CN100392399C - The Method of Detecting the Quality of Chinese Jinjiu with Fingerprint Technology - Google Patents

Abstract

The invention discloses a method for detecting the quality of Chinese Jinjiu by using fingerprint technology. The method comprises the following steps: the first step is to establish the comparative fingerprint of Chinese Jinjiu; the second step is to establish a common pattern of fingerprints of Chinese Jinjiu The third step is the detection of the test product; compare the measured spectrum of the test product with the control fingerprint, and input it into the computer-aided similarity evaluation system to calculate the similarity, obtain the correlation coefficient, and then make a detection conclusion; This method has the following characteristics: (1) it can control the quality of Chinese Jinjiu from an overall and macro perspective; (2) it is clear and intuitive, has good specificity and characteristics, good reproducibility, and strong operability; (3) It can be used to investigate the stability of production process and product; (4) It can be used to identify the authenticity of Chinese Jinjiu; (5) It can provide a demonstration for the monitoring of product quality in China's health wine industry.

Description

The Method of Detecting the Quality of Chinese Jinjiu with Fingerprint Technology

(1) Technical field:

The invention relates to a method for detecting the quality of traditional Chinese medicinal health wine, in particular to a method for detecting the quality of Chinese strong liquor prepared from various raw materials of traditional Chinese medicines by using high performance liquid phase-gas chromatography fingerprint technology.

(2) Existing technology: China Jinjiu is the health-care wine with invigorating qi, kidney-tonifying, and yin-nourishing effects produced by Hubei Jinpai Company. , cinnamon and other several kinds of Chinese herbal medicines. In terms of product quality control, before 2000, the main control indicators were color, solid content, alcohol content, and taste. After 2000, qualitative or quantitative indicators. However, as a traditional Chinese medicine compound health wine, its health function mechanism and functional factors are very complicated. It is impossible to completely control the product quality by controlling a few indicators, but must control its substance group as a whole. Therefore, in addition to microscopic analysis, it is also necessary to use some macroscopic analysis method to effectively characterize the internal quality of the product as a whole. Traditional Chinese medicine fingerprint refers to the chromatogram, spectrum, etc. that can mark the characteristics of the traditional Chinese medicinal material or Chinese patent medicine obtained by using certain analytical methods after proper processing of a certain or a certain place of origin of the Chinese medicinal material or Chinese patent medicine. As a quality control method of traditional Chinese medicine, research on fingerprints is in the ascendant at home and abroad. Internationally, atlas has become a consensus in the medical community as a quality control method for Chinese patent medicines, herbal extracts, and other mixed substance groups. The U.S. FDA (Food and Drug Administration) requires that the quality inspection of botanical drugs must formulate detection standards for fingerprints. ) and botanical drug products (Botanical Drug product) quality control to use fingerprints. The European Community has also adopted fingerprinting technology for quality control of herbal medicines. In Japan, the decoction obtained by decocting the formula of authentic medicinal pieces is used as the standard extract, and the fingerprint of the standard extract is the standard fingerprint. In this way, the raw materials, formula and process of production are strictly controlled, so that the fingerprint of the finished product is consistent with the standard fingerprint. The spectra are consistent. In addition, France, Germany, the United Kingdom, India, and WHO all use fingerprint technology to evaluate the quality of herbal medicines (herbal medicines). The State Food and Drug Administration issued the "Technical Requirements for Fingerprint Research of Traditional Chinese Medicine Injections" in 2000, requiring the implementation of fingerprint quality control methods for traditional Chinese medicine injections. It is also an inevitable trend in the development of traditional Chinese medicine to use fingerprint quality control technology of traditional Chinese medicine as the quality inspection standard for other Chinese patent medicine preparations. Therefore, the use of fingerprint quality control technology will play an important role in authenticity identification, quality control, product excellence and production process control of traditional Chinese medicine and related products.

(3) technical content: the purpose of the present invention is exactly to utilize the integrality, characteristic and quantification of Chinese medicine fingerprint, proposes a kind of method that can detect the quality of Chinese Jinjiu macroscopically and on the whole. At the same time, it also provides a demonstration for the product quality control of my country's health care wine industry.

