阅读说明：本技术 一种中药制剂的检测方法 (Detection method of traditional Chinese medicine preparation ) 是由 徐向平 夏新华 黄胜 朱丽 谢安 欧金秀 叶惠煊 于 2018-07-27 设计创作，主要内容包括：本发明涉及一种中药制剂的检测方法,所述制剂由制何首乌、金樱子肉、桑椹桑椹、女贞子、豨莶草、川牛膝和菟丝子等七种药味组成,具有补肝肾、益精髓的功效,所述检测方法包括以下步骤：对制剂中的制何首乌、桑椹桑椹、川牛膝和女贞子进行鉴别；测定制剂中制何首乌中二苯乙烯苷的含量。(The invention relates to a detection method of a traditional Chinese medicine preparation, the preparation consists of seven medicinal herbs of prepared fleece-flower root, cherokee rose fruit, mulberry, glossy privet fruit, siegesbeckia orientalis, medicinal cyathula root, dodder seed and the like, and has the effects of nourishing liver and kidney and benefiting essence, and the detection method comprises the following steps: identifying radix Polygoni Multiflori Preparata, Mori fructus, radix Cyathulae and fructus Ligustri Lucidi in the preparation; measuring the content of stilbene glucoside in radix Polygoni Multiflori Preparata.)

1. A detection method of a traditional Chinese medicine preparation is characterized in that the preparation is prepared from seven traditional Chinese medicines including radix polygoni multiflori preparata, cherokee rose fruit, mulberry, glossy privet fruit, herba siegesbeckiae, radix cyathulae and semen cuscutae, and the detection method comprises the following steps:

(1) identifying radix Polygoni Multiflori Preparata, Mori fructus, radix Cyathulae and fructus Ligustri Lucidi in the preparation by TLC;

(2) measuring the content of stilbene glucoside in radix Polygoni Multiflori Preparata by HPLC.

2. The assay of claim 1, wherein the identification of the prepared fleece flower root in the formulation comprises the steps of:

collecting 40ml of the preparation, placing in a separating funnel, adding dichloromethane, shaking and extracting for 3 times, adding dichloromethane 40ml for the first time, adding dichloromethane 30ml for the second time, adding dichloromethane 30ml for the third time, collecting combined dichloromethane solution, evaporating on water bath, and adding methanol 1ml into residue to dissolve to obtain sample solution; preparing negative control solution by the same method except that 40ml of negative sample lacking radix Polygoni Multiflori Preparata is prepared; taking 0.2g of radix Polygoni Multiflori reference material, adding dichloromethane 20ml, performing ultrasonic treatment for 30min, filtering, evaporating filtrate to dryness, and dissolving residue with methanol 1ml to obtain reference material solution; according to the test of thin-layer chromatography of 0502 of the four ministry of pharmacopoeia 2015 year edition, sucking 15 mul of the three solutions respectively, respectively dropping on the same silica gel G thin-layer plate, developing by using a toluene-ethyl acetate-formic acid mixed solution with a volume ratio of 9:1:0.1 as a developing agent, taking out, drying in the air, and placing under a 365nm ultraviolet lamp for inspection, wherein fluorescent spots with the same color appear on the chromatogram of the test sample at positions corresponding to the chromatogram of the reference medicinal material.

3. The assay of claim 1, wherein the identification of Cyathula officinalis root in the formulation comprises the steps of:

