阅读说明：本技术 一种具有改善睡眠功能的中药组合物的检测方法 (Detection method of traditional Chinese medicine composition with sleep improvement function ) 是由 郁晓艺 罗文光 欧阳道福 梁淑明 卜闪闪 陈晶 温馨 颜建章 孙静 胡瑞连 于 2018-08-23 设计创作，主要内容包括：本发明属于中药检测技术领域,公开了一种具有改善睡眠功能的中药组合物的检测方法,包括用薄层色谱法鉴别灵芝、首乌藤和酸枣仁及用高效液相色谱法测定斯皮诺素的含量,所述方法对灵芝及首乌藤采用相同的前处理方法,并选择特定的展开剂体系,使灵芝及首乌藤中的各种成分均能很好地分离,实现了两种药材的同时鉴别；各自供试品色谱中,在与阳性对照品色谱对应的位置上,显示相同颜色的斑点,而阴性对照品色谱中均无干扰斑点,表明本发明的方法具有较强的专属性。同时,本发明的方法简化了操作过程,减少了鉴别所需时间,提高了工作效率,降低了检测成本,同时也为中药组合物中各成分的薄层色谱法鉴别提供了新思路,有利于中药质量控制标准的提高。(The invention belongs to the technical field of traditional Chinese medicine detection, and discloses a detection method of a traditional Chinese medicine composition with a sleep improving function, which comprises the steps of identifying lucid ganoderma, multiflower knotweed stem and spina date seed by using a thin-layer chromatography and determining the content of spinosad by using a high performance liquid chromatography, wherein the lucid ganoderma and the multiflower knotweed stem are subjected to the same pretreatment method by using the method, and a specific developing agent system is selected, so that various components in the lucid ganoderma and the multiflower knotweed stem can be well separated, and the simultaneous identification of two medicinal materials is realized; in the chromatogram of each test sample, spots with the same color are displayed at the positions corresponding to the chromatograms of the positive reference substances, and no interference spots exist in the chromatograms of the negative reference substances, which shows that the method has stronger specificity. Meanwhile, the method simplifies the operation process, reduces the time required for identification, improves the working efficiency, reduces the detection cost, provides a new idea for the thin-layer chromatography identification of each component in the traditional Chinese medicine composition, and is beneficial to the improvement of the quality control standard of the traditional Chinese medicine.)

1. A detection method of a traditional Chinese medicine composition comprises the step of identifying lucid ganoderma, caulis polygoni multiflori and spina date seeds in the traditional Chinese medicine composition by using a thin-layer chromatography, and is characterized in that the thin-layer chromatography comprises the following steps:

(1) preparation of positive control solution and test solution

Respectively adding a first solvent into a ganoderma lucidum positive control, a polygonum multiflorum positive control and a spina date seed positive control, filtering, evaporating filtrate to dryness, and adding a second solvent to respectively serve as respective positive control solutions;

adding the first solvent into the Chinese medicinal composition, filtering, evaporating the filtrate, and adding the second solvent to obtain a sample solution;

(2) testing by thin layer chromatography

Sucking the Ganoderma positive control solution, caulis Polygoni Multiflori positive control solution and sample solution, respectively dropping on the same silica gel thin layer plate, developing with cyclohexane-ethyl acetate-formic acid as developing agent, taking out, air drying, placing under ultraviolet lamp for inspection and/or spraying with sulfuric acid ethanol solution, and oven drying until spots appear clearly; and the combination of (a) and (b),

sucking the positive control solution and the sample solution of semen Ziziphi Spinosae, respectively dropping on the same silica gel thin layer plate, developing with water saturated n-butanol as developing agent, taking out, air drying, and inspecting under ultraviolet lamp.

2. The detection method according to claim 1, wherein the positive control of Ganoderma lucidum is selected from at least one of Ganoderma lucidum control medicinal material or Ganoderma lucidum extract; the positive control of caulis Polygoni Multiflori is selected from at least one of caulis Polygoni Multiflori control medicinal materials or caulis Polygoni Multiflori extract; the positive control of the spina date seed is at least one of a spina date seed control medicinal material or a spina date seed extract.

