Construction method, detection method and application of characteristic spectra of different medicinal parts of wolfberry or their water extracts

Abstract

The present application relates to the technical field of Chinese medicine analysis, and specifically to a method for constructing a characteristic spectrum of different medicinal parts of wolfberry or its aqueous extract, a detection method and application. The conditions of the liquid chromatography analysis of the present application include: mobile phase A is a mixture of acetonitrile and methanol in a volume ratio of 1: (1.5~2.5), mobile phase B is a mixture of 0.2%~0.4% trifluoroacetic acid, 0.08%~0.12% triethylamine and water, and gradient elution includes: 0~8min, maintaining the volume percentage of the mobile phase A at 11%, 8min~18min, the volume percentage of the mobile phase A increases from 11% to 13%, 18min~28min, the volume percentage of the mobile phase A increases from 13% to 18%, 28min~33min, the volume percentage of the mobile phase A increases from 18% to 19%, 33min~38min, maintaining the volume percentage of the mobile phase A at 19%, 38min~65min, the volume percentage of the mobile phase A increases from 19% to 56%. The technical solution of the present application can effectively distinguish different medicinal parts of wolfberry and the medicinal parts of its water extract, and can be used to identify different medicinal parts of wolfberry and its water extract.

Description

Technical Field

[0001] The present application relates to the technical field of traditional Chinese medicine detection, and in particular to a method for constructing a characteristic map, a detection method and an application of different medicinal parts of wolfberry or their water extracts; the different medicinal parts of wolfberry or their water extracts include wolfberry root bark, wolfberry stem bark, wolfberry root wood core, wolfberry root bark water extract, wolfberry stem bark water extract and wolfberry root wood core water extract. Background Art

[0002] Lycium barbarum is a perennial woody plant of the genus Lycium in the Solanaceae family. It has different medicinal values ​​according to the medicinal parts. Lycium barbarum is the dried root bark of Lycium chinense Mill. or Lycium barbarum L. in the Solanaceae family. The roots are dug up in autumn, washed, the wood core is extracted, the root bark is peeled, and it is dried in the sun. It tastes sweet and cold. It enters the lung, liver, and kidney meridians. It has the effects of cooling blood and removing steam, clearing the lungs and reducing fire. It is often used for symptoms such as yin deficiency, hot flashes, bone steaming and night sweats, lung heat cough, hemoptysis, epistaxis, internal heat and thirst.

[0003] In ancient books, the bark of Rehmannia glutinosa is often listed under the entry for wolfberry. In the Western Han Dynasty, the roots, stems, leaves, and fruits of wolfberry could all be used as medicine, but the medicinal parts had not yet been clearly identified. The "Compendium of Materia Medica" first proposed "wolfberry" as the name of the medicinal material for the stem bark part, and distinguished it from the medicinal properties of wolfberry fruit and root: "Wolfberry should use the stem bark, Rehmannia glutinosa should use the root bark, and wolfberry fruit should use its red fruit, which is one thing with three uses." The "stalk" refers to the branch or stem of the plant, and the "stalk bark" is consistent with the modern term "stem (branch) bark." The wolfberry fruit can be clearly distinguished, while the root bark is more similar to the stem bark. According to investigations and studies, there are certain differences in their properties, but there are few studies on the specificity and universality of this difference, and no other methods of distinction have been reported.

[0004] The root bark, stem bark, and root wood core of wolfberry all contain effective compound components such as alkaloids, organic acids and their esters, and anthraquinones. However, due to differences in composition, they are differentiated in medicinal use. The 2020 edition of the Chinese Pharmacopoeia stipulates that the medicinal part of Lycium bark is the dried root bark. Lycium bark, stem bark, and root wood core should be used separately and should not be mixed or misused. It can be seen that it is necessary to provide an effective method for distinguishing different medicinal parts of Lycium barbarum.

[0005] Lan Ting et al. used the method of property identification and microscopic identification to distinguish the root bark of Lycium barbarum and its stem bark. However, this method did not study the differences in their chemical components, and its reliability needs to be improved. Qian Xin et al. studied the root of Lycium barbarum. The medicinal parts of the root of Lycium barbarum include the root wood core of Lycium barbarum. In the study, the main part of the main root of Lycium barbarum was qualitatively studied and the contents of chlorogenic acid, ferulic acid, betaine, etc. were determined. However, the specificity of these components is poor and they are not suitable as indicator components in different medicinal parts of Lycium barbarum. In addition, the content of lycium ethyl in the root bark of Lycium barbarum is often determined by UPLC or HPLC in literature reports. However, lycium ethyl is not only the main active ingredient of Lycium barbarum. The content of lycium ethyl in the stem bark and root wood core of Lycium barbarum is high. The determination of lycium ethyl alone cannot effectively distinguish the root bark, stem bark and root wood core.

[0006] Water extract is one of the important raw materials for Chinese medicine preparations such as Chinese patent medicines and Chinese medicine formula granules. Since it has lost the characteristics of medicinal materials, it cannot be effectively identified. Therefore, it is also necessary to establish a suitable differentiation method for water extracts of different medicinal parts of wolfberry. Summary of the invention

[0007] Based on this, the purpose of this application includes providing a method for constructing a characteristic map of different medicinal parts of wolfberry or their water extracts, a detection method and application, which can effectively distinguish different medicinal parts of wolfberry and their water extracts, and can be used to identify different medicinal parts of wolfberry and their water extracts.

[0008] In one aspect of the present application, a method for constructing a characteristic spectrum of different medicinal parts of wolfberry or their water extracts is provided, wherein the different medicinal parts of wolfberry or their water extracts include wolfberry root bark, wolfberry stem bark, wolfberry root wood core, wolfberry root bark water extract, wolfberry stem bark water extract and wolfberry root wood core water extract;

[0009] The method for constructing the characteristic spectrum of different medicinal parts of wolfberry or their water extracts comprises:

[0010] Take different medicinal parts of wolfberry or their water extracts, add extraction solvent for extraction, and prepare test solution;

[0011] Taking the test solution for liquid chromatography analysis to establish characteristic spectra of different medicinal parts of wolfberry or their water extracts;

[0012] Wherein, the conditions of the liquid chromatography analysis include:

[0013] (1) The mobile phase A is a mixture of acetonitrile and methanol, wherein the volume ratio of the acetonitrile to the methanol is 1:(1.5-2.5),

[0014] The mobile phase B is a mixture of trifluoroacetic acid, triethylamine and water, wherein the volume proportion of the trifluoroacetic acid in the mobile phase B is 0.2% to 0.4%, and the volume proportion of the triethylamine in the mobile phase B is 0.08% to 0.12%;

[0015] (2) Gradient elution is used, and the procedure of gradient elution includes:

[0016] 0-8min, maintaining the volume percentage of the mobile phase A at 11%,

[0017] From 8min to 18min, the volume percentage of the mobile phase A increased from 11% to 13%.

