteristic chromatogram was generated, such as... Figure 7 As shown in Table 5-6. According to... Figure 7 The characteristic chromatograms shown can be used to analyze and compare the detection results of the characteristic chromatograms of the lyophilized powder of the standard decoction, and are used for the quality control of the standard decoction. Through multiple batches of HPLC chromatograms, 15 common peaks with good resolution were identified in the characteristic chromatograms. Based on the peak localization results in the chromatogram of the reference solution, peak 1 was identified as uridine, peak 4 as adenosine, peak 5 as guanosine, peak 6 as 5-hydroxymethylfurfural, peak 12 as 4-coumaric acid, and peak 14 as ferulic acid. The peak corresponding to the adenosine reference standard peak is designated as peak S1. The relative retention times of peaks 1-6 with peak S1 are calculated. The peak corresponding to the 4-coumaric acid reference standard peak is designated as peak S2. The relative retention times of peaks 7-15 with peak S2 are calculated. These relative retention times are within ±10% of the specified values, which are: 0.63 (peak 1), 0.72 (peak 2), 0.85 (peak 3), 1.12 (peak 5), 1.15 (peak 6), 0.70 (peak 7), 0.72 (peak 8), 0.83 (peak 9), 0.86 (peak 10), 0.91 (peak 11), 1.03 (peak 13), 1.09 (peak 14), and 1.29 (peak 15). Mark peak fitting was used to fit the characteristic chromatograms of the reference standards.

[0100] Table 5. Relative retention times of common modes in freeze-dried powder of standard roasted barley sprout decoction.

[0101]

[0102] Table 6. Relative Peak Area of ​​Common Patterns in Standard Fried Germinated Rice Decoction Freeze-Dried Powder

[0103]

[0104] Example 3

[0105] This embodiment provides a method for constructing characteristic chromatograms of roasted germinated barley and its preparations. The construction method is basically the same as that in Example 1, except that the gradient elution program in the chromatographic conditions is different. The gradient elution program in this embodiment is shown in Table 7. Detection results are shown in... Figure 8 As can be seen from the results in the figure, the gradient elution program in this embodiment includes a post-equilibrium program that returns to the original gradient after peak elution. The peak separation effect in the chromatogram is good, and the peak elution occurs before 53 minutes.

[0106] Table 7 Gradient elution program

[0107]

[0108] Example 4

[0109] This embodiment provides a method for distinguishing between germinated rice and its preparations and roasted germinated rice and its preparations, including the following:

[0110] The roasted germinated barley test solution was prepared according to the preparation method of the test solution in Example 1.

[0111] Replace the freeze-dried powder of standard decoction of roasted barley sprouts with freeze-dried powder of standard decoction of barley sprouts, and prepare the barley sprout test solution according to the preparation method of the test solution in Example 1.

[0112] Take the roasted germinated rice test solution and the germinated rice test solution and determine them according to the liquid chromatography method in Example 1.

[0113] See results Figure 9 The results show that, compared with the chromatogram of the germinated barley sample solution, the chromatogram of the roasted germinated barley sample solution has significantly more peaks 2 and 3, and peaks 4, 5, 6, 7, 8, 11, and 13, with clear differences in peak size. Peak 6 is 5-hydroxymethylfurfural; under high temperature conditions, sugars are converted into 5-hydroxymethylfurfural, hence the significant increase in the elevation of this peak.

[0114] In summary, the method of the present invention can distinguish between germinated rice and its preparations and roasted germinated rice and its preparations.

[0115] Comparative Example 1

[0116] This comparative example provides a method for constructing characteristic chromatograms of roasted germinated barley and its preparations. The test solution prepared according to the method in Example 1 was analyzed by ultra-high performance liquid chromatography (UHPLC). The chromatographic conditions were as follows: Waters ACQUITY HSS T3 column, 100 mm × 2.1 mm, 1.8 μm; flow rate, 0.3 ml / min; column temperature, 25 °C; injection volume, 1 μl; and gradient elution program as shown in Table 8. The detection results are shown in Table 8. Figure 10The results in the figure show that the chromatographic peaks at 6-10 minutes are poorly separated, and the peaks at 6.9 minutes and 8.2 minutes are formed by the merging of several peaks.