3. The detection method according to claim 1 or 2, wherein the first solvent is one or more of methanol, ethanol and propanol; the second solvent is one or more of methanol, ethanol and propanol.

4. The detection method according to any one of claims 1 to 3, wherein the thin layer chromatography comprises the steps of:

(1) preparation of reference medicinal material solution, extract solution, negative reference substance solution and test solution

Taking 1.5-3 g of lucid ganoderma contrast medicinal material, 0.2-0.3 g of tuber fleeceflower stem contrast medicinal material and 0.5-1.5 g of spina date seed contrast medicinal material, respectively adding 20-40 mL of ethanol, carrying out ultrasonic treatment for 20-40 minutes, filtering, evaporating filtrate to dryness, adding 0.5-1.5 mL of ethanol for dissolving, and respectively taking the filtrate as respective contrast medicinal material solutions;

taking 0.5-1.5 g of lucid ganoderma extract, 0.5-1.5 g of tuber fleeceflower stem extract and 0.5-1.5 g of spina date seed extract, respectively adding 20-40 mL of ethanol, carrying out ultrasonic treatment for 20-40 minutes, filtering, evaporating filtrate to dryness, adding 0.5-1.5 mL of ethanol for dissolving, and respectively taking the filtrate as respective extract solution;

taking 4.5-5.5 g of a negative control product without a lucid ganoderma extract, 4.5-5.5 g of a negative control product without a vine of multiflower knotweed extract and 4.5-5.5 g of a negative control product without a spina date seed extract, respectively adding 20-40 mL of ethanol, carrying out ultrasonic treatment for 20-40 minutes, filtering, evaporating filtrate to dryness, adding 0.5-1.5 mL of ethanol for dissolving, and respectively taking the ethanol solution as a negative control product solution;

taking 4.5-5.5 g of the traditional Chinese medicine composition, adding 20-40 mL of ethanol into the traditional Chinese medicine composition, carrying out ultrasonic treatment for 20-40 minutes, filtering, evaporating the filtrate to dryness, and adding 0.5-1.5 mL of ethanol for dissolving to obtain a sample solution;

(2) testing by thin layer chromatography

Sucking 4-6 muL of the lucid ganoderma contrast medicinal material solution, 4-6 muL of the multiflower knotweed vine contrast medicinal material solution, 4-6 muL of the lucid ganoderma extract solution, 4-6 muL of the multiflower knotweed vine extract solution, 4-6 muL of the negative contrast product solution without the lucid ganoderma extract, 4-6 muL of the negative contrast product solution without the multiflower knotweed vine extract and 4-6 muL of the test product solution, respectively dropping on the same silica gel thin layer plate, developing by taking cyclohexane-ethyl acetate-formic acid with the volume ratio of 5: 0.2 as a developing agent, taking out, airing, placing under an ultraviolet lamp of 366nm to inspect and/or spray sulfuric acid ethanol solution, and drying until spots are clear in color; and the combination of (a) and (b),

sucking the spina date seed reference medicinal material solution 4-6 muL, the spina date seed extract solution 4-6 muL, the negative reference substance solution 4-6 muL without the spina date seed extract and the sample solution 4-6 muL, respectively dropping on the same silica gel thin layer plate, developing by using water saturated n-butyl alcohol as a developing agent, taking out, airing, and inspecting under an ultraviolet lamp of 366 nm.

5. The detection method according to any one of claims 1 to 4, wherein the thin layer chromatography comprises the steps of:

(1) preparation of reference medicinal material solution, extract solution, negative reference substance solution and test solution

