CN113777183A

CN113777183A - Method for constructing characteristic spectrum of glossy privet fruit medicinal material and processed product thereof and method for detecting content of multi-index components

Info

Publication number: CN113777183A
Application number: CN202110909888.0A
Authority: CN
Inventors: 邓李红; 姚晓璇; 林伟雄; 陈仕妍; 田清清; 钟文峰; 孙冬梅; 魏梅; 程学仁
Original assignee: Guangdong Yifang Pharmaceutical Co Ltd
Current assignee: Guangdong Yifang Pharmaceutical Co Ltd
Priority date: 2021-08-09
Filing date: 2021-08-09
Publication date: 2021-12-10
Anticipated expiration: 2041-08-09
Also published as: CN113777183B

Abstract

The invention relates to a glossy privet fruit medicinal material and a method for constructing a characteristic spectrum of a processed product thereof. The characteristic map construction method comprises the following steps: preparing a reference substance solution and a test solution, wherein the preparation method of the reference substance solution comprises the following steps: dissolving 5-hydroxymethylfurfural reference substance, salidroside reference substance, echinacoside reference substance, specnuezhenide reference substance and nuezhenide G13 reference substance with a first solvent; the preparation method of the test solution comprises the following steps: adding a second solvent into a glossy privet fruit medicinal material or a processed product thereof for extraction, and filtering an extracting solution to obtain a subsequent filtrate; carrying out ultra high performance liquid chromatography detection on the reference solution and the test solution, wherein the detection conditions of the ultra high performance liquid chromatography comprise: the adopted mobile phase A is acetonitrile, and the mobile phase B is water; the elution mode is gradient elution. The characteristic map construction method can efficiently identify the glossy privet fruit medicinal material and the processed products thereof with different processing degrees.

Description

Method for constructing characteristic spectrum of glossy privet fruit medicinal material and processed product thereof and method for detecting content of multi-index components

Technical Field

The invention relates to the technical field of quality detection of traditional Chinese medicinal materials, in particular to a method for constructing a characteristic spectrum of a glossy privet fruit medicinal material and a processed product thereof, a method for detecting the content of multiple index components, and a method for synchronously detecting the characteristic spectrum and the content of the multiple index components of the glossy privet fruit medicinal material and the processed product thereof.

Background

Fructus Ligustri Lucidi is dried mature fruit of Ligustrum lucidum ait. Harvesting in winter, removing branches and leaves, steaming or slightly scalding in boiling water, and drying; or directly dried. Sweet and bitter in taste and cool in nature. It enters liver and kidney meridians. Has effects of nourishing liver and kidney, improving eyesight and blackening hair, and can be used for treating yin deficiency of liver and kidney, giddiness tinnitus, soreness of waist and knees, early white hair, dim eyesight, internal heat, diabetes, hectic fever, etc. The Chinese pharmacopoeia 2020 edition contains glossy privet fruit and wine glossy privet fruit, and the theory of traditional Chinese medicine holds that glossy privet fruit is mainly used for clearing liver and improving vision, nourishing yin and moistening dryness, and is mainly used for symptoms such as liver heat dizziness, yin deficiency and intestine dryness constipation and the like; the wine can activate blood circulation to remove meridian obstruction, dispel wind and remove cold, so the processed wine glossy privet fruit has stronger liver and kidney nourishing effect than glossy privet fruit, has weakened cold and cool nature, is mainly used for treating liver and kidney yin deficiency, dizziness and tinnitus, blurred vision, early white beard and hair and other symptoms, and is mainly applied to the wine glossy privet fruit in the traditional Chinese medicine clinical practice.

The fructus Ligustri Lucidi mainly contains terpenes, iridoid glycosides, phenylethanoid glycosides, flavonoids, volatile oil, fatty acids, and also contains polysaccharide and trace elements; wherein the most active components and medicinal values belong to triterpenes, iridoid glycosides and phenethyl alcohol glycosides, and the representative components of the triterpenes, iridoid glycosides and phenethyl alcohol glycosides are oleanolic acid, specnuezhenide, salidroside and the like. Modern researches find that the chemical components and pharmacological actions of the glossy privet fruit are changed to a certain extent in the processing process, but the requirement of single index content limit that the content of specnuezhenide in the glossy privet fruit is not less than 0.7 percent and the content of salidroside in the wine glossy privet fruit is not less than 0.2 percent is only stipulated in the 2020 edition of Chinese pharmacopoeia, and the whole quality control method of the glossy privet fruit and the processed wine glossy privet fruit is lacked. Meanwhile, the difference of active ingredients in the glossy privet fruit medicinal materials with different processing degrees is also obvious, so that the exertion of the efficacy of the glossy privet fruit processed product is influenced, and therefore, the glossy privet fruit medicinal materials with different processing degrees need to be effectively identified.

The traditional method relates to the identification of glossy privet fruit raw products and wine products, and one or at least two of 10-hydroxy-oleanolic acid dimethyl ester, neoligustilide or isomer, 8-demethyl-7-ketologanin, elenolic acid, verbascoside and luteolin are used as markers; meanwhile, the method adopts ultra-high performance liquid chromatography/quadrupole-Orbitrap mass spectrometry (UHPLC/Q-Orbitrap-MS) to separate and detect the sample, can effectively separate and detect the privet fruit sample, and can accurately judge whether the sample is a raw product or a wine product. However, the traditional method can only identify the raw product and the commercial wine product, and the difference of the quality of the wine product caused by different processing degrees is difficult to reflect.

And the traditional method relates to the comparison research of the HPLC fingerprints of the glossy privet fruit and the wine glossy privet fruit, acetonitrile-0.1% phosphoric acid water is used as a mobile phase to carry out elution according to a certain elution gradient, the HPLC fingerprints of the glossy privet fruit and the wine glossy privet fruit are constructed and obtained, and 11 main differential substances are obtained by screening, wherein the main differential substances comprise hydroxytyrosol, tyrosol, specnuezhenoside, oleuropein and quercetin. However, the traditional method can only identify the raw product and the commercial wine finished product, the difference of the quality of the wine product caused by different processing degrees is difficult to reflect, the detection time is as long as 1 hour, the detection efficiency is low, and the high efficiency of processing production is difficult to adapt.

Disclosure of Invention

Based on the above, the invention provides the glossy privet fruit and the method for constructing the characteristic map of the processed product thereof, wherein the glossy privet fruit and the processed product thereof can efficiently identify the glossy privet fruit medicinal material and different processing degrees.

