CN113759065A

CN113759065A - Method for simultaneously identifying components of coptis chinensis and phellodendron amurense and application of method

Info

Publication number: CN113759065A
Application number: CN202110209280.7A
Authority: CN
Inventors: 张志强; 韩妮娜; 韩红梅; 邓海霞; 梁勇满; 刘利娟; 李纳纳; 付静
Original assignee: Beijing Tcmages Pharmaceutical Co Ltd
Current assignee: Beijing Tcmages Pharmaceutical Co Ltd
Priority date: 2021-02-24
Filing date: 2021-02-24
Publication date: 2021-12-07
Anticipated expiration: 2041-02-24
Also published as: CN113759065B

Abstract

The invention belongs to the technical field of traditional Chinese medicine detection, and particularly provides a method for identifying components of coptis chinensis and phellodendron amurense simultaneously, which comprises the following steps of (1) preparing a test solution, a reference solution and a reference medicinal material solution; (2) and (3) detection by thin-layer chromatography: comprises the steps of two times of unfolding and respectively inspecting; the method comprises the steps of carrying out primary development by using butyl acetate-methanol-isopropanol-water as a developing agent, carrying out secondary development by using n-butanol-water-acetic acid as a developing agent, respectively using developing agents with specific compositions by using a secondary thin layer development method for the first time, and qualitatively identifying the components of the coptis chinensis and the phellodendron bark simultaneously by using the same thin layer plate without carrying out secondary sample application, so that the improvement of resources and cost caused by separately and qualitatively identifying the components of the coptis chinensis and the phellodendron bark is avoided, the quality is ensured to be uniform, stable and reliable, and the method has the advantages of high accuracy, time and cost saving and easiness in popularization and application.

Description

Method for simultaneously identifying components of coptis chinensis and phellodendron amurense and application of method

Technical Field

The invention belongs to the technical field of traditional Chinese medicine detection, and particularly relates to a method for simultaneously identifying components of coptis chinensis and phellodendron amurense and application thereof.

Background

With the enhancement of health care consciousness of people, the quality and the safety of Chinese patent medicine products are required to be higher. The Dang Gui Liu Huang Tang is a famous prescription originated from Lidong Yuan, one of the four golden units, and is recorded in the book of lan Shi Mi (secret book of lan Shi). It is called as the "Sheng Yao for treating night sweat", and is mainly indicated for night sweat caused by yin deficiency with effulgent fire. The angelica six-yellow decoction comprises the following components: 7 kinds of Chinese medicinal materials including angelica, rehmannia root, prepared rehmannia root, coptis root, scutellaria root, phellodendron bark and astragalus root.

At present, the identification of effective components in the angelica six-yellow decoction is not published by standards, and for identifying coptis chinensis in the prescription of the angelica six-yellow decoction, the Huiying discloses a thin-layer chromatography in 'research on the material standard of the angelica six-yellow decoction', wherein epiberberine and berberine hydrochloride are adopted as reference substances, and n-butyl alcohol-acetic acid-water (2: 1: 2) is used as a developing agent for development and then is used for inspection. The results are shown in FIG. 1, in the chromatogram of the test sample, the positions corresponding to the chromatogram of the reference substance of epiberberine and berberine hydrochloride and the chromatogram of the reference medicinal material of coptis respectively show the fluorescent spots with the same color, while the negative chromatogram is free of interference at the same position, thus proving that coptis is added in the raw material. However, this method cannot identify phellodendron bark, which is one of the important raw materials of Dang Gui Liu Huang Tang.

Therefore, it is urgently needed to establish a method with high accuracy, time and cost saving and easy popularization to control the qualitative identification of the Chinese angelica six-yellow decoction compound preparation and the coptis chinensis and the phellodendron amurense of the substance reference real object so as to ensure the uniformity, stability and reliability of the quality.

Disclosure of Invention

Therefore, the invention aims to solve the problems in the prior art, and provides a method which has high accuracy, saves time and cost and is easy to popularize and is used for identifying the active ingredients of the golden cypress and the golden thread simultaneously, thereby ensuring the controllable and stable product quality.

Therefore, the invention discloses a method for simultaneously identifying the components of coptis chinensis and phellodendron amurense, which comprises the following steps,

(1) preparing a test solution, a reference solution and a reference medicinal material solution;

(2) and (3) detection by thin-layer chromatography: comprises the steps of two times of unfolding and respectively inspecting after the unfolding; carrying out primary development by adopting butyl acetate-methanol-isopropanol-water as a developing agent, and carrying out secondary development by adopting n-butyl alcohol-water-acetic acid as a developing agent.

Preferably, the thin layer chromatography detection comprises the following steps:

1) sample application of the sample solution on the thin layer plate is carried out;

2) carrying out first development by using butyl acetate-methanol-isopropanol-water as a developing agent, taking out, volatilizing the solvent, and placing the thin layer plate under a light source of 360-370nm for inspection;

3) and (3) adopting n-butyl alcohol-water-acetic acid as a developing agent to carry out secondary development on the thin-layer plate inspected in the step 2), taking out, volatilizing the solvent, developing by adopting a color developing agent and inspecting under visible light.

As a preferred embodiment, the solvent can be directly dried by evaporation or can be dried by other conventional auxiliary means, such as drying.

In a preferred embodiment, the thin layer plate used in step 1) is a silica gel G thin layer plate. The spotting volume is 1 to 10. mu.l, preferably 1 to 5. mu.l, more preferably 1 to 3. mu.l.

As a preferred embodiment, in step 2), the volume ratio of butyl acetate-methanol-isopropanol-water is 3-4: 3-4: 1-2: 0.6-1.

In a preferred embodiment, the step 2) further comprises pre-saturating the container for development with concentrated ammonia solution for 20-35min before the first development.

As a preferred embodiment, in the step 3), the volume ratio of n-butanol to water to acetic acid is 6 to 8: 1-3: 1-3.

In a preferred embodiment, in step 3), the color-developer is at least one selected from the group consisting of potassium bismuth iodide, phosphomolybdic acid, and dilute potassium bismuth iodide solution.

In a preferred embodiment, the step (1) comprises mixing the sample with methanol, performing ultrasonic extraction, and filtering to obtain a sample solution;

more preferably, the ratio of the mass of the sample to the volume of methanol is 1.5-2.5 g: 20ml of the solution;

more preferably, the extraction time is 20-40 min.

