Establishment method and application of liqueur characteristic chromatogram

Abstract

The invention discloses a liqueur characteristic chromatogram establishing method and application, the liqueur characteristic chromatogram establishing method is characterized in that a high performance liquid chromatograph is used for detecting the characteristic chromatogram of a liqueur sample, during detection, acetonitrile and a 0.1% phosphoric acid aqueous solution are selected as mobile phases, gradient elution is carried out, the volume ratio of acetonitrile in the mobile phases is 5-22%, and the collection wavelength is 210-330 nm. According to the method, acetonitrile and a 0.1% phosphoric acid aqueous solution are used as mobile phases for gradient elution of the liqueur sample, and the chromatogram of the sample is collected by adopting a variable wavelength means, so that various substance components in the liqueur sample can be accurately separated, and the characteristic chromatogram with obvious and complete characteristic peak pattern is obtained. According to the liqueur quality control method, whether the quality of the liqueur is stable or changed is determined by comparing the characteristic chromatograms of the blending liquid, the fine filtrate and the finished liqueur sample in the preparation process of any batch of liqueur, so that the liqueur and the quality of the liqueur in the preparation process are controlled.

Description

Technical Field

[0001] This invention relates to the field of liquid chromatography technology, specifically to a method for constructing characteristic chromatograms of spirits and its application. Background Technology

[0002] Liqueurs are beverages with a specific style made by using baijiu or huangjiu as a base, adding traditional food and medicinal materials or specific food ingredients, and then processing them through processes such as maceration or redistillation, or by directly adding specific components extracted from food.

[0003] Chinese medicinal herbs themselves have very complex components. Liquor is made from a variety of Chinese medicinal herbs and base liquor. Many substances in Chinese medicinal herbs have good solubility in ethanol, and many components can be effectively extracted. Moreover, the components affect each other, making it difficult to ensure complete separation of each component in liquid chromatography analysis, thus increasing the difficulty of quality control.

[0004] Existing technologies have studied the functional components or fingerprint spectra of certain liqueurs, but a comprehensive study of the characteristic spectra of liqueurs has not been conducted, and there is a lack of criteria for determining the corresponding chromatographic peaks of characteristic components in liqueurs and methods for quality control. The problem this invention aims to solve is how to establish a characteristic spectra detection method tailored to the characteristics of liqueurs. This method involves eluting liqueur samples with a specific mobile phase and gradient elution, and using variable wavelength technology to set reasonable acquisition wavelengths at different time periods. This allows for accurate and effective separation of various substances in the liqueur, ensuring that the characteristic components are fully presented and effectively separated on the chromatogram, resulting in characteristic spectra with the characteristics of liqueurs. Furthermore, the criteria for determining the corresponding chromatographic peaks of each characteristic component are clearly defined. By tracking and collecting characteristic spectra from multiple batches of qualified liqueur products, a representative control spectra have been established. The blending solution, filtrate, and finished liqueur products from actual production are collected according to the established method. The collected characteristic spectra are compared and analyzed with the control spectra. The analysis results are used to comprehensively judge the stability and quality of the liqueur, elucidate the quantitative and qualitative transfer laws in the liqueur preparation process, and provide a scientific basis for formulating process standards and quality standards for liqueurs. Summary of the Invention

[0005] The purpose of this invention is to provide a method and application for establishing characteristic chromatograms of liqueurs. This method can effectively separate various effective components in liqueurs using liquid chromatography technology to obtain complete characteristic chromatograms, and clarify the judgment criteria for the corresponding chromatographic peaks of each characteristic component and the similarity criteria of the sample characteristic chromatograms, thereby achieving the purpose of quality control.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides a method for establishing a characteristic spectrum of a liqueur, including detecting the characteristic spectrum of a liqueur sample using high performance liquid chromatography.

[0008] Acetonitrile and 0.1% phosphoric acid aqueous solution were used as the mobile phase for detection, and gradient elution was adopted. The volume ratio of acetonitrile in the mobile phase was 5% to 22%; the flow rate of the mobile phase was 1.0 mL / min.