[0117] Table 8 Gradient elution program

[0118]

[0119] Comparative Example 2

[0120] This comparative example provides a method for constructing characteristic chromatograms of roasted germinated barley and its preparations. The test solution prepared according to the method in Example 1 was analyzed by ultra-high performance liquid chromatography (UHPLC). The chromatographic conditions were as follows: Waters ACQUITY HSS T3 column, 150 mm × 2.1 mm, 1.8 μm; flow rate, 0.25 ml / min; column temperature, 25 °C; injection volume, 1 μl. The gradient elution program used in this comparative example is shown in Table 9. The detection results are shown in... Figure 11 The results in the figure show that the peak separation in the chromatogram at 10-15 minutes is poor.

[0121] Table 9 Gradient elution program

[0122]

[0123] Example 1: Establishment of Liquid Chromatography Conditions

[0124] The test solution was prepared according to the method described in Example 1, and the following chromatographic conditions were investigated.

[0125] 1. Selection of detection wavelength

[0126] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase (WatersACQUITY HSS T3, column length 150 mm, column inner diameter 2.1 mm, particle size 1.8 μm); methanol was used as mobile phase A, and 0.1% formic acid aqueous solution (V:V) was used as mobile phase B, with gradient elution according to the gradient specified in Table 1; the flow rate was 0.25 ml / min; the column temperature was 25 ℃; and the detection wavelengths were 250 nm, 270 nm, 310 nm, and 330 nm.

[0127] Test results are shown Figure 12 As shown in the figure, the peak response values ​​of nucleoside components are relatively large and clearly distinguishable at wavelengths of 250 nm and 270 nm. The chromatographic peak response values ​​of organic acids are relatively large at a wavelength of 310 nm. Considering all chromatographic peaks, the detection wavelength was determined to be 270 nm for the first 0–23 minutes, and then 310 nm for detection after 23 minutes.

[0128] 2. Selection of column temperature

[0129] Based on the above-determined detection wavelength, the test solution was tested at a column temperature of 25℃ or 30℃, and the other liquid phase conditions were consistent with the method under "Selection of Detection Wavelength".

[0130] Test results are shown Figure 13 As can be seen from the figure, the resolution of the chromatographic peaks at a column temperature of 25℃ is better than that at a column temperature of 30℃. Therefore, 25℃ is the preferred column temperature for the characteristic chromatogram.

[0131] 3. Selection of chromatographic column

[0132] Based on the determined detection wavelength and column temperature, the test solution was detected using Waters ACQUITY HSS T3, Waters CORTECS T3, and Agilent SB-SQ columns, respectively. The remaining liquid chromatography conditions were consistent with the method described in the "Selection of Detection Wavelength" section.

[0133] Test results are shown Figure 14 As can be seen from the figure, the chromatographic peaks under the Waters ACQUITY HSS T3 column have good resolution. Therefore, the Waters ACQUITY HSS T3 column is preferred as the column for characteristic chromatograms.

[0134] 4. Selection of injection volume

[0135] Based on the determined detection wavelength, column temperature, and chromatographic column, the test solution was detected in this study with injection volumes of 1 μl and 2 μl, respectively. The remaining liquid chromatography conditions were consistent with the method under the "Selection of Detection Wavelength" section.

[0136] Test results are shown Figure 15 As can be seen from the figure, compared with the injection volume of 2 μl, the peak shape of the chromatogram corresponding to the injection volume of 1 μl is better, and the peak height and peak width are relatively moderate. Therefore, the injection volume of 1 μl is preferred as the injection volume of the test solution.

[0137] 5. Determination of chromatographic conditions

[0138] Based on the above optimization results, the chromatographic conditions for the characteristic chromatograms are determined as follows:

[0139] Octadecylsilane-bonded silica gel was used as the packing material (column length 150 mm, column inner diameter 2.1 mm, particle size 1.8 μm); methanol was used as mobile phase A and 0.1% formic acid aqueous solution was used as mobile phase B, and gradient elution was performed according to the specifications in Table 10; the flow rate was 0.25 ml / min; the column temperature was 25 °C; and the detection wavelength was 270 nm from 0 to 23 minutes and 310 nm after 23 minutes.