collecting 40ml of the preparation, placing in a separating funnel, adding dichloromethane, shaking and extracting for 3 times, adding dichloromethane 40ml for the first time, adding dichloromethane 30ml for the second time, adding dichloromethane 30ml for the third time, collecting combined dichloromethane solution, evaporating on water bath, and adding methanol 1ml into residue to dissolve to obtain sample solution; preparing negative control solution by the same method with negative sample solution 40ml lacking radix Cyathulae; taking 1g of radix cyathulae control medicinal material, adding 50ml of water, decocting for 30 minutes, filtering, concentrating the filtrate to about 20ml, placing the filtrate in a separating funnel, adding 30ml of dichloromethane, shaking for extraction, separating dichloromethane solution, placing the dichloromethane solution on a water bath for drying by distillation, and adding 1ml of methanol into residues for dissolving to obtain a control medicinal material solution; according to the test of thin-layer chromatography of 0502 of the four ministerial rules of pharmacopoeia 2015 year edition, 20 mul of the control medicinal material solution and 10 mul of the test sample solution and the radix cyathulae-lacking negative control solution are respectively spotted on the same silica gel G thin-layer plate, a petroleum ether-ethyl acetate mixed solution with a boiling range of 60-90 ℃ in a volume ratio of 1:1 is taken as a developing solvent, the solution is saturated for 30min, the solution is developed, taken out, dried in the air and placed under a 365nm ultraviolet lamp for inspection, and as a result, fluorescent spots with the same color are displayed on the positions corresponding to the control medicinal material chromatogram in the test sample chromatogram.

4. The assay method according to claim 1, wherein the identification of morous alba in said preparation comprises the steps of:

collecting 40ml of the preparation, placing in a separating funnel, adding dichloromethane, shaking and extracting for 3 times, adding dichloromethane 40ml for the first time, adding dichloromethane 30ml for the second time, adding dichloromethane 30ml for the third time, collecting combined dichloromethane solution, evaporating on water bath, and adding methanol 1ml into residue to dissolve to obtain sample solution; preparing negative control solution by the same method with negative sample solution 40ml lacking Mori fructus; decocting fructus Mori 2g with water 60ml for 30min, filtering, concentrating the filtrate to about 20ml, placing in separating funnel, adding dichloromethane 30ml, shaking for extraction, collecting dichloromethane solution, evaporating on water bath, dissolving the residue with methanol 1ml to obtain control solution; according to the experiment of thin-layer chromatography of 0502 of the four ministerial rules of pharmacopoeia 2015 year edition, 10 mul of each of the test solution and the mulberry-lacking negative control solution and 20 mul of the control solution are respectively spotted on the same silica gel G thin-layer plate, a toluene-ethyl acetate-formic acid mixed solution with the volume ratio of 5:4:1 is taken as a developing agent, the developing agent is developed, taken out, dried in the air and placed under a 365nm ultraviolet lamp for inspection, and spots with the same color are displayed on the positions corresponding to the control solution chromatogram in the result of the test solution chromatogram.

5. The assay of claim 1, wherein the identification of fructus Ligustri Lucidi in the formulation comprises the steps of:

extracting 20ml of the preparation with diethyl ether under shaking for 2 times (20 ml each time), mixing diethyl ether solutions, volatilizing at low temperature, and dissolving the residue with 1ml of anhydrous ethanol to obtain test solution; preparing negative control solution from 20ml of negative sample solution lacking fructus Ligustri Lucidi by the same method; taking 0.5g fructus Ligustri Lucidi as reference material, adding 20ml 50% ethanol, performing ultrasonic treatment for 30min, filtering, and processing the filtrate according to the preparation method of the test solution to obtain reference material solution; according to the test of thin-layer chromatography of 0502 of the general rules of the four parts of the national pharmacopoeia 2015 edition, sucking the reference medicinal material solution, respectively dropping 10 mul of the test solution and the negative reference solution on the same silica gel G thin-layer plate, taking cyclohexane-acetone mixed solution with the volume ratio of 20:11 as a developing agent, saturating for 30min, developing, taking out, airing, spraying 10% sulfuric acid ethanol solution, blowing hot air until the spots are clearly developed, and displaying the spots with the same color in the test chromatogram at the position corresponding to the reference chromatogram.