[0018] From 18min to 28min, the volume percentage of the mobile phase A increased from 13% to 18%.

[0019] From 28min to 33min, the volume percentage of the mobile phase A increased from 18% to 19%.

[0020] 33min~38min, maintaining the volume percentage of the mobile phase A at 19%,

[0021] From 38 min to 65 min, the volume percentage of the mobile phase A increased from 19% to 56%.

[0022] In another aspect of the present application, a method for detecting different medicinal parts of wolfberry or their aqueous extracts is provided, wherein the different medicinal parts of wolfberry or their aqueous extracts include wolfberry root bark, wolfberry stem bark, wolfberry root wood core, wolfberry root bark aqueous extract, wolfberry stem bark aqueous extract and wolfberry root wood core aqueous extract;

[0023] The detection method of different medicinal parts of wolfberry or their water extracts comprises:

[0024] Taking different medicinal parts of wolfberry to be tested or their water extracts, adding extraction solvent for extraction, and preparing a solution of the test product;

[0025] Taking the test solution for liquid chromatography analysis to detect the content of index components in the test solution, the index components include caffeoyl butanediamine, N-feruloyl-1, 4-butanediamine, lycium barbarum, lycium barbarum A and lycium barbarum B;

[0026] Wherein, the conditions of the liquid chromatography analysis include:

[0027] (1) The mobile phase A is a mixture of acetonitrile and methanol, wherein the volume ratio of the acetonitrile to the methanol is 1:(1.5-2.5),

[0028] The mobile phase B is a mixture of trifluoroacetic acid, triethylamine and water, wherein the volume proportion of the trifluoroacetic acid in the mobile phase B is 0.2% to 0.4%, and the volume proportion of the triethylamine in the mobile phase B is 0.08% to 0.12%;

[0029] (2) Gradient elution is used, and the procedure of gradient elution includes:

[0030] 0-8min, maintaining the volume percentage of the mobile phase A at 11%,

[0031] From 8min to 18min, the volume percentage of the mobile phase A increased from 11% to 13%.

[0032] From 18min to 28min, the volume percentage of the mobile phase A increased from 13% to 18%.

[0033] From 28min to 33min, the volume percentage of the mobile phase A increased from 18% to 19%.

[0034] 33min~38min, maintaining the volume percentage of the mobile phase A at 19%,

[0035] From 38 min to 65 min, the volume percentage of the mobile phase A increased from 19% to 56%.

[0036] In another aspect of the present application, a characteristic spectrum constructed by the method for constructing a characteristic spectrum described in any of the above technical solutions and / or an application of the detection method described in any of the above technical solutions in identifying different medicinal parts of wolfberry or their aqueous extracts is provided. Identifying different medicinal parts of wolfberry or their aqueous extracts includes:

[0037] (1) Identify wolfberry root bark, wolfberry stem bark and wolfberry root wood core;

[0038] (2) Identification of the water extracts of wolfberry root bark, wolfberry stem bark, and wolfberry root wood core;

[0039] (3) Identify wolfberry root bark and its counterfeit products;

[0040] (4) Identify the water extract of wolfberry root bark and the water extract of counterfeit wolfberry root bark;

[0041] (5) Identify wolfberry stem bark and its counterfeit products;

[0042] (6) Identify the water extract of wolfberry stem bark and the water extract of the counterfeit wolfberry stem bark;

[0043] (7) Identify wolfberry root wood core and its counterfeit products;

[0044] (8) Identify the water extract of wolfberry root wood core and the water extract of counterfeit wolfberry root wood core.

[0045] The present application establishes appropriate chromatographic conditions to conduct liquid chromatography analysis on different medicinal parts of wolfberry or their water extracts, and constructs a characteristic spectrum thereof. The obtained characteristic spectrum has rich characteristic peak information, and the separation effect of each characteristic peak is good, which can well characterize the material transfer from the raw material to the decoction, and provide a rapid and comprehensive detection method for the quality of different medicinal parts of wolfberry and their water extracts.

[0046] The present application conducts a large amount of research on the characteristic components of different medicinal parts of wolfberry and their water extracts. By establishing appropriate chromatographic conditions, 8 common characteristic peaks are obtained: peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 7 and peak 8, and caffeoylbutyleneamine, N-feruloyl-1, 4-butyleneamine, lycium barbarum, lycium barbarum A and lycium barbarum B are identified. These common peaks and the peak information displayed in the characteristic spectrum can be used as the identification benchmark for different medicinal parts of wolfberry or their water extracts.

[0047] The characteristic spectrum construction method and / or detection method of the present application has strong reliability and good specificity, and can be used to detect the content of highly specific components in different medicinal parts of wolfberry or their water extracts, and can further be used as identification components and used as a basis for identification; preferably, the indicator components include caffeoylbutyleneamine, N-feruloyl-1, 4-butyleneamine, lycium barbarum ethyl, lycium barbarum A, and lycium barbarum B. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0049] Figure 1 It is an overlay of characteristic spectra of 13 batches of water extracts of Radix Lycii in Example 1 of the present application;

[0050] Figure 2 The reference characteristic spectrum of the water extract of Cortex Lycii in Example 1 of the present application;

[0051] Figure 3 This is an overlay of characteristic spectra of 10 batches of stem bark water extracts in Example 1 of the present application;

[0052] Figure 4 This is an overlay of characteristic spectra of 10 batches of root wood core water extracts in Example 1 of the present application;

[0053] Figure 5The characteristic spectrum reference material confirmation and specificity diagram of the water extract of Radix Lycii in Example 2 of the present application;

[0054] Figure 6 This is a comparison chart of the characteristic spectra of the water extract of Cortex Lycii in Example 3 of the present application and the water extracts of different counterfeits;

[0055] Figure 7 This is a comparison chart of the characteristic spectra of the water extract of Cortex Lycii of Example 4 of the present application and the water extracts of other different parts;

[0056] Figure 8 This is a comparison chart of characteristic spectra of water extract of Radix Lycii using different types of chromatographic columns in Comparative Example 1 of the present application;

[0057] Fig. 9 This is the characteristic spectrum of Radix Lycopodii with different mobile phases in Comparative Example 2 of the present application. DETAILED DESCRIPTION

[0058] The present application will be further described below in conjunction with the embodiments, examples and accompanying drawings. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. In addition, it should be understood that after reading the content taught in the present application, those skilled in the art may make various changes or modifications to the present application, and these equivalent forms also fall within the scope of protection of the claims attached to the present application.