Taking 2g of lucid ganoderma reference medicinal material, 0.25g of tuber fleeceflower stem reference medicinal material and 1g of spina date seed reference medicinal material, respectively adding 30mL of ethanol, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating filtrate to dryness, and then adding 1mL of ethanol for dissolving, wherein the ethanol is respectively used as respective reference medicinal material solutions;

respectively adding 30mL of ethanol into 1g of ganoderma lucidum extract, 1g of caulis polygoni multiflori extract and 1g of spina date seed extract, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating filtrate to dryness, and adding 1mL of ethanol for dissolving to obtain respective extract solutions;

taking 5g of a negative control product without a lucid ganoderma extract, 5g of a negative control product without a vine of multiflower knotweed extract and 5g of a negative control product without a spina date seed extract, respectively adding 30mL of ethanol into the negative control product, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating the filtrate to dryness, adding 1mL of ethanol into the filtrate to dissolve the filtrate, and respectively taking the filtrate as respective negative control product solutions;

taking 5g of the traditional Chinese medicine composition, adding 30mL of ethanol, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating the filtrate to dryness, and adding 1mL of ethanol for dissolving to obtain a test solution;

(2) testing by thin layer chromatography

Sucking the above Ganoderma contrast medicinal material solution 5 μ L, caulis Polygoni Multiflori contrast medicinal material solution 5 μ L, Ganoderma extract solution 5 μ L, caulis Polygoni Multiflori extract solution 5 μ L, negative control solution 5 μ L without Ganoderma extract, negative control solution 5 μ L without caulis Polygoni Multiflori extract and test sample solution 5 μ L, respectively dropping on the same silica gel thin layer plate, developing with cyclohexane-ethyl acetate-formic acid at volume ratio of 5: 0.2 as developing agent, taking out, air drying, inspecting and/or spraying with sulfuric acid ethanol solution under 366nm ultraviolet lamp, and drying until spots appear clearly; and the combination of (a) and (b),

sucking the above semen Ziziphi Spinosae control medicinal material solution 5 μ L, semen Ziziphi Spinosae extract solution 5 μ L, negative control solution 5 μ L without semen Ziziphi Spinosae extract and sample solution 5 μ L, respectively dropping on the same silica gel thin layer plate, developing with water saturated n-butanol as developing agent, taking out, air drying, and inspecting under 366nm ultraviolet lamp.

6. The detection method according to any one of claims 1 to 5, further comprising measuring the amount of spinosin in the Chinese medicinal composition by high performance liquid chromatography, wherein the high performance liquid chromatography comprises the following steps:

(1) preparation of standard solution of spinosin and sample solution of traditional Chinese medicine composition

Taking a spinosyn reference substance, and adding a third solvent to prepare a spinosyn standard solution;

and (3) adding a third solvent into the traditional Chinese medicine composition, and filtering to obtain filtrate, namely the sample solution of the traditional Chinese medicine composition.

(2) Absorbing the standard solution of the spinosyns, injecting the solution into a high performance liquid chromatograph, and drawing a peak area-solution concentration standard curve chart according to a measurement result;

(3) and (3) sucking the sample solution of the traditional Chinese medicine composition, injecting the sample solution into a high performance liquid chromatograph, and calculating the content of the spinosad in the traditional Chinese medicine composition according to the peak area of the sample and a standard curve graph.

7. The detection method according to claim 6, wherein the third solvent is methanol.

8. The detection method according to claim 6 or 7, wherein the high performance liquid chromatography comprises the steps of:

(1) chromatographic condition and system adaptability test

Octadecylsilane chemically bonded silica is used as a filler, and the particle size of the filler is not more than 2.5 microns; taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the column temperature is 25-35 ℃; the detection wavelength is 330-340 nm; the number of theoretical plates is not less than 5000 calculated according to the spinosad; the gradient elution process is as follows:

0-10 min, mobile phase A%: 12 → 19, mobile phase B: 88 → 81;

10-12 min, mobile phase A%: 19 → 100, mobile phase B: 81 → 0;

12-15 min, mobile phase A%: 100, mobile phase B: 0;

15-16 min, mobile phase A%: 100 → 12, mobile phase B: 0 → 88;

(2) preparation of standard solution of spinosad and sample solution of traditional Chinese medicine composition

Taking a spinosyn reference substance, adding methanol and diluting into at least 3 spinosyn standard solutions with different concentrations;

taking 1g of the traditional Chinese medicine composition, adding 25mL of methanol, carrying out ultrasonic treatment for 30 minutes, and filtering by using a 0.22-micrometer organic filter membrane to obtain a filtrate, namely a sample solution of the traditional Chinese medicine composition;

(3) sucking 1 mu L of the standard solution of the spinosad, injecting the solution into a high performance liquid chromatograph, and drawing a peak area-solution concentration standard curve chart according to a measurement result;

(4) and (3) sucking 1 mu L of the sample solution of the traditional Chinese medicine composition, injecting the sample solution into a high performance liquid chromatograph, and calculating the content of the spinosad in the traditional Chinese medicine composition according to the peak area of the sample and a standard curve graph.