As a preferred embodiment, the reference drug solution comprises a coptis reference drug solution and a phellodendron reference drug solution, and preferably, the preparation method of the reference drug solution comprises the following steps:

taking a coptis root reference medicinal material and a phellodendron bark reference medicinal material to prepare a coptis root reference medicinal material solution and a phellodendron bark reference medicinal material solution respectively according to the same method in the step (1) of the invention.

As a preferred embodiment, the reference solution comprises a palmatine hydrochloride reference solution and an phellodendrine hydrochloride reference solution.

Preferably, the preparation method of the palmatine hydrochloride reference solution comprises the steps of mixing the palmatine hydrochloride reference with methanol to prepare a reference solution with the concentration of 0.2-0.6 mg/ml;

the preparation method of palmatine hydrochloride reference solution comprises mixing phellodendrine hydrochloride reference with methanol to obtain reference solution with concentration of 0.2-0.6 mg/ml.

Preferably, the test sample is a traditional Chinese medicine composition containing the coptis chinensis and the phellodendron bark simultaneously or a traditional Chinese medicine preparation containing the coptis chinensis and the phellodendron bark simultaneously;

preferably, the test sample is angelica six-yellow soup or a compound preparation of the angelica six-yellow soup.

The compound preparation of the angelica six-yellow decoction is prepared by adding or not adding pharmaceutically conventional auxiliary materials into the angelica six-yellow decoction according to a pharmaceutically conventional process.

More preferably, the compound preparation of the angelica six-yellow decoction is selected from a solid preparation, a semi-solid preparation or a liquid preparation.

Wherein, the solid preparation can be but not limited to tablets, powders, granules, capsules and the like;

semisolid formulations can be, but are not limited to, ointments, pastes, creams, and the like;

the liquid preparation may be, but is not limited to, oral liquid, injection, emulsion, suspension, nano liquid preparation, etc.

According to the regulations of ancient classic famous prescription Chinese herbal compound preparations drafted by the food and drug administration and the material standard declaration data requirements (request for comments), the classic famous prescription material standard is the standard of Chinese herbal medicine materials prepared according to the ancient classic famous prescription preparation method recorded in ancient medical books, and except for the molding process, the other preparation methods are basically consistent with the records of the ancient medical books. Is used as a standard reference for judging whether the classical famous prescription compound preparation is basically consistent with clinical decoction. The product is prepared into the standard of the Chinese angelica six-yellow soup material by scientific research and reasonable processing on the basis of the Chinese angelica six-yellow soup ancient formula, and ensures the drug property and the drug effect of the original formula.

The invention determines a preparation method of a standard decoction of the angelica six-yellow decoction and a preparation method of a traditional decoction of the angelica six-yellow decoction through ancient documents, and the preparation method comprises the following steps:

raw materials for preparing standard decoction

The raw materials for research and preparation of standard decoction comprise traditional Chinese medicinal materials and decoction pieces thereof, and the specification is consistent with the requirements of the current edition of Chinese pharmacopoeia.

(II) preparation of standard decoction

Prescription: 1 part of angelica, 1 part of radix rehmanniae recen, 1 part of prepared rehmannia root, 1 part of scutellaria baicalensis, 1 part of coptis chinensis, 1 part of phellodendron and 2 parts of astragalus membranaceus.

The preparation method comprises the following steps: (1) pretreatment: except that the decoction pieces to be decocted meet the specification of clinical decoction, the decoction pieces are also subjected to necessary treatment according to the traditional experiences of Chinese medicine preparation such as 'pounding when meeting shell and breaking when meeting seed'.

(2) Soaking: weighing the angelica, the radix rehmanniae recen, the prepared rehmannia root, the scutellaria baicalensis, the coptis chinensis, the phellodendron amurense and the astragalus mongholicus according to the weight part of the prescription, crushing into coarse powder, adding water according to the ratio of the mass of the medicinal materials to the volume of the water of 1:20, soaking for 0-60 minutes.

(3) The decoction time is as follows: decocting until the amount of the liquid medicine is half of the added water amount.

(4) And (3) filtration: the solid-liquid separation should be carried out while hot.

(5) And (3) cooling: the separated liquid should be cooled rapidly to inhibit thermal decomposition of the components.

(6) Concentration: concentrating to obtain a fluid extract with the relative density of 1.02-1.04 (60 ℃).

(7) And (3) drying: drying under reduced pressure, spray drying or freeze drying.

The invention also provides application of the method for simultaneously identifying the components of the coptis chinensis and the phellodendron amurense in detecting the quality of a traditional Chinese medicine product.

The invention also provides a quality detection method of the traditional Chinese medicine product, which comprises the step of processing the product to be detected according to any one of the methods for identifying the components of the coptis chinensis and the phellodendron amurense.

After the first development and color development, comparing the chromatogram of the test sample with the chromatogram of the coptis chinensis reference medicinal material or the chromatogram of the palmatine hydrochloride reference substance, and judging whether the coptis chinensis exists in the product to be detected.

And after the second development and color development, comparing the chromatogram of the test sample with the chromatogram of a phellodendron reference medicinal material or the chromatogram of a phellodendrine hydrochloride reference substance to judge whether the phellodendron is present in the product to be detected.

If spots with the same color appear on the chromatogram of the test sample at the positions corresponding to the chromatograms of the coptis chinensis control medicinal material and the palmatine hydrochloride control, the existence of coptis chinensis in the product to be detected is proved.

If the chromatogram of the sample shows spots of the same color at the positions corresponding to the chromatogram of the cortex Phellodendri reference medicinal material and the chromatogram of the phellodendrine hydrochloride reference substance, the existence of cortex Phellodendri in the product to be detected is proved.

Preferably, the traditional Chinese medicine product is a traditional Chinese medicine composition, a traditional Chinese medicine compound decoction or a solid preparation prepared from the traditional Chinese medicine compound decoction, wherein the traditional Chinese medicine composition contains the coptis chinensis and the phellodendron amurense;

preferably, the test sample is selected from angelica six-yellow soup or a compound preparation of angelica six-yellow soup.

The angelica and six-yellow decoction can be obtained by directly decocting and extracting decoction pieces of various medicinal materials by adding a solvent, or can be obtained by crushing the decoction pieces and then decocting and extracting the decoction pieces by adding the solvent.