The present invention comprises the following steps:

The first step is to establish the comparative fingerprint of Chinese Jinjiu

1. Preparation of Chinese Jinjiu

Take 10-15 parts by weight of traditional Chinese medicine raw materials astragalus, 10-15 parts of Cistanche, 10-15 parts of curculigo, 10-15 parts of Epimedium, 10-15 parts of wolfberry, 10-15 parts of Chinese yam, 10-15 parts of angelica, and 3-5 parts of cloves , Cinnamon 3-5 percolate 12-15 days with 30-50 ° of Xiaoqu Baijiu of 50 times of total medicinal material amount to obtain percolate, get percolate 70-75 parts, add respectively 20 parts of 40% syrup, purified water 5- 10 parts, appropriate amount of caramel color, blending, filtering, aging at room temperature for 3-6 months, filtering to remove impurities, fine filtration with membrane filter, filling, and the product is ready;

2. Preparation of Chinese Jinjiu Liquid Phase Reference Substance Solution

a. Ether site

Take 100ml of Chinese Jinjiu, remove the sugar with a macroporous adsorption chromatographic column, extract 3 times with saturated diethyl ether, 20ml each time, combine the extracts, evaporate to dryness, dilute to 2ml with tetrahydrofuran, and filter through a membrane to obtain the product;

b. Ethyl acetate site of 5% isoamyl alcohol

Extract the test solution of the ether part with 5% isoamyl alcohol saturated with water three times, 20ml each time, combine the extracts, evaporate to dryness, dilute to 2ml with tetrahydrofuran, and filter through a membrane to obtain final product;

3. Preparation of Chinese Jinjiu Gas Phase Reference Substance Solution

Directly take 5ml of Chinese Jinjiu into a 10mi headspace sample bottle, cover and seal, and you get it;

4. Preparation of standard solution

a. Liquid phase part

Take an appropriate amount of ferulic acid standard substance and make a 1 mg/ml solution with 50% methanol; take an appropriate amount of icariin standard substance and make a 1 mg/ml solution with 50% methanol;

b. Gas phase part

Use a glass capillary to dip an appropriate amount of standard products of bornyl acetate and eugenol into a headspace vial, seal it, and obtain it;

5. Chromatographic conditions

a. Liquid phase part

Chromatographic column

Ether part: Hypersil C18 column,

Ethyl acetate part of 5% isoamyl alcohol: Aichrombond AQ C18,

mobile phase

Diethyl ether part: acetonitrile-0.01% phosphoric acid aqueous solution is gradient elution of mobile phase;

Ethyl acetate part of 5% isoamyl alcohol: acetonitrile-0.01% phosphoric acid aqueous solution is gradient elution of mobile phase;

Flow rate: 0.8ml/min

Detection wavelength: 280nm

Column temperature: 30°C

b. Gas phase part

Column Agilent HP-5

Carrier gas is N2

Sampler: G1888A headspace autosampler;

6. Control substance detection

a. Liquid phase part

Ether part: Accurately absorb 4 μl each of the reference solution and the standard solution of the ether part of the liquid phase of China Jinjiu, inject it into the liquid chromatograph, and measure it according to high-performance liquid chromatography to obtain the control fingerprint of the ether part of China Jinjiu and the corresponding The fingerprint of the standard product;

The ethyl acetate part of 5% isoamyl alcohol: 4 μl each of the reference solution and the standard solution of the ethyl acetate part of 5% isoamyl alcohol in the liquid phase of China Jinjiu were accurately drawn, injected into the liquid chromatograph, and followed by high-performance liquid chromatography. Chromatography is used to determine the different fingerprints of the ethyl acetate part of 5% amyl alcohol and the corresponding standard product fingerprints;

b. Gas phase part

The reference substance solution and the standard substance solution of the gas phase part were analyzed and measured by the automatic sampling of the headspace sampler, and the gas phase part control fingerprint and the corresponding standard fingerprint were obtained;

In the control fingerprint of Chinese Jinjiu, there are 10 characteristic peaks in the control fingerprint of the ether portion of the liquid phase, wherein peak No. 7 is ferulic acid, and there are 10 characteristic peaks in the control fingerprint of the ethyl acetate part of 5% amyl alcohol. There are 9 peaks, of which No. 11 peak is icariin, which is consistent with the fingerprint of the standard product; there are 8 characteristic peaks in the gas-phase partial control fingerprint, of which No. 7 peak is bornyl acetate, and No. 8 peak is clove Phenol, consistent with the fingerprint of the standard product;

The specific method for preparation of the reference substance solution of the ether part of the liquid phase part of the Chinese Jinjiu and the ethyl acetate part of 5% isoamyl alcohol is to take 100 ml of the Chinese Jinjiu, 40 ° C water bath rotary evaporation to nearly dry, and load the sample on an AB -8 on a macroporous adsorption chromatography column (12mm x 300mm), eluted with water until there is no sugar reaction (ie Molish reaction is negative), then eluted with 80ml of 95% ethanol, collected the eluate, and evaporated to dryness under reduced pressure. Add 20ml of water, sonicate to dissolve, and transfer to a separatory funnel. First extract 3 times with saturated diethyl ether, 20ml each time, combine the extracts, evaporate to dryness in a water bath, dilute to 2ml with tetrahydrofuran, shake well, and filter with a 0.45μ microporous membrane to obtain the diethyl ether reference solution; The solution was extracted 3 times with 5% ethyl acetate of isoamyl alcohol saturated with water, 20ml each time, the extracts were combined, evaporated to dryness under reduced pressure, fixed to 2ml with tetrahydrofuran, shaken well, and filtered with a 0.45μ microporous membrane, that is Obtain the ethyl acetate part reference substance solution of 5% isoamyl alcohol;

