CN113777183B

CN113777183B - Glossy privet fruit medicinal material and its processed product characteristic spectrum construction method and multi-index component content detection method

Info

Publication number: CN113777183B
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: 2023-06-30
Anticipated expiration: 2041-08-09
Also published as: CN113777183A

Abstract

The invention relates to a glossy privet fruit medicinal material and a characteristic spectrum construction method of a processed product thereof. The feature map construction method comprises the following steps: preparing a reference substance solution and a test substance solution, wherein the preparation method of the reference substance solution comprises the following steps: dissolving 5-hydroxymethylfurfural control, salidroside control, echinacoside control, terligustrin control and ligustrin G13 control in a first solvent; the preparation method of the sample solution comprises the following steps: extracting fructus Ligustri Lucidi or its processed product with a second solvent, filtering the extractive solution, and collecting filtrate; performing ultra-high performance liquid chromatography detection on the reference substance solution and the sample 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 spectrum construction method can effectively identify the glossy privet fruit medicinal materials and the processed products with different processing degrees.

Description

Glossy privet fruit medicinal material and its processed product characteristic spectrum construction method and multi-index component content detection method

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 multi-index components, and a method for synchronously detecting the characteristic spectrum and the content of the multi-index components of the glossy privet fruit medicinal material and the processed product thereof.

Background

Fructus Ligustri Lucidi is dry mature fruit of Ligustrum lucidum ait Ligustrum lucidum Ait. Harvesting in winter, removing branches and leaves, steaming or scalding in boiling water, and drying; or directly dried. Sweet and bitter in flavor 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 deficiency of liver-yin and kidney-yin, giddiness and tinnitus, soreness of waist and knees, premature graying hair, dim eyesight, internal heat and diabetes, bone steaming and hectic fever, etc. Fructus Ligustri Lucidi and fructus Ligustri Lucidi in the 2020 edition of Chinese pharmacopoeia are recorded, and the theory of traditional Chinese medicine holds that fructus Ligustri Lucidi is mainly used for removing liver heat, improving eyesight, nourishing yin, moistening dryness, and treating constipation due to liver heat, blurred vision, and yin deficiency and intestine dryness; the wine has the effects of activating blood and removing obstruction in collaterals, dispelling wind and dispelling cold, so the processed wine glossy privet fruit has stronger liver and kidney nourishing effect than glossy privet fruit, and weaker cold and cool property, and is mostly used for symptoms such as liver and kidney yin deficiency, dizziness and tinnitus, blurred vision, premature graying of beard and hair and the like, and the wine glossy privet fruit is mainly applied in the clinical of traditional Chinese medicine.

The fructus Ligustri Lucidi mainly comprises terpenes, iridoid glycosides, phenethyl alcohol glycosides, flavonoids, volatile oil, fatty acid, polysaccharides, and microelements; wherein the most active components and medicinal value belong to triterpenes, iridoid glycosides and phenethyl alcohol glycosides, and the representative components of the triterpene glycosides, the phenethyl alcohol glycosides are oleanolic acid, terligustroside, salidroside and the like. Modern researches show that the chemical components and pharmacological actions of the glossy privet fruit are changed to a certain extent in the processing process, but only the limit requirement of single index content that the terligustroside in the glossy privet fruit is not less than 0.7% and the content of the rhodiola rosea glycoside in the wine glossy privet fruit is not less than 0.2% is specified in the Chinese pharmacopoeia 2020 edition, and the whole quality control method of the glossy privet fruit medicinal material and the processed product wine glossy privet fruit is lacked. Meanwhile, the difference of active ingredients in the glossy privet fruit medicinal materials with different processing degrees is obvious, so that the exertion of the efficacy of the glossy privet fruit processed product is affected, and the glossy privet fruit medicinal materials with different processing degrees are required to be effectively identified.

The traditional method involves the identification of fructus ligustri lucidi and wine products, and takes one or at least two of 10-hydroxy-oleandrin dimethyl ester, neoligustrin or isomer, 8-methyl-7-ketologanin, elenolic acid, verbascoside and luteolin as markers; meanwhile, the ultra-high performance liquid chromatography/quadrupole rod-Orbitrap-MS (UHPLC/Q-Orbitrap-MS) is adopted to separate and detect the samples, so that the glossy privet fruit samples can be effectively separated and detected, and whether the samples are products or wine products can be accurately judged. However, the traditional method can only identify the raw products and the wine products sold in the market, and the quality difference of the wine products caused by different processing degrees is difficult to reflect.

The traditional method also relates to HPLC fingerprint comparison research of glossy privet fruit and 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 fingerprint of glossy privet fruit and wine glossy privet fruit is constructed and obtained, and 11 main differential substances are obtained by screening, wherein the substances comprise hydroxytyrosol, tyrosol, terglossy privet glycoside, oleuropein and quercetin. However, the traditional method can only identify the raw products and the wine products sold in the market, the quality difference of the wine products 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 method is difficult to adapt to the high efficiency of processing production.

Disclosure of Invention

Based on the above, the invention provides a method for constructing the characteristic spectrum of the glossy privet fruit and the processed products thereof, which can effectively identify the glossy privet fruit medicinal materials and the processed products with different processing degrees.