The technical scheme of the invention has the following advantages:

1. the method for simultaneously identifying the components of the coptis chinensis and the phellodendron amurense is characterized in that a secondary thin layer expansion method is utilized for the first time, namely, firstly, the first expansion is carried out and then the inspection is carried out, then, the second expansion is carried out and then the inspection is carried out, and in the first expansion and the second expansion, the developing agents with specific compositions are respectively adopted, the same thin layer plate is adopted, the second sample application is not needed, the components of the coptis chinensis and the phellodendron amurense can be simultaneously and qualitatively identified, the improvement of resources and cost caused by the independent qualitative identification of the components of the coptis chinensis and the phellodendron amurense is avoided, the qualitative identification of the coptis chinensis and the phellodendron amurense of a Chinese angelica six-huang decoction compound preparation and a substance reference real object can be simultaneously controlled, the quality is ensured to be uniform, stable and reliable, and the method has the advantages of high accuracy, time saving and cost and easiness in popularization and application.

2. According to the method for simultaneously identifying the components of the golden thread and the golden thread, the Rf value of phellodendrine hydrochloride is slightly small when the butyl acetate-methanol-isopropanol-water is singly adopted as a developing agent for developing, so that the components of the golden thread cannot be identified, and when the n-butyl alcohol-water-acetic acid is singly adopted as the developing agent, the separation effect of the phellodendrine hydrochloride and another spot is poor, so that the spot discrimination of the phellodendrine hydrochloride is poor, and the components of the golden thread and the golden thread cannot be simultaneously identified; the method adopts butyl acetate-methanol-isopropanol-water as a developing agent for development and color development to realize the identification of the components of the golden thread, and adopts n-butyl alcohol-water-acetic acid as the developing agent for secondary development and color development under the condition of not damaging a thin layer plate, so that the Rf value of phellodendrine hydrochloride is relatively small when the butyl acetate-methanol-isopropanol-water is developed once, the phellodendrine hydrochloride is prevented from being interfered by another spot and is difficult to identify when the n-butyl alcohol-water-acetic acid is developed once, and the phellodendrine hydrochloride and the n-butyl alcohol-water-acetic acid are in a complementary relationship, so that not only the components of the golden thread can be identified, but also the components of the golden thread can be identified.

3. The method for simultaneously identifying the components of the golden thread and the golden cypress adopts the phellodendrine hydrochloride as the thin-layer identification component of the golden cypress without negative interference, and adopts the palmatine hydrochloride as the thin-layer identification component of the golden thread without negative interference. Therefore, the palmatine hydrochloride is used as a characteristic spot for identifying the thin coptis layer, and the phellodendrine hydrochloride is used as a characteristic spot for identifying the thin phellodendron layer.

4. The method for simultaneously identifying the components of the coptis chinensis and the phellodendron amurense is characterized in that a sample solution is prepared by mixing a sample with methanol, ultrasonically extracting and filtering, and the preparation method is less in solvent, short in time, simple and easy to operate.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a thin-layer identification chromatogram of a standard of a Chinese angelica six-yellow decoction substance in the prior art; wherein the number 1 is epiberberine, 2 is coptisine hydrochloride, 3-4 are coptis root reference medicinal materials, 5 are test samples, and 6 are coptis root negatives;

FIG. 2 is a first developed chromatogram in example 1 of the present invention;

FIG. 3 is a second developed chromatogram in example 1 of the present invention;

FIG. 4 is a first developed chromatogram in example 2 of the present invention;

FIG. 5 is a second developed chromatogram in example 2 of the present invention;

FIG. 6 is a first developed chromatogram in example 3 of the present invention;

FIG. 7 is a second developed chromatogram in example 3 of the present invention;

FIG. 8 is a first developed chromatogram in example 4 of the present invention;

FIG. 9 is a second developed chromatogram in example 4 of the present invention;

FIG. 10 is a first developed chromatogram in example 5 of the present invention;

FIG. 11 is a second developed chromatogram in example 5 of the present invention;

FIG. 12 is a first expanded chromatogram of the spot locations of the control identified in Experimental example 1;

FIG. 13 is a chromatogram observed under visible light after air-drying in comparative example 1;

FIG. 14 is a chromatogram observed under visible light after spraying a diluted bismuth potassium iodide test solution in comparative example 1;

FIG. 15 is a chromatogram of the first color development of the method of comparative example 2;

FIG. 16 is a chromatogram of the color development of method two in comparative example 2;

FIG. 17 is a chromatogram of the third development of the method in comparative example 2;

FIG. 18 is a chromatogram for color development of comparative example 4;

FIG. 19 is a chromatogram of the color development of method two in comparative example 4.

Detailed Description

The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.

The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.

The specifications of the medicinal materials and the decoction pieces thereof adopted by the invention are consistent with the requirements of the current edition of Chinese pharmacopoeia.

The preparation method of the standard substance or compound preparation of the angelica six-yellow decoction and the angelica six-yellow decoction in the following examples, experimental examples and comparative examples is as follows:

(1) preparing the angelica six-yellow decoction: respectively mixing the Chinese angelica, the radix rehmanniae recen, the prepared rehmannia root, the baical skullcap root, the golden thread, the amur corktree bark and the astragalus root according to the mass ratio of 1: 1: 1: 1: 1: 1:2 weighing angelica, radix rehmanniae, prepared rehmannia root, scutellaria, coptis, phellodendron and astragalus, mixing, crushing into coarse powder, adding 20 times of water (the amount of water is milliliter per gram of the medicinal materials), decocting until the volume of the liquid medicine is half of the volume of the water, and filtering to obtain the angelica six-yellow soup.

(2) Preparing a standard substance of the angelica six-yellow decoction: preparing the angelica six-yellow soup according to the method, and drying to obtain the material reference substance real object powder of the angelica six-yellow soup.

(3) Preparing the Chinese angelica and six-yellow decoction compound preparation: taking the standard substance powder of the angelica six-yellow decoction, and preparing the tablet of the angelica six-yellow decoction by a direct powder tabletting process. And (3) taking the standard substance powder of the angelica six-yellow decoction, and filling the powder into capsules to prepare the capsules of the angelica six-yellow decoction.

