CN109212083B

CN109212083B - Quality detection method of compound endothelium corneum gigeriae galli chewable tablets

Info

Publication number: CN109212083B
Application number: CN201811181798.9A
Authority: CN
Inventors: 张立壮; 张志平; 其木格; 郭中华; 刘伟伟; 张冰冰; 韩芳芳; 胡宇
Original assignee: Henan Taloph Pharmaceutical Co ltd
Current assignee: Henan Taloph Pharmaceutical Co ltd
Priority date: 2018-10-11
Filing date: 2018-10-11
Publication date: 2022-02-11
Anticipated expiration: 2038-10-11
Also published as: CN109212083A

Abstract

A quality detection method of a compound endothelium corneum Gigeriae Galli chewable tablet comprises detecting the compound endothelium corneum Gigeriae Galli chewable tablet by high performance liquid chromatography to obtain fingerprint chromatogram thereof, wherein the mobile phase in the high performance liquid chromatography is one or more selected from water, phosphoric acid water solution, trichloroacetic acid water solution, phosphoric acid buffer solution, methanol, acetonitrile, isopropanol, acetone and tetrahydrofuran. The method can obtain the fingerprint with 9 common peaks, can reflect the multi-component information of the product, has simple detection method, good reproducibility and more characteristic peaks, thereby ensuring the relative uniformity and stability of the quality of the compound endothelium corneum gigeriae galli chewable tablet, more comprehensively and effectively controlling the internal quality of the product and ensuring the safety and effectiveness of the product.

Description

Quality detection method of compound endothelium corneum gigeriae galli chewable tablets

Technical Field

The invention relates to a quality detection method of compound chicken's gizzard-membrane chewable tablets.

Technical Field

A compound chewable tablet for treating infantile dyspepsia, emesis, and diarrhea due to incoordination between spleen and stomach is prepared from endothelium corneum Gigeriae Galli and Massa Medicata Fermentata. The endothelium corneum Gigeriae Galli in the prescription is the inner wall of dry sand bag of domestic animal chicken; medicated leaven is a leaven product prepared by fermenting polygonum hydropiper, xanthium sibiricum, sweet wormwood herb, bitter apricot seed, phaseolus calcaratus, wheat bran and flour, and has complex components, and the main active ingredients of the medicated leaven are chlorogenic acid, rutin, quercetin, amygdalin, artemisinin, scopoletin, xanthatin and the like. The compound chicken's gizzard-membrane chewable tablet for children belongs to a Chinese medicinal compound preparation, and the effective ingredients are undefined.

At present, the existing detection means for the compound chicken's gizzard-membrane chewable tablets for children mainly comprises the microscopic identification of powder loaded in quality standards and the content determination of nitrogen content in the product. The detection method is too simple, and the specificity of the content determination method is poor. CN 107478762B discloses a thin layer identification method and a content determination method of a compound chicken's gizzard-membrane chewable tablet for children, but the detection method still can not effectively control the inherent quality of the product.

The method reported by Liu Hui Min and the like in HPLC fingerprint spectrum research (Chinese medicine guide, 2015, 12 th period 24-26) of different producing areas of medicated leaven is used for measuring the pediatric compound chicken's gizzard-membrane chewable tablets, and the result always shows that the separation degree of 3 to 5 peaks can not meet the requirement and the reproducibility is poor.

The compound traditional Chinese medicine has complex components, and one or more components are taken as quality control indexes, so that the compound traditional Chinese medicine is not more and more suitable for the requirement of the quality control of the compound traditional Chinese medicine. In order to ensure the safe and effective medication of children, a detection method for comprehensively controlling the integral internal quality of the compound chicken's gizzard-membrane chewable tablets for children is urgently sought.

Disclosure of Invention

In order to solve the problems, the inventor provides a quality detection method of the compound endothelium corneum gigeriae galli chewable tablet, and the fingerprint spectrum of the compound endothelium corneum gigeriae galli chewable tablet is obtained by detecting the compound endothelium corneum gigeriae galli chewable tablet by a high performance liquid chromatography method, so that the internal quality of the product is more comprehensively and effectively controlled.