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

preparing a reference substance solution and a test substance solution, wherein the preparation method of the reference substance solution comprises the following steps: dissolving 5-hydroxymethylfurfural control, salidroside control, echinacoside control, terligustrin control and ligustrin G13 control in a first solvent; the preparation method of the sample 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;

Performing ultra-high performance liquid chromatography detection on the reference substance solution and the sample 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:

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

5-9 min, the volume percentage of the mobile phase A is increased from 10% to 18%, and the volume percentage of the mobile phase B is decreased from 90% to 82%;

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

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

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

In one embodiment, the detection conditions of the ultra performance liquid chromatography further include:

chromatographic column: octadecylsilane chemically bonded silica is used as a filler; detection wavelength: the detection wavelength is 268nm to 284 nm,3min to 25min, and 224nm to 275nm; flow rate: 0.2 mL/min-0.3 mL/min; column temperature: 25-32 ℃.

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

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

The dosage of the second solvent is 10-75 mL for each 0.3g of glossy privet fruit medicinal material or 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, and the characteristic peaks of the 10 common peaks comprise salidroside, echinacoside, terligustroside and ligustroside G13; the characteristic spectrum of the glossy privet fruit processed product is newly added with 1-4 common peaks on the basis of the characteristic spectrum of the glossy privet fruit medicinal material, and the newly added 1-4 common peaks comprise the characteristic peaks of 5-hydroxymethylfurfural.

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

preparing an index component reference substance solution and a solution to be measured, wherein the preparation method of the index component reference substance solution comprises the following steps: dissolving salidroside reference substance, terligustride reference substance and ligustrum lucidum glycoside G13 reference substance in third solvent to obtain index component reference substance solutions with different concentrations; the preparation method of the solution to be detected comprises the following steps: extracting fructus Ligustri Lucidi or its processed product with a second solvent, filtering the extractive solution, and collecting filtrate;

Performing ultra-high performance liquid chromatography detection on the index component reference substance solutions with different concentrations, and constructing a standard curve of the index component; 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 a 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 of preparing the test article solution,

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

constructing a standard curve of the index component according to the multi-index component content detection method;

preparing a sample solution to be tested: extracting fructus Ligustri Lucidi or its processed product with a second solvent, filtering the extractive solution, and collecting filtrate;

Performing 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 patterns of the glossy privet fruit medicinal materials and the processed products thereof and the standard curve of the index component;

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 of the invention can simultaneously establish the characteristic spectrum of the glossy privet fruit medicinal material and the processed products with different processing degrees by adopting the proper UPLC chromatographic condition, the characteristic spectrum of the glossy privet fruit contains 10 characteristic peaks, and the characteristic spectrum of the processed products is newly added with 1 to 4 characteristic peaks, so that the quality of the glossy privet fruit medicinal material and the processed products with different processing degrees can be comprehensively reflected, and the method is stable, strong in specificity, accurate and reliable, simple and convenient to operate and good in reproducibility, and provides a scientific detection and identification method for the quality control of the glossy privet fruit medicinal material and the processing process control of the processed products. Meanwhile, the construction method is short in time and high in detection efficiency, can adapt to the high efficiency of processing production, and is convenient for large-scale popularization and application.

According to the multi-index component content detection method, the appropriate UPLC chromatographic conditions are adopted to accurately quantify the index components of the salidroside, the terpriside and the ligustrside G13 in the glossy privet fruit medicinal materials and the processed products thereof, so that the result is reliable, the operation is simple and convenient, the blank that the content of the terpriside and the content of the salidroside of the glossy privet fruit medicinal materials and the processed products thereof cannot be detected simultaneously in the 2020 edition of Chinese pharmacopoeia is overcome, and the measurement of the G13 index of the glossy privet glycoside with higher content is increased simultaneously.

Furthermore, the invention can simultaneously realize the construction of the characteristic patterns of the glossy privet fruit medicinal material and the processed products with different processing degrees and the detection of the content of multi-index components therein by adopting the proper UPLC chromatographic conditions, is beneficial to the quality control of the glossy privet fruit medicinal material and the processed products with different processing degrees, is suitable for industrialized high-efficiency production, and has the advantages of 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 of 11 batches of glossy privet fruit medicinal materials and a comparison characteristic spectrum thereof in the embodiment 1 of the invention;

FIG. 2 is a characteristic map of the processing process of glossy privet fruit decoction pieces in the embodiment 1 of the invention within 8 hours of wine steaming after crushing;

FIG. 3 is a characteristic map of the processing process of the glossy privet fruit decoction pieces in the embodiment 1 of the invention within 8 hours without crushing and steaming;

FIG. 4 is a characteristic map of the processing process of the glossy privet fruit decoction pieces in the embodiment 1 of the invention within 8 hours of stewing without crushing wine;

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

FIG. 6 is a graph showing the PDA score of 14 characteristic peaks for the different wine-processed glossy privet fruit products of example 1;

FIG. 7 is a graph showing the OPLS-DA score of 14 characteristic peaks for the processed products of different wine-processed glossy privet fruits in example 1;

FIG. 8 is a specificity investigation map of the characteristic map of the glossy privet fruit medicinal material and the processed decoction pieces thereof in example 1;

FIG. 9 is a plot of elution gradient one and detection wavelength 275nm for the investigation of example 1;

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

FIG. 11 is a plot of elution gradient two and detection wavelength 275nm for the study of example 1;

FIG. 12 is a plot of elution gradient two and detection wavelength 224nm for example 1;

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

FIG. 14 is a chart of detection wavelengths 284nm (0 to 3 min) in example 1;