[0059] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0060] the term:

[0061] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0062] The terms "and / or", "or / and", and "and / or" used in this article include any one of two or more related listed items, and also include any and all combinations of related listed items, and the arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution that is all connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the combination of four of A, B, C, and D (that is, the technical solution that is all connected by "logical AND").

[0063] Herein, "preferred", "preferable", "better", etc. are only used to describe implementation methods or examples with better effects. It should be understood that they do not constitute limitations on the scope of protection of this application.

[0064] In the present application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0065] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0066] In this application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0067] In the present application, weight may be μg, mg, g, kg or other mass units known in the chemical industry.

[0068] In this application, "%" means percentage, which refers to the weight ratio; however, the percentage of a solution, unless otherwise specified, refers to the solute contained in 100 mL of the solution in grams; the percentage of an alcohol refers to the volume ratio at 20°C. In addition, the following symbols may be used as needed: % (g / g) means that a solution contains a solute in grams; % (mL / mL) means that a solution contains a solute in milliliters; % (mL / g) means that a solution contains a solute in milliliters; % (g / mL) means that a solution contains a solute in grams.

[0069] In one aspect of the present application, a method for constructing characteristic spectra of different medicinal parts of wolfberry or their water extracts is provided.

[0070] In the present application, different medicinal parts of wolfberry or their water extracts include wolfberry root bark, wolfberry stem bark, wolfberry root wood core, wolfberry root bark water extract, wolfberry stem bark water extract and wolfberry root wood core water extract.

[0071] The fruit, root bark, stem bark, and root wood core of wolfberry have different chemical compositions and pharmacological activities, so their medicinal values ​​are different, and it is necessary to identify and distinguish them. The wolfberry fruit and the root wood core of wolfberry can be distinguished from the other parts in appearance, while the wolfberry root bark and wolfberry stem bark are similar and difficult to identify from the appearance. However, the current research on different medicinal parts of wolfberry mainly focuses on chemical composition and pharmacological analysis, and few identification methods have been reported.

[0072] Different medicinal parts of wolfberry are often used for water extraction. The water extract is one of the important raw materials for Chinese medicine preparations such as Chinese patent medicines and Chinese medicine formula granules. Since the characteristics of the medicinal materials have been lost, it is impossible to effectively identify them. Therefore, it is urgent to provide a method that can effectively distinguish the water extract of Lycium bark that has lost its medicinal characteristics from the water extract of the root wood core and stem bark of wolfberry.

[0073] In order to solve the above problems, the present application provides a method for constructing characteristic spectra of different medicinal parts of wolfberry or their aqueous extracts, comprising:

[0074] Take different medicinal parts of wolfberry or their water extracts, add extraction solvent for extraction, and prepare test solution;

[0075] Take the test sample solution for liquid chromatography analysis to establish the characteristic spectrum of different medicinal parts of wolfberry or their water extracts;

[0076] Among them, the conditions for liquid chromatography analysis include:

[0077] (1) Mobile phase A is a mixture of acetonitrile and methanol, with a volume ratio of acetonitrile to methanol of 1:(1.5~2.5).

[0078] Mobile phase B is a mixture of trifluoroacetic acid, triethylamine and water, wherein the volume proportion of trifluoroacetic acid in mobile phase B is 0.2%-0.4%, and the volume proportion of triethylamine in mobile phase B is 0.08%-0.12%;

[0079] (2) Gradient elution is used. The procedure of gradient elution includes:

[0080] 0~8min, maintain the volume percentage of mobile phase A at 11%,

[0081] From 8min to 18min, the volume percentage of mobile phase A increased from 11% to 13%.

[0082] From 18min to 28min, the volume percentage of mobile phase A increased from 13% to 18%.

[0083] From 28min to 33min, the volume percentage of mobile phase A increased from 18% to 19%.

[0084] 33min~38min, maintain the volume percentage of mobile phase A at 19%,

[0085] From 38min to 65min, the volume percentage of mobile phase A increased from 19% to 56%.

[0086] In one embodiment, the conditions for liquid chromatography analysis further include at least one of the following:

[0087] (1) Liquid chromatography analysis is ultra-high performance liquid chromatography analysis;

[0088] (2) The flow rate of gradient elution is 0.7 mL / min~0.9 mL / min;

[0089] (3) The chromatographic column is an octadecylsilane bonded silica gel chromatographic column;

[0090] (4) Column temperature is 32°C~38°C;

[0091] (5) The injection volume is 4 μL~6 μL;

[0092] (6) The detection wavelength is 250nm~270nm.

[0093] Preferably, the parameters of the chromatographic column are: inner diameter 4.4 mm to 4.8 mm, column length 140 mm to 160 mm, and filler particle size 2.6 μm to 2.8 μm.

[0094] Preferably, the chromatographic column model is Agilent Poroshell 120 SB-C18.

[0095] In one embodiment, the extraction method includes ultrasound;

[0096] Preferably, the power of ultrasound is 200W~400W and / or the frequency of ultrasound is 30kHz~50kHz;

[0097] Preferably, the ultrasonication time is 20 min to 40 min.

[0098] In one embodiment, the extraction solvent is a mixture of methanol and an aqueous solution of acetic acid;

[0099] Preferably, the aqueous solution of acetic acid contains 0.4% to 0.6% acetic acid by volume;

[0100] Preferably, the volume ratio of methanol to the aqueous solution of acetic acid is 1:(0.8~1.2).