Technical Field

The invention belongs to the technical field of traditional Chinese medicine detection, and particularly relates to a detection method of a traditional Chinese medicine composition with a sleep improving function.

Background

Adequate sleep, balanced diet and proper exercise are three health standards recognized by international society. During a person's lifetime, one third of the time spent in sleep is an indispensable component of health as a necessary process for life. Persistent sleep disorders can lead to physical and mental damage, socioeconomic losses, and other adverse consequences, including cardiovascular disease, mental illness, poor immunity, and even increased mortality, have become serious public health problems.

Therefore, chinese patent document CN102188610A discloses a traditional Chinese medicine composition with sleep improving function, which comprises extracts of ganoderma lucidum, arillus longan, semen ziziphi spinosae, semen boitae, caulis polygoni multiflori, rhizoma polygonati, fructus ziziphi jujubae, etc., but only introduces the raw material composition and the preparation method of the traditional Chinese medicine composition, does not provide a corresponding product detection method, and is not beneficial to the control of the internal quality of the composition in the production, circulation and use processes. In the prior art, many detection and analysis methods for ganoderma lucidum and vine of multiflower knotweed are available, for example, the study of 'allergic health particle quality control method' in the publication of China journal literature discloses a thin-layer chromatography identification method for vine of multiflower knotweed and ganoderma lucidum, and the method has the disadvantages that the pretreatment modes of vine of multiflower knotweed and ganoderma lucidum are different, the finally selected developing agent system is quite different, the developing agent used in the former is a benzene-ethanol system, and the developing agent used in the latter is a petroleum ether (60-90 ℃) -ethyl formate-formic acid system, so that the operation is quite inconvenient, and particularly when large-scale detection is needed, the workload is quite large, and the working efficiency is low.

Disclosure of Invention

Therefore, the invention aims to provide a detection method of the traditional Chinese medicine composition with the function of improving sleep.

Therefore, the invention provides a detection method of a traditional Chinese medicine composition, which comprises the step of identifying lucid ganoderma, tuber fleeceflower stem and spina date seed in the traditional Chinese medicine composition by using a thin-layer chromatography, wherein the thin-layer chromatography comprises the following steps:

(1) preparation of positive control solution and test solution

Respectively adding a first solvent into a ganoderma lucidum positive control, a polygonum multiflorum positive control and a spina date seed positive control, filtering, evaporating filtrate to dryness, and adding a second solvent to respectively serve as respective positive control solutions;

adding the first solvent into the Chinese medicinal composition, filtering, evaporating the filtrate, and adding the second solvent to obtain a sample solution;

(2) testing by thin layer chromatography

Sucking the Ganoderma positive control solution, caulis Polygoni Multiflori positive control solution and sample solution, respectively dropping on the same silica gel thin layer plate, developing with cyclohexane-ethyl acetate-formic acid as developing agent, taking out, air drying, placing under ultraviolet lamp for inspection and/or spraying with sulfuric acid ethanol solution, and oven drying until spots appear clearly; and the combination of (a) and (b),

sucking the positive control solution and the sample solution of semen Ziziphi Spinosae, respectively dropping on the same silica gel thin layer plate, developing with water saturated n-butanol as developing agent, taking out, air drying, and inspecting under ultraviolet lamp.

Further, the positive reference substance of the lucid ganoderma is selected from at least one of lucid ganoderma reference medicinal materials or lucid ganoderma extracts; the positive control of caulis Polygoni Multiflori is selected from at least one of caulis Polygoni Multiflori control medicinal materials or caulis Polygoni Multiflori extract; the positive control of the spina date seed is at least one of a spina date seed control medicinal material or a spina date seed extract.