[0009] The liqueur is made from ingredients including 6-9 parts wolfberry, 3-5 parts eucommia, 4-6 parts dodder seed, 1-2 parts cinnamon, and light-aroma baijiu (Chinese white liquor).

[0010] In the above technical solution, this application uses high-performance liquid chromatography (HPLC) to detect the characteristic chromatograms of liqueur samples. The liqueur is made from raw materials including wolfberry, eucommia, dodder seed, cinnamon, and light-aroma baijiu (Chinese white liquor) using small-batch fermentation. This liqueur contains a variety of effective components. This application uses acetonitrile and 0.1% phosphoric acid aqueous solution as the mobile phase for gradient elution of the liqueur sample, which can accurately separate various components in the liqueur sample and obtain characteristic chromatograms with clear and complete characteristic peaks.

[0011] In conjunction with the first aspect, in a first possible example of the first aspect of this application, the chromatographic column used for the above detection is a C18 reverse chromatographic column.

[0012] In the example above, this chromatographic column facilitates the accurate and complete separation of various substances in the liqueur using high-performance liquid chromatography (HPLC), which are then displayed in the chromatogram.

[0013] In conjunction with the first aspect, in a second possible example of the first aspect of this application, the column temperature of the chromatographic column during the above detection is 25–35°C.

[0014] In the example above, the detection of column temperature in this chromatographic column is beneficial for the high-performance liquid chromatograph to accurately and completely separate the various components in the liqueur and display them in the chromatogram.

[0015] In conjunction with the first aspect, in a third possible example of the first aspect of this application, the gradient elution described above is performed using the following method:

[0016] During the period of 0–5 min, the volume percentage of acetonitrile in the mobile phase is 5%.

[0017] Within 5–20 minutes, the volume percentage of acetonitrile in the mobile phase increased from 5% to 9%.

[0018] Between 20 and 60 minutes, the volume percentage of acetonitrile in the mobile phase increased from 9% to 22%.

[0019] During the 60–62 min period, the volume ratio of acetonitrile in the mobile phase was 22%.

[0020] In the above example, eluting the mobile phase using the above method is beneficial for separating various effective components in the liqueur, thus allowing them to be fully and clearly presented in the spectrum.

[0021] In conjunction with the first aspect, in a fourth possible example of the first aspect of this application, the ultraviolet detector is set to a detection wavelength of 210–330 nm when performing the above-mentioned liquid phase detection.

[0022] In conjunction with the first aspect, in a fifth possible example of the first aspect of this application, the spectrum is acquired using the following variable wavelength method:

[0023] The acquisition wavelength is 220–240 nm from 0 to 20 minutes.

[0024] The acquisition wavelength is 320–330 nm during the period of 20–38 min.

[0025] The acquisition wavelength is 210–220 nm for 38–50 min;

[0026] The acquisition wavelength is 280-290 nm for 50-62 minutes.

[0027] In the above example, through experiments and analysis, it was found that the set detection wavelength of the ultraviolet detector for each detection time period is suitable for the presentation and effective separation of the corresponding chromatographic peaks of various effective substances in the liqueur, which can ensure that the effective substances in the liqueur can be fully presented in the spectrum and form a characteristic spectrum with product characteristics.

[0028] In conjunction with the first aspect, in a sixth possible example of the first aspect of this application, when the spectrum is acquired, the acquisition wavelength is 240 nm from 0 to 20 min; 330 nm from 20 to 38 min; 210 nm from 38 to 50 min; and 290 nm from 50 to 62 min.

[0029] In conjunction with the first aspect, in the seventh possible example of the first aspect of this application, the test solutions of the blending liquid sample, the filtrate sample, and the finished liqueur sample are obtained by the following means:

[0030] Take 2 mL each of the prepared solution sample, the fine filtrate sample, and the finished liquor sample, evaporate them to dryness under reduced pressure at 40℃, dissolve the residue in water, pass them through a solid phase extraction column, elute with 15 mL of water, remove the eluent, elute with 5 mL of 10% ethanol solution, remove the eluent, elute with 50% methanol, collect the eluent and make up to 5 mL.