The purpose of the invention is realized by the following technical scheme:

a quality detection method of compound endothelium corneum Gigeriae Galli chewable tablet is characterized in that a high performance liquid chromatography method is adopted to detect the compound endothelium corneum Gigeriae Galli chewable tablet and obtain the fingerprint thereof, wherein the mobile phase in the high performance liquid chromatography method is one or more selected from water, phosphoric acid aqueous solution, trichloroacetic acid aqueous solution, phosphoric acid buffer solution, methanol, acetonitrile, isopropanol, acetone and tetrahydrofuran.

Preferably, the high performance liquid phase method adopts mobile phase A, mobile phase B and mobile phase C to carry out gradient elution, wherein the mobile phase A is one or two selected from acetonitrile and methanol, and is more preferably acetonitrile; the mobile phase B is phosphoric acid aqueous solution or trichloroacetic acid aqueous solution, preferably, the mobile phase B is 0.5-2% (v/v), more preferably 0.8-1.5% (v/v), more preferably 1% (v/v) phosphoric acid aqueous solution, or 0.05-1%, preferably 0.1-0.8%, more preferably 0.3% (v/v) trichloroacetic acid aqueous solution; the mobile phase C is one or two selected from isopropanol and methanol, and isopropanol is more preferable.

Preferably, the elution time is 45-100min, preferably 60-90min, more preferably 75 min.

Preferably, the gradient elution procedure is:

(1) eluting with the mixed solution of the mobile phase A and the mobile phase B for 20-40 minutes, preferably for 25-35 minutes; more preferably for 34 minutes; preferably, the volume ratio of mobile phase a to mobile phase B is from 2 to 30:98 to 70, more preferably from 8 to 20:92 to 80.

(2) Eluting with the mixed solution of mobile phase A, mobile phase B and mobile phase C for 10-30 min, preferably for 20-28 min, and more preferably for 26 min; the volume ratio of the mobile phase A, the mobile phase B and the mobile phase C is preferably 15-50:80-34:5-16, preferably 20-41:74-45: 6-14.

(3) Optionally, eluting with the mixed solution of mobile phase A and mobile phase B for 5-15 min, preferably for 15 min; preferably, the volume ratio of mobile phase a to mobile phase B is 45-5:55-95, preferably 40-8: 60-92.

More preferably, the gradient elution procedure is:

time (minutes)	Mobile phase A (%)	Mobile phase B (%)	Mobile phase C (%)
				0	8	92	0
13	15	85	0
				22	20	80	0
34	20	74	6
				55	41	45	14
60	40	60	0
				75	8	92	0

Preferably, the high performance liquid chromatography column is: polystyrene-divinylbenzene (PS/DVB) column, C18 column; more preferably a polystyrene-divinylbenzene (PS/DVB) column.

Preferably, the flow rate is 0.8-1.2ml/min, more preferably 1.0 ml/min.

Preferably, the sample is taken in an amount of 10-30. mu.l, more preferably 20. mu.l.

Preferably, the detection wavelength is 210-280nm, more preferably 268 nm.

Preferably, the column temperature is 30-50 deg.C, more preferably 35 deg.C.

Preferably, the preparation method of the compound chicken's gizzard-membrane chewable tablet test solution comprises the following steps: grinding compound endothelium corneum Gigeriae Galli chewable tablet, precisely weighing, extracting with petroleum ether, adding methanol into residue, ultrasonic treating, filtering, passing filtrate through macroporous resin column, collecting eluate, evaporating under reduced pressure, and dissolving the residue with methanol to obtain sample solution.

Preferably, the preparation method of the compound chicken's gizzard-membrane chewable tablet test solution comprises the following steps: grinding compound endothelium corneum Gigeriae Galli chewable tablet, precisely weighing, mixing with petroleum ether, heating and refluxing, filtering, removing petroleum ether solution, adding methanol for ultrasonic treatment, filtering, passing filtrate through macroporous resin column, eluting with water, eluting with ethanol water solution, collecting ethanol water solution eluate, evaporating under reduced pressure, and dissolving residue with methanol to obtain sample solution.

Preferably, the preparation method of the compound chicken's gizzard-membrane chewable tablet test solution comprises the following steps: grinding compound endothelium corneum Gigeriae Galli chewable tablet, mixing powder and petroleum ether at a solid-liquid mass-volume ratio (g/ml) of 3:10, heating and refluxing for 1-2 hr, filtering, discarding petroleum ether solution, volatilizing residue, adding 80% methanol at a solid-liquid mass-volume ratio (g/ml) of 2:5, performing ultrasonic treatment for 20-40min, filtering, passing filtrate through macroporous resin column (preferably DM301), eluting with 2 column volumes of water, eluting with 2 column volumes of 70-80% ethanol, collecting ethanol eluate, evaporating under reduced pressure, and dissolving residue with methanol to obtain test solution.