The ether part of the liquid phase is eluted with acetonitrile-0.01% phosphoric acid aqueous solution as the mobile phase gradient, and the elution gradient is: when the retention time is 0min, 45min, 80min, 90min, 105min, the volume of acetonitrile and 0.01% phosphoric acid aqueous solution The ratios are 8:92, 30:70, 50:50, 80:20, 80:20;

Ethyl acetate part of 5% isoamyl alcohol: Acetonitrile-0.01% phosphoric acid aqueous solution is gradient elution of the mobile phase, and the elution gradient is: when the retention time is 0min, 50min, 80min, 100min, 110min, acetonitrile and 0.01% phosphoric acid The volume ratio of the aqueous solution is 15:85, 30:70, 55:45, 100:0, 100:0 respectively;

The computer-aided similarity evaluation system refers to the computer-aided similarity evaluation system recommended by the Pharmacopoeia Committee developed by the Modernization Research Center of Traditional Chinese Medicine of Hunan Central South University;

The establishment of China Jinjiu's comparative fingerprints has provided a very clear, intuitive and easy-to-operate standard for product quality control, and it is only necessary to compare the test results with it when using it.

The second step is to establish a common pattern map of Chinese Jinjiu fingerprints

According to the preparation method of Chinese Jinjiu, the preparation method of Chinese Jinjiu liquid phase part and gas phase part reference solution, the preparation method of standard solution, prepare 10 batches of Chinese Jinjiu and standard substance, test solution, and one by one Detected by the same method, the fingerprints of the ethyl ether part of each batch of Chinese Jinjiu, the fingerprints of the ethyl acetate part of 5% isoamyl alcohol, the fingerprints of the gas phase part and the fingerprints of the corresponding standard products were respectively obtained; respectively input into the computer-aided similarity evaluation The system can calculate the common pattern of the fingerprints of the ether part of China Jinjiu, the ethyl acetate part of 5% isoamyl alcohol and the gas phase part;

The role of establishing a common pattern map is to input the test results into the computer to calculate the correlation coefficient between the test product and the shared pattern map, and theoretically investigate the similarity between the test product and the shared pattern map, so as to control the quality of the product The purpose; the second is to facilitate the comparison between multiple batches of products to examine the stability of the production process and product stability to ensure that consumers are provided with high-quality products.

The third step, the test product detection

1. Preparation of the test solution

Prepare respectively the ethyl acetate need testing solution of liquid phase part ether part and 5% isoamyl alcohol and gas phase part need testing solution with above-mentioned same method;

2. Prepare liquid phase part and gas phase part standard solution respectively in the same method as above;

3. Determine the standard substance and the test substance to be tested by the same method as above;

4. Result analysis and judgment

10 characteristic peaks identical with the control fingerprint should be detected in the fingerprint of the test product in the ether part, wherein No. 7 peak is ferulic acid, which should be consistent with the fingerprint of the standard product; the ethyl acetate part of 5% isoamyl alcohol provides Nine characteristic peaks identical to those of the control fingerprint should be detected in the fingerprint of the test product; peak No. 11 is icariin, which should be consistent with the fingerprint of the standard product; Calculated by the computer-aided similarity evaluation system, the correlation coefficient should be greater than 0.9000;

The 8 characteristic peaks identical to those of the reference fingerprint should be detected in the fingerprint of the test product in the gas phase; wherein peak No. 7 is bornyl acetate and peak No. 8 is eugenol, which should be consistent with the fingerprint of the standard product; its fingerprint Calculated with the corresponding shared pattern map through the computer-aided similarity evaluation system, the correlation coefficient should be greater than 0.9000;

The principle of the present invention is based on the fact that Chinese Jinjiu contains a large amount of non-volatile components and some raw materials contain volatile components, and the non-volatile components have strong absorption in the ultraviolet region. According to the properties of its main chemical components, the finished product is separated into ether part and ethyl acetate part of 5% isoamyl alcohol, and the non-volatile components and volatile components are detected by liquid phase and gas chromatography respectively. ; During the research process, the extraction method, detection method, chromatographic conditions and system adaptability, precision, stability, reproducibility, etc. were investigated respectively. According to the detection results of more than 10 batches of pilot products, a product control fingerprint was established. Spectrum and common pattern atlas; And confirm that the correlation coefficient between the fingerprint spectrum of test product and corresponding common pattern atlas is 0.9000; Therefore, can grasp the quality situation of Chinese Jinjiu from the overall appearance characteristic of chromatogram.