[0101] In one embodiment, the preparation steps of the aqueous extracts of different medicinal parts of wolfberry or aqueous extracts thereof include:

[0102] The different medicinal parts of wolfberry for testing were processed according to the processing specifications to prepare the decoction pieces for testing;

[0103] Take the slices to be tested, soak for 20-40 minutes, decoct twice, add 8-12 times the volume of the slices of water for the first decoction, decoct for 10-30 minutes, add 6-10 times the volume of the slices of water for the second decoction, decoct for 10-20 minutes, filter the decoction with a 300-400 mesh sieve while hot, combine the two filtrates, and concentrate;

[0104] Further, the mixture is transferred to a vacuum freeze dryer for freeze drying and taken out to obtain the product.

[0105] In one embodiment, the method for constructing characteristic spectra of different medicinal parts of wolfberry or their aqueous extracts further includes: preparing a reference solution, and taking the reference solution for liquid chromatography analysis;

[0106] The conditions for liquid chromatography analysis are the same as those for liquid chromatography analysis described in any of the above technical solutions;

[0107] The reference solution includes a reference substance of lycium barbarum E, a reference substance of lycium barbarum A, a reference substance of lycium barbarum A and a reference substance of lycium barbarum B.

[0108] In one embodiment, appropriate amounts of lycium barbarum B reference substance, lycium barbarum A reference substance, and lycium barbarum B reference substance are taken, accurately weighed, and methanol is added to prepare a mixed solution containing 0.08 mg to 0.12 mg of each per 1 mL, which is used as a reference solution.

[0109] In one embodiment, the characteristic spectrum obtained by the construction method described in any of the above technical solutions includes the following 8 common peaks: peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 7 and peak 8;

[0110] Among them, peak 1 is caffeoyl butanediamine, peak 3 is N-feruloyl-1, 4-butanediamine, peak 4 is lycium barbarum, peak 6 is lycium barbarum, peak 7 is lycium barbarum A, and peak 8 is lycium barbarum B.

[0111] In another aspect of the present application, a method for detecting different medicinal parts of wolfberry or their aqueous extracts is provided, which can be used for qualitative and / or quantitative detection of highly specific components therein as indicator components and as a basis for identification of different medicinal parts of wolfberry or their aqueous extracts.

[0112] In one embodiment, the method for detecting different medicinal parts of wolfberry or their aqueous extracts includes:

[0113] Taking different medicinal parts of wolfberry to be tested or their water extracts, adding extraction solvent for extraction, and preparing a solution of the test product;

[0114] Taking the test solution for liquid chromatography analysis to detect the content of the index components in the test solution, the index components include caffeoyl butanediamine, N-feruloyl-1, 4-butanediamine, lycium barbarum, lycium barbarum A and lycium barbarum B;

[0115] Among them, the conditions for liquid chromatography analysis include:

[0116] (1) Mobile phase A is a mixture of acetonitrile and methanol, and the volume ratio of acetonitrile to the methanol is 1:(1.5-2.5).

[0117] Mobile phase B is a mixture of trifluoroacetic acid, triethylamine and water, wherein the volume proportion of trifluoroacetic acid in mobile phase B is 0.2%-0.4%, and the volume proportion of triethylamine in mobile phase B is 0.08%-0.12%;

[0118] (2) Gradient elution is used. The procedure of gradient elution includes:

[0119] 0-8min, maintaining the volume percentage of the mobile phase A at 11%,

[0120] From 8min to 18min, the volume percentage of the mobile phase A increased from 11% to 13%.

[0121] From 18min to 28min, the volume percentage of the mobile phase A increased from 13% to 18%.

[0122] From 28min to 33min, the volume percentage of the mobile phase A increased from 18% to 19%.

[0123] 33min~38min, maintaining the volume percentage of the mobile phase A at 19%,

[0124] From 38 min to 65 min, the volume percentage of the mobile phase A increased from 19% to 56%.

[0125] In one embodiment, the conditions for liquid phase analysis also include at least one of the following:

[0126] (1) Liquid phase analysis is ultra-high performance liquid chromatography analysis;

[0127] (2) The flow rate of gradient elution is 0.7 mL / min~0.9 mL / min;

[0128] (3) The chromatographic column is an octadecylsilane bonded silica gel chromatographic column;

[0129] (4) Column temperature is 32°C~38°C;

[0130] (5) The injection volume is 4 μL~6 μL;

[0131] (6) The detection wavelength is 250nm~270nm.

[0132] In one embodiment, the extraction method includes ultrasound;

[0133] Preferably, the power of ultrasound is 200W~400W and / or the frequency of ultrasound is 30kHz~50kHz;

[0134] Preferably, the ultrasonication time is 20 min to 40 min.

[0135] In one embodiment, the extraction solvent is a mixture of methanol and an aqueous solution of acetic acid;

[0136] Preferably, the aqueous solution of acetic acid contains 0.4% to 0.6% acetic acid by volume;

[0137] Preferably, the volume ratio of methanol to the aqueous solution of acetic acid is 1:(0.8~1.2).

[0138] In another aspect of the present application, it also relates to the application of the characteristic spectrum constructed by the method for constructing the characteristic spectrum described in any of the above technical solutions and / or the detection method described in any of the above technical solutions in identifying different medicinal parts of wolfberry or their aqueous extracts, including:

[0139] (1) Identify wolfberry root bark, wolfberry stem bark and wolfberry root wood core;

[0140] (2) Identification of the water extracts of wolfberry root bark, wolfberry stem bark, and wolfberry root wood core;

[0141] (3) Identify wolfberry root bark and its counterfeit products;

[0142] (4) Identify the water extract of wolfberry root bark and the water extract of counterfeit wolfberry root bark;

[0143] (5) Identify wolfberry stem bark and its counterfeit products;

[0144] (6) Identify the water extract of wolfberry stem bark and the water extract of the counterfeit wolfberry stem bark;

[0145] (7) Identify wolfberry root wood core and its counterfeit products;

[0146] (8) Identify the water extract of wolfberry root wood core and the water extract of counterfeit wolfberry root wood core.