[0031] In a second aspect, this application provides a method for quality control of liqueur, which includes taking samples of blending liquid, fine filtrate, and finished liqueur from different batches of liqueur during the preparation process, detecting the characteristic spectra using the aforementioned method for establishing characteristic spectra of liqueur, and clarifying the criteria for determining the chromatographic peaks corresponding to each characteristic component and the similarity criteria of the sample characteristic spectra, thereby achieving the purpose of quality control.

[0032] In the above technical solution, this application compares the characteristic chromatograms of the blending solution, filtrate, and finished liqueur samples of different batches of liqueur during the preparation process. By comparing the similarity of the number of common chromatographic peaks, retention times, and response values, it determines whether the various effective substances in the blending solution, filtrate, and finished liqueur samples during the preparation process are the same and stable, and whether the various effective substances in the liqueur change during the preparation process and in different batches, thereby achieving the control of the quality of the liqueur during the preparation process and the finished product.

[0033] In conjunction with the second aspect, in a first possible example of the second aspect of this application, the characteristic chromatogram includes 11 common chromatographic peaks, wherein peak 5 is chlorogenic acid, and peak 5 is used as a control peak. The relative retention times of each characteristic peak are as follows:

[0034] Peak 1: Relative retention time is 0.355;

[0035] Peak 2: Relative retention time is 0.488;

[0036] Peak 3: Relative retention time is 0.543;

[0037] Peak 4: Relative retention time is 0.662;

[0038] Peak 5: Relative retention time is 1.000;

[0039] Peak 6: Relative retention time is 1.029;

[0040] Peak 7: Relative retention time is 1.347;

[0041] Peak 8: Relative retention time is 1.380;

[0042] Peak 9: Relative retention time is 1.469;

[0043] Peak 10: Relative retention time is 1.617;

[0044] Peak 11: Relative retention time is 1.840;

[0045] Judgment criteria: The relative retention time of each characteristic peak should be within 5%;

[0046] In the above technical solution, through experiments and analysis, the chromatographic peaks corresponding to each characteristic component in the liqueur are stably presented on the characteristic spectrum, which can accurately identify each characteristic component in the liqueur.

[0047] In conjunction with the second aspect, in a second possible example of the second aspect of this application, the characteristic chromatograms of any batch of blending solution, fine filtrate, or finished liquor product from actual production are compared and analyzed with the control characteristic chromatograms. The similarity of the characteristic chromatograms is obtained by analysis based on the similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine (version 2012.130723). Multiple batches of qualified samples are selected, and the common pattern of HPLC characteristic chromatograms is established by using the median method and the Mark peak matching method of the above 11 common characteristic peaks to generate the control characteristic chromatogram. Then, any batch of samples is compared and analyzed with the control characteristic chromatograms to calculate the similarity of the samples. The similarity of the sample characteristic chromatograms is not less than 0.99.

[0048] In the above technical solution, by comparing and analyzing the characteristic spectra of semi-finished products and finished liqueurs, the method of the present invention can be used for overall control of the entire actual production process of liqueurs, with strong operability, and at the same time provides a scientific basis for formulating liqueur process standards and quality standards. Attached Figure Description

[0049] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly described below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation on the scope:

[0050] Figure 1 These are the HPLC characteristic chromatograms obtained in Example 1 and Comparative Examples 1-4 of this application;

[0051] Figure 2 This is the HPLC reference characteristic chromatogram obtained in Example 2 of this application;

[0052] Figure 3 This is a characteristic chromatogram of the HPLC sample obtained in Example 3 of this application. Detailed Implementation

[0053] The embodiments of this application will be described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are for illustrative purposes only and should not be considered as limiting the scope of this application. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0054] High-performance liquid chromatography (HPLC) is an important branch of chromatography. Using a liquid as the mobile phase, a high-pressure delivery system pumps single solvents or mixed solvents of different polarities, buffer solutions, and other mobile phases into a chromatographic column packed with a stationary phase. After separation within the column, the components are detected by a detector, thus enabling sample analysis. However, the methods for detecting different complex samples using HPLC vary. Inappropriate detection methods and conditions can lead to the appearance or incomplete separation of sample-specific components, resulting in chromatograms that lack typical sample characteristics. The following describes in detail a method for establishing characteristic chromatograms of a spirit or liqueur and a method for quality control of a spirit or liqueur, based on embodiments of this application.