The following are some results of the investigation of this method:

(1) Reproducibility test

Take the same sample solution, operate 6 times according to the preparation method and detection method of the test sample, and calculate the similarity. The results show that the liquid phase fingerprints and gas phase fingerprints of the ether part of China Jinjiu and the ethyl acetate part of 5% isoamyl alcohol The similarity of the six test results of the spectrum is greater than 0.9000, and the method has good reproducibility;

(2) Investigation on the Similarity of Fingerprints of Ten Batches of Samples

The similarity of 10 batches of Chinese Jinjiu fingerprints collected randomly was investigated, and the calculation results showed that the similarity of the 10 batches was greater than 0.9000, suggesting that this method can not only control the internal quality of Jinjiu products as a whole, but also Can effectively monitor the stability of the production process and the stability of the product;

(3) Comparison of similar products on the market

Using this method to detect 8 kinds of similar products on the market, and compare the fingerprints of each product, it can be clearly seen that the liquid phase fingerprints of the ether part of these 8 kinds of health wines and the ethyl acetate part of 5% isoamyl alcohol are different from those of the ethyl acetate parts of 5% isoamyl alcohol. There are great differences in the Chinese Jinjiu produced by our company, which shows that the detection method of the present invention has good specificity and characteristics; it can not only control the product quality, but also effectively identify the products of our company;

4. Description of drawings

Figure 1 is the comparative fingerprint of the ether part of Chinese Jinjiu

Figure 2 is the control fingerprint of the ethyl acetate part of 5% isoamyl alcohol in Chinese Jinjiu

Figure 3 is the fingerprint of Chinese Jinjiu finished product gas phase comparison

Figure 4 is the common pattern map of the ether part of Chinese Jinjiu

Figure 5 is the common pattern map of the ethyl acetate part of 5% isoamyl alcohol in Chinese Jinjiu

Figure 6 is a part of the shared model map of the gas phase of Chinese Jinjiu

Figure 7 is the fingerprint of the ether part of China Jinjiu with batch number 20041220

Figure 8 is the fingerprint of the ethyl acetate part of 5% isoamyl alcohol in China Jinjiu with batch number 20041220

Figure 9 is the fingerprint of the gas phase part of the batch number 20041220 Chinese Jinjiu

Figure 10 is the fingerprint of the ether part of China Jinjiu with batch number 20050425

Figure 11 is the fingerprint of the ethyl acetate portion of 5% isoamyl alcohol in China Jinjiu with batch number 20050425

Figure 12 is the fingerprint of the gas phase part of the batch number 20050425 Chinese Jinjiu

Figure 13 is the fingerprint of the ether part of Chinese Jinjiu with batch number 20050723

Figure 14 is the fingerprint of the ethyl acetate portion of 5% isoamyl alcohol in China Jinjiu with batch number 20050723

Figure 15 is the fingerprint of the gas phase part of the batch number 20050723 Chinese Jinjiu

Figure 16 is the similarity evaluation results of the fingerprints of the ether parts of China Jinjiu with three batch numbers of 20041220, 20050425 and 20050723

Figure 17 is a graph showing the similarity evaluation results of the fingerprints of the ethyl acetate part of 5% isoamyl alcohol in three batches of China Jinjiu 20041220, 20050425 and 20050723

Figure 18 is a graph of the similarity evaluation results of the fingerprints of the gas phase part of the three batches of 20041220, 20050425, and 20050723 Chinese Jinjiu

Figure 19 is the fingerprint of ferulic acid standard product

Figure 20 is the fingerprint of icariin standard product

Figure 21 is the fingerprint of bornyl acetate standard product

Figure 22 is the fingerprint of eugenol standard product

(5) Specific implementation methods:

The batch number of embodiment 1 is the quality inspection of 20041220 Chinese Jinjiu

1. Sample source Jinpai company warehouse

2. Sample batch number 20041220

3. Preparation of standard solution

a. Liquid phase part

Take an appropriate amount of ferulic acid standard substance and make a 1mg/ml solution with 50% methanol; take an appropriate amount of icariin standard substance and make a 1mg/ml solution with 50% methanol;

b. Gas phase part

Use a glass capillary to dip an appropriate amount of standard products of bornyl acetate and eugenol into a headspace vial, seal it, and obtain it;