In the following examples, experimental examples and comparative examples, the preparation methods of the coptis single-negative test sample powder, the phellodendron single-negative test sample powder and the coptis and phellodendron double-negative test sample powder are as follows:

(1) preparing coptis chinensis single negative test sample powder: the Chinese angelica six-yellow decoction is prepared by the same method as the standard substance real object powder of the Chinese angelica six-yellow decoction, and is only characterized in that no coptis is added in the Chinese angelica six-yellow decoction, and the other raw materials and the step parameters are the same.

(2) Preparing a cortex phellodendri single negative test sample powder: the Chinese angelica six-yellow decoction is prepared by the same method as the standard substance real object powder of the Chinese angelica six-yellow decoction, and is only characterized in that no phellodendron is added in the Chinese angelica six-yellow decoction, and the other raw materials and the step parameters are the same.

(3) Preparing a coptis and phellodendron double-negative test sample powder: the Chinese angelica six-yellow decoction is prepared by the same method as the standard substance real object powder of the Chinese angelica six-yellow decoction, and is only characterized in that no phellodendron and coptis are added in the Chinese angelica six-yellow decoction, and the other raw materials and the step parameters are the same.

Example 1

The embodiment provides a method for simultaneously identifying components of coptis chinensis and phellodendron amurense, which comprises the following steps:

(1) preparation of the solution

Preparing a test solution: adding methanol 20ml into 2g of radix Angelicae sinensis Liuhuang decoction reference substance powder, performing ultrasonic treatment for 30min, and filtering to obtain filtrate as sample solution.

Preparing a reference medicinal material solution: taking 2g of Coptidis rhizoma and cortex Phellodendri reference medicinal material, adding 20ml of methanol respectively, performing ultrasonic treatment for 30min, and filtering to obtain Coptidis rhizoma reference medicinal material solution and cortex Phellodendri reference medicinal material solution respectively.

Negative control solution preparation: taking Coptidis rhizoma single negative sample powder, cortex Phellodendri single negative sample powder, and Coptidis rhizoma cortex Phellodendri double negative sample powder 2g, adding methanol 20ml respectively, ultrasonic treating for 30min, filtering, and respectively getting Coptidis rhizoma single negative solution, cortex Phellodendri single negative solution, and Coptidis rhizoma cortex Phellodendri double negative solution.

Preparation of a reference solution: taking phellodendrine hydrochloride and palmatine hydrochloride reference substances, and adding methanol to prepare a reference substance solution (phellodendrine hydrochloride reference substance solution) containing 0.5mg of phellodendrine hydrochloride and a reference substance solution (palmatine hydrochloride reference substance solution) containing 0.5mg of palmatine hydrochloride per lml respectively.

(2) Thin layer chromatography detection

1) Sucking 2 mul of each group of solution prepared in the step (1), and respectively dropping the solution on the same silica gel G thin-layer plate.

2) Taking butyl acetate-methanol-isopropanol-water as developing agent at volume ratio of 4: 3: 1.5: 1, placing in developing cylinder pre-saturated with concentrated ammonia solution for 20min, developing, taking out, air drying, and observing under 365nm light source, as shown in FIG. 2. The result shows that the chromatogram of the test sample shows spots with the same color at the corresponding positions of the chromatogram of the coptis chinensis control drug and the chromatogram of the palmatine hydrochloride control.

3) And 2) observing the thin-layer plate, wherein the volume ratio of the thin-layer plate to the thin-layer plate is 7: 2: 1, taking n-butanol-water-acetic acid as a developing agent, developing, taking out and airing. Spraying diluted potassium bismuth iodide solution, and observing under visible light, as shown in FIG. 3. The result shows that the chromatogram of the test sample shows spots with the same color at the corresponding positions of the chromatogram of the cortex phellodendri reference material and the chromatogram of the phellodendrine hydrochloride reference material.

Example 2

(1) preparation of the solution

Preparing a test solution: drying the decoction to obtain powder, adding methanol 20ml into 2g of the powder, performing ultrasonic treatment for 20min, and filtering to obtain filtrate as sample solution.

Preparing a reference medicinal material solution: taking 2g of the reference medicinal materials of the coptis chinensis and the phellodendron, and preparing the reference medicinal material solution according to the same method of preparing the test solution.

Negative control solution preparation: taking 1g of Coptidis rhizoma single negative, cortex Phellodendri single negative test sample powder, and Coptidis rhizoma cortex Phellodendri double negative test sample powder, and making into negative control solution according to the same method as above for preparing test sample solution.

Preparation of a reference solution: taking phellodendrine hydrochloride, palmatine hydrochloride, coptisine hydrochloride and berberine hydrochloride reference substances, and adding methanol respectively to prepare solutions containing 0.5mg of reference substance per lml as reference substance solutions.

(2) Thin layer chromatography detection

1) Sucking 1 mu l of each group of solution prepared in the step (1), and respectively dropping the solution on the same silica gel G thin-layer plate.

2) Taking butyl acetate-methanol-isopropanol-water (volume ratio of 3: 1.5: 0.8) as developing agent, placing in a developing tank pre-saturated with concentrated ammonia test solution for 25min, developing, taking out, air drying, observing related characteristics of Coptidis rhizoma under 360nm light source, and displaying spots of the same color at positions corresponding to reference medicinal material chromatogram and reference substance chromatogram, as shown in FIG. 4.

3) The thin-layer plate is prepared by mixing n-butanol-water-acetic acid (volume ratio is 6: 2: 1) developing with developer, taking out, and air drying. Spraying diluted potassium bismuth iodide solution, observing related characteristics of cortex Phellodendri under visible light, and displaying spots of the same color at the positions corresponding to the chromatogram of the reference material and the chromatogram of the reference product, as shown in FIG. 5.

In fig. 4 and 5, the number 1 is berberine hydrochloride, 2 is coptisine hydrochloride, 3 is palmatine hydrochloride, 4 is phellodendrine hydrochloride, 5 is a sample, 6 is a coptis root control medicinal material, 7 is a phellodendron bark control medicinal material, 8 is a coptis root single negative control solution, 9 is a phellodendron bark single negative control solution, and 10 is a coptis root and phellodendron bark double negative control solution.

Example 3

(1) preparation of the solution

Preparing a test solution: taking the Angelica sinensis Liuhuang decoction substance tablet, crushing to obtain powder, taking 1.5g of the powder, adding 20ml of methanol, carrying out ultrasonic treatment for 40min, and filtering to obtain filtrate as a test solution.