[0140] Table 10 Gradient elution program

[0141]

[0142] Example 2: Establishment of a method for preparing the test solution

[0143] Based on the chromatographic conditions determined in Experiment 1, “Establishment of Liquid Chromatography Conditions”, the preparation method of the test sample solution was further investigated.

[0144] The prepared test solution was tested according to the chromatographic conditions determined under Experimental Example 2, "Establishment of Liquid Chromatography Conditions".

[0145] 1. Selection of processing method

[0146] The effects of different extraction methods on the extraction efficiency of freeze-dried powder of standard roasted barley sprout decoction were investigated, with the information content of chromatographic peaks as the main evaluation index.

[0147] A. Direct ultrasonic extraction: Take about 0.5g of freeze-dried powder of standard roasted barley sprout decoction, accurately weigh it, place it in a stoppered conical flask, accurately add 10ml of 70% methanol aqueous solution, seal tightly, weigh, ultrasonically treat (power 250W, frequency 40kHz) for 30 minutes, cool, and make up the lost weight with 70% methanol aqueous solution, shake well, filter, and take the filtrate to obtain the product.

[0148] B. Extraction: Accurately weigh approximately 0.5g of freeze-dried powder of standard roasted barley sprout decoction, place it in a stoppered conical flask, accurately add 10ml of 70% methanol aqueous solution, seal tightly, weigh, sonicate (power 250W, frequency 40kHz) for 30 minutes, cool, filter, evaporate the filtrate to dryness, sonicate with 25ml of pure water (power 250W, frequency 40kHz) for 30 minutes, then extract with 25ml of ethyl acetate, collect the aqueous phase and ethyl acetate phase separately, evaporate to dryness, dissolve with 5ml of 50% methanol aqueous solution, filter, and collect the filtrate to obtain the final product.

[0149] C. Column separation: Accurately weigh approximately 0.5g of freeze-dried powder of roasted barley sprout standard decoction, place it in a stoppered conical flask, accurately add 10ml of 70% methanol aqueous solution, seal tightly, weigh, sonicate (power 250W, frequency 40kHz) for 30 minutes, cool, and replenish the lost weight with 70% methanol aqueous solution, shake well, filter, and collect the filtrate. Pass the filtrate through an acidic alumina column (inner diameter 1.5cm, packing weight 5g, column height 5cm; activate the acidic alumina column with approximately 3 column volumes of 70% methanol aqueous solution before loading), collect the sample solution after column separation, and then elute with methanol containing 1% formic acid (approximately 3 column volumes). Continue to collect the eluent, evaporate the solvent at low temperature (55℃) until dry, dissolve the residue in 50% methanol aqueous solution, transfer to a 5ml volumetric flask, add 50% methanol aqueous solution to the mark, shake well, filter, and collect the filtrate.

[0150] Test results are shown Figure 16 As shown in the figure, compared with direct ultrasonic extraction and extraction, the chromatogram of the test sample solution prepared by column separation has a flatter baseline, better separation of each chromatographic peak, and higher response of active ingredients. Therefore, column separation extraction is the preferred method for processing the test sample.

[0151] 2. Examination of the height of the separated columns

[0152] Based on the above-determined treatment method, the test solution was prepared using column heights of 5 cm or 12 cm.

[0153] Test results are shown Figure 17 As can be seen from the figure, compared with the chromatogram corresponding to a column height of 12cm, the chromatographic peak separation effect is better when the separation column height is 5cm, indicating that the column height of 5cm has achieved the effect of impurity removal. Therefore, the separation column height of 5cm is preferred.

[0154] 3. Investigation of solvent evaporation methods

[0155] Based on the above-determined processing method and column height, the evaporation methods selected were water bath (100℃) evaporation or low-temperature rotary evaporation (55℃) to prepare the test solution.

[0156] Test results are shown Figure 18 As shown in the figure, since the column eluent is a methanol solvent containing 1% formic acid, the low-temperature rotary evaporation process produces more components compared to water bath evaporation. Therefore, the low-temperature rotary evaporation method is preferred.

[0157] Experiment Example 3: Methodological Validation

[0158] 1. Precision test

[0159] The test solution was prepared according to the preparation method of the test solution in Example 1. The test solution was injected repeatedly 6 times under the chromatographic conditions in Example 1 for detection. Adenosine was selected as the S1 peak and 4-coumaric acid was selected as the S2 peak. The relative retention times of peaks 1, 2, 3, 5, and 6 with the S1 peak and the relative retention times of peaks 7, 8, 9, 10, 11, 13, 14, and 15 with the S2 peak were calculated.