[0055] This invention provides a method for establishing characteristic spectra of liqueurs, which uses HPLC to detect liqueur samples.

[0056] The liqueur is made from ingredients including 6-9 parts wolfberry, 3-5 parts eucommia, 4-6 parts dodder seed, 1-2 parts cinnamon, and light-aroma baijiu.

[0057] Based on the effective components of the above-mentioned traditional Chinese medicine ingredients, and considering the need to analyze and eliminate interference between different substances, selecting appropriate chromatographic columns, mobile phases, and elution conditions, as well as choosing suitable acquisition wavelengths at different sampling time points, is beneficial for separating various effective components in the liqueur. This ensures that the corresponding chromatographic peaks of each effective component are fully presented in the chromatogram, resulting in accurate chromatographic data and forming the unique characteristic chromatogram of the liqueur.

[0058] This application uses 5-10 μL of sample for HPLC analysis. Acetonitrile and 0.1% phosphoric acid aqueous solution are used as the mobile phase, gradient elution is adopted, the volume ratio of acetonitrile in the mobile phase is 5%-22%, the flow rate of the mobile phase is 1.0 mL / min, and the acquisition wavelength is 210-330 nm.

[0059] Preferably, the chromatographic column used in this application is a C18 reverse-phase chromatographic column, and the column temperature during detection is 25–35°C. The chromatographic column is selected based on the active ingredients and impurities in the sample, as well as their type, structure, polarity, acidity / alkalinity, and molecular weight. Correct selection of the chromatographic column and setting of the detection column temperature facilitate the accurate and complete separation of various substances in the liqueur and their display in the chromatogram.

[0060] Preferably, when performing gradient elution in this application, the following elution method is adopted: From 0 to 5 min, the volume percentage of acetonitrile in the mobile phase is maintained at 5%, and the volume percentage of 0.1% phosphoric acid aqueous solution is maintained at 95%; from 5 to 20 min, the volume percentage of acetonitrile in the mobile phase increases from 5% to 9%, and the volume percentage of 0.1% phosphoric acid aqueous solution decreases from 95% to 91%; from 20 to 60 min, the volume percentage of acetonitrile in the mobile phase increases from 9% to 22%, and the volume percentage of 0.1% phosphoric acid aqueous solution decreases from 91% to 78%; from 60 to 62 min, the volume percentage of acetonitrile in the mobile phase is maintained at 22%, and the volume percentage of 0.1% phosphoric acid aqueous solution is maintained at 78%.

[0061] Optionally, the measurement wavelength of the ultraviolet detector is set to 210–330 nm, wherein: from 0 to 20 min, the acquisition wavelength is 220–240 nm; from 20 to 38 min, the acquisition wavelength is 320–330 nm; from 38 to 50 min, the acquisition wavelength is 210–220 nm; and from 50 to 62 min, the acquisition wavelength is 280–290 nm. Selecting this wavelength for the acquisition spectrum ensures that the effective components in the liqueur exhibit specific peaks in the spectrum.

[0062] Preferably, the measurement wavelength of the ultraviolet detector is set to 210–330 nm, wherein the acquisition wavelength is 240 nm from 0 to 20 min; 330 nm from 20 to 38 min; 210 nm from 38 to 50 min; and 290 nm from 50 to 62 min.

[0063] Preferably, the preparation method of the test solution of the blending solution sample, the fine filtrate sample and the finished liquor sample of this application is as follows: Take 2 mL of each of the blending solution sample, the fine filtrate sample and the finished liquor sample, evaporate to dryness under reduced pressure at 40℃, dissolve the residue in water, pass through a solid phase extraction column, elute with 15 mL of water, remove the eluent, elute with 5 mL of 10% ethanol solution, remove the eluent, elute with 50% methanol, collect the eluent and make up to 5 mL.