Preparing a reference medicinal material solution: taking 1.5g of rhizoma Coptidis and cortex Phellodendri as reference materials, and preparing the reference material solutions according to the same method as above.

(2) Thin layer chromatography detection

2) Taking butyl acetate-methanol-isopropanol-water (volume ratio of 4: 3.5: 1: 0.6) as developing agent, placing in a developing tank pre-saturated with concentrated ammonia test solution for 30min, developing, taking out, air drying, observing related characteristics of Coptidis rhizoma under 370nm light source, and displaying spots of the same color at positions corresponding to reference medicinal material chromatogram and reference substance chromatogram, as shown in FIG. 6.

3) The thin-layer plate is prepared by mixing n-butanol-water-acetic acid (volume ratio is 7: 1: 1.5) developing agent, taking out, and drying. Spraying diluted potassium bismuth iodide solution, observing related characteristics of cortex Phellodendri under visible light, and displaying spots of the same color at the positions corresponding to the chromatogram of the reference material and the chromatogram of the reference product, as shown in FIG. 7.

In fig. 6 and 7, number 1 is berberine hydrochloride; 2 is coptisine hydrochloride; 3 is palmatine hydrochloride; 4 is phellodendrine hydrochloride; 5 is a test sample; 6 is a reference medicinal material of rhizoma Coptidis; 7 is cortex Phellodendri reference material; 8 is a coptis single negative control solution; 9 is a cortex phellodendri single negative control solution; 10 is a double negative control solution of coptis and phellodendron bark.

Example 4

(1) preparation of the solution

Preparing a test solution: adding methanol 20ml into 2g of radix Angelicae sinensis Liuhuang decoction reference substance powder, performing ultrasonic treatment for 25min, and filtering to obtain filtrate as sample solution.

Negative control solution preparation: taking Coptidis rhizoma single negative, cortex Phellodendri single negative sample powder, and Coptidis rhizoma cortex Phellodendri double negative sample powder 1g, and preparing negative control solution according to the same method as above.

(2) Thin layer chromatography detection

1) Pipetting 3. mu.l of each solution, and spotting on the same silica gel G thin layer plate

2) Taking butyl acetate-methanol-isopropanol-water (volume ratio of 4: 3: 2: 1) as developing agent, placing in a developing tank pre-saturated with concentrated ammonia test solution for 35min, developing, taking out, air drying, observing related characteristics of Coptidis rhizoma under 368nm light source, and displaying spots of the same color at positions corresponding to control material chromatogram and control product chromatogram, as shown in FIG. 8.

3) The thin-layer plate is prepared by mixing n-butanol-water-acetic acid (volume ratio is 8: 2.5: 3) developing with developer, taking out, and air drying. Spraying diluted potassium bismuth iodide solution, observing related characteristics of cortex Phellodendri under visible light, and displaying spots of the same color at the positions corresponding to the chromatogram of the reference material and the chromatogram of the reference product, as shown in FIG. 9.

In fig. 8 and 9, number 1 is berberine hydrochloride; 2 is coptisine hydrochloride; 3 is palmatine hydrochloride; 4 is phellodendrine hydrochloride; 5 is a test sample; 6 is a reference medicinal material of rhizoma Coptidis; 7 is cortex Phellodendri reference material; 8 is a coptis single negative control solution; 9 is a cortex phellodendri single negative control solution; 10 is a double negative control solution of coptis and phellodendron bark.

Example 5

(1) preparation of the solution

Preparing a reference medicinal material solution: taking 2g of coptis and phellodendron reference medicinal materials, and preparing the reference medicinal material solution according to the same method of preparing the test solution.

Preparation of a reference solution: taking phellodendrine hydrochloride and palmatine hydrochloride reference substances, and adding methanol respectively to prepare solutions each containing 0.5mg per lml as reference substance solutions.

(2) Thin layer chromatography detection

1) 2. mu.l of each solution was pipetted onto the same silica gel G thin layer plate

2) Taking butyl acetate-methanol-isopropanol-water (volume ratio of 4: 3: 2: 1) as developing agent, placing in a developing tank pre-saturated with concentrated ammonia test solution for 30min, developing, taking out, air drying, observing related characteristics of Coptidis rhizoma under 360nm light source, and displaying spots of the same color at positions corresponding to reference medicinal material chromatogram and reference substance chromatogram, as shown in figure 10.

3) The thin-layer plate is prepared by mixing n-butanol-water-acetic acid (volume ratio is 7: 2: 1) developing with developer, taking out, and air drying. Spraying diluted potassium bismuth iodide solution, observing related characteristics of cortex Phellodendri under visible light, and displaying spots of the same color at positions corresponding to control material chromatogram and control product chromatogram, as shown in FIG. 11.

In FIGS. 10 and 11, reference numeral 1 denotes a phellodendrine hydrochloride control solution; 2 is a palmatine hydrochloride reference substance solution; 3 is a phellodendron bark reference medicinal material solution; 4 is a reference medicinal solution of rhizoma Coptidis; 5 is a test solution; 6 is a single negative solution of Chinese angelica, six yellow decoction and coptis root; 7 is a single negative solution of radix Angelicae sinensis, Liuhuang decoction and cortex Phellodendri; 8 is a double-yin solution of Chinese angelica, six-yellow decoction, coptis and phellodendron bark.

Experimental example 1 screening of control and determination of characteristic spots

(1) Determining spot location of control

A test solution, a Coptidis rhizoma control solution and a Coptidis rhizoma single negative control solution were prepared as in step (1) of example 1.

Preparation of control solutions: taking phellodendrine hydrochloride, jateorhizine hydrochloride, palmatine hydrochloride, epiberberine hydrochloride, coptisine hydrochloride and berberine hydrochloride reference substances respectively, and adding methanol respectively to prepare solutions containing 0.5mg of each reference substance per lml.

The test solution, the coptis chinensis control solution, the coptis chinensis single negative control solution and each control solution prepared in the embodiment are detected by the method in the step (2) in the embodiment 1, and the first developed chromatogram is shown in fig. 12.

Sample numbers represent respectively: 1. phellodendrine hydrochloride; 2. jatrorrhizine hydrochloride; 3. palmatine hydrochloride; 4. epiberberine hydrochloride; 5. coptisine hydrochloride; 6. berberine hydrochloride; 7. rhizoma Coptidis as reference material; 8. a test article; 9. coptis chinensis single negative control solution.