Further, the first solvent is one or more of methanol, ethanol and propanol; the second solvent is one or more of methanol, ethanol and propanol.

Further, the thin layer chromatography comprises the following steps:

(1) preparation of reference medicinal material solution, extract solution, negative reference substance solution and test solution

Taking 1.5-3 g of lucid ganoderma contrast medicinal material, 0.2-0.3 g of tuber fleeceflower stem contrast medicinal material and 0.5-1.5 g of spina date seed contrast medicinal material, respectively adding 20-40 mL of ethanol, carrying out ultrasonic treatment for 20-40 minutes, filtering, evaporating filtrate to dryness, adding 0.5-1.5 mL of ethanol for dissolving, and respectively taking the filtrate as respective contrast medicinal material solutions;

taking 0.5-1.5 g of lucid ganoderma extract, 0.5-1.5 g of tuber fleeceflower stem extract and 0.5-1.5 g of spina date seed extract, respectively adding 20-40 mL of ethanol, carrying out ultrasonic treatment for 20-40 minutes, filtering, evaporating filtrate to dryness, adding 0.5-1.5 mL of ethanol for dissolving, and respectively taking the filtrate as respective extract solution;

taking 4.5-5.5 g of a negative control product without a lucid ganoderma extract, 4.5-5.5 g of a negative control product without a vine of multiflower knotweed extract and 4.5-5.5 g of a negative control product without a spina date seed extract, respectively adding 20-40 mL of ethanol, carrying out ultrasonic treatment for 20-40 minutes, filtering, evaporating filtrate to dryness, adding 0.5-1.5 mL of ethanol for dissolving, and respectively taking the ethanol solution as a negative control product solution;

taking 4.5-5.5 g of the traditional Chinese medicine composition, adding 20-40 mL of ethanol into the traditional Chinese medicine composition, carrying out ultrasonic treatment for 20-40 minutes, filtering, evaporating the filtrate to dryness, and adding 0.5-1.5 mL of ethanol for dissolving to obtain a sample solution;

(2) testing by thin layer chromatography

Sucking 4-6 muL of the lucid ganoderma contrast medicinal material solution, 4-6 muL of the multiflower knotweed vine contrast medicinal material solution, 4-6 muL of the lucid ganoderma extract solution, 4-6 muL of the multiflower knotweed vine extract solution, 4-6 muL of the negative contrast product solution without the lucid ganoderma extract, 4-6 muL of the negative contrast product solution without the multiflower knotweed vine extract and 4-6 muL of the test product solution, respectively dropping on the same silica gel thin layer plate, developing by taking cyclohexane-ethyl acetate-formic acid with the volume ratio of 5: 0.2 as a developing agent, taking out, airing, placing under an ultraviolet lamp of 366nm to inspect and/or spray sulfuric acid ethanol solution, and drying until spots are clear in color; and the combination of (a) and (b),

sucking the spina date seed reference medicinal material solution 4-6 muL, the spina date seed extract solution 4-6 muL, the negative reference substance solution 4-6 muL without the spina date seed extract and the sample solution 4-6 muL, respectively dropping on the same silica gel thin layer plate, developing by using water saturated n-butyl alcohol as a developing agent, taking out, airing, and inspecting under an ultraviolet lamp of 366 nm.

Further, the thin layer chromatography comprises the following steps:

(1) preparation of reference medicinal material solution, extract solution, negative reference substance solution and test solution