The invention provides a method for constructing a characteristic spectrum of a glossy privet fruit medicinal material and a processed product thereof, which comprises the following steps:

preparing a reference substance solution and a test solution, wherein the preparation method of the reference substance solution comprises the following steps: dissolving 5-hydroxymethylfurfural reference substance, salidroside reference substance, echinacoside reference substance, specnuezhenide reference substance and nuezhenide G13 reference substance with a first solvent; the preparation method of the test solution comprises the following steps: extracting fructus Ligustri Lucidi or its processed product with a second solvent, filtering the extractive solution, and collecting the filtrate;

carrying out ultra high performance liquid chromatography detection on the reference solution and the test solution, wherein the detection conditions of the ultra high performance liquid chromatography comprise: the adopted mobile phase A is acetonitrile, and the mobile phase B is water; the elution mode is gradient elution.

In one embodiment, the gradient elution comprises the following procedure:

the volume percentage of the mobile phase A is increased to 10% from 2% in 0-5 min, and the volume fraction of the mobile phase B is decreased to 90% from 98%;

5-9 min, increasing the volume percentage of the mobile phase A from 10% to 18%, and decreasing the volume fraction of the mobile phase B from 90% to 82%;

the volume percentage of the mobile phase A is increased from 18% to 24% and the volume fraction of the mobile phase B is decreased from 82% to 76% in 9-11 min;

11-24 min, the volume percentage of the mobile phase A is increased from 24% to 28%, and the volume fraction of the mobile phase B is decreased from 76% to 72%;

and 24-25 min, the volume percentage of the mobile phase A is reduced from 28% to 2%, and the volume fraction of the mobile phase B is increased from 72% to 98%.

In one embodiment, the detection conditions of the ultra-high performance liquid chromatography further comprise:

a chromatographic column: octadecylsilane chemically bonded silica is used as a filling agent; detection wavelength: the detection wavelength is 268nm to 285nm, 3min to 25min and 224nm to 275nm when the time is 0min to 3 min; flow rate: 0.2mL/min to 0.3 mL/min; column temperature: 25-32 ℃.

In one embodiment, in the method for preparing a test solution,

the second solvent is methanol or a mixed solution of methanol and water; and/or

The dosage of the second solvent is 10 mL-75 mL per 0.3g of the glossy privet fruit medicinal material or the processed product thereof; and/or

The extraction method is ultrasonic extraction or heating to reflux extraction, and the extraction time is 30-60 min.

In one embodiment, the characteristic spectrum of the glossy privet fruit medicinal material comprises 10 common peaks, wherein the 10 common peaks comprise characteristic peaks of salidroside, echinacoside, specnuezhenide and nuezhenide G13; the feature map of the processed product of the glossy privet fruit is additionally provided with 1-4 common peaks on the basis of the feature map of the glossy privet fruit medicinal material, and the additionally provided 1-4 common peaks comprise the feature peak of 5-hydroxymethylfurfural.

The second aspect of the invention provides a method for detecting the content of multi-index components in glossy privet fruit medicinal materials and processed products thereof, which comprises the following steps:

preparing an index component reference solution and a to-be-detected sample solution, wherein the preparation method of the index component reference solution comprises the following steps: dissolving salidroside reference substance, specnuezhenide reference substance and nuezhenide G13 reference substance with a third solvent to obtain reference substance solutions with different concentrations of index components; the preparation method of the solution to be tested comprises the following steps: taking a glossy privet fruit medicinal material to be detected or a processed product thereof, adding a second solvent for extraction, filtering an extracting solution, and taking a subsequent filtrate;

carrying out ultra-high performance liquid chromatography detection on the index component reference substance solutions with different concentrations to construct a standard curve of the index components; carrying out ultra-high performance liquid chromatography detection on the solution of the sample to be detected, and calculating the content of the index component according to the detection result and the standard curve of the index component;

the detection conditions of the ultra-high performance liquid chromatography comprise: the adopted mobile phase A is acetonitrile, and the mobile phase B is water; the elution mode is gradient elution.

In one embodiment, the gradient elution comprises the following procedure:

In one embodiment, in the method for preparing the solution of the sample to be tested,

The third aspect of the invention provides a method for synchronously detecting the characteristic spectrum and the content of multi-index components of glossy privet fruit medicinal materials and processed products thereof, which comprises the following steps:

constructing the characteristic map of the glossy privet fruit medicinal material and the processed product thereof according to the characteristic map construction method;

constructing a standard curve of the index components according to the method for detecting the content of the multiple index components;

preparing a sample solution to be tested: taking a glossy privet fruit medicinal material to be detected or a processed product thereof, adding a second solvent for extraction, filtering an extracting solution, and taking a subsequent filtrate;

carrying out ultra-high performance liquid chromatography detection on the sample solution to be detected, and obtaining a detection result according to the detection result, the characteristic spectrum of the glossy privet fruit medicinal material and the processed product thereof and the standard curve of the index components;

In one embodiment, the gradient elution comprises the following procedure:

In one embodiment, in the method for preparing the sample solution to be tested,

The construction method can simultaneously establish the characteristic maps of the glossy privet fruit medicinal materials and the processed products with different processing degrees by adopting proper UPLC chromatographic conditions, the glossy privet fruit characteristic maps contain 10 characteristic peaks, 1-4 characteristic peaks are additionally arranged on the processed product characteristic maps, the quality of the glossy privet fruit medicinal materials and the processed products with different processing degrees can be comprehensively reflected, the method is stable and strong in specificity, accurate and reliable, simple and convenient to operate and good in reproducibility, and a scientific detection and identification method is provided for quality control of the glossy privet fruit medicinal materials and processing process control of the processed products. Meanwhile, the construction method is short in time consumption and high in detection efficiency, can adapt to the high efficiency of processing production, and is convenient for large-scale popularization and application.

The method for detecting the content of the multi-index components can accurately quantify the salidroside, specnuezhenide and nuezhenide G13 index components in the glossy privet fruit medicinal material and the processed product thereof by adopting a proper UPLC chromatographic condition, has reliable result and simple and convenient operation, makes up the blank that the 2020 edition of Chinese pharmacopoeia cannot simultaneously detect the content of the salidroside and the content of the specnuezhenide of the glossy privet fruit medicinal material and the processed variety thereof, and increases the index determination of the ligustilide G13 with higher content.

Furthermore, by adopting a proper UPLC chromatographic condition, the invention can simultaneously realize the construction of the characteristic spectrums of the glossy privet fruit medicinal materials and the processed products with different processing degrees and the detection of the contents of multiple index components in the glossy privet fruit medicinal materials, is favorable for comprehensively carrying out quality control on the glossy privet fruit medicinal materials and the processed products with different processing degrees, is suitable for industrial high-efficiency production, has simple and convenient operation, high precision, high accuracy, good stability and good reproducibility, and is convenient for large-scale popularization and application.