As can be seen from fig. 12, under the chromatographic conditions of this experimental example,

samples

1 and 2 were not detected, and spots of epiberberine hydrochloride, coptisine hydrochloride, and berberine hydrochloride were determined by comparing the spot positions (Rf values) and the colors of the

samples

3, 4, and 5 with those of the control.

(2) Selecting suitable reference substance

The chromatogram obtained after the two developments of example 1 was used as the subject.

TABLE 1 thin-layer chromatography test results table

As can be seen from fig. 3, 8 spots are detected in the chromatogram of the test sample, and compared with

samples

6 and 8, except for palmatine hydrochloride, the negatives of other 7 spots are interfered, so palmatine hydrochloride in the test sample is a characteristic spot for identifying the thin layer of coptis chinensis, and the palmatine hydrochloride reference is used as a reference for identifying the components of coptis chinensis.

As can be seen from fig. 4, 5 points are detected in the chromatogram of the test sample, and compared with

samples

6 and 8, the palmatine hydrochloride is different, but the spot trailing at the position of the test sample is serious, and the identification effect is poorer than that of the test sample which is developed for the 1 st time under ultraviolet; compared with

samples

7 and 8, the phellodendrine hydrochloride is different, so that the phellodendrine hydrochloride in the test sample is a characteristic spot for identifying the phellodendron thin layer, and the phellodendrine hydrochloride is used as a reference substance for identifying the phellodendron components.

In addition, as can be seen from fig. 3 and 4, only phellodendrine hydrochloride can be used as a thin layer identification component of phellodendron amurense without negative interference, and palmatine hydrochloride can be used as a thin layer identification component of coptis chinensis without negative interference. If only the 1 st expansion is adopted, only the coptis chinensis component can be identified, and the phellodendron amurense component cannot be identified.

Experimental example 2 methodological verification

1. Instruments, reagents and reagents

The instrument comprises the following steps: a numerical control ultrasonic cleaner, an electric heating constant temperature water bath, a GoodLook-1000 thin layer chromatography imaging system (Shanghai science and technology Co., Ltd.), a JJ500 electronic balance (Shuangjie testing apparatus factory in Hei), and an MSA6 & 6S & 0CE-DM electronic balance (Sidolisi scientific apparatus (Beijing) Co., Ltd.).

Trade name of thin layer plate: silica gel G thin layer plate, manufacturer: the institute of chemical industry of cigarette tai city; specification: 100 x 200 cm; the model is as follows: HSG; batch number: 20190808.

reagent: acetic acid, methanol, bismuth subnitrate, potassium iodide, purified water, chloroform, cyclohexane, ethyl acetate, isopropanol, triethylamine, n-butanol, petroleum ether (60-90 ℃), butyl acetate and ammonia water are all analytically pure.

Reagent testing: the standard substance of the Chinese angelica six-yellow decoction, the Chinese angelica six-yellow decoction coptis single-negative test sample powder, the Chinese angelica six-yellow decoction phellodendron single-negative test sample powder and the Chinese angelica six-yellow decoction phellodendron double-negative test sample powder are respectively prepared according to the method.

The coptis root reference medicinal material, the phellodendron bark reference medicinal material, the phellodendrine hydrochloride and the palmatine hydrochloride are purchased from China food and drug testing research institute.

2. Preparation of the solution

Preparing a test solution: adding methanol 20ml into 2g of radix Angelicae sinensis Liuhuang decoction reference substance, performing ultrasonic treatment for 30min, and filtering to obtain filtrate as sample solution.

Negative control solution preparation: taking Coptidis rhizoma single negative, cortex Phellodendri single negative test sample powder, and Coptidis rhizoma cortex Phellodendri double negative test sample powder 2g, adding methanol 20ml respectively, ultrasonic treating for 30min, filtering, and respectively getting Coptidis rhizoma single negative solution, cortex Phellodendri single negative solution, and Coptidis rhizoma cortex Phellodendri double negative solution.

Preparation of a reference solution: taking phellodendrine hydrochloride and palmatine hydrochloride reference substances, and adding methanol to prepare a reference substance solution containing 0.5mg of phellodendrine hydrochloride and a reference substance solution containing 0.5mg of palmatine hydrochloride per lml respectively.

3. Conditions of thin layer chromatography

1) Sucking 3 mul of each group of solution prepared in the step (1), and respectively dropping the solution on the same silica gel G thin-layer plate.

2) Taking butyl acetate-methanol-isopropanol-water (volume ratio of 4: 3: 1.5: 0.6) as developing agent, placing in a developing tank pre-saturated with concentrated ammonia test solution for 20min, developing, taking out, air drying, observing related characteristics of Coptidis rhizoma under 365nm light source, and displaying spots of the same color at positions corresponding to the reference medicinal material chromatogram and the reference substance chromatogram.

3) The thin-layer plate is prepared by mixing n-butanol-water-acetic acid (volume ratio is 7: 2: 1) developing with developer, taking out, and air drying. Spraying diluted potassium bismuth iodide solution, observing the related characteristics of cortex Phellodendri under visible light, and displaying spots of the same color at the positions corresponding to the chromatogram of the reference material and the chromatogram of the reference substance.

4. Investigation of different sample weights

1.5g, 2.0g and 2.5g of the reference substance of the Angelica sinensis Liuhuang decoction were weighed, and each sample solution was prepared according to item 2 of this experimental example. Considering that the chromatographic spots of the coptis control medicinal material and the phellodendron control medicinal material are clear and the dosage is reasonable, the reference medicinal material is not examined for the weighing amount.

The sample solution, the control solution and the control solution were pipetted in 3. mu.l each, spotted onto the same silica gel G thin layer plate, and the procedure was performed as in item 3 of this example.

And (3) knotting: through the investigation of the symmetrical sample amount, when the sample amount of the sample is 2.0g, the thin-layer chromatography has clear spots, moderate spot size, good separation degree and moderate Rf.

5. Investigation of different dot sizes

A sample solution, a control solution and a reference solution were prepared according to item 2 of this example, and 1. mu.l, 2. mu.l and 3. mu.l of each of the sample solution, the control solution and the reference solution were pipetted onto the same silica gel G thin layer plate, and the rest were performed according to item 3 of this example.

And (3) knotting: through the investigation of the sample amount, when the sample amount of the sample is 1-3 mu l, the thin-layer chromatography has clear spots, moderate spot size, good separation degree and moderate Rf.