[0064] This application also provides a method for quality control of liqueur, which includes using the above-mentioned method for establishing characteristic chromatograms of liqueur to detect the characteristic chromatograms of the blended liquid, the finely filtered liquid and the finished liqueur samples during the production process. All characteristic chromatograms include 11 common characteristic chromatographic peaks, and the criteria for determining the corresponding chromatographic peaks of each characteristic component and the similarity criteria of the sample characteristic chromatograms are clearly defined.

[0065] This application compares the similarity of the number, retention time, and response value of common chromatographic peaks in the characteristic spectra of the blending solution, the fine filtrate, and the finished liqueur samples during the preparation process of the liqueur. By comparing the differences in the spectra, the differences in various effective substances in the liqueur can be reflected, thereby verifying whether the various effective substances in the liqueur have changed during the preparation process and in different batches, thus achieving the goal of controlling the quality of the liqueur during the preparation process.

[0066] Characteristic chromatographic peaks are identified and determined based on their relative retention times:

[0067] The characteristic chromatogram includes 11 characteristic chromatographic peaks, with peak 5 representing chlorogenic acid and serving as the reference peak. The relative retention times of each characteristic peak should meet the following requirements: Peak 1: relative retention time 0.355; Peak 2: relative retention time 0.488; Peak 3: relative retention time 0.543; Peak 4: relative retention time 0.662; Peak 5: relative retention time 1.000; Peak 6: relative retention time 1.029; Peak 7: relative retention time 1.347; Peak 8: relative retention time 1.380; Peak 9: relative retention time 1.469; Peak 10: relative retention time 1.617; Peak 11: relative retention time 1.840. The relative retention times of each characteristic peak should be within ±5%.

[0068] The similarity of the feature maps is calculated using the following method:

[0069] All characteristic chromatogram similarities were obtained using the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System Software (version 2012.130723). All characteristic chromatograms to be compared were imported into the software. Multiple batches of qualified samples were selected, and a common pattern of HPLC characteristic chromatograms was established using the median method and the Mark peak matching method based on the 11 common characteristic peaks mentioned above to generate a reference characteristic chromatogram. Then, any batch of samples was compared with the reference characteristic chromatogram to calculate the sample similarity. The sample characteristic chromatogram similarity was not less than 0.99.

[0070] The following describes in further detail a method for establishing a characteristic spectrum of liqueur and a method for quality control of liqueur, based on embodiments of the present application.

[0071] Example 1

[0072] This embodiment provides a method for establishing a characteristic spectrum of liqueurs, including the following steps:

[0073] 1. Sample preparation

[0074] Take 2 mL of the liqueur sample, evaporate it to dryness under reduced pressure at 40℃, dissolve the residue in water, pass it through a solid phase extraction column, elute with 15 mL of water, remove the eluent, then elute with 5 mL of 10% ethanol solution, remove the eluent, and then elute with 50% methanol. Collect the eluent and make up to 5 mL.

[0075] 2. Establish characteristic fingerprints of liqueurs

[0076] In this embodiment, 10 μL of liqueur sample was taken for HPLC analysis. A C18 reverse-phase column was selected. The column temperature was 25–35 °C. The wavelength of the UV detector was set to 210–330 nm. Acetonitrile and 0.1% phosphoric acid aqueous solution were used as the mobile phase, and the flow rate was 1.0 mL / min.

[0077] The mobile phase was eluted in the following manner, as shown in Table 1:

[0078] Table 1: Gradient elution method of mobile phase in Example 1

[0079]

[0080] The wavelength settings for each time period are shown in Table 2:

[0081] Table 2: Acquisition wavelengths corresponding to different detection time periods in Example 1

[0082]

[0083] The obtained HPLC characteristic chromatograms are as follows Figure 1 As shown, the chromatographic peaks corresponding to various substances are presented completely with high separation and signal-to-noise ratio.