Preferably, the preparation method of the compound chicken's gizzard-membrane chewable tablet test solution comprises the following steps: grinding compound chicken's gizzard-membrane chewable tablets, precisely weighing 15g of powder, mixing the powder with petroleum ether (60-90 ℃) according to the mass-to-volume ratio (g/ml) of solid to liquid (3: 10), heating and refluxing for 1 hour, filtering, removing petroleum ether liquid, volatilizing residues, adding 80% methanol according to the mass-to-volume ratio (g/ml) of solid to liquid (2: 5), carrying out ultrasonic treatment for 30min, filtering, enabling filtrate to pass through a DM301 macroporous resin column, eluting with 2 column volumes of water, eluting with 3 column volumes of 75% ethanol, collecting ethanol eluate, drying by evaporation under reduced pressure, dissolving the residues with methanol to obtain a test solution.

Preferably, rutin is used as a control. Preferably, the control solution is prepared as follows: accurately weighing appropriate amount of rutin control, placing into a measuring flask, adding methanol to obtain control solution with mass concentration of 10-60ug/ml, preferably 30 ug/ml.

Injecting the test solution and the reference solution into a high performance liquid chromatograph to obtain a fingerprint spectrum, wherein the spectrum has more than 9 common peaks.

The invention has the following beneficial effects:

the quality detection method of the compound chicken's gizzard-membrane chewable tablet adopts a high performance liquid chromatography method to detect the compound chicken's gizzard-membrane chewable tablet, can obtain the fingerprint with 9 common peaks, can reflect the multi-component information of the product, has simple detection method, good reproducibility and more characteristic peaks, and thus ensures the relatively uniform and stable quality of the compound preparation. Especially, under the condition that the effective components are not completely definite, the internal quality of the product can be more comprehensively and effectively controlled, and the safety and effectiveness of the product are ensured. The method has the characteristics of simplicity, convenience, stability, high precision and good reproducibility.

Description of the drawings:

FIG. 1: HPLC chromatogram of rutin control solution in example 1

FIG. 2: example 1 HPLC fingerprint of test solution of Compound endothelium corneum Gigeriae Galli chewable tablet for infants

FIG. 3 is a fingerprint overlay of 22 batches of the infant compound chicken's gizzard-membrane chewable tablet test solutions in example 3

Detailed Description

The invention is further illustrated by the following examples. It should be understood that the examples herein are for illustrative purposes only and are not intended to limit the scope of the present invention in any way.

The following examples used instruments and reagents:

the instrument comprises the following steps: ultimate 3000 high performance liquid chromatography (siemmer fei, germany); a DAD-3000 diode array detector; an electronic analytical balance model MS105 DU; KQ-300E type ultrasonic cleaning machine; HH-S26S model digital display constant temperature water bath; RE-52AA rotary evaporator, chromatographic column, etc.

Reagent: acetonitrile, isopropanol and methanol are used as chromatographic purities, and ethanol and petroleum ether are used as analytical purities.

The reference and sample sources used in the following examples:

comparison products: rutin control (batch number: 100080-201811, China institute for food and drug testing);

a sample to be tested: the compound chicken's gizzard-membrane chewable tablets for children are 1.2 g/tablet, provided by Henan Tailong pharmaceutical industry GmbH, and the batch number information is shown in Table 1.

TABLE 1

Numbering	Batch number	Numbering	Batch number
				S1	170401	S12	171003
S2	170402	S13	171004
				S3	170403	S14	171005
S4	170601	S15	171006
				S5	170602	S16	180101
S6	170603	S17	180102
				S7	170604	S18	180103
S8	170905	S19	180401
				S9	170906	S20	180402
S10	170907	S21	180403
				S11	170908	S22	180404

Example 1: determination of finger print of compound chicken's gizzard-membrane chewable tablet for children

1. Chromatographic conditions and system adaptability

(1) Preparation of control solutions

Accurately weighing an appropriate amount of rutin reference substance, placing into a measuring flask, and adding methanol to prepare reference substance solution with mass concentration of 30 μ g/ml.