Drawings

FIG. 1 is a characteristic spectrum and a reference characteristic spectrum of 11 batches of fructus Ligustri Lucidi material in example 1 of the present invention;

FIG. 2 is a characteristic map of a processing process in which fructus Ligustri Lucidi decoction pieces are crushed and then steamed with wine for 8 hours in example 1 of the present invention;

FIG. 3 is a characteristic diagram of a processing process of fructus Ligustri Lucidi decoction pieces steamed without crushing wine for 8 hours in example 1 of the present invention;

FIG. 4 is a characteristic map of a processing process of fructus Ligustri Lucidi decoction pieces stewed with unburnt wine for 8 hours in example 1 of the present invention;

FIG. 5 is a chromatogram of the mixed control of example 1 (Peak 11: 5-hydroxymethylfurfural; Peak 3: salidroside; Peak 4: echinacoside; Peak 7: specnuezhenide; Peak 9: nuezhenide G13);

FIG. 6 is a PDA score chart of 14 characteristic peaks of different processed glossy privet fruits in example 1;

FIG. 7 is an OPLS-DA score chart of 14 characteristic peaks of different processed glossy privet fruits in example 1;

FIG. 8 is a specificity study map of the characteristic maps of the fructus Ligustri Lucidi and its processed decoction pieces in example 1;

FIG. 9 is a study of the elution gradient one and the detection wavelength 275nm in example 1;

FIG. 10 is a plot of the elution gradient one and detection wavelength 224nm investigation of example 1;

FIG. 11 is a two elution gradients and a 275nm detection spectrum from example 1;

FIG. 12 is a second elution gradient and a detection wavelength of 224nm as investigated in example 1;

FIG. 13 is a graph of a test spectrum at a detection wavelength of 275nm (0-3 min) in example 1;

FIG. 14 is a spectrum of the detection wavelength of 284nm (0-3 min) in example 1;

FIG. 15 is a map of the examination of the extraction solvent in example 1;

FIG. 16 is a specificity investigation map for determining the content of target components in fructus Ligustri Lucidi and processed decoction pieces thereof in example 2;

FIG. 17 is a linear investigation spectrum of the index component salidroside of fructus Ligustri Lucidi and processed decoction pieces thereof in example 2;

FIG. 18 is a linear inspection spectrum of the index component specnuezhenide of fructus Ligustri Lucidi and its processed decoction pieces in example 2;

FIG. 19 is a linear survey map of ligustrum lucidum medicinal material and ligustilide G13 as an index component of processed decoction pieces thereof in example 2.

Detailed Description

The method for constructing the characteristic spectrum of the glossy privet fruit medicinal material and the processed product thereof is further described in detail with reference to specific examples. The present invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

As used herein, "one or more" refers to any one, any two, or any two or more of the listed items.

In the present invention, "first aspect", "second aspect", "third aspect" and the like are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or quantity, nor are they to be construed as implicitly indicating the importance or quantity of the technical feature indicated. Also, "first," "second," "third," etc. are for non-exhaustive enumeration description purposes only and should not be construed as constituting a closed limitation to the number.

In the present invention, the technical features described in the open type include a closed technical solution composed of the listed features, and also include an open technical solution including the listed features.

In the present invention, the numerical range is defined to include both end points of the numerical range unless otherwise specified.

The percentage contents referred to in the present invention mean, unless otherwise specified, mass percentages for solid-liquid mixing and solid-solid mixing, volume percentages for liquid-liquid mixing, and water as a solvent base.

The percentage concentrations referred to in the present invention refer to the final concentrations unless otherwise specified. The final concentration refers to the ratio of the additive component in the system to which the component is added.

The temperature parameter in the present invention is not particularly limited, and may be a constant temperature treatment or a treatment within a certain temperature range. The constant temperature process allows the temperature to fluctuate within the accuracy of the instrument control.

In some of these examples, the gradient elution includes the following procedure:

increasing the volume percentage of the mobile phase A from 2% to 10% and decreasing the volume fraction of the mobile phase B from 98% to 90% in 0-5 min;

9-11 min, increasing the volume percentage of the mobile phase A from 18% to 24%, and decreasing the volume fraction of the mobile phase B from 82% to 76%;

the volume percentage of the mobile phase A is increased to 28% from 24% and the volume fraction of the mobile phase B is decreased to 72% from 76% in 11-24 min;

and 24-25 min, the volume percentage of the mobile phase A is reduced to 2% from 28%, and the volume fraction of the mobile phase B is increased to 98% from 72%.

In some examples, the detection conditions of ultra-high performance liquid chromatography further comprise:

In some examples, the detection wavelength of the ultra-high performance liquid chromatography is: 0min to 3min, the detection wavelength is 282nm to 285nm, 3min to 25min, and the detection wavelength is 224nm to 230 nm.

In some examples, the flow rate of the ultra high performance liquid chromatography is 0.2mL/min to 0.3 mL/min. Specifically, flow rates include, but are not limited to: 0.2mL/min, 0.21mL/min, 0.22mL/min, 0.23mL/min, 0.24mL/min, 0.25mL/min, 0.26mL/min, 0.27mL/min, 0.28mL/min, 0.29mL/min, 0.3 mL/min.

In some of these examples, the column temperature of ultra performance liquid chromatography is 25 ℃ to 32 ℃. Specifically, column temperatures include, but are not limited to: 25 ℃, 26 ℃, 27 ℃, 28 ℃, 29 ℃, 30 ℃, 31 ℃ and 32 ℃.

In some examples, the sample solution is prepared by a method in which the second solvent is methanol or a mixture of methanol and water. Furthermore, the volume fraction of the methanol in the mixed liquid of the methanol and the water is 25 to 80 percent. Specifically, the volume fraction of methanol in the mixed solution of methanol and water includes, but is not limited to: 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 68%, 70%, 72%, 75%, 80%.

In some examples, in the method for preparing the test solution, the amount of the second solvent is 10mL to 75mL per 0.3g of the fructus ligustri lucidi medicinal material or the processed product thereof. Furthermore, the dosage of the second solvent is 20mL to 30mL per 0.3g of the glossy privet fruit medicinal material or the processed product thereof.

In some examples, the sample solution is prepared by ultrasonic extraction or heating to reflux extraction. Furthermore, the power of the ultrasonic wave is 450W-500W, and the frequency is 35 kHz-45 kHz.