FIG. 15 is an extraction solvent investigation map of example 1;

FIG. 16 is a plot of the specific investigation of the content of index components of the glossy privet fruit medicinal material and its processed decoction pieces in example 2;

FIG. 17 is a linear investigation map of the index component salidroside of the glossy privet fruit medicinal material and the processed decoction pieces in example 2;

FIG. 18 is a linear investigation map of the index component of the glossy privet fruit medicinal material and the processed decoction pieces thereof in example 2;

fig. 19 is a linear investigation map of glossy privet fruit medicinal material and its processed decoction piece index ingredient glossy privet glycoside G13 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 below by combining specific embodiments. 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 herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

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", etc. are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or quantity, nor as implying an importance or quantity of the indicated technical features. Moreover, "first," "second," "third," etc. are for non-exhaustive list description purposes only, and it should be understood that no closed limitation on the number is made.

In the invention, the technical characteristics described in an open mode comprise a closed technical scheme composed of the listed characteristics and also comprise an open technical scheme comprising the listed characteristics.

In the present invention, the numerical range is referred to, and both ends of the numerical range are included unless otherwise specified.

The percentage content referred to in the present invention refers to mass percentage for both solid-liquid and solid-solid phase mixtures and water for volume percentage for liquid-liquid phase mixtures unless otherwise specified.

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 after 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 predetermined temperature range. The constant temperature process allows the temperature to fluctuate within the accuracy of the instrument control.

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:

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

In some examples, the detection conditions of the ultra performance liquid chromatography further include:

In some examples, the detection wavelength of the ultra performance liquid chromatography is: the detection wavelength is 282 nm-284 nm,3 min-25 min and 224 nm-230 nm.

In some examples, the flow rate of the ultra performance liquid chromatography is 0.2mL/min to 0.3mL/min. In particular, 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, 0.3mL/min.

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

In some examples, the second solvent is methanol or a mixture of methanol and water. Further, the volume fraction of the methanol in the mixed solution of the methanol and the water is 25-80%. Specifically, the volume fraction of methanol in the mixture 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, the second solvent is used in an amount of 10-75 mL per 0.3g of fructus Ligustri Lucidi or processed product thereof. Further, the dosage of the second solvent is 20-30 mL for each 0.3g of glossy privet fruit medicinal material or processed product thereof.

In some examples, the method of extraction is ultrasonic extraction or heating to reflux extraction. Further, the power of the ultrasonic wave is 450-500W, and the frequency is 35-45 kHz.

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

In some examples, the control solution is prepared by a process wherein the first solvent is an alcoholic solvent, such as methanol.

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

In some examples, the characteristic spectrum of the glossy privet fruit medicinal material comprises 10 common peaks, and the characteristic peaks of the 10 common peaks comprise salidroside, echinacoside, terligustroside and ligustroside G13; the characteristic spectrum of the glossy privet fruit processed product is newly added with 1-4 common peaks on the basis of the characteristic spectrum of the glossy privet fruit medicinal material, and the newly added 1-4 common peaks comprise the characteristic peaks 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 substance solution and a solution to be detected, wherein the preparation method of the index component reference substance solution comprises the following steps: dissolving salidroside reference substance, terligustride reference substance and ligustrum lucidum glycoside G13 reference substance in third solvent to obtain index component reference substance solutions with different concentrations; the preparation method of the solution to be tested comprises the following steps: extracting fructus Ligustri Lucidi or its processed product with a second solvent, filtering the extractive solution, and collecting filtrate;

Performing ultra-high performance liquid chromatography detection on index component reference substance solutions with different concentrations, and constructing a standard curve of 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 a standard curve of the index component;

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

It can be understood that the technical scheme of the method for detecting the content of the multi-index components of the glossy privet fruit medicinal material and the processed product thereof is similar to the method for constructing the characteristic spectrum of the glossy privet fruit medicinal material and the processed product thereof, wherein the preparation of the solution of the to-be-detected product is the same as that of the solution of the to-be-detected product, and the description is omitted.

The invention also provides a 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, which comprises the following steps:

It can be understood that the technical scheme of the method for synchronously detecting the characteristic patterns and the multi-index component content of the glossy privet fruit medicinal materials and the processed products thereof is similar to the method for constructing the characteristic patterns of the glossy privet fruit medicinal materials and the processed products thereof, wherein the preparation of the sample solution to be detected is the same as the preparation of the sample solution to be detected, and the description is omitted.

The instruments and reagents employed in the examples of the present invention are as follows:

1. instrument for measuring and controlling the intensity of light

Waters ultra-high Performance liquid chromatograph (Waters H-class), PDA detector (Waters); an Empower workstation; one ten thousandth balance (mertrer-tolidol, ME 204E); parts per million flat (mertrer-tolidol, XP 26); ultrapure water machine (merck, milli-Q Direct 8/16 system); ultrasonic cleaners (kunshan ultrasonic instruments limited, KQ5500 DE); 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 remaining reagents were all analytically pure.

5-hydroxymethylfurfural control (lot number: wkq20041311; source: vickers biotechnology Co., ltd., sichuan province); rhodiola rosea glycoside reference (lot number: 110818-201507; source: china food and drug inspection institute; content%: 99.4); echinacoside control (lot number: 111670-201907; china food and drug inspection institute; content%: 91.8); a terligustroside reference substance (batch number: 111926-201906; chinese food and drug verification institute; content%: 95.0); glossy privet glycoside G13 reference substance (batch number: 8857; shanghai Shiadad Standard technical service Co., ltd.; content%: 95.0).