4. Preparation of the test solution

Take 100ml of Chinese Jinjiu with batch number 20041220, and evaporate in a water bath at 40°C to nearly dryness, load the sample on a macroporous adsorption chromatography column (12mm×300mm) equipped with AB-8, and elute with water until there is no sugar reaction, (ie Molish reaction is negative), then elute with 80ml 95% ethanol, collect eluent, evaporate to dryness under reduced pressure. Add 20ml of water, sonicate to dissolve, and transfer to a separatory funnel. First extract 3 times with saturated diethyl ether, 20ml each time, combine the extracts, evaporate to dryness in a water bath, dilute to 2ml with tetrahydrofuran, shake well, and filter with a 0.45μ microporous membrane to obtain the test solution of the diethyl ether part; This solution was extracted 3 times with 5% ethyl acetate of isoamyl alcohol saturated with water, 20ml each time, the extracts were combined, evaporated to dryness under reduced pressure, fixed to 2ml with tetrahydrofuran, shaken up, and filtered with a 0.45μ microporous membrane. Promptly get the need testing solution of the ethyl acetate part of 5% isoamyl alcohol;

5. Main instruments and reagents

a. Instrument

Chromatography instrument Varian company instrument (230 pump 330 detector 7725i manual injector AT-830 column thermostat) and Agilent6890

Japan EYELA1000 rotary evaporator

b. Test drug

Diethyl ether, produced in Tianjin, analytically pure

Isoamyl alcohol, produced in Tianjin, analytically pure

Ethyl acetate, produced in Beijing, analytically pure

Acetonitrile USAJ.T.Baker chromatographically pure

Phosphoric acid, produced in Beijing, analytically pure

Water Robust purified water once distilled

N2 high-purity nitrogen, purity 99.999%

Sample of Chinese Jinjiu produced by Hubei Jinpai Company

6. Chromatographic conditions

a. Liquid phase part

Column

Ether site: Hypersil BDS C18, 4.6×250mm, 5μm

Ethyl acetate part of 5% isoamyl alcohol: Aichrombond AQ C18, 4.6×250mm, 5um

mobile phase

Diethyl ether part: Acetonitrile-0.01% phosphoric acid aqueous solution is gradient elution of the mobile phase, when the retention time is 0min, 45min, 80min, 90min, 105min, the volume ratio of acetonitrile and 0.01% phosphoric acid aqueous solution is 8:92, 30:70, respectively, 50:50, 80:20, 80:20;

Ethyl acetate part of 5% isoamyl alcohol: acetonitrile-0.01% phosphoric acid aqueous solution is gradient elution of mobile phase. 15:85, 30:70, 55:45, 100:0, 100:0;

Flow rate: 0.8ml/min

Detection wavelength: 280nm

Column temperature: 30°C

b. Gas phase part

Chromatographic column Agilent HP-5, 30m×320m×0.25μm

Inlet temperature: 230°C

FID detector temperature: 260°C

Heating program: Initially 60°C, keep for 12min, rise to 210°C at 6°C/min, keep for 3min

The carrier gas is N2, the flow rate is 1.0ml/min, the constant flow mode, the split ratio is 5:1

Injector: G1888A headspace autosampler, headspace vial temperature 85°C, injection valve temperature 100°C, transfer line temperature 115°C, headspace vial equilibration time 15min, headspace vial pressurization time 0.5min, quantitative loop filling Time 0.2min, quantitative loop equilibrium time 0.5min, sampling time 1.0min;

7. Detection

a. Liquid phase part

Ether part: Precisely draw 4 μl of the test solution and standard solution of the ether part respectively and inject it into the liquid chromatograph, measure according to the high-performance liquid chromatography in the first appendix of "Chinese Pharmacopoeia" 2000 edition, and obtain the fingerprint of the ether part and the fingerprint of the standard product Atlas

5% ethyl acetate part of isoamyl alcohol: 4 μl each of the test sample and standard solution of the ethyl acetate part of 5% isoamyl alcohol were accurately absorbed, injected into the liquid chromatograph, according to the appendix of "Chinese Pharmacopoeia" 2000 edition Determination by high-performance liquid chromatography, the fingerprints of the ethyl acetate part of 5% isoamyl alcohol and the fingerprints of standard products are obtained;

b. Gas phase part

Directly take the Chinese Jinjiu of the above batch number in the headspace sample bottle, cover and seal; analyze and measure according to the automatic sample injection of the headspace sampler, and obtain the fingerprint of the gas phase part and the fingerprint of the standard product;

8. Results Analysis and Judgment

a. The obtained batch number is 20041220 China Jinjiu Ether part. In the fingerprint spectrum of the test product, 10 characteristic peaks identical to those of the photographic fingerprint spectrum were detected; among them, peak No. 7 was ferulic acid, which was consistent with the fingerprint spectrum of the standard product; for the test The product fingerprint and the corresponding shared pattern were calculated by the computer-aided similarity evaluation system, and the correlation coefficient was 0.9899, which was greater than 0.9000, which was in line with the relevant quality standards formulated by the company.