In some examples, the sample solution is prepared by a method in which the extraction time is 30min to 60 min. Specifically, the time of extraction includes, but is not limited to: 30min, 35min, 40min, 45min, 50min, 53min, 55min, 56min, 57min, 58min, 59min, 60 min.

In some examples, the control solution is prepared by a method in which the first solvent is an alcohol solvent, such as methanol.

In some examples, the sample volumes of the control solution and the test solution are 0.5 μ L to 1.5 μ L.

In some examples, the characteristic spectrum of the fructus ligustri lucidi medicinal material comprises 10 common peaks, wherein the 10 common peaks comprise characteristic peaks of salidroside, echinacoside, specnuezhenide and ligustilide G13; the feature map of the processed product of the glossy privet fruit is additionally provided with 1-4 common peaks on the basis of the feature map of the glossy privet fruit medicinal material, and the additionally provided 1-4 common peaks comprise the feature peak of 5-hydroxymethylfurfural.

The invention also provides a method for detecting the content of the multi-index components of the glossy privet fruit medicinal material and the processed product thereof, which comprises the following steps:

preparing an index component reference solution and a solution of a sample to be tested, wherein the preparation method of the index component reference solution comprises the following steps: dissolving salidroside reference substance, specnuezhenide reference substance and nuezhenide G13 reference substance with a third solvent to obtain reference substance solutions with different concentrations of index components; the preparation method of the solution of the to-be-detected object comprises the following steps: taking a glossy privet fruit medicinal material to be detected or a processed product thereof, adding a second solvent for extraction, filtering an extracting solution, and taking a subsequent filtrate;

carrying out ultra-high performance liquid chromatography detection on the index component reference substance solutions with different concentrations to construct a standard curve of the index components; performing ultra-high performance liquid chromatography detection on the solution to be detected, and calculating the content of the index component according to the detection result and the standard curve of the index component;

It can be understood that the technical scheme of the method for detecting the content of the multiple index components of the fructus ligustri lucidi medicinal material and the processed product thereof is similar to the method for constructing the characteristic map of the fructus ligustri lucidi medicinal material and the processed product thereof, wherein the preparation of the solution of the to-be-detected product is the same as the preparation of the solution of the test product, and the details are not repeated herein.

The invention also provides a method for synchronously detecting the characteristic spectrum and the multi-index component content of the glossy privet fruit medicinal material and the processed product thereof, which comprises the following steps:

It can be understood that the technical scheme of the above-mentioned method for synchronously detecting the characteristic spectrum and the multi-index component content of the fructus ligustri lucidi medicinal material and the processed product thereof is similar to the aforementioned method for constructing the characteristic spectrum of the fructus ligustri lucidi medicinal material and the processed product thereof, wherein the preparation of the sample solution to be detected is the same as the preparation of the test sample solution, and the details are not repeated herein.

The instruments and reagents used in the examples of the invention were as follows:

1. instrument for measuring the position of a moving object

Waters ultra high performance liquid chromatograph (Waters H-class), PDA detector (Waters); an Empower workstation; a ten thousandth balance (mettler-toledo, ME 204E); one millionth (mettler-toledo, XP 26); ultra pure water machines (Merck, Milli-Q Direct 8/16 system); an ultrasonic cleaner (KQ 5500DE, ultrasonic instruments, Inc. of Kunshan); an ACQUITY UPLC BEH Shield RP18 Column (100 mm. times.2.1 mm,1.7 μm).

2. Reagent

Acetonitrile is chromatographic pure, and water is ultrapure water; the other reagents are analytically pure.

5-hydroxymethylfurfural control (batch No. wkq 20041311; source: Vickqi Biotech, Inc., Sichuan); salidroside reference (batch No. 110818-; echinacoside reference (batch number: 111670-; a specnuezhenide reference substance (batch number: 111926-; ligustrum lucidum G13 reference substance (batch No. 8857; Shanghai Shidande Standard technology service Co., Ltd.; content%: 95.0).

Example 1

The embodiment is a UPLC characteristic spectrum establishing method of a glossy privet fruit medicinal material and processed decoction pieces thereof.

Glossy privet fruit medicinal materials: 11 batches of glossy privet fruit medicinal materials which are from Henan, Jiangsu, Shandong and other production places and meet the regulation of 'Chinese pharmacopoeia' of 2020 edition are collected.

Processing decoction pieces wine and glossy privet fruit: taking a glossy privet fruit medicinal material, removing impurities, uniformly dividing into three parts, crushing one part to obtain a glossy privet fruit crushed decoction piece, and crushing the other two parts to obtain a glossy privet fruit uncrushed decoction piece; putting the glossy privet fruit decoction pieces into a container, diluting the yellow wine with drinking water which is 1 time of the weight of the yellow wine, and taking out the glossy privet fruit decoction pieces after moistening the glossy privet fruit decoction pieces for 2-4 hours in 20kg of the yellow wine per 100kg of glossy privet fruit decoction pieces; one part of the glossy privet fruit crushed decoction pieces are steamed with wine for 8.0 hours, and the other two parts of the glossy privet fruit crushed decoction pieces are respectively steamed with wine and stewed with wine for 8 hours; sampling every 0.5 hours, and drying in an oven to obtain the glossy privet fruit decoction pieces processed with the wine in different decoction piece specifications and different processing times. Wherein, the wine steaming label of the glossy privet fruit crushed decoction pieces is YP-JZ, the wine steaming label of the glossy privet fruit not crushed decoction pieces is BYP-JZ, and the wine stewing label of the glossy privet fruit not crushed decoction pieces is BYP-JD.

1. Chromatographic conditions and System suitability test

Octadecylsilane chemically bonded silica was used as a filler (ACQUITY UPLC BEH Shield RP18 Column, Column length 100mm, inner diameter 2.1mm, particle diameter 1.7 μm); acetonitrile is taken as a mobile phase A, water is taken as a mobile phase B, and gradient elution is carried out according to the specification in the table 1; the flow rate is 0.3mL per minute; the column temperature is 30 ℃; the detection wavelength is 284nm for 0min to 3min and 224nm for 3min to 25 min. The number of theoretical plates should not be less than 30000 calculated according to specnuezhenide peak.

TABLE 1 gradient elution Table

2. Preparation of control solutions

Taking appropriate amount of 5-hydroxymethyl reference substance, salidroside reference substance, echinacoside reference substance, specnuezhenide reference substance, and nuezhenide G13 reference substance, precisely weighing, and adding methanol to obtain solution containing 0.1mg per 1mL as reference solution.