Example 1

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

Fructus ligustri lucidi medicinal material: collecting 11 batches of glossy privet fruit medicinal materials meeting the requirements of Chinese pharmacopoeia of 2020 edition from places of origin such as Henan, jiangsu and Shandong.

Processing decoction piece wine glossy privet fruit: taking glossy privet fruit medicinal materials, removing impurities, dividing the glossy privet fruit medicinal materials into three parts, crushing one part to obtain glossy privet fruit crushed decoction pieces, and crushing the other two parts to obtain glossy privet fruit non-crushed decoction pieces; placing 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, moistening each 100kg of glossy privet fruit decoction pieces with 20kg of yellow wine for 2-4 hours, and taking out; one part of glossy privet fruit is steamed with wine for 8.0 hours, and the other two parts of glossy privet fruit are not steamed with wine and stewed with wine for 8 hours respectively; sampling every 0.5 hour, and oven drying to obtain the fructus Ligustri Lucidi decoction pieces with different decoction piece specifications and different processing times. Wherein the glossy privet fruit crushed decoction piece wine steaming mark is YP-JZ, the glossy privet fruit non-crushed decoction piece wine steaming mark is BYP-JZ, and the glossy privet fruit non-crushed decoction piece wine stewing mark is BYP-JD.

1. Chromatographic condition and System applicability test

Octadecylsilane chemically bonded silica is used as filler (ACQUITY UPLC BEH Shield RP Column, column length of 100mm, inner diameter of 2.1mm, particle diameter of 1.7 μm); acetonitrile as mobile phase A and water as mobile phase B, and performing gradient elution according to the specification in Table 1; the flow rate was 0.3mL per minute; the column temperature is 30 ℃; the detection wavelength is 284nm, 3-25 min and 224nm. The theoretical plate number is not less than 30000 calculated according to the peak of the terprivet glycoside.

TABLE 1 gradient elution table

2. Preparation of control solution

Taking appropriate amount of 5-hydroxymethyl reference substance, salidroside reference substance, echinacoside reference substance, terprivet glycoside reference substance, and ligustrum japonicum glycoside G13 reference substance, precisely weighing, and adding methanol to make into solution containing 0.1mg per 1mL as reference substance solution.

3. Preparation of test solutions

Taking fructus Ligustri Lucidi medicinal material or fructus Ligustri Lucidi decoction pieces (YP-JZ, BYP-JD), making into powder (sieving with a third sieve), taking about 0.3g of powder, precisely weighing, placing into conical flask with plug, precisely adding 70% methanol 25mL, weighing, processing with ultrasound (power: 500W, frequency: 40 kHz) for 60 min, cooling, weighing again, supplementing the reduced weight with 70% methanol, shaking, filtering, and collecting the subsequent filtrate.

4. Measurement

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

5. Measurement results

(1) Measurement results of glossy privet fruit medicinal materials

The characteristic patterns of 11 batches of glossy privet fruit medicinal materials are subjected to common peak matching by adopting a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, a comparison characteristic pattern is generated, a total of 10 characteristic peaks are determined, the chemical components of the 10 common peaks are identified, a No. 3 peak is a rhodioside peak, a No. 4 peak is a echinacoside peak, a No. 7 peak is a terligustrin peak, a No. 9 peak is a glossy privet glycoside G13 peak, a No. 7 peak terligustrin peak is a reference peak S, reference data of a glossy privet fruit medicinal material characteristic pattern analysis method is prepared, characteristic peaks are selected from characteristic pattern angles of glossy privet fruit medicinal materials, and relative retention time and relative peak area of each characteristic peak are calculated to form the overall appearance of the glossy privet fruit medicinal material characteristic pattern. Specifically, the peak 7 terprivet glycoside 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 2 relative retention time of characteristic spectra of glossy privet fruit batches 11

TABLE 3 relative peak areas of characteristic maps of glossy privet fruit batches 3

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

In the 8-hour processing process of YP-JZ, BYP-JZ and BYP-JD, the processed decoction piece wine glossy privet fruit samples sampled every 0.5 hour are subjected to characteristic spectrum analysis by adopting a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, 1 to 4 characteristic peaks are newly added along with the prolongation of processing time, see figures 2, 3 and 4, chemical components of 1 to 14 common peaks are identified, the No. 3 peak is a rhodioside peak, the No. 4 peak is an echinacoside peak, the No. 7 peak is a terligustroside peak, the No. 9 peak is a glossy privet glycoside G13 peak, and the No. 11 peak is a 5-hydroxymethylfurfural peak, see figure 5.

And (3) carrying out principal component analysis (PDA) and partial orthogonal least squares discriminant analysis (OPLS-DA) on the peak areas of chromatographic peaks in the UPLC characteristic spectrum of the measured wine glossy privet fruit decoction pieces, recording the peak areas of 1-4 newly added characteristic peaks which are not detected in the wine glossy privet fruit decoction pieces by 0, and introducing the peak areas of 14 common peaks calibrated by the wine glossy privet fruit decoction pieces in different decoction piece specifications and different processing modes into SIMCA14.0 software for analysis, wherein the score diagrams of the PDA and the OPLS-DA are shown in fig. 6 and 7. The wine glossy privet fruit decoction pieces YP-JZ and BYP-JZ or BYP-JD are obviously classified and gathered in the PDA unsupervised model, and the wine glossy privet fruit decoction pieces BYP-JZ and BYP-JD are gathered together. Further analysis by OPLS-DA supervised model, R thereof ₂ Y (representing the fitting degree of the OPLS-DA model) reaches 0.924, Q ² (representing the model predictive power) reaches 0.889. The method shows that the wine glossy privet fruit decoction pieces in different decoction piece specifications and different processing modes are obviously classified and aggregated in an OPLS-DA model.