6. The detection method according to claim 1, wherein the content of stilbene glucoside in radix Polygoni Multiflori Preparata in the preparation is determined by high performance liquid chromatography according to 0512 of the four-part general rules of China pharmacopoeia 2015, and the steps are as follows:

(1) chromatographic conditions are as follows: the chromatographic column is a PhenomenexC18 column (250mm multiplied by 4.6mm,5 μm); the detection wavelength is 310-330 nm; (ii) a The mobile phase A is acetonitrile, the mobile phase B is water, the analysis time is 30 minutes, isocratic elution is carried out, and the volume ratio of the mobile phase A to the mobile phase B is 20: 80; the flow rate is 0.8-1.2 mL/min^-1(ii) a The column temperature is 25-40 ℃; the sample amount is 10 mul; the theoretical plate number is not less than 2000;

(2) preparing a reference solution, namely precisely weighing 13mg of a stilbene glucoside (2,3,5, 4' -tetrahydroxystilbene-2-O- β -D-glucoside) reference, placing the reference in a 50mL volumetric flask, adding diluted ethanol to dissolve the reference and fixing the volume to the scale, shaking up to obtain a reference solution mother solution, precisely weighing 1mL of the reference solution mother solution, placing the reference solution mother solution in a 10mL volumetric flask, adding diluted ethanol to dilute the reference solution and fixing the volume to the scale, and shaking up to obtain a reference solution with the concentration of 0.0260 mg/mL;

(3) preparation of test sample and negative control solution: precisely measuring 3.0ml of preparation sample, placing in a 100ml measuring flask, adding appropriate amount of diluted ethanol, shaking for dissolving, adding diluted ethanol to scale, shaking, standing, filtering with 0.45 μm microporous membrane, and collecting filtrate as sample solution; taking negative samples lacking the prepared fleece flower root, and preparing the negative control solution lacking the prepared fleece flower root by the same method as the test solution.

7. The detection method according to claim 6, wherein the detection wavelength in the step (1) is 320 nm; flow rate 1.0 mL/min^-1(ii) a The column temperature was 30 ℃.

Technical Field

The invention relates to a detection method of a traditional Chinese medicine preparation, and belongs to the technical field of traditional Chinese medicines.

Background

Yilingjing is collected in the sixth volume of the pharmaceutical Standard Chinese medicinal prescription preparation of the Ministry of public health of the people's republic of China, and is a unique production variety of Jiuzhitang GmbH. The preparation is prepared from seven medicinal herbs of prepared fleece-flower root, cherokee rose fruit, mulberry, glossy privet fruit (steamed with wine), siegesbeckia orientalis (steamed with honey wine), medicinal cyathula root (steamed with wine) and dodder seed (steamed with wine), has the effects of nourishing liver and kidney and benefiting essence and marrow, and is clinically used for treating dizziness, blurred vision, tinnitus, palpitation, hypodynamia, dry throat, insomnia and the like caused by liver and kidney deficiency.

In the existing quality standard, only thin-layer identification methods of prepared fleece flower root and mulberry are adopted, and no content determination item exists. The expanded system in the radix polygoni multiflori preparata identification item contains benzene with high toxicity, and has great influence on the health of inspectors; the water adding amount of the mulberry contrast medicinal material in the mulberry identification item during pretreatment is not clear, and in the actual inspection work, different inspectors have different understandings, so that the result repeatability is easy to cause; the polygonum multiflorum which is a monarch drug in the formula has the effects of tonifying liver and kidney, benefiting essence and blood, blackening beard and hair, strengthening bones and muscles, eliminating turbid pathogen and reducing blood fat, and the stilbene glucoside is a main active ingredient of the polygonum multiflorum, has the effects of resisting oxidation and tumors, regulating blood fat, protecting nerves and the like, is used as an index ingredient for content measurement, and can objectively evaluate the quality of the life-prolonging essence.

Disclosure of Invention

The invention aims to provide a detection method of Yilingjing of a traditional Chinese medicine preparation, which can be used for simply, effectively, qualitatively and quantitatively detecting the active ingredients of the traditional Chinese medicine preparation, ensuring the effective control of the medicine quality,

in order to achieve the purpose, the invention adopts the following technical scheme:

a detection method of a traditional Chinese medicine preparation Yilingjing is prepared from seven traditional Chinese medicines including radix polygoni multiflori preparata, cherokee rose fruit, mulberry, glossy privet fruit, herba siegesbeckiae, radix cyathulae, semen cuscutae and the like, and is characterized by comprising the following steps:

(1) identifying radix Polygoni Multiflori Preparata, Mori fructus, radix Cyathulae and fructus Ligustri Lucidi in the preparation by TLC;

(2) measuring the content of stilbene glucoside in radix Polygoni Multiflori Preparata by HPLC.