Taking 2g of lucid ganoderma reference medicinal material, 0.25g of tuber fleeceflower stem reference medicinal material and 1g of spina date seed reference medicinal material, respectively adding 30mL of ethanol, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating filtrate to dryness, and then adding 1mL of ethanol for dissolving, wherein the ethanol is respectively used as respective reference medicinal material solutions;

respectively adding 30mL of ethanol into 1g of ganoderma lucidum extract, 1g of caulis polygoni multiflori extract and 1g of spina date seed extract, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating filtrate to dryness, and adding 1mL of ethanol for dissolving to obtain respective extract solutions;

taking 5g of a negative control product without a lucid ganoderma extract, 5g of a negative control product without a vine of multiflower knotweed extract and 5g of a negative control product without a spina date seed extract, respectively adding 30mL of ethanol into the negative control product, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating the filtrate to dryness, adding 1mL of ethanol into the filtrate to dissolve the filtrate, and respectively taking the filtrate as respective negative control product solutions;

taking 5g of the traditional Chinese medicine composition, adding 30mL of ethanol, carrying out ultrasonic treatment for 30 minutes, filtering, evaporating the filtrate to dryness, and adding 1mL of ethanol for dissolving to obtain a test solution;

(2) testing by thin layer chromatography

Sucking the above Ganoderma contrast medicinal material solution 5 μ L, caulis Polygoni Multiflori contrast medicinal material solution 5 μ L, Ganoderma extract solution 5 μ L, caulis Polygoni Multiflori extract solution 5 μ L, negative control solution 5 μ L without Ganoderma extract, negative control solution 5 μ L without caulis Polygoni Multiflori extract and test sample solution 5 μ L, respectively dropping on the same silica gel thin layer plate, developing with cyclohexane-ethyl acetate-formic acid at volume ratio of 5: 0.2 as developing agent, taking out, air drying, inspecting and/or spraying with sulfuric acid ethanol solution under 366nm ultraviolet lamp, and drying until spots appear clearly; and the combination of (a) and (b),

sucking the above semen Ziziphi Spinosae control medicinal material solution 5 μ L, semen Ziziphi Spinosae extract solution 5 μ L, negative control solution 5 μ L without semen Ziziphi Spinosae extract and sample solution 5 μ L, respectively dropping on the same silica gel thin layer plate, developing with water saturated n-butanol as developing agent, taking out, air drying, and inspecting under 366nm ultraviolet lamp.

Further, the detection method also comprises the step of measuring the content of the spinosad in the traditional Chinese medicine composition by using a high performance liquid chromatography, wherein the high performance liquid chromatography comprises the following steps:

(1) preparation of standard solution of spinosin and sample solution of traditional Chinese medicine composition

Taking a spinosyn reference substance, and adding a third solvent to prepare a spinosyn standard solution;

and (3) adding a third solvent into the traditional Chinese medicine composition, and filtering to obtain filtrate, namely the sample solution of the traditional Chinese medicine composition.

(2) Absorbing the standard solution of the spinosyns, injecting the solution into a high performance liquid chromatograph, and drawing a peak area-solution concentration standard curve chart according to a measurement result;

(3) and (3) sucking the sample solution of the traditional Chinese medicine composition, injecting the sample solution into a high performance liquid chromatograph, and calculating the content of the spinosad in the traditional Chinese medicine composition according to the peak area of the sample and a standard curve graph.

Further, the third solvent is methanol.

Further, the high performance liquid chromatography comprises the following steps:

(1) chromatographic condition and system adaptability test

Octadecylsilane chemically bonded silica is used as a filler, and the particle size of the filler is not more than 2.5 microns; taking acetonitrile as a mobile phase A and water as a mobile phase B for gradient elution; the column temperature is 25-35 ℃; the detection wavelength is 330-340 nm; the number of theoretical plates is not less than 5000 calculated according to the spinosad; the gradient elution process is as follows:

0-10 min, mobile phase A%: 12 → 19, mobile phase B: 88 → 81;

10-12 min, mobile phase A%: 19 → 100, mobile phase B: 81 → 0;

12-15 min, mobile phase A%: 100, mobile phase B: 0;

15-16 min, mobile phase A%: 100 → 12, mobile phase B: 0 → 88;

(2) preparation of standard solution of spinosad and sample solution of traditional Chinese medicine composition

Taking a spinosyn reference substance, adding methanol and diluting into at least 3 spinosyn standard solutions with different concentrations;

taking 1g of the traditional Chinese medicine composition, adding 25mL of methanol, carrying out ultrasonic treatment for 30 minutes, and filtering by using a 0.22-micrometer organic filter membrane to obtain a filtrate, namely a sample solution of the traditional Chinese medicine composition;

