Method for processing leeches processed with wine and vinegar

Abstract

The invention discloses a method for processing leeches processed with wine and vinegar, and belongs to the technical field of traditional Chinese medicine processing technologies. The invention aims to solve the problems of low dissolution rate of effective components, low anticoagulant property and fishy smell in the existing leech processing method. The method comprises the following steps: adding short-section leeches into a wine-vinegar mixed solution, fully shaking to uniformly mix, sealing, moistening at room temperature, putting into a preheated herbal medicine roaster, continuously stir-frying, taking out when stir-frying to a specified degree, cooling, crushing and sieving to obtain the wine-vinegar-processed leeches. According to the method, the vinegar and the wine are used as auxiliary materials to jointly process the leeches, so that the dissolution of effective components in the leeches can be remarkably improved, the toxicity of the leeches is reduced, meanwhile, the peculiar smell of the leeches is reduced, and a relatively strong scientific basis is provided for establishing a leech processing method.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine processing technology, and particularly relates to a method for processing leeches with wine and vinegar. Background Art

[0002] Leeches are commonly used animal traditional Chinese medicines in my country. They are salty, bitter, flat, slightly toxic, and belong to the liver meridian. They have the effects of breaking blood, promoting menstruation, removing blood stasis and eliminating symptoms. Leeches contain chemical components such as protein polypeptides, amino acids, pteridine components, lipids, inorganic trace elements and nucleosides. Among them, protein polypeptide macromolecules are the main physiologically active components of leeches and are the hotspot of current research. At present, more than 40 active ingredients such as hirudin, hirudin, and histamine have been discovered. Among them, hirudin is the most active natural thrombin inhibitor discovered so far, which can directly act on the coagulation system. Amino acids are the basic units that make up proteins. There are 20 kinds of amino acids that make up proteins in organisms. The test results show that the total amino acid content of medicinal leeches is above 58%, and it contains 18 kinds of amino acids, 7 of which are essential amino acids for the human body. Except for cysteine ​​which was not detected in the report, the highest content of hydrolyzed amino acids is glutamic acid, with a content range of 7.26-18.36 mg / g; the highest content of free amino acids is alanine, with a content range of 1.43-4.39 mg / g.

[0003] According to the Shennong's Herbal Classic, leeches are highly effective and slightly toxic, and are classified as inferior. Traditional Chinese medicine theory believes that processing can reduce the toxicity of leeches. At the same time, leeches have a special fishy smell, so in clinical practice, leeches are usually processed before use. The modern processing specifications for Chinese herbal medicine slices collected in various provinces (cities, autonomous regions) mainly focus on the two processing methods of "talcum powder scalding" and "wine roasting". Some regions also retain processing methods with regional characteristics. For example, the "Hunan Province Chinese Herbal Medicine Slice Processing Specifications" records the "oil processing" processing method, and the "Gansu Province Chinese Herbal Medicine Processing Specifications" records the processing method of leech "ultrafine slices". However, these processing methods are mainly concentrated on one kind of auxiliary material processing, which has the disadvantages of low anticoagulant activity, fishy smell, and is not easily accepted by patients. Summary of the invention

[0004] The invention aims to solve the problems of low effective component dissolution rate, low anticoagulant activity and fishy smell in the existing leech processing method, and to provide a wine vinegar processing method for leeches.

[0005] A method for preparing leeches with wine and vinegar is specifically completed by the following steps:

[0006] Add short pieces of leeches into the wine and vinegar mixed solution, shake thoroughly to mix, seal, and let it sit at room temperature, then put it into a preheated medicine stir-frying machine, stir-fry continuously, take it out when it is stir-fried to a specified degree, cool it, crush it, and sieve it to obtain leeches made with wine and vinegar.

[0007] Principle of the present invention:

[0008] Wine itself is fermented and has a certain aroma after fermentation. It is used as an auxiliary material as a fermentation solvent, which can change the pharmacological properties of the drug to a certain extent. In the application of leeches, the main function of auxiliary material wine is to remove the fishy smell and reduce the toxicity of the drug. In addition, leeches have a strong fishy smell and are difficult to take raw. Auxiliary material wine can be used as a flavor corrector to remove the fishy smell and make the drug easier to take.