3. Preparation of test solution

Taking a glossy privet fruit medicinal material or glossy privet fruit decoction pieces (YP-JZ, BYP-JZ and BYP-JD) and preparing the glossy privet fruit medicinal material or glossy privet fruit decoction pieces into powder (passing through a third sieve), taking about 0.3g of the powder, precisely weighing the powder, placing the powder into a conical flask with a plug, precisely adding 25mL of 70 percent methanol, weighing the weight, carrying out ultrasonic treatment (power: 500W and frequency: 40kHz) for 60 minutes, cooling the mixture, weighing the weight again, complementing the lost weight with 70 percent methanol, shaking the mixture evenly, filtering the mixture, and taking a subsequent filtrate to obtain the glossy privet fruit decoction pieces.

4. Measurement of

Precisely sucking 1 μ L of each of the reference solution and the sample solution, injecting into Waters H-class ultra high performance liquid chromatograph, and measuring.

5. Measurement results

(1) Measurement of fructus Ligustri Lucidi

The method comprises the steps of performing common peak matching on characteristic spectrums of 11 batches of glossy privet fruit medicinal materials by using a traditional Chinese medicine chromatography fingerprint similarity evaluation system to generate a comparison characteristic spectrum, determining 10 characteristic peaks in total as shown in figure 1, performing chemical component identification on the 10 common peaks, determining that the peak 3 is a salidroside peak, the peak 4 is an echinacoside peak, the peak 7 is a specnuezhenoside peak, the peak 9 is a privet glycoside G13 peak, and using the peak 7 and the specnuezhenoside peak as a reference peak S to prepare reference data of the glossy privet fruit medicinal material characteristic spectrum analysis method. Specifically, the peak 7 specnuezhenide peak is taken as a reference peak, the relative retention time and the relative peak area of the other characteristic peaks are calculated, and the experimental results are shown in tables 2 and 3.

TABLE 211 relative retention times of characteristic spectra of glossy privet fruit

TABLE 311 relative peak area of characteristic spectrum of glossy privet fruit

(2) Determination result of processing process of wine glossy privet fruit

In the 8-hour processing process of YP-JZ, BYP-JZ and BYP-JD, a processed decoction piece wine glossy privet sample sampled every 0.5 hour is subjected to characteristic map analysis by adopting a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, 1-4 characteristic peaks are newly added along with the prolonging of processing time, as shown in figures 2, 3 and 4, and 1-14 common peaks are subjected to chemical component identification, so that the peak 3 is determined to be a salidroside peak, the peak 4 is determined to be an echinacoside peak, the peak 7 is determined to be a specnuezhenide peak, the peak 9 is determined to be a rhodinin G13 peak, and the peak 11 is determined to be a 5-hydroxymethylfurfural peak, as shown in figure 5.

Performing main component analysis (PDA) and partial orthogonal least squares discriminant analysis (OPLS-DA) on the peak areas of chromatographic peaks in UPLC characteristic maps of the determined wine fructus Ligustri Lucidi decoction pieces, recording the peak areas of 1-4 newly added characteristic peaks which are not detected in the wine fructus Ligustri Lucidi decoction pieces by 0, introducing the peak areas of 14 common peaks calibrated in different decoction piece specifications and different processing modes into SIMCA14.0 software for analysis, and showing the PDA and OPLS-DA score maps in FIGS. 6 and 7. The classification and aggregation of the YP-JZ and the BYP-JZ or the BYP-JD of the glossy privet fruit decoction pieces are obvious in a PDA unsupervised model, and the BYP-JZ and the BYP-JD of the glossy privet fruit decoction pieces are aggregated together. Further analysis by OPLS-DA supervised model, R₂Y (representing the degree of fitting of OPLS-DA model) reaches 0.924, Q²(representing the predictive power of the model) reaches 0.889. The results show that the glossy privet fruit decoction pieces with different decoction piece specifications and different processing modes are obviously clustered and collected in an OPLS-DA model.

The UPLC characteristic spectrum determination result of the processing process of the wine and glossy privet fruit shows that the glossy privet fruit is added with yellow wine after being crushed and moistened, and new chromatographic peaks appear in a short time along with the prolonging of the wine steaming time, namely chemical components are converted and generated, and 14 characteristic peaks are generated after the wine is steamed for 1 hour; directly adding yellow wine into fructus Ligustri Lucidi without crushing, moistening, steaming with wine for 2 hr to generate 14 characteristic peaks, and stewing with wine for 7.5 hr to generate 14 characteristic peaks; the main influencing factor for generating the difference is whether water vapor is in direct contact in the processing process, namely, the seed coat of the fructus ligustri lucidi is damaged after being crushed, the water vapor in the wine steaming process can be directly in contact with the fructus ligustri lucidi, the chemical components are easy to convert and generate under the high humidity and high heat conditions, the direct contact of the high humidity water vapor is weakened under the protection of the seed coat of the non-crushed fructus ligustri lucidi, the conversion and generation time of the chemical components is prolonged, the direct contact of the water vapor is further isolated by stewing with wine, and the conversion and generation time of the chemical components is further prolonged. Further analysis by combining PDA and OPLS-DA models shows that the proportion of chemical components of glossy privet fruit is different under different processing conditions.

In summary, 10 common peaks are found in the more stable characteristic spectrum with good separation degree among 11 batches of fructus ligustri lucidi medicinal materials. The retention time of 4 peaks should correspond to the retention time of the reference peak of the corresponding reference substance, the peak corresponding to the specnuezhenide reference substance is the S peak, the relative retention time of each characteristic peak and the S peak should be calculated to be within ± 10% of the specified value, and the specified values are 0.32 (peak 1), 0.35 (peak 2), 0.93 (peak 5), 0.96 (peak 6), 1.11 (peak 8) and 1.46 (peak 10). And calculating the relative retention time of each characteristic peak and the S peak, wherein the relative retention time is within +/-10% of a specified value, and the specified values are 0.80 (peak 12), 1.05 (peak 13) and 1.18 (peak 14).

6. Methodology validation

(1) Specificity test

Taking the sample solution, blank solvent and reference solution prepared in the same batch, detecting under the condition of the liquid chromatography, injecting 1 mu L of sample each time, and recording the chromatogram. The result is shown in fig. 8, and no chromatographic peak is present in the blank solvent chromatogram at the retention time position corresponding to the characteristic peak, indicating that the extraction solvent has no interference with the measurement of each characteristic peak and has good specificity.

(2) Precision test

And taking the same sample solution, repeatedly injecting sample for 6 times under the liquid chromatography condition, injecting 1 mu L of sample each time, taking the specnuezhenide peak as an S peak, and calculating the relative retention time of each characteristic peak and the RSD value of the relative peak area. The RSD of the relative retention time of each characteristic peak is less than 2%, the RSD of the relative peak area is less than 3%, and the result shows that the precision of the instrument is good.