(3) sucking 1 mu L of the standard solution of the spinosad, injecting the solution into a high performance liquid chromatograph, and drawing a peak area-solution concentration standard curve chart according to a measurement result;

(4) and (3) sucking 1 mu L of the sample solution of the traditional Chinese medicine composition, injecting the sample solution into a high performance liquid chromatograph, and calculating the content of the spinosad in the traditional Chinese medicine composition according to the peak area of the sample and a standard curve graph.

In the invention, the traditional Chinese medicine composition has the function of improving sleep, and the raw materials of the traditional Chinese medicine composition comprise a lucid ganoderma extract, a spina date seed extract and a caulis polygoni multiflori extract.

The technical scheme of the invention has the following advantages:

1. the method comprises the steps of identifying the lucid ganoderma, the vine of multiflower knotweed and the spina date seed in the traditional Chinese medicine composition by using a thin-layer chromatography, and determining the content of the spinosad in the traditional Chinese medicine composition by using a high performance liquid chromatography. Meanwhile, the method simplifies the operation process, reduces the time required for identification, improves the working efficiency, reduces the detection cost, provides a new idea for the thin-layer chromatography identification of each component in the traditional Chinese medicine composition, and is beneficial to the improvement of the quality control standard of the traditional Chinese medicine.

2. When the method is used for identifying the spina date seed extract by using the thin-layer chromatography, the pretreatment conditions are the same as those of lucid ganoderma and tuber fleeceflower stem, so that the operation process is further simplified, and the working efficiency is improved.

3. The detection method of the traditional Chinese medicine composition provided by the invention has the characteristics of good stability, strong specificity, high accuracy, good repeatability and the like, and can effectively control the product quality of the traditional Chinese medicine composition in production, circulation, use and other links.

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 chromatogram for simultaneous identification of Ganoderma lucidum and caulis Polygoni Multiflori in example 1 of the present invention;

FIG. 2 is a thin layer chromatogram for identification of wild jujube seeds in example 2 of the present invention;

FIG. 3 is a chromatogram for determining the content of spinosad in the Chinese medicinal composition by high performance liquid chromatography;

FIG. 4 is a special investigation chromatogram for determining the content of spinosad in the Chinese medicinal composition by high performance liquid chromatography;

FIG. 5 is a linear relationship investigation diagram for measuring the content of spinosad in the Chinese medicinal composition by high performance liquid chromatography.

Description of reference numerals:

1-negative control containing no caulis Polygoni Multiflori extract; 2-caulis polygoni multiflori extract; 3-caulis polygoni multiflori contrast medicinal materials; 4-negative control containing no Ganoderma extract; 5-ganoderma lucidum extract; 6-ganoderma lucidum reference medicinal material; 7-9 test articles (20170204-1, 20170204-2, 20170204-3); 10-negative control without spina date seeds; 11-wild jujube seed extract; 12-semen Ziziphi Spinosae control drug.

Detailed Description

The technical solutions of the present invention will be described clearly and completely below, and it should be apparent that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention. In addition, the technical features involved in the different embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

In the following examples, the traditional Chinese medicine composition for improving sleep function is provided by perfection (china) ltd; the negative control containing no caulis Polygoni Multiflori extract, the negative control containing no Ganoderma extract, and the negative control containing no semen Ziziphi Spinosae extract are provided by Perfect (Chinese) Limited company; ganoderma extract, caulis Polygoni Multiflori extract and semen Ziziphi Spinosae extract are provided by Guangdong pharmaceutical limited; the ganoderma lucidum reference medicinal material, the caulis polygoni multiflori reference medicinal material, the spina date seed reference medicinal material and the spinosin reference substance are purchased from China food and drug testing research institute.

In the following examples, methanol, ethanol and propanol were analytically pure, with a volume concentration of 95% or more; acetonitrile is chromatographic grade, and the volume concentration is more than or equal to 99.9 percent.