[0160] The results are shown in Table 11 below. The results show that the RSD of the relative retention time of each characteristic peak and the reference S peak is less than 2%, indicating that the instrument has good precision.

[0161] Table 11 Results of Instrument Precision and Relative Retention Time Tests

[0162]

[0163]

[0164] 2. Method repeatability test

[0165] Six test solutions were prepared using the same batch of freeze-dried roasted barley sprout standard decoction powder according to the preparation method of the test solution in Example 1. The solutions were measured under the liquid chromatography conditions in Example 1. Adenosine was selected as the S1 peak and 4-coumaric acid as the S2 peak. The relative retention times of peaks 1, 2, 3, 5, and 6 with the S1 peak, and the relative retention times of peaks 7, 8, 9, 10, 11, 13, 14, and 15 with the S2 peak were calculated.

[0166] The results are shown in Table 12 below. The results show that the RSD of the relative retention times of each characteristic peak and the reference S peak is less than 2%, indicating that the method has good repeatability.

[0167] Table 12 Results of the method repeatability relative retention time test

[0168]

[0169]

[0170] 3. Intermediate precision test (by different operators)

[0171] Three inspectors at different times prepared test solutions using the same batch of freeze-dried roasted barley sprout standard decoction powder according to the preparation method of the test solution in Example 1. The prepared test solutions were measured using the same equipment. Adenosine was selected as the S1 peak and 4-coumaric acid as the S2 peak. The relative retention times of peaks 1, 2, 3, 5, and 6 with peak S1, and the relative retention times of peaks 7, 8, 9, 10, 11, 13, 14, and 15 with peak S2 were calculated.

[0172] The results are shown in Table 13 below. The results show that the RSD of the relative retention times of each characteristic peak and the reference S peak is less than 2%, indicating good intermediate precision of this method.

[0173] Table 13 Intermediate Precision Relative Retention Time Test Results (Different Operators)

[0174]

[0175]

[0176] 4. Specificity test

[0177] A blank solvent (50% methanol aqueous solution) was selected as the negative control solution. The test solution was prepared according to the preparation method of the test solution in Example 1. The negative control solution and the test solution were detected according to the chromatographic conditions in Example 1.

[0178] Test results are shown Figures 19-20 As can be seen from the figure, there is no interference from negative results, indicating that the method has good specificity.

[0179] 5. Stability test

[0180] Following the preparation method of the test solution in Example 1, the same batch of roasted germinated barley standard decoction lyophilized powder was used to prepare the test solution, and the test solution was detected according to the liquid phase conditions in Example 1. The test solution was injected at 0, 2, 4, 8, 12 and 24 hours after preparation, and the retention times of 15 common peaks were recorded and the relative retention times were calculated.

[0181] The results are shown in Table 14 below. The results show that the relative retention times of each characteristic peak and the reference peak S are less than 2%, indicating that the test solution is stable within 24 hours and meets the determination requirements.

[0182] Table 14 Results of the relative retention time test for stability

[0183]

[0184]

[0185] 6. Durability test

[0186] A. Investigation of different flow velocities

[0187] The test solution was prepared according to the preparation method of the test solution in Example 1. The test solution was injected and detected under the chromatographic conditions in Example 1, except that the determination was carried out at flow rates of 0.23 ml / min, 0.25 ml / min and 0.27 ml / min to examine the separation effect of each characteristic peak when the flow rate changed.

[0188] The results are shown in Tables 15-16 below. The results indicate that all chromatographic peaks were detected at different flow rates, demonstrating good separation performance.

[0189] Table 15 Results of relative retention time for different flow velocities

[0190]

[0191] Table 16 Results of relative peak area at different flow velocities

[0192]

[0193]

[0194] B. Investigation at different column temperatures

[0195] The test solution was prepared according to the preparation method of the test solution in Example 1. The test solution was injected and detected under the chromatographic conditions in Example 1. The only difference was that the determination was carried out at column temperatures of 23℃, 25℃ and 27℃ to examine the separation effect of each characteristic peak when the column temperature changed.