(3) Repeatability test

Taking the same batch of samples, preparing 6 test sample solutions according to the test sample preparation method, carrying out sample injection measurement under the liquid chromatography condition, carrying out sample injection for 1 mu L each time, taking the specnuezhenide peak as an S peak, and calculating the relative retention time of each characteristic peak and the RSD value of the relative peak area. The RSD of the relative retention time of each characteristic peak is less than 2%, the RSD of the relative peak area is less than 3%, and the result shows that the analysis method is good in repeatability.

(4) Stability test

Taking the sample solution, placing at room temperature, detecting under the above liquid chromatography conditions for 0 hr, 2 hr, 4 hr, 8 hr, 12 hr and 24 hr, respectively, sampling 1 μ L each time, taking the specnuezhenide peak as S peak, and calculating relative retention time of each characteristic peak and RSD value of relative peak area. The RSD of the relative retention time of each characteristic peak is less than 2%, and the RSD of the relative peak area is less than 5%, and the result shows that the stability of the test solution is good within 24 hours.

7. Inspection of chromatographic conditions

(1) Elution gradient and detection wavelength

1.1 elution gradient one

The preparation of the test sample is carried out by using the wine glossy privet fruit YP-JZ (8 hours) according to the item of 'preparation of test sample solution', according to the same conditions as 'chromatographic conditions and system applicability test', the detection wavelengths adopted are dual-wavelength 224nm and 275nm, the elution gradient is the elution gradient shown in the following table 4, and the elution gradient investigation is carried out.

TABLE 4

As a result, referring to fig. 9 and fig. 10, it can be seen that the peak appearance time of each spectral peak becomes shorter as the polarity of the mobile phase becomes larger, and the peak appearance density of each spectral peak is poor, indicating that a chromatogram with good separation and adapted to the retention time of each spectral peak cannot be obtained well under this gradient condition.

1.2 elution gradient two

The preparation of the test sample is carried out by using the wine glossy privet fruit YP-JZ (8 hours) according to the item of 'preparation of test sample solution', according to the same conditions as 'chromatographic conditions and system applicability test', the detection wavelengths adopted are dual-wavelength 224nm and 275nm, the elution gradient is the elution gradient shown in the following table 5, and the elution gradient investigation is carried out.

TABLE 5

Referring to fig. 11 and 12, it can be seen that the peak emergence time of each chromatographic peak is moderate and the separation degree is good in comparison with the chromatograms of gradient elution one; in addition, as the chromatograms at the wavelengths of 224nm and 275nm are compared, the absorption of each chromatographic peak at the wavelength of 224nm is large, the peak type is good, but a part of chromatographic peaks at the wavelength of 275nm is lacked within 0-3 min, and the lacked peaks comprise 5-hydroxymethylfurfural (namely, no absorption exists at the wavelength of 224 nm) through comparison of a reference substance, so that the influence of the detection wavelength on the absorption of 5-hydroxymethylfurfural needs to be further examined.

1.3 detection wavelength

The preparation of the test sample is carried out by using the wine glossy privet fruit YP-JZ (8 hours) according to the item of 'preparation of test sample solution', and the detection wavelengths in the process of elution for 0-3 min are only changed according to the same conditions as 'chromatographic conditions and system applicability test', and are 275nm and 284nm respectively.

The results are shown in FIGS. 13 to 14. Therefore, the absorption (peak area) of the No. 11 peak 5-hydroxymethylfurfural within 0-3 min at the detection wavelength of 284nm is larger than the wavelength of 275nm, and the following table is shown in 6-7.

TABLE 6 Retention time

TABLE 7 Peak area

(2) Investigation of extraction solvent in preparation of test sample

Preparing the test sample by using the wine-processed glossy privet fruit YP-JZ (8 hours) according to the item of 'preparation of test sample solution' under the same conditions as 'chromatographic conditions and system applicability test', and only changing the extraction solvent in the preparation of the test sample to carry out extraction solvent investigation. The extraction solvents considered were 30% methanol, 50% methanol, 70% methanol, 50% ethanol, and 70% ethanol, respectively.

The results are shown in tables 8 to 9 below and FIG. 15.

TABLE 8 Retention time

TABLE 9 Peak area

The results show that different extraction solvents are adopted for extraction, an ethanol system is adopted as the solvent, and the number of chromatographic peaks is lacked; a methanol system is adopted as an extraction solvent, the number of chromatographic peaks has no obvious difference, and the main chromatographic peaks can reach baseline separation; methanol and methanol-water are used as extraction solvents, and the ratio of the total peak area/the sample weighing amount of 14 common characteristic peaks is increased along with the increase of the concentration of the methanol.

(3) Investigation of extraction mode in preparation of test sample

Preparing the test sample by wine glossy privet fruit YP-JZ (8 hours) according to the item of 'preparation of test sample solution' under the same conditions as 'chromatographic conditions and system applicability test', and observing the extraction mode by only changing the extraction mode in the preparation of the test sample. The examined extraction modes are ultrasonic extraction and reflux extraction respectively.

The results are shown in tables 10 to 11 below.

TABLE 10 Retention time

TABLE 11 Peak area

The results show that the two extraction modes of reflux and ultrasonic have no obvious difference in the aspects of chromatographic peak number, chromatographic peak separation effect and 'total peak area/sample weighing', and the ultrasonic extraction can be selected in consideration of convenient operation.

(3) Investigation of extraction solvent dosage in sample preparation

Preparing the test sample by wine fructus Ligustri Lucidi YP-JZ (8 hr) according to the item of "preparation of test sample solution" under the same conditions as "chromatographic conditions and System suitability test", and only changing the amount of extraction solvent in the preparation of the test sample. The amount of the extraction solvent was 10mL, 25mL, 50mL, and 75mL, respectively.

The results are shown in tables 12 to 13 below.

TABLE 12 Retention time

TABLE 13 Peak area

The result shows that the use amount of the extraction solvent is the largest by adopting the ratios of the total peak area/sample weighing amount of the 14 common characteristic peaks to add 25mL of the extraction solvent, and 25mL is selected as the use amount of the extraction solvent in consideration of saving the solvent.

(4) Investigation of extraction time in sample preparation

Preparing the test sample by wine fructus Ligustri Lucidi YP-JZ (8 hr) according to the item of "preparation of test sample solution" under the same conditions as "chromatographic conditions and System suitability test", and only changing the extraction time in the preparation of the test sample. The extraction time is 30min, 45min, and 60min respectively.

The results are shown in tables 14 to 15 below.