6. Investigation of different humiture

Preparing a test solution, a control solution and a reference solution according to the item 2 in the experimental example, sucking 3 μ l of each test solution, control solution and reference solution, respectively spotting the solutions on the same silica gel G thin-layer plate, and respectively inspecting the development conditions under three relative humidity conditions of 5-35 ℃ high and low temperatures, 15-35%, 35-75% and 75-90% high, medium and low temperatures: firstly, selecting high and low temperatures within the range of 5-35 ℃, wherein the temperature is 7.0 ℃ and the humidity is 46 percent respectively; (2) the temperature is 25.3 ℃, and the humidity is 46%; (II) selecting 1 humidity condition within the range of 15-35% and 75-90% of relative humidity respectively, wherein the humidity conditions are as follows: (1) the temperature is 25.1 ℃, and the humidity is 17%; (2) the temperature was 25.2 ℃ and the humidity was 84%. Taken out, dried and the rest are operated according to the item 3 of the experimental example.

And (3) knotting: the test samples can be well separated under different temperature and humidity conditions, and the thin-layer chromatography identification method has good durability for different temperatures and humidity.

7. Investigation of different thin layer panels

Preparing a sample solution, a reference medicinal material solution and a reference substance solution according to the item 2 of the experimental example, respectively adopting silica gel G thin-layer plates produced by Nicoti chemical industry research institute, Nicoti Chiense silica gel development and test factory and Merck KGaA, sucking 2 mu l of each sample solution, reference medicinal material solution and reference substance solution, respectively spotting the sample solutions on the three thin-layer plates, and respectively operating according to the item 3 of the experimental example.

And (3) knotting: the silica gel G thin-layer plates produced by different manufacturers can obtain good separation effect on the standard substance of the angelica six-yellow decoction, and the thin-layer chromatography identification method has better durability on the thin-layer plates produced by different manufacturers.

Comparative example 1

The comparative example provides a thin layer chromatography identification method, comprising the following steps:

(1) preparation of the solution

Preparing a test solution: adding methanol 20ml into 2.5g of radix Angelicae sinensis Liuhuang decoction reference substance powder, performing ultrasonic treatment for 30min, and filtering to obtain filtrate as sample solution.

Preparation of a reference solution: taking phellodendrine hydrochloride, berberine hydrochloride and coptisine hydrochloride reference substances, and adding methanol respectively to obtain solutions containing 0.5mg of phellodendrine hydrochloride reference substance per lml.

(2) Thin layer chromatography detection

1) Sucking 4 mul of each group of solution prepared in the step (1), and respectively dropping the solution on the same silica gel G thin-layer plate.

2) The volume ratio is 3: 3.5: 1: 1.5: 0.5: 1, taking cyclohexane-ethyl acetate-isopropanol-methanol-water-triethylamine as a developing agent, placing the developing agent into a developing cylinder pre-saturated with concentrated ammonia test solution for 20min, developing, taking out, and respectively inspecting according to the following two methods, namely a method I: inspecting under visible light after drying; the second method comprises the following steps: after air drying, spraying diluted bismuth potassium iodide solution, and observing under visible light, as shown in FIGS. 15 and 16, respectively.

In FIGS. 13 to 14, the number 1 is phellodendrine hydrochloride; 2 is berberine hydrochloride; 3 is coptisine hydrochloride; 4 is a coptis root reference medicinal material; 5 is cortex Phellodendri reference material; 6, a test article; 7 is a coptis single negative control solution; 8 is a cortex phellodendri single negative control solution; 9 is a negative control solution of rhizoma Coptidis and cortex Phellodendri.

The results show that after drying, the glass is inspected under visible light: the phellodendrine hydrochloride shows no spots, and the phellodendrine hydrochloride is not developed and is still at the origin when the phellodendrine hydrochloride is detected under visible light after being sprayed with diluted bismuth potassium iodide test solution, which indicates that the method cannot identify the components of the phellodendron.

Comparative example 2

(1) preparation of the solution

Preparing a test solution: preparing radix Angelicae sinensis Liuhuang decoction reference substance powder according to the above method, adding methanol 20ml into 2g of the powder, performing ultrasonic treatment for 30min, and filtering to obtain filtrate as sample solution.

(2) Thin layer chromatography detection

2) The volume ratio of the components is 1: 1, developing by using petroleum ether-ethyl acetate as a developing agent, taking out, and airing.

3) The color is developed according to the following three ways respectively:

the method comprises the following steps: inspecting under visible light after drying;

the second method comprises the following steps: after drying, spraying and developing 5% vanillin concentrated sulfuric acid, heating at 105 ℃, and inspecting under visible light;

the third method comprises the following steps: after air drying, spraying diluted bismuth potassium iodide solution, and inspecting under visible light, the results are shown in FIGS. 17-19.

In FIGS. 15 to 17, the number 1 is phellodendrine hydrochloride; 2 is berberine hydrochloride; 3 is coptisine hydrochloride; 4 is a coptis root reference medicinal material; 5 is cortex Phellodendri reference material; 6, a test article; 7 is a coptis single negative control solution; 8 is a cortex phellodendri single negative control solution; 9 is a negative control solution of rhizoma Coptidis and cortex Phellodendri.

The results show that only yellow spots are observed at the origin after the first method is developed, and other samples except

samples

4 and 5 are not developed after the second method is developed, which indicates that the identification condition does not reach the purpose of thin layer development identification, and that the samples are not developed after the third method is developed, which indicates that the identification condition does not reach the purpose of thin layer development identification.

Comparative example 3

The experimental result is found by adopting a method developed in the existing literature, namely research on thin-layer identification of coptis chinensis and phellodendron amurense in patent prescription: the combined prescription totally displays 3 bright yellow fluorescent spots, wherein 3 spots corresponding to the coptis control medicinal material and 2 spots corresponding to the phellodendron control medicinal material are provided; the 2 spots displayed by the cortex phellodendri contrast medicinal material can be observed in the rhizoma coptidis contrast medicinal material, the negative mutual interference of the rhizoma coptidis and the cortex phellodendri is realized, the cortex phellodendri has no specific identification component, and whether the cortex phellodendri is fed or not can not be determined when the cortex phellodendri is identified in a combined prescription.