(2) Preparation of test solution

Grinding compound chicken's gizzard-membrane chewable tablets for children (batch No. 170401), precisely weighing 15g of powder of the chewable tablets, mixing the powder with petroleum ether (60-90 ℃) according to the mass-to-volume ratio (g/ml) of solid to liquid (3: 10), heating and refluxing for 1 hour, filtering, discarding petroleum ether liquid, volatilizing residues, adding 80% methanol according to the mass-to-volume ratio (g/ml) of solid to liquid (2: 5), carrying out ultrasonic treatment for 30min, filtering, passing filtrate through a DM301 macroporous resin column, eluting with 2 column volumes of water, eluting with 3 column volumes of 75% ethanol, collecting ethanol eluate, carrying out reduced pressure evaporation to dryness, dissolving the residues with 10ml of chromatographic grade methanol, and filtering with a 0.45 mu m filter membrane to obtain a test solution.

(3) Detection of

Performing gradient elution by using polystyrene divinyl benzene as a stationary phase, acetonitrile as an A mobile phase, 0.1% (v/v) trichloroacetic acid solution as a B mobile phase and isopropanol as a C mobile phase, wherein the flow rate is as follows: 1.0ml/min, detection wavelength: 268 nm; column temperature: 35 ℃; sample introduction amount: 20 ul; the theoretical plate number is not lower than 2500 calculated according to rutin peak.

The control solution and the test solution were injected into a high performance liquid chromatograph, and the gradient elution procedure is shown in table 2.

TABLE 2

Obtaining the reference solution chromatogram and the sample solution fingerprint chromatogram as shown in figure 1 and figure 2.

Example 2 examination of the methodology for detecting pediatric Compound endothelium corneum Gigeriae Galli chewable tablets

1. Precision test

1.1 precision test of control solution: by using the same operation and conditions as in example 1, the rutin control solution was sampled continuously for 5 times, the chromatogram was recorded, and the RSD of the peak area of the control solution was calculated as shown in Table 3.

TABLE 3

Numbering	Peak area
		1	12296.736
2	12247.265
		3	12263.041
4	12272.522
		5	12203.997
Average	12256.7122
		RSD	0.28％

1.2 precision test of test solution by using the same operation and conditions as in example 1, continuously feeding 5 times of test solution of the same batch of compound chicken's gizzard-membrane chewable tablets for children (batch No. 170401), using rutin as a reference peak, investigating the consistency of relative retention time and relative peak area of each common peak, and evaluating the stability of the apparatus, the results are shown in tables 4 and 5.

TABLE 49 relative retention time of common peaks

Peak number	1	2	3	4	5	RSD
							1	0.155	0.156	0.154	0.155	0.155	0.46％
2	0.286	0.288	0.286	0.287	0.286	0.31％
							3	0.344	0.345	0.345	0.345	0.346	0.20％
4	0.756	0.753	0.754	0.755	0.754	0.15％
							5	0.802	0.801	0.800	0.803	0.801	0.14％
6	0.855	0.859	0.857	0.855	0.856	0.20％
							7(S)	1.000	1.000	1.000	1.000	1.000	--
8	1.124	1.123	1.123	1.121	1.124	0.11％
							9	1.152	1.154	1.150	1.154	1.151	0.16％

TABLE 59 relative peak area of common peaks

Peak number	1	2	3	4	5	RSD
							1	0.126	0.124	0.130	0.120	0.125	2.88％
2	0.148	0.150	0.142	0.146	0.145	2.07％
							3	0.057	0.053	0.056	0.055	0.054	2.88％
4	0.164	0.162	0.163	0.164	0.160	1.03％
							5	0.140	0.142	0.145	0.144	0.143	1.35％
6	0.171	0.170	0.173	0.169	0.168	1.13％
							7(S)	1	1	1	1	1
8	0.080	0.083	0.082	0.086	0.082	2.65％
							9	0.084	0.083	0.080	0.083	0.085	2.25％

1.3 the result shows that the reference substance has a solution peak area RSD less than 2 percent, which indicates good precision; the relative standard deviation RSD of 9 common peaks of the test solution relative to the retention time is less than 1 percent, the RSD of the relative peak area is less than 3 percent, the precision is good, the instrument is stable, and the requirement of the fingerprint spectrum is met.