[0196] The results are shown in Tables 17-18 below. The results indicate that all chromatographic peaks were detected at different column temperatures, demonstrating good separation performance.

[0197] Table 17 Comparison of relative retention time results at different column temperatures

[0198]

[0199] Table 18 Results of relative peak areas at different column temperatures

[0200]

[0201]

[0202] C. Investigation using different instruments

[0203] The test solution was prepared according to the preparation method of the test solution in Example 1. The test solution was injected and detected under the chromatographic conditions in Example 1. The only difference was that the detection was carried out on Waters instrument and Thermo ultra-high performance liquid chromatography to examine the separation effect of each characteristic peak of different instruments.

[0204] The results are shown in Tables 19-20 below. The results indicate that the chromatographic peaks were detected on all instruments, demonstrating good peak separation.

[0205] Table 19 Comparison of Relative Retention Time Measurement Results from Different Instruments

[0206]

[0207] Table 20 Relative peak area results for different instruments

[0208]

[0209] D. Investigation of different chromatographic columns

[0210] The test solution was prepared according to the preparation method of the test solution in Example 1. The test solution was injected and detected under basically the chromatographic conditions in Example 1. The only difference was that Waters ACQUITYUPLC HSS T3 (2.1×150mm, 1.8μm) and Luna Omega PS C18100A (150mm×2.1mm, 1.6μm) columns were used for injection analysis.

[0211] The results are shown in Tables 21-22 below. The results show that all characteristic peaks in the chromatograms corresponding to different columns were detected and the separation effect was good. However, the retention times of the chromatographic peaks varied significantly among different columns, with peak 2 showing the largest difference in relative retention time. Therefore, the Waters ACQUITYUPLC HSS T3 (2.1×150mm 1.8μm) was the preferred column.

[0212] Table 21 Comparison of relative retention time determination results for different chromatographic columns

[0213]

[0214] Table 22 Results of relative peak areas for different chromatographic columns

[0215]

[0216] The above methodological investigation results show that among the 15 common peaks in the characteristic chromatogram of the freeze-dried powder of the standard decoction of roasted barley sprouts established in Example 1, the retention time of the chromatographic peaks is affected by different flow rates, column temperatures, etc. In order to increase the applicability of the method, the specified value range of the relative retention time is controlled within ±10%.

[0217] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A method for constructing a characteristic spectrum of germinated rice or roasted germinated rice and its preparations, characterized in that, Includes the following steps: (1) Preparation of test solution: 1) Take the analyte, add solvent and extract ultrasonically to obtain the extract; 2) Separate the solid and liquid of the extract, take the liquid, pass the liquid through the separation column, add the eluent for elution, collect the eluent and dry and dissolve it; this is the test solution; the solvent is selected from methanol; the extraction solvent is ethyl acetate; the eluent is a methanol solution containing 0.5-2% formic acid by volume; the separation column is an acidic alumina column; the construction method also includes the step of preparing a reference solution using uridine, adenosine, guanosine, 5-hydroxymethylfurfural, 4-coumaric acid and ferulic acid reference standards; (2) The test solution and the reference solution were detected by ultra-high performance liquid chromatography. The packing material was octadecylsilane bonded silica gel. The mobile phase included methanol and formic acid aqueous solution. Gradient elution was performed. The gradient elution program included: 0 → 8 minutes → 38 minutes → 53 minutes. The volume percentage of methanol in the mobile phase was 0% → 3% → 32% → 32%.

2. The construction method according to claim 1, characterized in that, Step (1) also satisfies any one or more of the following AD: A. In step 1), the mass ratio of the analyte to the volume of the solvent is 0.2-0.7g:10-40mL; B. In step 1), the extraction time is 10 min to 1 h; C. In step 2), the solid-liquid separation is performed by centrifugation or filtration; D. In step 1), the test substance is selected from one or more of the following: germinated rice medicinal materials, germinated rice or roasted germinated rice slices, germinated rice or roasted germinated rice granules, and germinated rice or roasted germinated rice powder.

3. The construction method according to claim 1, characterized in that, The height of the separation column is 5cm-12cm; and / or the drying is carried out by evaporation at 55-100°C.