[0084] Comparative Example 1

[0085] This comparative example provides a method for establishing the characteristic spectrum of liqueurs, including the following steps:

[0086] The liqueur sample obtained in Example 1 was subjected to HPLC analysis. A C18 reverse-phase column was used. The column temperature was 25–35 °C. The wavelength of the UV detector was set to 210 nm. Acetonitrile and 0.1% phosphoric acid aqueous solution were used as the mobile phase. The flow rate was 1.0 mL / min, and the injection volume was 10 μL. Elution was performed using the mobile phase as shown in Table 5.

[0087] Table 5. Comparative Example 1: HPLC Detection of Mobile Phase and Flow Rate

[0088]

[0089] The obtained HPLC characteristic chromatograms are as follows Figure 1 As shown.

[0090] Comparative Example 2

[0091] This comparative example provides a method for establishing the characteristic spectrum of liqueurs, including the following steps:

[0092] The liqueur samples prepared in Example 1 were subjected to HPLC analysis. A C18 reverse-phase column was used. The column temperature was 25–35°C. Acetonitrile and 0.1% phosphoric acid aqueous solution were used as the mobile phase, and the elution method was the same as in Example 1. The flow rate was 1.0 mL / min, and the injection volume was 10 μL. The wavelengths selected for each time period are shown in Table 6.

[0093] Table 6 shows the acquisition wavelengths corresponding to different detection time periods in Comparative Example 2.

[0094]

[0095] The obtained HPLC characteristic chromatograms are as follows Figure 1 As shown.

[0096] Comparative Example 3

[0097] This comparative example provides a method for establishing the characteristic spectrum of liqueurs, including the following steps:

[0098] CN117092252A discloses a method for constructing a fingerprint spectrum of a compound health wine. This method uses a compound health wine made from ginseng, polygonatum, eucommia male flowers, and baijiu (Chinese white liquor) as the sample, and octadecylsilane-bonded silica gel as the packing material; the detection wavelength is 270 nm; and elution is performed using a gradient of acetonitrile and 0.1% formic acid aqueous solution. This comparative example processes a liqueur sample according to the method provided in Example 3 of this patent and detects the chromatogram. The obtained HPLC characteristic spectrum is shown below. Figure 1 As shown.

[0099] Comparative Example 4

[0100] CN109709250A discloses a method for detecting the fingerprint spectrum of ginseng and deer antler wine. This method uses ginseng and deer antler wine, made from raw materials such as ginseng, deer antler, prepared rehmannia root, and baijiu (Chinese white liquor), as the sample, and the detection wavelength is 203 nm; elution is performed using a gradient of acetonitrile and water. This comparative example uses the method provided in Example 1 of this patent to process a liqueur sample and detect the chromatogram. The obtained HPLC characteristic chromatogram is shown below. Figure 1 As shown.

[0101] Comparison Figure 1Of the five spectra obtained in Example 1, the characteristic spectrum showed complete chromatographic peaks of all components, good separation, and a high signal-to-noise ratio, with each active ingredient corresponding to its characteristic peak. In contrast, the spectrum obtained in Comparative Example 1 showed poor sample separation, with almost all substances exiting the column together, failing to display the characteristic peaks corresponding to each component, indicating that the gradient elution method in Comparative Example 1 was unsuitable. The spectrum obtained in Comparative Example 2 only showed 5-6 characteristic peaks, and some peaks had low response values, indicating that the acquisition wavelength set in Comparative Example 2 could not completely and reasonably present the characteristic peaks of each component of the liqueur, and therefore could not obtain a representative characteristic spectrum. The spectra obtained by the methods provided in Comparative Examples 3 and 4 showed large baseline fluctuations and could not effectively separate the various components in the sample.

[0102] Example 2

[0103] This embodiment provides a method for quality control of liqueur, including the following steps:

[0104] 1. Sample preparation

[0105] Ten batches of liqueur samples were randomly selected from the production process, and the samples were prepared according to the sample preparation method in Example 1.