The UPLC characteristic spectrum measurement result of the processing process of the glossy privet fruit shows that after the glossy privet fruit is crushed, yellow wine is added for moistening, and along with the extension of the wine steaming time, new chromatographic peaks appear in a short time, namely, chemical components are converted and generated, and 14 characteristic peaks are generated after the wine steaming is carried out for 1 hour; directly adding yellow wine without crushing fructus Ligustri Lucidi, moistening, steaming for 2 hr to generate 14 characteristic peaks, and stewing for 7.5 hr to generate 14 characteristic peaks; the main influencing factors of the difference are that whether water vapor is in direct contact or not in the processing process, namely, the seed coats of the glossy privet fruits are destroyed after being crushed, the water vapor in the wine steaming process can be directly contacted with the glossy privet fruits, the conversion and the generation of chemical components are easy to occur under the high-humidity and high-heat conditions, the direct contact of the high-humidity water vapor is weakened by the non-crushed glossy privet fruits under the protection of peel, the conversion and the generation time of the chemical components are prolonged, the direct contact of the water vapor is further isolated by the wine stewing, and the conversion and the generation time of the chemical components are further prolonged. Further analysis by combining PDA and OPLS-DA model can show that the conversion and generation ratio of the chemical components of fructus Ligustri Lucidi under different processing conditions are different.

In conclusion, the characteristic patterns with good separation degree and stability in 11 batches of glossy privet fruit medicinal materials have 10 common peaks. Wherein 4 peaks correspond to the retention time of the corresponding reference peak of the reference substance, the peak corresponding to the terligustrum lucidum glycoside reference is S peak, the relative retention time of each characteristic peak and the S peak is calculated, the relative retention time is 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). Among 1-4 characteristic peaks newly added in the processed decoction piece wine glossy privet fruit, 1 peak is corresponding to the retention time of the corresponding reference substance peak of the reference substance, the peak corresponding to the terligustride reference substance is S peak, the relative retention time of each characteristic peak and the S peak is calculated, 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. Methodological verification

(1) Specificity test

Taking the same batch of prepared sample solution, blank solvent and reference substance solution, detecting under the condition of liquid chromatography, feeding 1 mu L of sample each time, and recording a chromatogram. As shown in fig. 8, the blank solvent chromatography showed no chromatographic peak at the retention time position corresponding to the characteristic peak, indicating that the extraction solvent had no interference with measurement of each characteristic peak and had good specificity.

(2) Precision test

Taking the same sample solution, repeatedly sampling for 6 times under the condition of liquid chromatography, sampling 1 mu L each time, taking the terprivet glycoside 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 relative retention time of each characteristic peak has RSD less than 2% and relative peak area has RSD less than 3%, and the result shows that the instrument precision is good.

(3) Repeatability test

Taking the same batch of samples, preparing 6 parts of sample solution according to the preparation method of the sample, carrying out sample injection measurement under the condition of liquid chromatography, carrying out sample injection for 1 mu L each time, taking a terprivet glycoside 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 relative retention time of each characteristic peak has an RSD of less than 2% and the relative peak area has an RSD of less than 3%, and the result shows that the analysis method has good repeatability.

(4) Stability test

Taking a sample solution, placing at room temperature, detecting under the liquid chromatography conditions for 0 hour, 2 hours, 4 hours, 8 hours, 12 hours and 24 hours respectively, injecting 1 mu L of sample each time, taking a terprivet glycoside 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 relative retention time of each characteristic peak is less than 2% and the relative peak area is less than 5%, and the result shows that the stability of the test sample solution is better within 24 hours.

7. Chromatographic condition investigation

(1) Elution gradient and detection wavelength

1.1 elution gradient one

The preparation of the test article was carried out with the use of wine glossy privet fruit YP-JZ (8 hours) according to the item "preparation of test article solution", and the elution gradient was examined by using the detection wavelength of 224nm and 275nm with the same conditions as the "chromatographic condition and the system applicability test" as shown in the following Table 4.

TABLE 4 Table 4

As a result, referring to fig. 9 and 10, it can be seen that the peak time of each chromatographic peak becomes shorter as the polarity of the mobile phase becomes larger, and the peak density and the separation degree of each chromatographic peak are poor, which indicates that the chromatograms with good separation degree and adaptive retention time of each chromatographic peak cannot be obtained well under the gradient condition.

1.2 elution gradient two

The preparation of the test article was carried out with the use of wine glossy privet fruit YP-JZ (8 hours) according to the item "preparation of test article solution", and the elution gradient was examined by using the detection wavelength of 224nm and 275nm with the same conditions as the "chromatographic condition and the system applicability test" as shown in the following Table 5.