4. The construction method according to claim 1, characterized in that, Step (2) The chromatographic conditions for ultra-high performance liquid chromatography also include: detection wavelength of 250-330 nm, flow rate of 0.23-0.27 ml / min, column temperature of 23-30 ℃, and injection volume of 1-2 μl.

5. The construction method according to claim 4, characterized in that, The detection wavelength is 270nm from 0 to 23 minutes, and 310nm after 23 minutes.

6. The construction method according to claim 4, characterized in that, The column temperature is 24-26 ℃.

7. The construction method according to claim 4, characterized in that, The injection volume is 1-1.5 μl.

8. The construction method according to claim 1, characterized in that, The chromatographic column has a length of 100-150 mm, an inner diameter of 2.1 mm, and a particle size of 1.6-1.8 μm; and / or, the volume percentage of formic acid in the formic acid aqueous solution in the mobile phase is 0.08-0.12%.

9. The construction method according to claim 1, characterized in that, The gradient elution program further includes 53 minutes → 54 minutes, with the volume percentage of methanol in the mobile phase changing from 32% to 0%; or, the gradient elution program further includes 53 minutes → 54 minutes → 60 minutes, with the volume percentage of methanol in the mobile phase changing from 32% to 0% to 0%.

10. The construction method according to any one of claims 1-9, characterized in that, The characteristic spectra of germinated rice or roasted germinated rice and their preparations obtained by the construction method are selected from any one of the following (1)-(2): (1) The characteristic chromatogram of the roasted germinated barley and its preparation has 15 characteristic peaks. Peak 4 is used as reference peak 1, and peak 12 is used as reference peak 2. The relative retention times of peaks 1-3 and 5-6 with reference peak 1, and the relative retention times of peaks 7-11 and 13-15 with reference peak 2 are within ±10% of the specified values. The specified values ​​of peaks 1-3, 5-6, 7-11, and 13-15 are 0.63, 0.72, 0.85, and 1.1, respectively. 2, 1.15, 0.70, 0.72, 0.83, 0.86, 0.91, 1.03, 1.09, 1.29; or, the characteristic chromatogram of the germinated rice and its preparation has 13 characteristic peaks, with peak 4 as reference peak 1 and peak 12 as reference peak 2, and the relative retention times of characteristic peaks 1 and 5-6 with reference peak 1 and the relative retention times of peaks 7-11 and 13-15 with reference peak 2 are within ±10% of the specified values; the specified values ​​are: 0.63、1.12、1.15、0.70、0.72、0.83、0.86、0.91、1.03、1.09、1.29； (2) The characteristic chromatogram of the roasted germinated barley and its preparation has 15 characteristic peaks. The peak corresponding to the adenosine reference standard peak is reference peak 1, and the peak corresponding to the 4-coumaric acid reference standard peak is reference peak 2. The relative retention times of peaks 1-3, 5-6 and reference peak 1, and peaks 7-11, 13-15 and reference peak 2 are within ±10% of the specified values. The specified values ​​of peaks 1-3, 5-6, 7-11, and 13-15 are 0.63, 0.72, 0.85, and 1.1, respectively. 2, 1.15, 0.70, 0.72, 0.83, 0.86, 0.91, 1.03, 1.09, 1.29; or, the characteristic chromatogram of the germinated rice and its preparation has 13 characteristic peaks, with the peak corresponding to the adenosine reference standard peak as reference peak 1, and the peak corresponding to the 4-coumaric acid reference standard peak as reference peak 2. The relative retention times of characteristic peaks 1 and 5-6 with reference peak 1 and the relative retention times of peaks 7-11 and 13-15 with reference peak 2 are within ±10% of the specified values; the specified values ​​are: 0.63、1.12、1.15、0.70、0.72、0.83、0.86、0.91、1.03、1.09、1.29。 11. A method for quality testing of germinated rice or roasted germinated rice and its preparations, characterized in that, This includes the step of constructing a feature map of the product to be tested using the product to be tested as a test sample according to any one of the construction methods described in claims 1-10.

12. A method for distinguishing between germinated rice and its preparations and roasted germinated rice and its preparations, characterized in that, include: The characteristic spectrum of the product to be identified is constructed using the test sample according to any one of the construction methods described in claims 1-10; The obtained feature map of the product to be identified is compared with the feature map constructed by any of the construction methods described in claims 1-10, and identification is performed based on the comparison results.

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