b. The obtained batch number is 20041220 China Jinjiu 5% isoamyl alcohol in the ethyl acetate portion of the test product fingerprints, 9 characteristic peaks identical to those of the control fingerprints are detected; wherein No. 11 peak is icariin, which is the same as that of the control fingerprint. The fingerprints of the standard product are consistent, and the fingerprints of the test product and the corresponding shared pattern are calculated by the computer-aided similarity evaluation system. The correlation coefficient should be 0.9617, which is greater than 0.9000; it meets the relevant quality standards formulated by our company.

c. The obtained batch number is 20041220 China Jinjiu gas phase part of the test product fingerprints detected 8 characteristic peaks identical to the control fingerprints; wherein peak No. 7 is borneol acetate, peak No. 8 is eugenol, which is consistent with the standard The fingerprints of the products are consistent; the fingerprints of the test products and the corresponding shared pattern are calculated by the computer-aided similarity evaluation system, and the correlation coefficient is 0.9746, which is greater than 0.9000; it meets the relevant quality standards formulated by our company.

Example 2 The batch number is the quality inspection of 20050425 Chinese Jinjiu

1. Sample source Jinpai company warehouse

2. Sample batch number 20050425

3. The preparation of standard solution is the same as in Example 1

4, the preparation of need testing solution is the same as embodiment 1

5. The main instruments and reagents are the same as in Example 1

6. Chromatographic conditions are the same as in Example 1

7. Detection is carried out by the same steps and methods as in Example 1 to detect the liquid phase part and the gas phase part of the standard solution, the test solution of this batch number respectively, and obtain the acetic acid of the corresponding standard product, ether part and 5% isoamyl alcohol respectively. Fingerprints of ethyl ester part and gas phase part;

8. Results Analysis and Judgment

a. The obtained batch number is 20050425 China Jinjiu ether part. In the fingerprint spectrum of the test product, 10 characteristic peaks identical to those of the control fingerprint were detected; among them, peak No. 7 was ferulic acid, which was consistent with the fingerprint of the standard product; the test product The fingerprint and the corresponding shared pattern were calculated by the computer-aided similarity evaluation system, and the correlation coefficient was 0.9890, greater than 0.9000, which was in line with the relevant quality standards formulated by our company.

b. The resulting batch number is 20050425 China Jinjiu 5% isoamyl alcohol in the ethyl acetate portion of the test product fingerprints, 9 characteristic peaks identical to those of the control fingerprints are detected; wherein No. 11 peak is icariin, which is the same as The fingerprints of the standard product are consistent; the fingerprints of the test product and the corresponding shared pattern are calculated by the computer-aided similarity evaluation system, and the correlation coefficient is 0.9837, which is greater than 0.9000; it meets the relevant quality standards formulated by our company.

c. The obtained batch number is 20050425 China Jinjiu gas phase part of the test product fingerprints detected 8 characteristic peaks identical to the control fingerprints; wherein No. 7 peak is borneol acetate, and No. 8 peak is eugenol, which is consistent with the standard The fingerprints of the products are consistent; the fingerprints of the test products and the corresponding shared pattern are calculated by the computer-aided similarity evaluation system, and the correlation coefficient is 0.9953, which is greater than 0.9000; it meets the relevant quality standards formulated by our company.

Example 3 The batch number is the quality inspection of 20050723 Chinese Jinjiu

1. Sample source Jinpai company warehouse

2. Sample batch number 20050723

3. The preparation of standard solution is the same as in Example 1

4, the preparation of need testing solution is the same as embodiment 1

5. The main instruments and reagents are the same as in Example 1

6. Chromatographic conditions are the same as in Example 1

7. Detection is carried out by the same steps and methods as in Example 1 to detect the liquid phase part and the gas phase part of the standard solution, the test solution of this batch number respectively, and obtain the acetic acid of the corresponding standard product, ether part and 5% isoamyl alcohol respectively. Fingerprints of ethyl ester part and gas phase part;

8. Results Analysis and Judgment

a. The obtained batch number is 20050723 China Jinjiu Ether part. In the fingerprint of the test product, the same 10 characteristic peaks as the control fingerprint were detected; the No. 7 peak was ferulic acid, which was consistent with the fingerprint of the standard product; the test product The fingerprint and the corresponding shared pattern were calculated by the computer-aided similarity evaluation system, and the correlation coefficient was 0.9972, greater than 0.9000, which was in line with the relevant quality standards formulated by our company.

b. The obtained batch number is 20050723 China Jinjiu 5% isoamyl alcohol in the ethyl acetate portion of the test product fingerprints, 9 characteristic peaks identical to those of the control fingerprints are detected; wherein No. 11 peak is icariin, which is the same as that of the control fingerprint. The fingerprints of the standard product are consistent; the fingerprints of the test product and the corresponding common pattern are calculated by the computer-aided similarity evaluation system, and the correlation coefficient is 0.9427, which is greater than 0.9000; it meets the relevant quality standards formulated by our company.

c. The batch number obtained is 20050723 China Jinjiu gas phase part of the test product fingerprints detected 8 characteristic peaks identical to the control fingerprints; wherein peak No. 7 is borneol acetate and peak No. 8 is eugenol, which is consistent with the standard The fingerprints of the products are consistent; the fingerprints of the test products and the corresponding shared pattern are calculated by the computer-aided similarity evaluation system, and the correlation coefficient is 0.9930, which is greater than 0.9000; it meets the relevant quality standards formulated by our company.