[0009] Vinegar has a long history of being used as an auxiliary material in the preparation of traditional Chinese medicine. The main purposes of using vinegar as an auxiliary material in the preparation of medicinal materials are as follows: 1. To guide the medicine into the liver, and enhance the blood circulation, blood stasis, qi circulation, and pain relief effects of the medicine; 2. To increase the dissolution of the active ingredients of the medicine, especially to enhance the dissolution and absorption of fat-soluble ingredients; 3. To reduce the toxicity of the medicine; 4. To correct odor and taste for ease of taking. Vinegar can combine with trimethylamine components with a fishy smell to form salts, thereby removing the fishy smell of the medicine; 5. Preservative effect.

[0010] According to the above analysis, 1. Leeches enter the liver meridian, and vinegar roasting can lead drugs into the liver. The two promote each other and synergistically exert the medicinal effects of leeches. 2. Roasting can increase the dissolution and absorption of active ingredients of drugs, especially fat-soluble ingredients. Vinegar-soaked drugs have the effects of leading drugs into the liver and promoting the dissolution of active ingredients. Therefore, wine and vinegar co-production can simultaneously play the role of "wine production lifting" and "vinegar production entering the liver", providing experimental basis for the rational development and utilization of wine and vinegar roasting of leeches.

[0011] Advantages of the present invention:

[0012] The invention provides a wine and vinegar leech preparation method. The method has reasonable process design, strong operability, can significantly enhance the dissolution of effective components, reduce the toxicity of leeches, and reduce the peculiar smell of leeches.

[0013] Unprocessed leeches have a distinct fishy smell, mainly carboxylic acids, aldehydes, phenols, hydrocarbons, etc. The main substances that provide the fishy smell of leeches are propionic acid, butyric acid, isovaleric acid, acetic acid, 4-methylphenol, benzaldehyde, 2-methylbutyric acid, etc., accounting for 34.667%, 14.093%, 12.559%, 11.387%, 7.219%, 2.955%, and 2.324%, respectively. After being roasted with wine, vinegar, and wine and vinegar, the contents of these volatile components changed to varying degrees, which improved the fishy smell of leeches. The results of acute toxicity experiments showed that when the dosage was far higher than the clinically recommended dosage, the processed leeches had not yet shown toxicity. This shows that there is no risk of severe acute poisoning after freezing and processing of leeches, and the short-term commonly used dosage in clinical practice is safe and reliable. DETAILED DESCRIPTION

[0014] Specific implementation method 1: This implementation method is a method for preparing leeches with wine and vinegar, which is specifically completed according to the following steps:

[0015] Add short pieces of leeches into the wine and vinegar mixed solution, shake thoroughly to mix, seal, and let it sit at room temperature, then put it into a preheated medicine stir-frying machine, stir-fry continuously, take it out when it is stir-fried to a specified degree, cool it, crush it, and sieve it to obtain leeches made with wine and vinegar.

[0016] Specific embodiment 2: This embodiment differs from specific embodiment 1 in that the short leeches are washed and then hung to dry, and the length is 10 mm to 15 mm. The other steps are the same as those of specific embodiment 1.

[0017] Specific implementation method three: This implementation method is different from specific implementation method one or two in that the mass ratio of the short-segment leech to the wine vinegar mixed solution is (10-30): 100. The other steps are the same as those of specific implementation method one or two.

[0018] Specific embodiment 4: This embodiment differs from specific embodiments 1 to 3 in that the volume ratio of wine to vinegar in the wine-vinegar mixed solution is (1-3):(1-3). The other steps are the same as those of specific embodiments 1 to 3.

[0019] Specific implementation mode 5: This implementation mode is different from specific implementation modes 1 to 4 in that the wine in the wine-vinegar mixed solution is yellow wine, and the alcohol content of the yellow wine is 15 to 20 degrees. The other steps are the same as those of specific implementation modes 1 to 4.

[0020] Specific implementation method 6: This implementation method is different from specific implementation methods 1 to 5 in that the vinegar in the wine vinegar mixed solution is rice vinegar, and the degree of rice vinegar is 3.5 to 9 degrees. The other steps are the same as those of specific implementation methods 1 to 5.

[0021] Specific implementation method 7: This implementation method is different from specific implementation methods 1 to 6 in that the time of the soaking is 30 minutes to 150 minutes. The other steps are the same as those of specific implementation methods 1 to 6.

[0022] Specific implementation eight: This implementation differs from specific implementations one to seven in that the power of the stir-frying machine is 400W to 800W, and the stir-frying time is 20min to 30min. The other steps are the same as those of specific implementations one to seven.

[0023] Specific embodiment 9: This embodiment differs from specific embodiments 1 to 8 in that the shaking process is: shaking at room temperature for 5 to 7 hours, with a shaking frequency of 100 times / min. The other steps are the same as those of specific embodiments 1 to 8.