2. Stability survey

2.1 adopting the same operation and conditions as in example 1, sample injection analysis is carried out on the same batch of test solution of the compound pediatric chicken's gizzard-membrane chewable tablets (batch No. 170401) at 0, 3, 6, 8, 12 and 24 hours respectively, and rutin is taken as a reference peak to calculate the relative retention time and the relative peak area of 9 common peaks.

2.1 results the RSD of the relative retention time of each common peak is less than 1%, and the RSD of the relative peak area is less than 4%, indicating that the test solution is stable within 24 hours.

3. Reproducibility study

3.1 by adopting the same operation and conditions as in example 1, 6 test sample solutions were prepared in parallel from the same batch of compound chicken's gizzard-membrane chewable tablets for children (batch No. 170401), and were subjected to sample injection analysis, and a fingerprint was recorded, and the relative retention time and the relative peak area of each common peak were calculated using rutin as a reference peak.

3.2 results show that the RSD of the relative retention time of all the shared peaks is less than 1 percent, and the RSD of the relative peak area is less than 4 percent, which indicates that the method has good repeatability.

Example 322 batch fingerprint determination and fingerprint analysis of Compound endothelium corneum Gigeriae Galli chewable tablets for infants

1. Taking 22 batches of compound chicken gizzard-membrane chewable tablets for children, determining by adopting the same operation and conditions in the embodiment 1 to obtain 22 batches of HPLC (high performance liquid chromatography) spectra, matching chromatographic peaks by adopting software of 'three batches of traditional Chinese medicine fingerprint similarity evaluation system 2004A edition', generating a superposition spectrum (figure 3), determining 9 common peaks, taking rutin (the number of the chromatographic peak is 7) as a reference peak, and calculating the relative retention time and the relative peak area of each peak in the chromatogram.

TABLE 6 relative retention time of common peaks

TABLE 7 relative peak area of common peaks

2. Results show that the RSD of the relative retention time of 9 common peaks of 22 batches of the pediatric compound chicken's gizzard-membrane chewable tablets is less than 1 percent, and the RSD of the relative peak area is less than 4 percent.

HPLC fingerprint similarity analysis of 3.22 batches of compound chicken's gizzard-membrane chewable tablets for children

The similarity calculation results are shown in Table 8 by using software of a Chinese medicine three-batch fingerprint similarity evaluation system 2004A.

Similarity evaluation between Table 8 and S1

Numbering	Batch number	Degree of similarity	Numbering	Batch number	Degree of similarity
						S2	170402	0.965	S13	171004	0.982
S3	170403	0.971	S14	171005	0.974
						S4	170601	0.987	S15	171006	0.960
S5	170602	0.964	S16	180101	0.975
						S6	170603	0.985	S17	180102	0.970
S7	170604	0.954	S18	180103	0.968
						S8	170905	0.967	S19	180401	0.974
S9	170906	0.973	S20	180402	0.969
						S10	170907	0.981	S21	180403	0.981
S11	170908	0.970	S22	180404	0.956
						S12	171003	0.966

The 1.4 result shows that compared with S1, the similarity of 21 batches of compound chicken 'S gizzard-membrane chewable tablets for children is more than 0.95, the main chromatographic peaks in each sample are basically consistent, the established fingerprint has stability and controllability, and can be used as an evaluation basis for controlling the quality of the compound chicken' S gizzard-membrane chewable tablets for children, so as to ensure the stable and uniform product quality and ensure the safe and effective medication for children.

Claims

1. A quality detection method of a compound endothelium corneum Gigeriae Galli chewable tablet is characterized in that a high performance liquid chromatography is adopted to detect the compound endothelium corneum Gigeriae Galli chewable tablet and obtain a fingerprint spectrum thereof, wherein a chromatographic column of the high performance liquid chromatography adopts: a polystyrene-divinylbenzene column; the high performance liquid phase method adopts a mobile phase A, a mobile phase B and a mobile phase C to carry out gradient elution, wherein the mobile phase A is acetonitrile; the mobile phase B is trichloroacetic acid aqueous solution, and the mobile phase C is isopropanol;

the gradient elution procedure is as follows:

time in minutes Mobile phase A,% Mobile phase B,% Mobile phase C,% 0 8 92 0 13 15 85 0 22 20 80 0 34 20 74 6 55 41 45 14 60 40 60 0 75 8 92 0

The preparation method of the test solution of the compound chicken's gizzard-membrane chewable tablets comprises the following steps: grinding compound endothelium corneum Gigeriae Galli chewable tablet, precisely weighing, extracting with petroleum ether, adding methanol into residue, performing ultrasonic treatment, filtering, passing filtrate through macroporous resin column, collecting eluate, evaporating under reduced pressure, and dissolving the residue with methanol to obtain sample solution;

the fingerprint of the test solution of the compound chicken's gizzard-membrane chewable tablet obtained by the high performance liquid chromatography detection has 9 common peaks.