[0147] The conditions for liquid chromatography detection in the present application are based on a reasonable analysis of the chemical composition of different medicinal parts of wolfberry and their water extracts. The characteristic spectrum construction method and detection method provided in the present application can well reflect the intrinsic chemical information of different medicinal parts of wolfberry and their water extracts, so that different medicinal parts have distinguishable peak information on the characteristic spectrum. At the same time, the characteristic spectrum can also reflect the transfer of chemical information from raw materials to decoctions. Therefore, the characteristic spectrum and detection method of the present application can not only effectively identify different medicinal parts of wolfberry, including the identification of their material benchmarks, but also effectively identify the water extracts that have been processed into preparations.

[0148] The following are some specific embodiments.

[0149] For experimental parameters not specified in the following specific embodiments, reference is made to the instructions given in the present application document, and reference may also be made to experimental manuals in the art or other experimental methods known in the art, or to experimental conditions recommended by manufacturers.

[0150] The raw materials and reagents involved in the following specific examples can be obtained from commercial sources, or can be prepared by those skilled in the art according to known methods.

[0151] Instruments and reagents

[0152] Instruments: Waters ultra-high performance liquid chromatograph (H-Class, Waters (China) Co., Ltd.), Agilent ultra-high performance liquid chromatograph (1290, Agilent Co., Ltd.), Agilent Poroshell 120 SB-C18 chromatographic column (4.6 mm × 150 mm, 2.7 μm), one-ten-thousandth balance (ME204E, Mettler-Toledo), one-millionth balance (XP26, Mettler-Toledo).

[0153] Reagents: ethanol (Tianjin Fuyu Fine Chemical Co., Ltd.) and methanol (Tianjin Fuyu Fine Chemical Co., Ltd.) were of analytical grade; trifluoroacetic acid (Shanghai Aladdin Biochemical Technology Co., Ltd., chromatographic grade, ≥99.5%), triethylamine (Tianjin Komiou Chemical Reagent Co., Ltd., analytical grade), acetonitrile (Merck Co., Ltd.), and methanol (Merck Co., Ltd.) were of HPLC chromatographic grade; water was from a laboratory ultrapure water system (Merck Co., Ltd., Milli-Q Direct).

[0154] Lycium barbarum B reference substance (batch number: D04102204013, content 98.0%, Chengdu Ruifensi Biotechnology Co., Ltd.); Lycium barbarum A reference substance (batch number: D04002203022, content: 98%, Chengdu Ruifensi Biotechnology Co., Ltd.); Lycium barbarum A reference substance (batch number: PS012737, content: 99.60%, Chengdu Pusi Biotechnology Co., Ltd.); Lycium barbarum B reference substance (batch number: PS012738, content: 99.43%, Chengdu Pusi Biotechnology Co., Ltd.); Lycium barbarum reference medicinal material (batch number: 121087-201707, China Food and Drug Inspection Institute); Information on each batch of Lycium barbarum (wolfberry root bark) and medicinal materials from different parts is shown in Table 1.

[0155] Table 1 Information of Radix Lycii and its different parts

[0156]

[0157] Example 1 Establishment of a characteristic spectrum construction method

[0158] 1.1 Preparation of samples

[0159] 1.1.1 Preparation of aqueous extracts

[0160] After the different parts of the wolfberry plants from different origins in Table 1 are processed according to the processing specifications, they are made into pieces, and the water extract is prepared according to the following method. Preparation method of water extract: Take the pieces of Lycium bark, stem bark, and root wood core, soak for 30 minutes, decoct twice, add 10 times the amount of water of the pieces for the first decoction, decoct for 20 minutes, add 8 times the amount of water of the pieces for the second decoction, decoct for 15 minutes, filter the decoction with a 350-mesh sieve while hot, combine the two filtrates, concentrate, transfer to a vacuum freeze dryer for freeze drying, take out, and obtain.

[0161] 1.1.2 Preparation of test solution

[0162] Grind the water extract, take about 0.2g, weigh accurately, put it in a stoppered conical flask, accurately add 25ml of a mixed solution of 50% methanol-0.5% acetic acid (1:1), weigh, and ultrasonically treat (power 300W, frequency 40KHz) for 30 minutes. Cool, make up the lost weight with a mixed solution of 50% methanol-0.5% acetic acid (1:1), shake well, filter, and take the filtrate to obtain the product.

[0163] 1.1.3 Preparation of reference solution

[0164] Take appropriate amount of lycium barbarum B reference substance, lycium barbarum A reference substance, lycium barbarum A reference substance, and lycium barbarum B reference substance, weigh them accurately, and add methanol to prepare a mixed solution containing 0.1 mg of each substance per 1 mL as the reference solution.

[0165] 1.2 Liquid chromatography analysis

[0166] Agilent Poroshell 120 SB-C18 column (150 mm × 4.6 mm, 2.7 μm) was used; acetonitrile-methanol (1:2) was used as mobile phase A, 0.3% trifluoroacetic acid (containing 0.1% triethylamine) solution was used as mobile phase B, and gradient elution was performed according to the provisions in Table 2; the flow rate was 0.8 ml per minute; the column temperature was 35 °C; the detection wavelength was 260 nm, and the injection volume was 5 μL.

[0167] Table 2 Gradient elution program

[0168]

[0169] 1.3 Establishing feature maps

[0170] The “Chinese Medicine Chromatographic Characteristic Spectrum Similarity Evaluation System (2012.0 Version)” was used to process the data of 13 batches of Radix Lycii water extract samples and generate superimposed spectra. Figure 1 , select 8 common peaks with relatively stable relative retention time and relative peak area RSD as characteristic peaks, and generate a control characteristic spectrum. Figure 2 .

[0171] The data of 10 batches of stem bark water extract samples were processed to generate superimposed maps. Figure 3 .

[0172] The data of 10 batches of root wood core water extract samples were processed to generate superimposed maps. Figure 4 .