The identification of the prepared fleece-flower root in the preparation comprises the following steps: collecting 40ml of the preparation, placing in a separating funnel, adding dichloromethane, shaking and extracting for 3 times, adding dichloromethane 40ml for the first time, adding dichloromethane 30ml for the second time, adding dichloromethane 30ml for the third time, collecting combined dichloromethane solution, evaporating on water bath, and adding methanol 1ml into residue to dissolve to obtain sample solution; preparing negative control solution by the same method except that 40ml of negative sample lacking radix Polygoni Multiflori Preparata is prepared; taking 0.2g of radix Polygoni Multiflori reference material, adding dichloromethane 20ml, performing ultrasonic treatment for 30min, filtering, evaporating filtrate to dryness, and dissolving residue with methanol 1ml to obtain reference material solution; according to the test of thin-layer chromatography of 0502 of the four ministry of pharmacopoeia 2015 year edition, sucking 15 mul of the three solutions respectively, respectively dropping on the same silica gel G thin-layer plate, developing by using a toluene-ethyl acetate-formic acid mixed solution with a volume ratio of 9:1:0.1 as a developing agent, taking out, drying in the air, and placing under a 365nm ultraviolet lamp for inspection, wherein fluorescent spots with the same color appear on the chromatogram of the test sample at positions corresponding to the chromatogram of the reference medicinal material.

The identification of the radix cyathulae in the preparation comprises the following steps: collecting 40ml of the preparation, placing in a separating funnel, adding dichloromethane, shaking and extracting for 3 times, adding dichloromethane 40ml for the first time, adding dichloromethane 30ml for the second time, adding dichloromethane 30ml for the third time, collecting combined dichloromethane solution, evaporating on water bath, and adding methanol 1ml into residue to dissolve to obtain sample solution; preparing negative control solution by the same method with negative sample solution 40ml lacking radix Cyathulae; taking 1G of radix cyathulae control medicinal material, adding 50ml of water, decocting for 30 minutes, filtering, concentrating the filtrate to about 20ml, placing the mixture in a separating funnel, adding 30ml of dichloromethane, shaking for extraction, separating dichloromethane solution, placing the dichloromethane solution on a water bath for drying, adding 1ml of methanol into residues for dissolving, using the residues as the control medicinal material solution, performing a thin-layer chromatography test according to 0502 general rules of the four parts of pharmacopoeia 2015 edition of China, sucking 20 mu l of the control medicinal material solution, respectively placing 10 mu l of a test sample solution and 10 mu l of a radix cyathulae-lacking negative control solution on the same silica gel G thin-layer plate, using a petroleum ether-ethyl acetate mixed solution with a boiling range of 60-90 ℃ in a volume ratio of 1:1 as a developing agent, saturating for 30min, developing, taking out, drying, placing the test sample solution under an ultraviolet lamp with 365nm, and displaying fluorescent spots with the same color in the position of the test sample chromatogram.

The identification of morous alba in the preparation comprises the following steps: collecting 40ml of the preparation, placing in a separating funnel, adding dichloromethane, shaking and extracting for 3 times, adding dichloromethane 40ml for the first time, adding dichloromethane 30ml for the second time, adding dichloromethane 30ml for the third time, collecting combined dichloromethane solution, evaporating on water bath, and adding methanol 1ml into residue to dissolve to obtain sample solution; preparing negative control solution by the same method with negative sample solution 40ml lacking Mori fructus; decocting fructus Mori 2g with water 60ml for 30min, filtering, concentrating the filtrate to about 20ml, placing in separating funnel, adding dichloromethane 30ml, shaking for extraction, collecting dichloromethane solution, evaporating on water bath, dissolving the residue with methanol 1ml to obtain control solution; according to the experiment of thin-layer chromatography of 0502 of the four ministerial rules of pharmacopoeia 2015 year edition, 10 mul of each of the test solution and the mulberry-lacking negative control solution and 20 mul of the control solution are respectively spotted on the same silica gel G thin-layer plate, a toluene-ethyl acetate-formic acid mixed solution with the volume ratio of 5:4:1 is taken as a developing agent, the developing agent is developed, taken out, dried in the air and placed under a 365nm ultraviolet lamp for inspection, and spots with the same color are displayed on the positions corresponding to the control solution chromatogram in the result of the test solution chromatogram.