[0106] 2. Establish a quality control system for liqueurs.

[0107] Following the HPLC method described in Example 1, characteristic chromatograms of 10 batches of liquor samples were collected. All characteristic chromatograms were imported into the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System software (version 2012.130723). HPLC characteristic chromatograms were established using the median method and Mark peak matching method, generating a control characteristic chromatogram consisting of 11 common characteristic peaks. Figure 2 As shown.

[0108] Using peak 5 as the control peak, the relative retention times of each characteristic peak of 10 batches of liqueur were calculated and the average value was calculated. The results are shown in Table 3 below.

[0109] Table 3. Relative retention times of various characteristic components in the characteristic chromatograms of 10 batches of liquor.

[0110]

[0111]

[0112] As shown in Table 3, the average relative retention times are: peak 1: 0.355, peak 2: 0.488, peak 3: 0.543, peak 4: 0.662, peak 5: 1.000, peak 6: 1.029, peak 7: 1.347, peak 8: 1.380, peak 9: 1.469, peak 10: 1.617, and peak 11: 1.840. The relative retention times of the 11 characteristic peaks in each sample are all within ±5%, which can be used as the criterion for determining the corresponding chromatographic peaks of each characteristic component.

[0113] Example 3

[0114] This embodiment provides a method for quality control of liqueur, including the following steps:

[0115] 1. Sample preparation

[0116] Samples were prepared by taking the blending liquid, the fine filtrate, and the finished liqueur, respectively, according to the sample preparation method in Example 1.

[0117] 2. Establish a quality control system for liqueurs.

[0118] Following the HPLC method established in Example 1, the HPLC characteristic chromatograms of the above-mentioned blending solution, fine filtrate, and finished liquor samples were obtained. The chromatographic peaks of each characteristic chromatogram were determined according to the criteria for determining the corresponding chromatographic peaks of characteristic components in Example 2. There should be 11 identical characteristic peaks at the corresponding positions. Furthermore, the similarity was calculated using the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System software (version 2012.130723) using the median method and the Mark peak matching method based on the above 11 common characteristic peaks. The chromatograms and data are as follows: Figure 3 And as shown in Table 4:

[0119] Table 4. Results of Feature Map Similarity Calculation

[0120]

[0121] Note: The above are all average values ​​calculated from multiple batches of samples.

[0122] As shown in Table 2, the similarity of each chromatogram is greater than 0.99.

[0123] In summary, the method for establishing characteristic spectra of liqueur provided in this application involves detecting the characteristic spectra of liqueur samples using high-performance liquid chromatography (HPLC). The liqueur is made from raw materials including wolfberry, eucommia, dodder seed, cinnamon, and light-aroma baijiu (Chinese white liquor) using small-batch elution. This liqueur contains various effective components. This application uses acetonitrile and a 0.1% phosphoric acid aqueous solution as the mobile phase, eluting the liqueur sample using a specific gradient elution method. By employing a variable wavelength technique and setting appropriate acquisition wavelengths at different time periods, various components in the liqueur sample can be accurately and effectively separated, resulting in a characteristic spectrum with distinct and complete peak shapes.

[0124] A method for quality control of liqueurs involves comparing the characteristic chromatograms of the blending solution, filtrate, and finished liqueur samples from different batches during the preparation process. By comparing the similarity of the number of common chromatographic peaks, retention times, and response values, the method determines whether the various effective components in the blending solution, filtrate, and finished liqueur samples are the same, and whether the various effective components change during the preparation process and between different batches, thereby controlling the quality of the liqueur during the preparation process.