TABLE 14 Retention time

TABLE 15 Peak area

The results show that the ultrasonic extraction time is different, the number and the separation degree of characteristic spectrum chromatographic peaks and the total peak area/sample weighing amount of 14 characteristic peaks of the characteristic spectrum have no obvious difference, the ultrasonic extraction is basically complete within 30 minutes, and the ultrasonic time can be properly prolonged to 60 minutes.

Example 2

The embodiment is a method for detecting the content of index components of a glossy privet fruit medicinal material and processed decoction pieces thereof.

The main effective components of the glossy privet fruit comprise triterpenes, iridoid glycosides and phenethyl alcohol glycosides, and the main representative components comprise salidroside, specnuezhenide, privet glycoside, ligustilide G13, Oleonezhezhezhenzhenide and the like. The invention evaluates the glossy privet fruit medicinal material and the processing decoction pieces thereof by taking salidroside, specnuezhenide and nuezhenide G13 as content measurement indexes.

1. Chromatographic conditions and System suitability test

TABLE 1 gradient elution Table

2. Preparation of reference substance solution of index component

Taking appropriate amount of salidroside reference substance, specnuezhenide reference substance, and nuezhenide G13 reference substance, precisely weighing, and adding methanol to obtain 1mL salidroside reference substance solution containing 200 μ G, specnuezhenide reference substance solution containing 200 μ G, and privet glycoside G13 reference substance solution containing 200 μ G.

3. Preparation of test solution

Taking a glossy privet fruit medicinal material or a processed decoction piece thereof, preparing into powder (sieving by a No. three sieve), taking about 0.3g of powder, precisely weighing, placing into a conical flask with a plug, precisely adding 25mL of 70% methanol, weighing, ultrasonically treating for 60 minutes (power: 500W, frequency: 40kHz), cooling, weighing again, complementing the lost weight with 70% methanol, shaking uniformly, filtering, and taking a subsequent filtrate to obtain the traditional Chinese medicine composition.

4. Measurement of

Precisely sucking 1 μ L of each of the reference solution and the sample solution, and injecting into Waters H-class ultra high performance liquid chromatograph.

5. Measurement results

The results of measuring the contents of 11 batches of glossy privet fruit medicinal materials and the glossy privet fruit decoction pieces processed in different processing times are shown in the following tables 16 and 17:

TABLE 1611 determination of the amount of glossy privet fruit in batches (%)

TABLE 17 determination of the processing procedure of glossy privet fruit (in%) with wine

6. Methodology validation

(1) Specialization inspection

Taking the sample solution, index component reference solution and blank solvent prepared in the same batch, detecting under the liquid chromatography condition, injecting 1 uL sample each time, and recording chromatogram. The results are shown in FIG. 16. The result shows that no chromatographic peak exists in the blank solvent chromatogram at the retention time positions corresponding to salidroside, specnuezhenide and nuezhenide G13, which indicates that the extraction solvent has no interference to the content measurement of salidroside, specnuezhenide and nuezhenide G13 and has good specificity.

(2) Investigation of linear relationships

Firstly, the linear inspection of salidroside

Precisely weighing salidroside reference substances, placing into a 5mL measuring flask, adding methanol to prepare into each 1mL of serial reference substance solutions containing salidroside 301.1820 μ g, salidroside 120.4728 μ g, salidroside 60.2364 μ g, salidroside 30.2176 μ g and salidroside 12.1268 μ g, respectively, detecting under the above liquid chromatogram condition, introducing 1 μ L of sample each time, performing linear regression calculation with salidroside concentration as abscissa and measured peak area as ordinate, and drawing standard curve, wherein the regression equation of salidroside is as follows: 4797.6x-1697.5, and its correlation coefficient R²0.9996, the salidroside has good linear relation with peak area in the range of 12.1268 mu g/ml to 301.1820 mu g/ml. The results are shown in FIG. 17.

② linear investigation of specnuezhenide

Precisely weighing the specnuezhenide reference substance, placing the specnuezhenide reference substance into a 5mL measuring flask, adding methanol to prepare each 1mL of serial reference substance solutions containing 247.7600 μ g of specnuezhenide, 100.1300 μ g of specnuezhenide, 49.5900 μ g of specnuezhenide, 24.7000 μ g of specnuezhenide and 9.8800 μ g of specnuezhenide respectively, detecting under the liquid chromatography condition, carrying out linear regression calculation by taking the concentration of the specnuezhenide as a horizontal coordinate and the measured peak area as a vertical coordinate, drawing a standard curve, wherein the regression equation of the specnuezhenide is as follows: 4794.0x-3785.6, and its correlation coefficient R²0.9992, the linear relation between the injection concentration and the peak area is good in the range of 9.8800 mu g/ml to 247.7600 mu g/ml of specnuezhenide. The results are shown in FIG. 18.

(iii) Linear examination of Ligustrum lucidum G13

Precisely weighing ligustrin G13 reference substance, placing into 5mL measuring flask, adding methanol, and making into ligustrin containing 271.1300 μ G per 1mLDetecting serial reference solutions of glycoside G13, 101.0800 mu G of ligustilide G13, 59.2800 mu G of ligustilide G13, 27.1700 mu G of ligustilide G13 and 10.8300 mu G of ligustilide G13 under the liquid chromatography condition, carrying out linear regression calculation by taking the concentration of the ligustilide G13 as an abscissa and the measured peak area as an ordinate for each injection, and drawing a standard curve, wherein the regression equation of the ligustilide G13 is as follows: 3817.2x +25885.8, and its correlation coefficient R²When the injection concentration is in the range of 10.8300 mu G/ml to 271.1300 mu G/ml, the linear relation between the injection concentration and the peak area is good for ligustrazine G13 which is 0.9991. The results are shown in FIG. 19.

(3) Precision test

And taking the same batch of samples, repeatedly injecting samples for 6 times under the liquid chromatography condition, injecting 1 mu L of sample each time, and calculating the peak area RSD value. The result shows that the RSD value of the rhodiola glycoside peak area is 0.60%, the RSD value of the specnuezhenide peak area is 0.30%, and the RSD value of the privet glycoside G13 peak area is 0.23%, which indicates that the precision of the instrument is good.

(4) Repeatability test

Taking the same batch of samples, preparing 6 parts of sample solution according to the sample preparation method, and performing sample injection measurement under the condition of the liquid chromatography, wherein the sample injection amount is 1 mu L each time. The result shows that the average value of the salidroside content is 0.23 percent, and the RSD value is 0.62 percent; the mean value of the content of specnuezhenide is 2.88 percent, and the RSD value is 0.73 percent; the content average value of the ligustilide G13 is 1.80 percent, and the RSD value is 0.76 percent; the analysis method is proved to have good repeatability.