The identification of the glossy privet fruit in the preparation comprises the following steps: extracting 20ml of the preparation with diethyl ether under shaking for 2 times (20 ml each time), mixing diethyl ether solutions, volatilizing at low temperature, and dissolving the residue with 1ml of anhydrous ethanol to obtain test solution; preparing negative control solution from 20ml of negative sample solution lacking fructus Ligustri Lucidi by the same method; taking 0.5g fructus Ligustri Lucidi as reference material, adding 20ml 50% ethanol, performing ultrasonic treatment for 30min, filtering, and processing the filtrate according to the preparation method of the test solution to obtain reference material solution; according to the test of thin-layer chromatography of 0502 of the general rules of the four parts of the national pharmacopoeia 2015 edition, sucking the reference medicinal material solution, respectively dropping 10 mul of the test solution and the negative reference solution on the same silica gel G thin-layer plate, taking cyclohexane-acetone mixed solution with the volume ratio of 20:11 as a developing agent, saturating for 30min, developing, taking out, airing, spraying 10% sulfuric acid ethanol solution, blowing hot air until the spots are clearly developed, and displaying the spots with the same color in the test chromatogram at the position corresponding to the reference chromatogram.

The content of stilbene glucoside in the prepared fleece-flower root in the preparation is tested by high performance liquid chromatography according to the four-part general rule 0512 in 2015 edition of Chinese pharmacopoeia, and the steps are as follows:

(1) chromatographic conditions are as follows: color(s)The spectrum column is a PhenomenexC18 column (250mm multiplied by 4.6mm,5 μm); the detection wavelength is 310-330 nm; the mobile phase A is acetonitrile, the mobile phase B is water, the analysis time is 30 minutes, isocratic elution is carried out, and the volume ratio of the mobile phase A to the mobile phase B is 20: 80; the flow rate is 0.8-1.2 mL/min^-1(ii) a The column temperature is 25-40 ℃; the sample amount is 10 mul; the theoretical plate number is not less than 2000;

(2) precisely weighing 13mg of stilbene glucoside (2,3,5, 4' -tetrahydroxystilbene-2-O- β -D-glucoside) reference substance, placing in a 50mL volumetric flask, adding diluted ethanol for dissolving, fixing the volume to scale, shaking up to obtain reference substance solution mother liquor, precisely weighing 1mL of reference substance solution mother liquor, placing in a 10mL volumetric flask, adding diluted ethanol for diluting, fixing the volume to scale, and shaking up to obtain reference substance solution with the concentration of 0.0260 mg/mL;

(3) preparation of test sample and negative control solution: precisely measuring 3.0ml of the preparation sample, placing in a 100ml measuring flask, adding appropriate amount of diluted ethanol, shaking to dissolve, adding diluted ethanol to scale, shaking, standing, filtering with 0.45 μm microporous membrane, and collecting the filtrate as sample solution. Taking negative samples lacking the prepared fleece flower root, and preparing the negative control solution lacking the prepared fleece flower root by the same method as the test solution.