[0173] 1.4 Comparison of water extracts of Lycium bark and stem bark and root core

[0174] Comparison of the water extract of Lycium bark with the water extract of stem bark and root wood core showed that the relative retention times of peaks 1, 2, 3 and 5 of each batch of samples were shown in Table 3, with Lycium barbarum A as the reference peak S2, and the relative peak areas of peak 2 of each batch of samples were shown in Table 4. The results in Table 4 show that the relative peak area fluctuation range of peak 2 of 13 batches of Lycium bark water extracts was 0.096-0.927, while the relative peak area fluctuation range of peak 2 of 10 batches of stem bark water extracts was 1.858-13.333, and the relative peak area fluctuation range of peak 2 of 10 batches of root wood core water extracts was 1.692-23.032. The relative peak areas of the water extracts of the three parts are quite different, so the three parts can be distinguished according to the relative peak area ratio of peak 2. According to the measured value of the relative peak area of ​​Peak 2, based on the measured minimum value of the root wood core, 70% of the minimum value, i.e. 1.18, was taken and rounded to 1.1, and the basis for distinguishing the water extract of the bark of Lycium bark from the water extract of the stem bark and the root wood core was formulated:

[0175] Water extract of Lycium barbarum bark: ① There should be 8 characteristic peaks in the chromatogram of the test sample, of which 4 peaks should correspond to the retention times of the corresponding reference peaks; the peak corresponding to the Lycium barbarum ethyl reference is the S1 peak, and the relative retention time of each characteristic peak and the S1 peak is calculated, and the relative retention time should be within ±10% of the specified value, and the specified value is: 0.35 (peak 1), 0.40 (peak 2), 0.77 (peak 3), 1.03 (peak 5); ② The peak corresponding to the Lycium barbarum A reference peak is the S2 peak, and the relative peak area of ​​Peak 2 and S peak is calculated, and the relative peak area should be within the specified value range, and the specified value is: not greater than 1.1.

[0176] Aqueous extracts of root core and stem bark: ① 8 or less corresponding characteristic peaks are present in the chromatogram of the sample; ② The relative peak area between peak 2 and peak S2 is greater than or equal to 1.1.

[0177] Table 3 Relative retention time results of some characteristic peaks of water extracts from different parts of wolfberry plants and characteristic peaks of lycium barbarum

[0178]

[0179] Table 4 Comparison of relative peak areas of characteristic spectrum peak 2 and Lycium barbarum A in aqueous extracts from different parts of wolfberry plants

[0180]

[0181] Example 2 Liquid phase confirmation and specificity investigation of reference substance

[0182] This example is to confirm the reference substance of the water extract of Lycium bark and verify the specificity of the chromatographic detection method.

[0183] 2.1 Sample preparation

[0184] The instruments and reagents used are the same as those in Example 1.

[0185] The test drugs used are as follows: Lycium barbarum E-reference substance (batch number: D04102204013, content 98.0%, Chengdu Ruifensi Biotechnology Co., Ltd.); Lycium barbarum A reference substance (batch number: D04002203022, content: 98%, Chengdu Ruifensi Biotechnology Co., Ltd.); Lycium barbarum A reference substance (batch number: PS012737, content: 99.60%, Chengdu Pusi Biotechnology Co., Ltd.); Lycium barbarum B reference substance (batch number: PS012738, content: 99.43%, Chengdu Pusi Biotechnology Co., Ltd.); Lycium barbarum reference medicinal material (batch number: 121087-201707, China Food and Drug Inspection Institute); Caffeic acid butanediamine reference substance (batch number: CFS202101, content: 98%, Chem Faces); N-feruloyl-1,4-butanediamine reference substance (batch number: wkq21072603, content: 98%, Sichuan Weikeqi Biotechnology Co., Ltd.);

[0186] The preparation of the water extract and the test sample were the same as in Example 1.

[0187] The preparation method of the reference solution is as follows: take appropriate amounts of the lycopene ethyl reference substance and the lycopene A reference substance, accurately weigh them, and add methanol to make a mixed solution A containing 0.1 mg of the lycopene ethyl reference substance and 0.05 mg of the lycopene A reference substance per 1 mL; take appropriate amounts of the caffeoyl butanediamine reference substance, N-feruloyl-1,4-butanediamine reference substance, wolfberry extract A reference substance, and wolfberry extract B reference substance, accurately weigh them, and add methanol to make a mixed solution containing 0.1 mg of each reference substance per 1 mL, as the reference substance mixed solution B.

[0188] 2.2 Testing and Verification

[0189] 5 μl of each of the test solution, reference solution and blank solvent of 1 part of the Chinese wolfberry bark was accurately drawn and injected into the ultra-high performance liquid chromatograph. The chromatographic conditions under "Example 1" were referred to for determination to obtain the chromatograms of the test solution, reference solution and blank solvent of 1 part of the Chinese wolfberry bark, as shown in FIG. Figure 5 .

[0190] The results showed that the chromatogram of the test sample had the same chromatographic peaks at the corresponding retention time of the chromatogram of the reference sample, and six components in the characteristic spectrum of the water extract of Lycium barbarum were confirmed, including caffeoyl butanediamine (peak 1), N-feruloyl-1,4-butanediamine (peak 3), lycium barbarum ethyl (peak 4), lycium barbarum A (peak 6), lycium barbarum A (peak 7), and lycium barbarum B (peak 8), and the negative result was without interference, indicating that the method has good specificity.

[0191] Example 3 Comparison of similar products

[0192] The counterfeit products of Lycium bark are mainly the root bark of Cynanchum chinense R. Br., a plant of the genus Cynanchum in the Apocynaceae family, and Lycium bark.

[0193] The preparation of instruments and reagents, test drugs, chromatographic conditions, reference solution, test solution and water extract is the same as that in Example 1. The source information of Anemarrhena asphodeloides and Radix Glechomae is shown in Table 5.

[0194] Table 5 Information on the sources of medicinal materials of Tudigupi and Goose Down Vine

[0195]

[0196] Take the water extract of Radix Lycii and the water extract of the counterfeit products in Table 5, make the test solution and then inject it for determination. The results are shown in Figure 6 .

[0197] The results showed that the characteristic spectrum of the water extract of Lycium bark had 8 common peaks, but the characteristic spectrum of Lycium bark and Anemarrhena asphodeloides differed greatly from that of Lycium bark. Different varieties can be distinguished according to the number of chromatographic peaks. Therefore, the characteristic spectrum of Lycium bark established by the method of this application can effectively characterize its chemical composition, can be effectively distinguished from other counterfeits, and has a certain degree of specificity.