Comparative example 4

Preparation of a reference solution: taking phellodendrine hydrochloride, berberine hydrochloride and coptisine hydrochloride reference substances, and adding methanol to prepare a reference substance solution containing 0.5mg of phellodendrine hydrochloride, a reference substance solution containing 0.5mg of berberine hydrochloride and a reference substance solution containing 0.5mg of coptisine hydrochloride per lml respectively.

3. Conditions of thin layer chromatography

2) The method comprises the following steps of (1) mixing n-butanol-acetic acid-water (volume ratio is 2: 1: 2) developing with developer, taking out, and air drying.

3) The following two ways are respectively adopted for color development:

the first method is as follows: when the method is directly detected under visible light, the phellodendrine hydrochloride does not show spots, and the phellodendron cannot be detected, but only the berberine hydrochloride in the coptis chinensis can be identified.

The second method comprises the following steps: spraying diluted bismuth potassium iodide test solution, observing under visible light, and observing related characteristics of cortex Phellodendri, wherein the result shows that phellodendrine hydrochloride is interfered by another spot and is not easily distinguished in the test sample, so that cortex Phellodendri cannot be detected, and only berberine hydrochloride in Coptidis rhizoma can be distinguished.

As shown in FIGS. 18-19, number 1 is phellodendrine hydrochloride; 2 is berberine hydrochloride; 3 is coptisine hydrochloride; 4 is a coptis root reference medicinal material; 5 is cortex Phellodendri reference material; 6, a test article; 7 is a coptis single negative control solution; 8 is a cortex phellodendri single negative control solution; 9 is a negative control solution of rhizoma Coptidis and cortex Phellodendri.

Comparative example 5

3. Conditions of thin layer chromatography

2) The method comprises the following steps of (1) mixing n-butanol, water and acetic acid (volume ratio is 7: 2: 1) developing with developer, taking out, and air drying.

3) The following two ways are respectively adopted for color development:

the first method is as follows: when the sample is directly detected under visible light, phellodendrine hydrochloride does not show spots, and phellodendron cannot be detected.

The second method comprises the following steps: spraying diluted potassium bismuth iodide solution, and observing under visible light, wherein phellodendrine hydrochloride is interfered by another spot and is difficult to distinguish, and the thin layer plate can not be developed for the second time, and thus, phellodendron and coptis chinensis can not be detected simultaneously.

It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.

Claims

1. A method for simultaneously identifying components of rhizoma Coptidis and cortex Phellodendri, which comprises the following steps,

(2) and (3) detection by thin-layer chromatography: comprises the steps of two times of unfolding and respectively inspecting after the unfolding; wherein, butyl acetate-methanol-isopropanol-water is used as a developing solvent for the first development, and n-butanol-water-acetic acid is used as a developing solvent for the second development.

2. The method for simultaneously identifying ingredients of coptis chinensis and phellodendron amurense according to claim 1, wherein the step (2) comprises:

2) carrying out first development by using butyl acetate-methanol-isopropanol-water as a developing agent, taking out, volatilizing a solvent, and placing the thin-layer plate under a light source of 360-370nm for inspection;

3. The method for simultaneously identifying ingredients of coptis chinensis and phellodendron amurense according to claim 2, wherein any one of the following a to E is satisfied:

A. in the step 1), the adopted thin layer plate is a silica gel G thin layer plate, and the sample application volume is 1-10 mu l;

B. in the step 2), the volume ratio of butyl acetate-methanol-isopropanol-water is 3-4: 3-4: 1-2: 0.6 to 1;

C. in the step 2), a container for unfolding is pre-saturated by concentrated ammonia test solution before the first unfolding, and the pre-saturation time is preferably 20-35 min;

D. in the step 3), the volume ratio of n-butanol to water to acetic acid is 6-8: 1-3: 1-3;

E. in the step 3), the color developing agent is selected from at least one of potassium bismuth iodide, phosphomolybdic acid and dilute potassium bismuth iodide test solution.

4. The method for simultaneously identifying ingredients of Coptidis rhizoma and Phellodendri cortex as claimed in any one of claims 1-3, wherein step (1) comprises mixing the sample with methanol, ultrasonic extracting, and filtering to obtain sample solution;

preferably, the ratio of the mass of the sample to the volume of methanol is 1.5-2.5 g: 20ml of the solution;

preferably, the extraction time is 20-40 min.

5. The method for simultaneously identifying coptis chinensis and phellodendron amurense ingredients according to any one of claims 1 to 4, wherein the reference medicinal material solution comprises a coptis chinensis reference medicinal material solution and a phellodendron amurense reference medicinal material solution, and preferably, the preparation method of the reference medicinal material solution comprises the step of preparing the coptis chinensis reference medicinal material solution and the phellodendron amurense reference medicinal material solution respectively according to the method of any one of claims 1 to 4.

6. The method for simultaneously identifying ingredients of Coptis chinensis Franch and phellodendron amurense according to any one of claims 1 to 5, wherein the reference solution comprises a palmatine hydrochloride reference solution and a phellodendrine hydrochloride reference solution;

preferably, the preparation method of the reference solution comprises mixing the palmatine hydrochloride reference and the phellodendrine hydrochloride reference with methanol to obtain the palmatine hydrochloride reference solution with a concentration of 0.2-0.6mg/ml and the phellodendrine hydrochloride reference solution with a concentration of 0.2-0.6 mg/ml.

7. The method for simultaneously identifying the components of Coptis chinensis Franch and phellodendron amurense according to any one of claims 1 to 6, wherein the sample is a Chinese medicinal composition containing Coptis chinensis Franch and phellodendron amurense or a Chinese medicinal preparation containing Coptis chinensis Franch and phellodendron amurense together;

preferably, the test sample is angelica six-yellow soup or a compound preparation of the angelica six-yellow soup;

8. Use of the method of any one of claims 1-7 for simultaneously identifying the components of Coptidis rhizoma and cortex Phellodendri in the detection of quality of Chinese medicinal product.

9. A quality detection method of a Chinese medicinal product, which is characterized by comprising the step of detecting the product to be detected according to the method for simultaneously identifying the components of coptis chinensis and phellodendron amurense as claimed in any one of claims 1 to 7.

10. The quality detection method according to claim 9, wherein the Chinese medicinal product is a Chinese medicinal composition containing Coptidis rhizoma and cortex Phellodendri simultaneously or a Chinese medicinal preparation containing Coptidis rhizoma and cortex Phellodendri simultaneously;