(5) Accuracy test

And (3) carrying out sample adding recovery rate determination by adopting a reference substance, designing 3 groups of experiments according to the condition that the ratio of the adding amount of the salidroside reference substance, the specnuezhenide reference substance and the ligustrin G13 reference substance to the content of the components to be detected, namely salidroside, specnuezhenide and ligustrin G13 in the test solution is 0.5:1, 1:1 and 1.5:1, precisely transferring appropriate amounts of salidroside reference substance storage solution, specnuezhenide reference substance storage solution and ligustrin G13 reference substance storage solution in parallel of 3 parts of each group, and blowing and drying under a nitrogen blowing instrument. Taking about 0.15g of a same batch of samples with known content, precisely weighing, placing in 3 groups of conical flasks, each group containing 3 parts in parallel, preparing a sample solution according to the sample preparation method, performing sample injection measurement under the liquid chromatography condition, performing sample injection of 1 mu L each time, and calculating the accuracy respectively, see tables 18, 19 and 20. The result shows that the recovery rate of the salidroside content determination analysis method is within the range of 98.02-103.02%, and the RSD is 1.51%; the recovery rate of the specnuezhenide content determination analysis method is 96.01-101.03%, and the RSD is 1.73%; the recovery rate of the glossy privet fruit glycoside G13 content determination analysis method is in the range of 96.97-105.75%, and the RSD is 2.78%; meets the requirement of the verification guiding principle of the quality standard analysis method of the drug of 9101 in 2020 edition of Chinese pharmacopoeia.

TABLE 18 survey of the accuracy of the determination of salidroside content in wine Ligustrum lucidum

TABLE 19 examination of the accuracy of the determination of specnuezhenide content in the wine Ligustrum lucidum samples

TABLE 20 Ligustrum lucidum ait G13 content determination accuracy in Ligustrum lucidum ait samples

(6) Stability test

Taking the test solution, placing at room temperature, detecting under the above liquid chromatography conditions at 0 hr, 2 hr, 4 hr, 8 hr, 12 hr and 24 hr, injecting 1 μ L sample each time, recording peak area, and calculating RSD value. The result shows that the RSD value of the salidroside peak area measured by injection in 24 hours is 4.84 percent; the specific privet glycoside peak area RSD value is 2.94%; the RSD value of the peak area of the ligustrazine G13 is 2.43%, which shows that the stability of the test solution is better within 24 hours.

Meanwhile, the UPLC spectra of the fructus ligustri lucidi medicinal material and the processed decoction pieces thereof can be respectively compared with the characteristic spectra of the fructus ligustri lucidi medicinal material and the processed decoction pieces thereof established in example 1, so that the characteristic spectrum detection results of the corresponding fructus ligustri lucidi medicinal material and the processed decoction pieces thereof and the content results of the index components of salidroside, specnuezhenide and ligustilide G13 can be synchronously obtained, and the evaluation of the fructus ligustri lucidi medicinal material and the processed decoction pieces thereof can be comprehensively carried out.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, so as to understand the technical solutions of the present invention specifically and in detail, but not to be understood as the limitation of the protection scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. It should be understood that the technical solutions provided by the present invention, which are obtained by logical analysis, reasoning or limited experiments, are within the scope of the appended claims. Therefore, the protection scope of the present invention should be subject to the content of the appended claims, and the description and the drawings can be used for explaining the content of the claims.

Claims

1. A method for constructing a characteristic spectrum of a glossy privet fruit medicinal material and a processed product thereof is characterized by comprising the following steps:

2. The method for constructing the feature map of the glossy privet fruit medicinal material and the processed product thereof according to claim 1, wherein the gradient elution comprises the following procedures:

3. The method for constructing the characteristic spectrum of the glossy privet fruit medicinal material and the processed product thereof according to claim 1, wherein the detection conditions of the ultra-high performance liquid chromatography further comprise:

4. The method for constructing the characteristic spectrum of the glossy privet fruit medicinal material and the processed product thereof according to claim 1, wherein in the method for preparing the test solution, the second solvent is methanol or a mixed solution of methanol and water; and/or

5. The method for constructing the characteristic spectrum of the glossy privet fruit medicinal material and the processed product thereof according to any one of claims 1 to 4, wherein the characteristic spectrum of the glossy privet fruit medicinal material comprises 10 common peaks, and the 10 common peaks comprise characteristic peaks of salidroside, echinacoside, specnuezhenide and nuezhenoside G13; the feature map of the processed product of the glossy privet fruit is additionally provided with 1-4 common peaks on the basis of the feature map of the glossy privet fruit medicinal material, and the additionally provided 1-4 common peaks comprise the feature peak of 5-hydroxymethylfurfural.

6. A method for detecting the content of multi-index components in glossy privet fruit medicinal materials and processed products thereof is characterized by comprising the following steps:

7. The method for detecting the content of the multi-index components in the glossy privet fruit medicinal material and the processed product thereof according to claim 6, wherein the gradient elution comprises the following steps:

8. The method for detecting the content of the multi-index components in the glossy privet fruit medicinal material and the processed product thereof according to claim 6, wherein the detection conditions of the ultra-high performance liquid chromatography further comprise:

9. The method for detecting the content of the multi-index components in the fructus ligustri lucidi medicinal material and the processed product thereof according to any one of claims 6 to 8, wherein in the preparation method of the solution to be detected, the second solvent is methanol or a mixed solution of methanol and water; and/or

10. A method for synchronously detecting the characteristic spectrum and the content of multi-index components of a glossy privet fruit medicinal material and a processed product thereof is characterized by comprising the following steps:

constructing a characteristic map of the glossy privet fruit medicinal material and the processed product thereof according to the characteristic map construction method of any one of claims 1 to 5;

constructing a standard curve of the index components according to the method for detecting the content of the multi-index components of any one of claims 6 to 8;

11. The method for synchronously detecting the characteristic spectrum and the content of multi-index components of the glossy privet fruit medicinal material and the processed product thereof according to claim 10, wherein the gradient elution comprises the following procedures:

12. The method for synchronously detecting the characteristic spectrum and the content of multi-index components of the glossy privet fruit medicinal material and the processed product thereof according to claim 10, wherein the detection conditions of the ultra-high performance liquid chromatography further comprise:

13. The method for synchronously detecting the characteristic spectrum and the content of multi-index components of the glossy privet fruit medicinal material and the processed product thereof according to any one of claims 10 to 12, wherein in the preparation method of the sample solution to be detected, the second solvent is methanol or a mixed solution of methanol and water; and/or