[0198] Example 4 Comparison of aqueous extracts from different parts of wolfberry plants

[0199] The preparation of instruments and reagents, test drugs, chromatographic conditions, reference solutions and water extracts is the same as in Example 1. Information on different parts of wolfberry branches, wolfberry fruits and wolfberry leaves from the same plant is shown in Table 6, and information on the source of Lycium barbarum root bark (wolfberry root bark) is shown in Example 1.

[0200] Table 6 Information on the sources of medicinal materials from different parts of wolfberry plants

[0201]

[0202] Take the water extract of Radix Lycii and the water extracts of different parts in Table 6, make the test solution and then inject it for determination. The results are shown in Figure 7 .

[0203] The results showed that the characteristic spectrum of Lycium barbarum (wolfberry root bark) water extract had 8 common peaks, but the characteristic spectrum of Lycium barbarum branches, Lycium barbarum, and Lycium barbarum leaves was quite different from that of Lycium barbarum (wolfberry root bark) water extract. Different parts of the same plant can be distinguished by the number of chromatographic peaks. Therefore, the characteristic spectrum of Lycium barbarum established by the method of this application can effectively characterize its chemical components and can be effectively distinguished from other parts, and has a certain degree of specificity.

[0204] Comparative Example 1

[0205] The characteristic spectrum of the water extract of Lycium bark was detected using different types of chromatographic columns, and the other detection conditions were the same as those in Example 1, including:

[0206] Take the test solution of Lycium bark in Example 1; use different chromatographic columns Waters Xbridge C18 chromatographic column (4.6mm×150mm, 5μm), Agilent ZORBAX SB-C18 chromatographic column (4.6mm×150mm, 5μm), Agilent Poroshell 120 SB-C18 chromatographic column (4.6mm×150mm, 2.7μm), and detect according to the chromatographic conditions and elution gradient in Example 1. The results are shown in Figure 8 .

[0207] The results showed that when using chromatographic columns of specifications such as Waters Xbridge C18 (4.6mm×150mm, 5μm) and Agilent ZORBAXSB-C18 (4.6mm×150mm, 5μm), the spectra were quite different and many main characteristic peaks were missing. When the chromatographic column Agilent Poroshell 120 SB-C18 column (4.6mm×150mm, 2.7μm) specified in this application was used, 8 chromatographic peaks could be eluted within 60 minutes, the overall distribution of retention time was relatively appropriate, and each chromatographic peak achieved baseline separation. Therefore, Agilent Poroshell 120 SB-C18 (4.6mm×150mm, 2.7μm) was selected as the chromatographic column for the characteristic spectrum of the water extract of Radix Rehmanniae.

[0208] Comparative Example 2

[0209] The characteristic spectrum of the water extract of Lycium bark was detected, using different mobile phase conditions, and the other detection conditions were the same as those in Example 1, including:

[0210] Take the test solution of Lycium barbarum in Example 1; use acetonitrile-methanol (1:2)-0.3% trifluoroacetic acid and acetonitrile-methanol (1:1)-0.3% trifluoroacetic acid (containing 0.1% triethylamine) as mobile phases, and detect according to the chromatographic conditions and elution gradient in Example 1. The results are shown in Fig. 9 .

[0211] The results showed that the purity of peak 1 and the chromatographic peak of Lycium barbarum A in the chromatogram obtained by using acetonitrile-methanol (1:2)-0.3% trifluoroacetic acid solution, as well as the separation of Lycium barbarum A and Lycium barbarum B were poor; using acetonitrile-methanol (1:1)-0.3% trifluoroacetic acid (containing 0.1% triethylamine) solution, the peak purity of individual chromatographic peaks was poor, affecting the durability of the characteristic spectrum. Using the method established in this application, the separation of the chromatographic peaks was good, the purity was high, and the stability and durability were good. Therefore, acetonitrile-methanol (1:2)-0.3% trifluoroacetic acid (containing 0.1% triethylamine) solution was selected as the mobile phase for the characteristic spectrum of Lycium barbarum water extract.

[0212] All documents mentioned in this application are cited as references in this application, just as each document is cited as reference separately. Unless they conflict with the application purpose and / or technical solution of this application, the cited documents involved in this application are cited with all contents and all purposes. When the cited documents are involved in this application, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited. When the cited documents are involved in this application, the examples and preferred methods of the cited relevant technical features can also be incorporated into this application as references, but are limited to the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, the present application shall prevail or be modified adaptively according to the description of this application.

[0213] The technical features of the above-mentioned implementation modes and examples can be combined in any appropriate manner. To make the description concise, not all possible combinations of the technical features in the above-mentioned implementation modes and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0214] The above-described embodiments only express several implementation methods of the present application, but they cannot be understood as limiting the scope of the patent application. It should be pointed out that, for those of ordinary skill in the art, without departing from the concept of the present application, several variations and improvements can also be made, which all belong to the protection scope of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by those skilled in the art on the basis of the technical solutions provided in the present application through logical analysis, reasoning or limited experiments are all within the protection scope of the claims attached to the present application. Therefore, the protection scope of the patent of the present application shall be based on the attached claims, and the description and drawings may be used to explain the contents of the claims.