Example 4 Three batches of Chinese Jinjiu (batch numbers 20041220, 20050425, 20050723) fingerprint similarity evaluation

The fingerprints of each part of the three batch numbers 20041220, 20050425, and 20050723 measured in Example 1, Example 2, and Example 3 are respectively input into the computer-aided similarity evaluation system for calculation, and the correlation coefficients are respectively ether parts: 0.9899 0.9890 0.9972, ethyl acetate part of 5% isoamyl alcohol: 0.9617 0.9837 0.942 7 gas phase part: 0.9746 0.9953 0.9930, all greater than 0.9000, indicating that the quality difference between the three batches is not large, and the product quality is stable.

Claims (4)

1. detect Chinese strength wine method for quality with fingerprint pattern technology, may further comprise the steps:

The first step is set up Chinese strength wine reference fingerprint

(1) preparation of Chinese strength wine

Take off and state parts by weight of Chinese traditional medicine raw material Radix Astragali 10-15, saline cistanche 10-15, thizoma curculiginis 10-15, barrenwort 10-15, matrimony vine 10-15, Chinese yam 10-15, Radix Angelicae Sinensis 10-15, cloves 3-5, Chinese cassia tree 3-5,30-50 ° of distilled liquor diacolation with 50 times of total medicinal material amounts got percolate in 12-15 days, get percolate 70-75 part, it is an amount of to add 20 parts in 40% syrup, pure water 5-10 part, burnt sugar coloring respectively, allotment, filter, put under the normal temperature filtering and impurity removing, the smart filter of filter membrane ageing 3-6 month, can, promptly;

(2) preparation of Chinese strength wine liquid phase part reference substance solution

A. ether position

Get Chinese strength wine 100ml,, use water saturation extracted with diethyl ether 3 times with macroporous absorption chromatographic column desaccharification, each 20ml, combining extraction liquid, evaporate to dryness is settled to 2ml with tetrahydrofuran, membrane filtration, promptly;

B.5% the ethyl acetate extract of isoamylol

With ether position reference substance solution with the ethyl acetate extraction of water saturated 5% isoamylol 3 times, each 20ml, combining extraction liquid, evaporate to dryness is settled to 2ml with tetrahydrofuran, membrane filtration, promptly;

(3) preparation of Chinese strength fume phase part reference substance solution

Directly get Chinese strength wine 5ml in 10ml headspace sampling bottle, seal, promptly;

(4) preparation of standard solution

A. liquid phase part

It is an amount of to get the forulic acid standard items, makes the solution of 1mg/ml with 50% methyl alcohol; It is an amount of to get the icariin standard items, makes the solution of 1mg/ml with 50% methyl alcohol;

B. gas phase part

Dip in glass capillary and to get Bronyl acetate, eugenol standard items in right amount in the headspace sampling bottle, sealing, promptly;

(5) chromatographic condition

A. liquid phase part

Chromatographic column

The ether position: Hypersil C18 post,

The ethyl acetate extract of 5% isoamylol: Aichrombond AQ C18,

Moving phase

The ether position: acetonitrile-0.01% phosphate aqueous solution is the eluent gradient wash-out;

The ethyl acetate extract of 5% isoamylol: acetonitrile-0.01% phosphate aqueous solution is the eluent gradient wash-out;

Flow velocity: 0.8ml/ minute

Detect wavelength: 280nm

Column temperature: 30 ℃

B. gas phase part

Chromatographic column Agilent HP-5

Injector temperature: 230 ℃

Fid detector temperature: 260 ℃

Carrier gas is N2, flow velocity 1.0ml/min, split ratio 5: 1

Injector: G1888A head space automatic sampler;

(6) reference substance detects

A. liquid phase part

The ether position:

Accurate respectively Chinese strength wine liquid phase part ether position reference substance solution and each 4 μ l of standard solution of drawing inject liquid chromatograph, according to high effective liquid chromatography for measuring, promptly get Chinese strength wine ether position reference fingerprint and corresponding standard product finger-print;

The ethyl acetate extract of 5% isoamylol: accurate respectively ethyl acetate extract reference substance solution and each 4 μ l of standard solution that draw Chinese strength wine liquid phase part 5% isoamylol, inject liquid chromatograph, according to high effective liquid chromatography for measuring, promptly get the ethyl acetate extract reference fingerprint and the corresponding standard product finger-print of 5% isoamylol;