Further, the content of stilbene glucoside in the prepared fleece-flower root in the preparation is measured by high performance liquid chromatography according to the four-part general regulation 0512 of the national pharmacopoeia 2015 edition, and the chromatographic conditions in the step (1) are preferably as follows: the chromatographic column is a PhenomenexC18 column (250mm multiplied by 4.6mm,5 μm); the detection wavelength is 320 nm; the mobile phase A is acetonitrile, the mobile phase B is water, the analysis time is 30 minutes, isocratic elution is carried out, the volume ratio of the mobile phase A to the mobile phase B is 20:80, and the flow rate is 1.0 mL/min^-1(ii) a The column temperature is 30 ℃; the sample amount is 10 mul; the theoretical plate number is not less than 2000.

For a better understanding of the invention, the following further illustrates the advantageous effects thereof by means of test examples, which are intended to illustrate the invention without limiting it.

The inventor carries out intensive research on the method for measuring the content of stilbene glucoside in the Yilingjing of the traditional Chinese medicine preparation, and details are as follows:

1.1 instruments and materials

shimadzu high performance liquid chromatograph (shimadzu corporation, shimadzu, japan); PhenomenexC18 column (4.6X 250mm,5 μm); SK3300H ultrasonic cleaner (shanghai koku ultrasonic instruments ltd); DK-S22 model electric heating constant temperature water bath (Shanghai Jing Macro experimental facilities, Co., Ltd.).

Yilingjing 3 batches, pilot products produced by Jiuzhitangtang limited company with the batch numbers of 20090821, 20090901 and 20090903, fleece-flower root reference medicinal material (the batch number of 120934-201009), radix cyathulae reference medicinal material (the batch number of 121065-200503), mulberry reference medicinal material (the batch number of 121158-201103), glossy privet fruit reference medicinal material (the batch number of 121041-200604) and 2,3,5, 4' -tetrahydroxystilbene-2-O- β -D-glucoside (the batch number of 110844-200607 for content determination) are purchased from Chinese medicine biological product assay places, methanol is a chromatographic pure reagent, water is redistilled water, and other reagents are analytical pure.

1.2 methods

1.2.1 chromatographic conditions: the chromatographic column is a PhenomenexC18 column (250mm multiplied by 4.6mm,5 μm); the mobile phase is acetonitrile-water (20: 80); isocratic elution, analysis time 30 minutes; the detection wavelength is 320 nm; flow rate 1.0 mL/min^-1(ii) a The column temperature is 30 ℃; the sample amount is 10 mul; the theoretical plate number is not less than 2000;

1.2.2 preparation of reference solution, namely precisely weighing 13mg of stilbene glucoside (2,3,5, 4' -tetrahydroxystilbene-2-O- β -D-glucoside) reference, placing the reference in a 50mL volumetric flask, adding diluted ethanol for dissolving and fixing the volume to the scale, shaking up to obtain reference solution mother liquor, precisely weighing 1mL of reference solution mother liquor, placing the reference solution mother liquor in a 10mL volumetric flask, adding diluted ethanol for diluting and fixing the volume to the scale, and shaking up to obtain reference solution with the concentration of 0.0260 mg/mL;

1.2.3 preparation of test article, negative control solution: precisely measuring 3.0ml of sample, placing in a 100ml measuring flask, adding appropriate amount of diluted ethanol, shaking to dissolve, adding diluted ethanol to scale, shaking, standing, filtering with 0.45 μm microporous membrane, and collecting filtrate as sample solution. Taking negative samples lacking the prepared fleece flower root, and preparing the negative control solution lacking the prepared fleece flower root by the same method as the test solution.

1.3 specificity test

Precisely sucking 10 μ l of each of the reference solution, the sample solution and the negative reference solution under the chromatographic conditions of item 1.2.1, and injecting into a liquid chromatograph. The result shows that the sample chromatogram has no same chromatographic peak at the position corresponding to the negative control chromatogram, which indicates that the negative control sample has no interference, and the method has good specificity, and the result is shown in the attached figures 1-3.

1.4 Linear relationship examination, precisely sucking 1, 2, 4, 8, 12 and 16 mu l of 2,3,5, 4' -tetrahydroxystilbene-2-O- β -D-glucoside reference substance solution, injecting the solution into a high performance liquid chromatograph, measuring the peak area integral value, drawing a standard curve by taking the sample amount as a horizontal coordinate and the peak area integral value as a vertical coordinate, and obtaining a regression equation of Y1498.7X +4077.2 and r 0.9999, wherein the results show that the stilbene glucoside has a good linear relationship in the range of 26.0-416.0 ng, and the chart is shown in FIG. 4.