[0125] The above description is merely a specific embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A method for establishing a characteristic spectrum of spirits, characterized in that, The method for establishing the characteristic spectrum of the liqueur includes detecting the characteristic spectrum of the liqueur sample using a high-performance liquid chromatograph. For the detection, acetonitrile and a 0.1% phosphoric acid aqueous solution were used as the mobile phase, with the volume ratio of acetonitrile in the mobile phase being 5%–22%; the flow rate of the mobile phase was 1.0 mL / min; gradient elution was performed using the following method: During the period of 0–5 min, the volume percentage of acetonitrile in the mobile phase is 5%. Within 5–20 minutes, the volume percentage of acetonitrile in the mobile phase increased from 5% to 9%. Between 20 and 60 minutes, the volume percentage of acetonitrile in the mobile phase increased from 9% to 22%. During the 60–62 min period, the volume percentage of acetonitrile in the mobile phase was 22%. The ultraviolet detector is set to collect wavelengths of 210–330 nm.

2. The method for establishing the characteristic spectrum of liqueurs according to claim 1, characterized in that, The chromatographic column used for the detection was a C18 reverse-phase column.

3. The method for establishing the characteristic spectrum of liqueurs according to claim 2, characterized in that, The column temperature of the chromatographic column during detection is 25–35°C.

4. The method for establishing the characteristic spectrum of liqueurs according to claim 1, characterized in that, The liqueur is made from raw materials including 6-9 parts of wolfberry, 3-5 parts of eucommia, 4-6 parts of dodder seed, 1-2 parts of cinnamon, and light-aroma baijiu.

5. The method for establishing the characteristic spectrum of liqueur according to claim 1, wherein the spectrum is acquired using the following variable wavelength method: The acquisition wavelength is 220–240 nm from 0 to 20 minutes. The acquisition wavelength is 320–330 nm during the period of 20–38 min. The acquisition wavelength is 210–220 nm for 38–50 min; The acquisition wavelength is 280-290 nm for 50-62 minutes.

6. The method for establishing the characteristic spectrum of liqueurs according to claim 5, characterized in that, During the acquisition of the spectrum, the acquisition wavelength is 240 nm from 0 to 20 min; 330 nm from 20 to 38 min; 210 nm from 38 to 50 min; and 290 nm from 50 to 62 min.

7. The method for quality control of liqueur according to claim 1, characterized in that, The blending solution sample, the filtrate sample, and the finished liqueur sample were obtained through the following methods: Take 2 mL each of the prepared solution sample, the fine filtrate sample, and the finished liquor sample, evaporate them to dryness under reduced pressure at 40℃, dissolve the residue in water, pass them through a solid phase extraction column, elute with 15 mL of water, remove the eluent, elute with 5 mL of 10% ethanol solution, remove the eluent, elute with 50% methanol, collect the eluent and make up to 5 mL.

8. A method for quality control of liqueur, characterized in that, Includes the following steps: (1) Take samples of the blending liquid, the fine filtrate and the finished liqueur from different batches of liqueur during the preparation process and collect characteristic spectra using the liqueur characteristic spectra establishment method described in any one of claims 1 to 7. (2) Compare and analyze the characteristic chromatograms obtained in step (1) with the control characteristic chromatograms. The similarity of the characteristic chromatograms is obtained by analysis based on the similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine (version 2012.130723). Select multiple batches of qualified samples, and use the median method and the Mark peak matching method of the above 11 common characteristic peaks to establish the common pattern of HPLC characteristic chromatograms to generate the control characteristic chromatograms. Then compare and analyze any batch of samples with the control characteristic chromatograms to calculate the similarity of the samples. The similarity of the sample characteristic chromatograms is not less than 0.

99.

9. The method for quality control of liqueur according to claim 8, characterized in that, The characteristic spectrum includes 11 common characteristic peaks, of which peak 5 is chlorogenic acid, and peak 5 is used as the control peak. The relative retention times of each characteristic peak are as follows: Peak 1: Relative retention time is 0.355; Peak 2: Relative retention time is 0.488; Peak 3: Relative retention time is 0.543; Peak 4: Relative retention time is 0.662; Peak 5: Relative retention time is 1.000; Peak 6: Relative retention time is 1.029; Peak 7: Relative retention time is 1.347; Peak 8: Relative retention time is 1.380; Peak 9: Relative retention time is 1.469; Peak 10: Relative retention time is 1.617; Peak 11: Relative retention time is 1.840; The relative retention time of each characteristic peak should be within 5%.

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