TABLE 5

As shown in fig. 11 and 12, the chromatograms of gradient elution one are compared, and the peak-out time of each chromatographic peak is moderate and the separation degree is good; in addition, as can be seen from comparing chromatograms at 224nm and 275nm, the absorption of each chromatographic peak at 224nm is large, the peak type is good, but within 0-3 min, part of chromatographic peaks at 275nm are deleted, and by comparison of the comparison products, the deleted peaks contain 5-hydroxymethylfurfural (i.e. have no absorption at 224 nm), and 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 article was carried out with the wine glossy privet fruit YP-JZ (8 hours) according to the item "preparation of the test article solution", and the detection wavelength in the elution process was changed to 275nm and 284nm respectively according to the same conditions as the "chromatographic condition and the system applicability test".

The results are shown in FIGS. 13 to 14. It can be seen that the absorption (peak area) of No. 11 peak 5-hydroxymethylfurfural at 284nm detection wavelength is greater than 275nm wavelength within 0 to 3min, see tables 6 to 7 below.

TABLE 6 retention time

TABLE 7 peak area

(2) Investigation of extraction solvent in sample preparation

According to the same conditions as the "chromatographic conditions and the system applicability test", the preparation of the test article was performed with the wine glossy privet fruit YP-JZ (8 hours) according to the "preparation of the test article solution", and the extraction solvent was only changed in the preparation of the test article to perform the extraction solvent investigation. The solvents examined were 30% methanol, 50% methanol, 70% methanol, 50% ethanol, 70% ethanol, respectively.

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

TABLE 8 retention time

TABLE 9 peak area

The result shows that the extraction is carried out by adopting different extraction solvents, an ethanol system is adopted as a solvent, and the number of chromatographic peaks is missing; the methanol system is used as an extraction solvent, the number of chromatographic peaks is not obviously different, 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 to the sample amount of 14 common characteristic peaks increases with the increase of the concentration of the methanol.

(3) Investigation of extraction mode in sample preparation

According to the same conditions as the "chromatographic conditions and the system applicability test", the preparation of the test article was carried out with the wine glossy privet fruit YP-JZ (8 hours) according to the "preparation of the test article solution", and the extraction mode was examined by changing only the extraction mode in the preparation of the test article. 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 reflux and ultrasonic extraction modes have no obvious difference in terms of chromatographic peak number, chromatographic peak separation effect and total peak area/sample amount, and the ultrasonic extraction can be selected in consideration of convenient operation.

(3) Investigation of extraction solvent dosage in sample preparation

According to the same conditions as the "chromatographic conditions and the system applicability test", the preparation of the test article was carried out with the wine glossy privet fruit YP-JZ (8 hours) according to the "preparation of the test article solution", and only the extraction solvent amount in the preparation of the test article was changed. The extraction solvent was used in amounts of 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 results show that the use amount of the extraction solvent is the maximum by adding 25mL of the extraction solvent by adopting the ratio of the total peak area to the sample amount of 14 common characteristic peaks, and the use amount of 25mL is selected as the use amount of the extraction solvent in consideration of solvent saving.

(4) Investigation of extraction time in sample preparation

According to the same conditions as the "chromatographic conditions and the System applicability test", the preparation of the test article was performed with the wine fructus Ligustri Lucidi YP-JZ (8 hours) according to the "preparation of the test article solution", and only the extraction time in the preparation of the test article was changed. 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 result shows that the ultrasonic extraction is carried out for different time, the number of chromatographic peaks of the characteristic spectrum, the separation degree and the total peak area/sample weighing amount of 14 characteristic peaks of the characteristic spectrum have no obvious difference, the ultrasonic extraction is carried out for 30 minutes, the ultrasonic extraction is basically complete, 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 glossy privet fruit medicinal materials and processed decoction pieces thereof.

The triterpenes, iridoid glycosides and phenethyl alcohol glycosides contained in fructus Ligustri Lucidi are main effective components, and the main representative components are salidroside, terligustroside, ligustroside G13, oleonuezhenide, etc. The invention evaluates the glossy privet fruit medicinal material and the processed decoction pieces by taking salidroside, terglossy privet glycoside and glossy privet glycoside G13 as content measurement indexes.

1. Chromatographic condition and System applicability test

TABLE 1 gradient elution table

2. Preparation of index ingredient reference substance solution

Taking appropriate amounts of salidroside reference substance, terligustride reference substance and ligustride G13 reference substance, precisely weighing, and respectively adding methanol to obtain 200 μg of salidroside reference substance solution, 200 μg of terligustride reference substance solution and 200 μg of ligustride G13 reference substance solution per 1 mL.

3. Preparation of test solutions

Collecting fructus Ligustri Lucidi or its processed decoction pieces, making into powder (sieving with No. three sieve), collecting powder about 0.3g, precisely weighing, placing into conical flask with plug, precisely adding 70% methanol 25mL, weighing, ultrasonic treating (power: 500W, frequency: 40 kHz) for 60 min, cooling, weighing again, supplementing the lost weight with 70% methanol, shaking, filtering, and collecting the filtrate.

4. Measurement

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

5. Measurement results

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

table 16 results of measurement of the content of glossy privet fruit medicinal materials (percent)

Table 17 determination of the content of glossy privet fruit processed with wine (%)

6. Methodological verification

(1) Investigation of specificity

Taking the same batch of prepared sample solution, index component reference substance solution and blank solvent, detecting under the condition of liquid chromatography, feeding 1 mu L each time, and recording the chromatogram. The results are shown in FIG. 16. The results show that the blank solvent chromatography has no chromatographic peak at the retention time positions corresponding to the rhodiola rosea glycoside, the terprivet glycoside and the privet glycoside G13, and the extraction solvent has no interference to the content measurement of the rhodiola rosea glycoside, the terprivet glycoside and the privet glycoside G13 and has good specificity.