Claims

1. A method for constructing characteristic spectra of different medicinal parts of wolfberry or their aqueous extracts, characterized in that: The different medicinal parts of wolfberry or their water extracts include wolfberry root bark, wolfberry stem bark, wolfberry root wood core, wolfberry root bark water extract, wolfberry stem bark water extract and wolfberry root wood core water extract; The method for constructing the characteristic spectrum of different medicinal parts of wolfberry or their water extracts comprises: Taking different medicinal parts of wolfberry or their water extracts for testing, adding an extraction solvent for extraction to prepare a test solution; the extraction solvent is a mixture of methanol and an aqueous solution of acetic acid, the aqueous solution of acetic acid contains 0.4% to 0.6% acetic acid by volume, and the volume ratio of the methanol to the aqueous solution of acetic acid is 1:(0.8 to 1.2); Taking the test solution for liquid chromatography analysis to establish characteristic spectra of different medicinal parts of wolfberry or their water extracts; Prepare a reference solution, and take the reference solution for liquid chromatography analysis; the reference solution includes a lycium barbarum E-reference substance, a lycium barbarum A-reference substance, a lycium barbarum A-reference substance, and a lycium barbarum B-reference substance; Wherein, the conditions of the liquid chromatography analysis include: (1) Mobile phase A is a mixture of acetonitrile and methanol, wherein the volume ratio of the acetonitrile to the methanol is 1:2, and mobile phase B is a mixture of trifluoroacetic acid, triethylamine and water, wherein the volume proportion of the trifluoroacetic acid in the mobile phase B is 0.2% to 0.4%, and the volume proportion of the triethylamine in the mobile phase B is 0.08% to 0.12%; (2) Gradient elution is used, wherein the procedure of the gradient elution comprises: 0-8min, maintaining the volume percentage of the mobile phase A at 11%, 8min~18min, the volume percentage of the mobile phase A increases from 11% to 13%, 18min~28min, the volume percentage of the mobile phase A increases from 13% to 18%, 28min~33min, the volume percentage of the mobile phase A increases from 18% to 19%, 33min~38min, maintain the volume percentage of the mobile phase A at 19%, From 38min to 65min, the volume percentage of the mobile phase A increases from 19% to 56%; (3) The liquid chromatography analysis is ultra-high performance liquid chromatography analysis; (4) The column model is Agilent Poroshell 120SB-C18 column, with a length of 150 mm, a diameter of 4.6 mm, and a filler particle size of 2.7 μm; (5) The detection wavelength is 250nm~270nm.

2. The method for constructing a characteristic spectrum according to claim 1, wherein: The conditions of the liquid chromatography analysis also include at least one of the following: (1) The flow rate of the gradient elution is 0.7 mL / min to 0.9 mL / min; (2) Column temperature is 32°C to 38°C; (3) The injection volume is 4 μL to 6 μL.

3. The method for constructing a characteristic spectrum according to claim 1 or 2, characterized in that: Methods of extraction include ultrasound; The power of ultrasound is 200W to 400W and / or the frequency of ultrasound is 30kHz to 50kHz; The ultrasound time is 20 minutes to 40 minutes.

4. The method for constructing a characteristic spectrum according to claim 1 or 2, characterized in that: The characteristic spectrum includes the following 8 common peaks: peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 7 and peak 8; Among them, peak 1 is caffeoyl butanediamine, peak 3 is N-feruloyl-1,4-butanediamine, peak 4 is lycium barbarum et al., peak 6 is lycium barbarum et al., peak 7 is lycium barbarum A, and peak 8 is lycium barbarum B.

5. A method for detecting different medicinal parts of wolfberry or their aqueous extracts, characterized in that: The different medicinal parts of wolfberry or their water extracts include wolfberry root bark, wolfberry stem bark, wolfberry root wood core, wolfberry root bark water extract, wolfberry stem bark water extract and wolfberry root wood core water extract; The detection method of different medicinal parts of wolfberry or their water extracts comprises: Taking different medicinal parts of wolfberry to be tested or their aqueous extracts, adding an extraction solvent for extraction to prepare a test solution; the extraction solvent is a mixture of methanol and an aqueous solution of acetic acid, the aqueous solution of acetic acid contains 0.4% to 0.6% acetic acid by volume, and the volume ratio of the methanol to the aqueous solution of acetic acid is 1:(0.8 to 1.2); Taking the test solution for liquid chromatography analysis to detect the content of index components in the test solution, the index components include caffeoyl butanediamine, N-feruloyl-1,4-butanediamine, lycium barbarum, lycium barbarum A and lycium barbarum B; Wherein, the conditions of the liquid chromatography analysis include: (1) The mobile phase A is a mixture of acetonitrile and methanol, wherein the volume ratio of the acetonitrile to the methanol is 1:

2. The mobile phase B is a mixture of trifluoroacetic acid, triethylamine and water, wherein the volume proportion of the trifluoroacetic acid in the mobile phase B is 0.2% to 0.4%, and the volume proportion of the triethylamine in the mobile phase B is 0.08% to 0.12%; (2) Gradient elution is used, wherein the procedure of the gradient elution comprises: 0-8min, maintaining the volume percentage of the mobile phase A at 11%, 8min~18min, the volume percentage of the mobile phase A increases from 11% to 13%, 18min~28min, the volume percentage of the mobile phase A increases from 13% to 18%, 28min~33min, the volume percentage of the mobile phase A increases from 18% to 19%, 33min~38min, maintain the volume percentage of the mobile phase A at 19%, From 38min to 65min, the volume percentage of the mobile phase A increases from 19% to 56%; (3) The liquid chromatography analysis is ultra-high performance liquid chromatography analysis; (4) The column model is Agilent Poroshell 120SB-C18 column, with a length of 150 mm, a diameter of 4.6 mm, and a filler particle size of 2.7 μm; (5) The detection wavelength is 250nm~270nm.

6. The detection method according to claim 5, characterized in that The conditions of the liquid chromatography analysis also include at least one of the following: (1) The flow rate of the gradient elution is 0.7 mL / min to 0.9 mL / min; (2) Column temperature is 32°C to 38°C; (3) The injection volume is 4 μL to 6 μL.

7. The detection method according to claim 5 or 6, characterized in that: Methods of extraction include ultrasound; The power of ultrasound is 200W to 400W and / or the frequency of ultrasound is 30kHz to 50kHz; The ultrasound time is 20 minutes to 40 minutes.

8. Use of the characteristic spectrum constructed by the method for constructing the characteristic spectrum according to any one of claims 1 to 4 and / or the detection method according to any one of claims 5 to 7 in identifying different medicinal parts of wolfberry or their aqueous extracts, characterized in that: Identification of different medicinal parts of wolfberry or their water extracts include: (1) Identify wolfberry root bark, wolfberry stem bark and wolfberry root wood core; (2) Identification of the water extracts of wolfberry root bark, wolfberry stem bark, and wolfberry root wood core; (3) Identify wolfberry root bark and its counterfeits; (4) Identify the water extract of wolfberry root bark and the water extract of the counterfeit wolfberry root bark; (5) Identify the wolfberry stem bark and its counterfeit products; (6) Identify the water extract of wolfberry stem bark and the water extract of the counterfeit wolfberry stem bark; (7) Identify wolfberry root wood core and its counterfeit products; (8) Identify the water extract of wolfberry root wood core and the water extract of the counterfeit wolfberry root wood core.

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