B. gas phase part

Respectively gas phase part reference substance solution and standard solution are carried out assay determination by the head-space sampler auto injection, promptly get gas phase part reference fingerprint and corresponding standard product finger-print;

In second step, set up Chinese strength wine finger-print common pattern collection of illustrative plates

Prepare 10 batches Chinese strength wine and standard items, need testing solution to be measured respectively by the preparation method of the preparation method of Chinese strength wine, Chinese strength wine liquid phase part and gas phase part reference substance solution, the preparation method of standard solution, and detect by same procedure one by one, obtain ethyl acetate extract finger-print, gas phase part finger-print and the corresponding standard product finger-print of the Chinese strength wine of each batch ether position finger-print, 5% isoamylol respectively; Import computer aided similarity evaluation system respectively and calculate, can draw the ethyl acetate extract of Chinese strength wine ether position, 5% isoamylol and the finger-print common pattern collection of illustrative plates of gas phase part;

In the 3rd step, test sample detects

1. the preparation of need testing solution

The ethyl acetate need testing solution and the gas phase part need testing solution that prepare liquid phase part ether position and 5% isoamylol with above-mentioned same procedure respectively;

2. prepare liquid phase part and gas phase part of standards product solution respectively with above-mentioned same procedure;

3. measure standard items and test sample to be measured with above-mentioned same procedure;

4, interpretation of result is judged

Should detect 10 characteristic peaks identical with reference fingerprint in the test sample finger-print of ether position, wherein No. 7 peaks are forulic acid, should be consistent with the standard items finger-print; Should detect 9 characteristic peaks identical in the ethyl acetate extract test sample finger-print of 5% isoamylol with reference fingerprint; Wherein No. 11 peaks are the pubeseent epimedium herb glycosides, should be consistent with the standard items finger-print; The finger-print of two parts and corresponding common pattern the collection of illustrative plates auxiliary similarity evaluation system of machine as calculated calculate, and related coefficient should be greater than 0.9000.

Should detect 8 characteristic peaks identical in the gas phase part test sample finger-print with reference fingerprint; Wherein No. 7 peaks are acetic acid bornyl ester, and No. 8 peaks are eugenol, should be consistent with the standard items finger-print; Its finger-print and corresponding common pattern the collection of illustrative plates auxiliary similarity evaluation system of machine as calculated calculate, and related coefficient should be greater than 0.9000.

2. according to claim 1ly detect Chinese strength wine method for quality with fingerprint pattern technology, it is characterized in that: the concrete grammar of the ethyl acetate extract reference substance solution preparation of described Chinese strength wine liquid phase part ether position and 5% isoamylol is, get Chinese strength wine 100ml, 40 ℃ of water-bath rotary evaporations are done near, be splined on 12mm * 300mm macroporous absorption chromatographic column that AB-8 is housed, water is eluted to the sugar-free reaction---and be that the Molish reaction is negative, back 80ml95% ethanol elution is collected eluent, evaporated under reduced pressure, add 20ml water, ultrasonicly make dissolving, and be transferred in the separating funnel, use water saturation extracted with diethyl ether 3 times earlier, each 20ml, combining extraction liquid, water bath method, be settled to 2ml with tetrahydrofuran, shake up, use 0.45 filtering with microporous membrane, promptly get ether position reference substance solution; The ethyl acetate extraction of 5% again that this solution with water is saturated isoamylol 3 times, each 20ml, combining extraction liquid, evaporated under reduced pressure is settled to 2ml with tetrahydrofuran, shakes up, and uses 0.45 filtering with microporous membrane, promptly gets the ethyl acetate extract reference substance solution of 5% isoamylol.

3. according to claim 1ly detect Chinese strength wine method for quality with fingerprint pattern technology, it is characterized in that: described liquid phase part ether position is the eluent gradient wash-out with acetonitrile-0.01% phosphate aqueous solution, its gradient is: when retention time is 0min, 45min, 80min, 90min, 105min, the volume ratio of acetonitrile and 0.01% phosphate aqueous solution was respectively 8: 92,30: 70,50: 50,80: 20,80: 20;

The ethyl acetate extract of 5% isoamylol: acetonitrile-0.01% phosphate aqueous solution is the eluent gradient wash-out, its gradient is: when retention time is 0min, 50min, 80min, 100min, 110min, the volume ratio of acetonitrile and 0.01% phosphate aqueous solution was respectively 15: 85,30: 70,55: 45,100: 0,100: 0.

4. according to claim 1ly detect Chinese strength wine method for quality with fingerprint pattern technology, it is characterized in that: described computer aided similarity evaluation system is meant the computer aided similarity evaluation system by the modernization of cmm center exploitation of Hunan Central South University that pharmacopoeia commission is recommended.

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