(2) Linear relationship investigation

(1) Linear investigation of salidroside

Precisely weighing salidroside reference substance, placing into 5mL measuring flask, adding methanol to obtain a series of reference substance solutions containing 301.1820 μg, 120.4728 μg, 60.2364 μg, 30.2176 μg and 12.1268 μg of salidroside, detecting under the condition of liquid chromatography, sampling 1 μl each time, performing linear regression calculation with the concentration of salidroside as abscissa and the measured peak area as ordinate, and drawing standard curve, wherein the regression equation of salidroside is: y= 4797.6x-1697.5, its correlation coefficient R ² = 0.9996, salidroside in the sample concentration range of 12.1268 μg/ml to 301.1820 μg/ml, sample concentrationThe linear relation with the peak area is good. The results are shown in FIG. 17.

(2) Linear investigation of terprivet glycoside

Precisely weighing a special ligustrum glycoside reference substance, placing the special ligustrum glycoside reference substance into a 5mL measuring flask, adding methanol to prepare a series of reference substance solutions containing 247.7600 mug of special ligustrum glycoside, 100.1300 mug of special ligustrum glycoside, 49.5900 mug of special ligustrum glycoside, 24.7000 mug of special ligustrum glycoside and 9.8800 mug of special ligustrum glycoside in each 1mL of special ligustrum glycoside reference substance solution, detecting under the condition of liquid chromatography, carrying out linear regression calculation by taking the special ligustrum glycoside concentration as an abscissa and the measured peak area as an ordinate, and drawing a standard curve, wherein the regression equation of the special ligustrum glycoside is as follows: y= 4794.0x-3785.6, its correlation coefficient R ² The specnuezhenide is in the range of 9.8800-247.7600 mug/ml in sample concentration, and the linear relation between the sample concentration and the peak area is good. The results are shown in FIG. 18.

(3) Ligustrin G13 linear investigation

Precisely weighing the ligustrum glycoside G13 reference substance, placing the ligustrum glycoside G13 reference substance into a 5mL measuring flask, adding methanol to prepare a series of reference substance solutions containing 271.1300 mug ligustrum glycoside G13, 101.0800 mug ligustrum glycoside G13, 59.2800 mug ligustrum glycoside G13, 27.1700 mug ligustrum glycoside G13 and 10.8300 mug ligustrum glycoside G13 in each 1mL, detecting under the condition of the liquid chromatography, carrying out linear regression calculation by taking the concentration of ligustrum glycoside G13 as an abscissa and the measured peak area as an ordinate, and drawing a standard curve, wherein the regression equation of the ligustrum glycoside G13 is as follows: y=3817.2x+25885.8, its correlation coefficient R ² The sample concentration of the ligustrum lucidum glycoside G13 is in the range of 10.8300-271.1300 mug/ml, and the linear relation between the sample concentration and the peak area is good. The results are shown in FIG. 19.

(3) Precision test

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

(4) Repeatability test

Taking the same batch of samples, preparing 6 parts of sample solution according to the preparation method of the sample, and carrying out sample injection measurement under the condition of liquid chromatography, wherein 1 mu L of sample is injected each time. The result shows that the average value of the content of the salidroside is 0.23%, and the RSD value is 0.62%; the average value of the content of the terprivet glycoside is 2.88 percent, and the RSD value is 0.73 percent; the average value of the content of the ligustrum lucidum glycoside G13 is 1.80 percent, and the RSD value is 0.76 percent; the analytical method is shown to have good repeatability.

(5) Accuracy test

And (3) carrying out sample recovery rate measurement by adopting a reference substance, designing 3 groups of experiments according to the ratio of the added amounts of the rhodioside reference substance, the terprivet glycoside reference substance and the privet glycoside G13 reference substance to the contents of the components to be detected in the sample solution, namely, the rhodioside, the terprivet glycoside and the privet glycoside G13, wherein 3 parts of each group are parallel, respectively precisely transferring a proper amount of rhodioside reference substance stock solution, terprivet glycoside reference substance stock solution and privet glycoside G13 reference substance stock solution, and drying under a nitrogen blowing instrument. Taking about 0.15g of the same batch of samples with known content, precisely weighing, placing the samples into 3 groups of conical flasks, preparing test sample solutions according to a test sample preparation method, carrying out sample injection measurement under the condition of liquid chromatography, carrying out sample injection at each time by 1 mu L, and calculating the accuracy respectively, wherein the accuracy is shown in tables 18, 19 and 20. The result shows that the recovery rate of the rhodiola rosea glycoside content determination analysis method is 98.02-103.02%, and the RSD is 1.51%; the recovery rate of the terprivet glycoside content determination analysis method is within the range of 96.01-101.03%, and the RSD is 1.73%; the recovery rate of the glossy privet glycoside G13 content determination analysis method is in the range of 96.97-105.75%, and the RSD is 2.78%; meets the requirements of the verification and guidance principle of the quality standard analysis method of 9101 medicine in the 2020 edition of Chinese pharmacopoeia.