[0024] Specific embodiment 10: This embodiment differs from specific embodiments 1 to 9 in that the sieving is through a No. 3 sieve. The other steps are the same as those of specific embodiments 1 to 9.

[0025] The following examples are used to verify the beneficial effects of the present invention:

[0026] Embodiment 1: A method for preparing leeches with wine and vinegar, which is specifically completed by the following steps:

[0027] Add the short leeches into the wine-vinegar mixed solution, shake thoroughly to mix, seal, and steam at room temperature, then put into a preheated medicine stir-frying machine, stir-fry continuously, and keep the bottom of the pot shining until the leeches are completely dry and the surface color deepens, then remove from the pot, take out, cool, crush, and sieve to obtain wine-vinegar leeches;

[0028] The short-segment leeches are washed and then hung to dry, and have a length of 15 mm;

[0029] The mass ratio of the short-segment leech to the wine-vinegar mixed solution is 20:100;

[0030] The volume ratio of wine to vinegar in the wine-vinegar mixed solution is 1.1:1;

[0031] The wine in the wine-vinegar mixed solution is yellow rice wine, and the degree of the yellow rice wine is 15 degrees;

[0032] The vinegar in the wine-vinegar mixed solution is rice vinegar, and the degree of the rice vinegar is 6 degrees;

[0033] The preheating method of the preheated medicine frying machine is as follows: preheat the medicine frying machine, heat the frying pan at an outer ring firepower of 2100W for 1 to 2 minutes until the temperature of the bottom of the pan rises to about 150°C (slow fire);

[0034] The time of moistening is 90 minutes;

[0035] The power of the medicine frying machine is 800W, and the frying time is 20min;

[0036] The sieving is through a No. 3 sieve;

[0037] The shaking process is as follows: shaking at room temperature for 6 hours, with a shaking frequency of 100 times / min.

[0038] Embodiment 2: This embodiment differs from Embodiment 1 in that the volume ratio of wine to vinegar in the wine-vinegar mixed solution is 1:2. Other steps and parameters are the same as those in Embodiment 1.

[0039] Example 3: The difference between this example and Example 1 is that the volume ratio of wine to vinegar in the wine-vinegar mixed solution is 3:1; the mass ratio of the short-segment leech to the wine-vinegar mixed solution is 25:100. The other steps and parameters are the same as those in Example 1.

[0040] Example 4: The difference between this example and Example 1 is that the volume ratio of wine to vinegar in the wine-vinegar mixed solution is 3:1; the soaking time is 180 minutes. The other steps and parameters are the same as those in Example 1.

[0041] Comparative Example 1: The difference between this comparative example and Example 1 is that the wine vinegar mixed solution is replaced with yellow rice wine, and the degree of the yellow rice wine is 15 degrees. The other steps and parameters are the same as those in Example 1.

[0042] Comparative Example 2: The difference between this comparative example and Example 1 is that the wine vinegar mixed solution is replaced with rice vinegar, and the degree of rice vinegar is 6 degrees. The other steps and parameters are the same as those in Example 1.

[0043] Comparative Example 3: The difference between this comparative example and Example 1 is that the wine vinegar mixed solution is replaced by salt water, and the mass fraction of the salt water is 5%. The other steps and parameters are the same as those in Example 1.

[0044] The total amino acid content of the wine vinegar leeches obtained in Examples 1 to 4 and the processed leeches obtained in Comparative Examples 1 to 3 was tested, and the specific method is as follows:

[0045] Determination of total amino acid content:

[0046] Chromatographic conditions:

[0047] Chromatographic column: kromasil 100-5-C8 (4.6×250mm, 5μm)

[0048] Mobile phase: Mobile phase A: 0.06 mol / L sodium acetate (adjusted to pH 6.40 with glacial acetic acid)-acetonitrile (200:30); Mobile phase B: acetonitrile-water (50:50), Detection wavelength: UV detector (360 nm), Flow rate: 0.8 ml / min, Column temperature: 40 °C;

[0049]

[0050] Preparation of reference solution:

[0051] Accurately weigh 10 reference substances including aspartic acid, glutamic acid, threonine, proline, alanine, valine, isoleucine, leucine, lysine and tyrosine respectively, place them in a 200ml volumetric flask, add 0.1% hydrochloric acid solution to dissolve them and make up to the scale.