2. The quality detection method of the compound chewable tablet of chicken's gizzard-membrane according to claim 1, wherein the mobile phase B is trichloroacetic acid aqueous solution with a volume ratio of 0.05-1%.

3. The method for detecting the quality of the compound chewable tablet of chicken's gizzard-membrane according to claim 1, wherein the mobile phase B is trichloroacetic acid aqueous solution with the volume ratio of 0.1 to 0.8 percent.

4. The method for detecting the quality of the compound chewable tablet of chicken's gizzard-membrane according to claim 1, wherein the mobile phase B is 0.1% trichloroacetic acid aqueous solution by volume.

5. The method for detecting the quality of the compound chicken's gizzard-membrane chewable tablet according to any one of claims 1 to 4, wherein the preparation method of the test solution of the compound chicken's gizzard-membrane chewable tablet comprises the following steps: grinding compound endothelium corneum Gigeriae Galli chewable tablet, precisely weighing, mixing with petroleum ether, heating and refluxing, filtering, removing petroleum ether solution, adding methanol for ultrasonic treatment, filtering, passing filtrate through macroporous resin column, eluting with water, eluting with ethanol water solution, collecting ethanol water solution eluate, evaporating under reduced pressure, and dissolving residue with methanol to obtain sample solution.

6. The method for detecting the quality of the compound chicken's gizzard-membrane chewable tablet according to any one of claims 1 to 4, wherein the preparation method of the test solution of the compound chicken's gizzard-membrane chewable tablet comprises the following steps: grinding compound endothelium corneum Gigeriae Galli chewable tablet, mixing powder and petroleum ether at a solid-liquid mass volume ratio of 3:10, heating and refluxing for 1-2 hr, filtering, discarding petroleum ether solution, volatilizing residue, adding 80% methanol at a solid-liquid mass volume ratio of 2:5, performing ultrasonic treatment for 20-40min, filtering, passing filtrate through macroporous resin column, eluting with 2 column volumes of water, eluting with 2 column volumes of 70-80% ethanol, collecting ethanol eluate, evaporating under reduced pressure, and dissolving residue with methanol to obtain sample solution.

7. The method for detecting the quality of the compound chicken's gizzard-membrane chewable tablet according to any one of claims 1 to 4, wherein the preparation method of the test solution of the compound chicken's gizzard-membrane chewable tablet comprises the following steps: grinding compound chicken's gizzard-membrane chewable tablets, precisely weighing 15g of powder, mixing the powder with petroleum ether at the temperature of 60-90 ℃ according to the mass-to-volume ratio of solid to liquid of 3:10, heating and refluxing for 1 hour, filtering, removing petroleum ether liquid, volatilizing residues, adding 80% methanol according to the mass-to-volume ratio of solid to liquid of 2:5, carrying out ultrasonic treatment for 30min, filtering, enabling filtrate to pass through a DM301 macroporous resin column, eluting with 2 column volumes of water, eluting with 3 column volumes of 75% ethanol, collecting ethanol eluate, drying by distillation under reduced pressure, and dissolving the residues with methanol to obtain a sample solution.

8. The method for detecting the quality of the compound chewable tablet of chicken's gizzard-membrane according to any one of claims 1 to 4, wherein rutin is used as a reference substance.

9. The quality detection method of the compound chewable tablet of chicken's gizzard-membrane according to claim 8, wherein the rutin control solution is prepared as follows: accurately weighing appropriate amount of rutin control, placing into a measuring flask, and adding methanol to obtain control solution with mass concentration of 10-60 ug/ml.

10. The method for detecting the quality of the compound chewable tablet of chicken's gizzard-membrane according to claim 9, wherein the concentration of rutin in reference to the quality of the chewable tablet is 30 ug/ml.