Table 18 accuracy of determination of salidroside in wine glossy privet fruit sample

Table 19 investigation of accuracy of determination of the content of terligustroside in a wine glossy privet fruit sample

Table 20 investigation of accuracy of determination of the content of ligustroside G13 in wine glossy privet fruit samples

(6) Stability test

Sample solutions were taken and allowed to stand at room temperature, and under the above-mentioned liquid chromatography conditions, detection was performed for 0 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours, respectively, 1 μl was injected each time, peak areas were recorded, and RSD values were calculated. The result shows that the RSD value of the peak area of the salidroside measured by sample injection within 24 hours is 4.84%; the peak area RSD value of the terprivet glycoside is 2.94%; the peak area RSD value of the ligustrum lucidum glycoside G13 is 2.43%, which shows that the stability of the test sample solution is better within 24 hours.

Meanwhile, the UPLC spectrum of the glossy privet fruit medicinal material and the processed decoction pieces thereof can be respectively compared with the characteristic spectrum of the glossy privet fruit medicinal material and the processed decoction pieces thereof which are established in the embodiment 1, the characteristic spectrum detection result of the glossy privet fruit medicinal material and the processed decoction pieces thereof which are corresponding samples and the content result of the index components of salidroside, terglossy privet glycoside and glossy privet glycoside G13 are synchronously obtained, and the evaluation of the glossy privet fruit medicinal material and the processed decoction pieces thereof is comprehensively carried out.

The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.

The above examples merely represent a few embodiments of the present invention, which facilitate a specific and detailed understanding of the technical solutions of the present invention, but are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. It should be understood that, based on the technical solutions provided by the present invention, those skilled in the art may obtain technical solutions through logical analysis, reasoning or limited experiments, which are all within the scope of protection of the appended claims. The scope of the patent is therefore intended to be covered by the appended claims, and the description and drawings may be interpreted as illustrative of the contents of the claims.

Claims

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

preparing a reference substance solution and a test substance solution, wherein the preparation method of the reference substance solution comprises the following steps: dissolving 5-hydroxymethylfurfural control, salidroside control, echinacoside control, terligustrin control and ligustrin G13 control in a first solvent; the preparation method of the sample 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; the second solvent is methanol or a mixed solution of methanol and water;

performing ultra-high performance liquid chromatography detection on the reference substance solution and the sample 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 chromatographic column takes octadecylsilane chemically bonded silica gel as a filler;

the gradient elution included the following procedure:

2. 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:

detection wavelength: the detection wavelength is 268nm to 284 nm,3min to 25min, and 224nm to 275nm; flow rate: 0.2 mL/min-0.3 mL/min; column temperature: 25-32 ℃.

3. The method for constructing a characteristic spectrum of a glossy privet fruit medicinal material and a processed product thereof according to claim 1, wherein the dosage of the second solvent is 10-75 mL for every 0.3g of the glossy privet fruit medicinal material or the processed product thereof.

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

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 characteristic peaks of salidroside, echinacoside, terligustroside and glossy privet glycoside G13 are included in the 10 common peaks; the characteristic spectrum of the glossy privet fruit processed product is newly added with 1-4 common peaks on the basis of the characteristic spectrum of the glossy privet fruit medicinal material, and the newly added 1-4 common peaks comprise the characteristic peaks of 5-hydroxymethylfurfural.

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

preparing an index component reference substance solution and a solution to be measured, wherein the preparation method of the index component reference substance solution comprises the following steps: dissolving salidroside reference substance, terligustride reference substance and ligustrum lucidum glycoside G13 reference substance in third solvent to obtain index component reference substance solutions with different concentrations; the preparation method of the solution to be detected comprises the following steps: extracting fructus Ligustri Lucidi or its processed product with a second solvent, filtering the extractive solution, and collecting filtrate; the second solvent is methanol or a mixed solution of methanol and water;

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 chromatographic column takes octadecylsilane chemically bonded silica gel as a filler;

the gradient elution included the following procedure:

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

8. The method for detecting the content of the multi-index components of the glossy privet fruit medicinal material and the processed product thereof according to any one of claims 6 to 7, wherein the dosage of the second solvent is 10mL to 75mL per 0.3g of the glossy privet fruit medicinal material or the processed product thereof.

9. The method for detecting the content of the multi-index components of the glossy privet fruit medicinal material and the processed product thereof according to any one of claims 6 to 7, wherein the extraction method is ultrasonic extraction or heating to reflux extraction, and the extraction time is 30 to 60 minutes.

10. A synchronous detection method for the characteristic spectrum and the content of multi-index components of glossy privet fruit medicinal materials and processed products thereof is characterized by comprising the following steps:

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

Constructing a standard curve of index components according to the multi-index component content detection method of the glossy privet fruit medicinal material and the processed product thereof according to any one of claims 6 to 8;

preparing a sample solution to be tested: extracting fructus Ligustri Lucidi or its processed product with a second solvent, filtering the extractive solution, and collecting filtrate; the second solvent is methanol or a mixed solution of methanol and water;

the gradient elution included the following procedure:

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 detection conditions of the ultra-high performance liquid chromatography further comprise:

12. The method for synchronously detecting the characteristic patterns and the multi-index component content of the glossy privet fruit medicinal material and the processed product thereof according to any one of claims 10 to 11, wherein the dosage of the second solvent is 10mL to 75mL per 0.3g of the glossy privet fruit medicinal material or the processed product thereof.

13. The 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 according to any one of claims 10 to 11, wherein the extraction method is ultrasonic extraction or heating to reflux extraction, and the extraction time is 30 to 60 minutes.