1.5 precision test: 10 μ l of the same sample solution (lot 20090821) was precisely aspirated, and sample introduction was performed 6 times, and the peak area integral value was measured, resulting in an RSD value of 1.82%, indicating that the instrument precision was good.

1.6 stability test: respectively and precisely sucking 10 mu l of the same test solution, injecting samples for 0, 2, 4, 8, 12 and 24 hours, and measuring the peak area integral value, wherein the result shows that the RSD value is 0.93 percent, which indicates that the test solution is basically stable within 24 hours.

1.7 repeatability tests: precisely measuring 3.0ml and 6 parts of the same batch of samples (batch number 20090821), and according to the preparation method of the test solution under item 1.2.3, according to the chromatographic condition under item 1.2.1, and the sample injection measurement, the result shows that the average value of the content of the stilbene glucoside is 0.67mg/ml, and the RSD value is 1.99 percent, which indicates that the method has good repeatability.

1.8 recovery test: 1.5ml of a sample with a known content (the content of the stilbene glucoside is 0.67mg/ml) is precisely measured, 6 parts of the sample are placed in a 50ml measuring flask, 3.9ml (namely 1.014mg) of a stilbene glucoside reference substance solution (0.26mg/ml) is precisely added respectively, then the sample solution is prepared according to the method, the content is measured, the recovery rate is calculated, the average recovery rate of the stilbene glucoside is 100.13 percent, and the RSD value is 1.93 percent, and the results are shown in Table 1.

TABLE 1 recovery test results

1.9 determination of samples: the test solutions were prepared according to the methods of the test solution preparation, and the content of stilbene glucoside in the three samples was determined, the results are shown in Table 2.

TABLE 2 results of the assay of three samples

The invention has the beneficial effects that: (1) thin-layer chromatography identification is carried out on the prepared fleece-flower root, the medicinal cyathula root, the mulberry and the glossy privet fruit, and a sample solution for thin-layer identification of the prepared fleece-flower root, the medicinal cyathula root and the mulberry can be prepared by the same treatment method, namely, one sample solution is used for thin-layer identification of three medicinal materials, so that the operation steps, the operation time and the solvent consumption are simplified;

(2) the detection method provided by the invention is a quality control method which has more comprehensive detection items, more scientific and reasonable quality control indexes and simpler operation and can further ensure the safety and curative effect of the medicine. The qualitative and quantitative detection method can be used for qualitative and quantitative detection of the yieldingjing and qualitative and quantitative detection of other dosage forms such as capsules, granules and the like with the same prescription.

Drawings

FIG. 1 HPLC chromatogram of stilbene glucoside reference substance

FIG. 2 is HPLC chromatogram of Yilingjing sample solution

FIG. 3 is HPLC chromatogram of solution of negative sample lacking radix Polygoni Multiflori Preparata

FIG. 4 is a graph of stilbene glycoside standard curve

FIG. 5 identification of thin layer prepared radix Polygoni Multiflori with reference to reference solution, 2-test solution (20090821), 3-test (20090901), 4-test solution (20090903), and 5-negative reference solution in FIG. 1

FIG. 6 identification of radix Cyathulae by thin layer chromatography with reference to FIG. 1-control solution, 2-test solution (20090821), 3-test solution (20090901), 4-test solution (20090903), and 5-negative control solution

FIG. 7 thin-layer identification of Mori fructus in FIG. 1, test solution (20090821), 5-test solution (20090901), 3-test solution (20090903), 4-control solution, and 5-negative control solution

FIG. 8 thin layer identification of fructus Ligustri Lucidi in FIG. 1, including control solution, 2-test solution (20090821), 3-test solution (20090901), 4-test solution (20090903), and 5-negative control solution

Detailed description of the preferred embodiments