[0052] Preparation of test solution

[0053] Take about 1g of this product, weigh it accurately, put it in a stoppered conical bottle, accurately add 25ml of 0.1mol / L hydrochloric acid solution, weigh it, extract it in a water bath under reflux for 1.5 hours, place it at room temperature, weigh it, make up the weight loss with 0.1mol / L hydrochloric acid solution, centrifuge it to get the supernatant, and filter it. Accurately measure 2ml of the filtrate, accurately add 2ml of hydrochloric acid, and place it at 105℃ for 24 hours. After taking it out and cooling it, transfer it to a 25ml volumetric bottle, add 4ml of 6mol / L sodium hydroxide solution, and dilute it to the scale with 0.1mol / L hydrochloric acid solution. Shake it well, filter it, and you have it.

[0054] Linear relationship investigation

[0055] Accurately take 0.1ml, 0.25ml, 0.5ml, 1ml, 2ml, and 3ml of the reference solution, accurately draw 5μl of the reference solution, inject it into the liquid chromatograph, and record the peak area. Perform linear regression with the peak area (Y) of each amino acid as the ordinate and the concentration (X, μg) as the abscissa. The concentrations of the 10 amino acids obtained are all in the range of 0.05μg to 0.18μg and show a good linear relationship with their respective peak areas.

[0056] Precision test

[0057] Accurately pipette 5 μl of the reference solution, inject it into the liquid chromatograph, and record the peak area. The results show that the RSD values ​​of the peak areas of the 10 amino acids are all less than 0.5%, indicating that the instrument has good precision.

[0058] Repeatability test

[0059] Weigh 1g of sample accurately, weigh 6 portions in parallel, prepare the test solution according to the test solution preparation method and determination method, accurately draw 5μl of reference solution, inject into liquid chromatograph, record the peak area, and calculate the amino acid content. The results show that the RSD of the 10 amino acid contents and the total amino acid content are all less than 2.0%, with good repeatability.

[0060] Stability test

[0061] Take 1g of sample, prepare the test solution according to the test solution preparation method and determination method, accurately draw 5μl of reference solution, inject at 0h, 8h, 15h, 26h, 35h, 44h, 53h, 62h, 71h, 78h, record the chromatographic peak area. The results show that the RSD of the peak area of ​​10 amino acids in the sample within 0-78 hours is less than 2.0%, and the stability is good.

[0062] Recovery test

[0063] Weigh 0.5g of sample accurately and weigh 6 portions in parallel. Prepare the test solution according to the test solution preparation method and the determination method, accurately draw 5μl of the reference solution, inject it into the liquid chromatograph, record the peak area, calculate the amino acid content, and calculate the recovery rate of each amino acid. The results showed that the average recovery rate of the 10 amino acids was between 93% and 100%, and the RSD was less than 3%, which was a good recovery rate.

[0064] The performance of the wine vinegar leeches obtained in Examples 1 to 4 and the processed leeches obtained in Comparative Examples 1 to 3 were tested, as shown in the following table;

[0065]

[0066] The OD values ​​in the above table are determined according to the corresponding contents determined by the "total amino acid determination" and "antithrombin activity" content determination methods. The content determination data are normalized using the Hassan method, and the overall evaluation index (OD value) is obtained after processing.

[0067] From Table 1, we can see that the leech preparation prepared in this experiment has obvious anticoagulant activity and can increase the outflow of total amino acid components. Compared with the preparation of single auxiliary materials, the co-preparation of wine and vinegar can achieve the effect of "1+1>2".

Claims

1. A method for preparing leeches with wine and vinegar, characterized in that The method is specifically completed according to the following steps: Add short pieces of leeches into the wine and vinegar mixed solution, shake thoroughly to mix, seal, and let it sit at room temperature, then put it into a preheated medicine stir-frying machine, stir-fry continuously, take it out when it is stir-fried to a specified degree, cool it, crush it, and sieve it to obtain leeches made with wine and vinegar.

2. The method for preparing leeches with wine and vinegar according to claim 1, characterized in that The short-segment leeches are washed and then hung to dry, and have a length of 10 mm to 15 mm.

3. The method for preparing leeches with wine and vinegar according to claim 1, characterized in that The mass ratio of the short-segment leech to the wine-vinegar mixed solution is (10-30):

100.

4. The method for preparing leeches with wine and vinegar according to claim 1, characterized in that The volume ratio of wine to vinegar in the wine-vinegar mixed solution is (1-3):(1-3).

5. The method for preparing leeches with wine and vinegar according to claim 4, characterized in that The wine in the wine-vinegar mixed solution is yellow rice wine, and the degree of the yellow rice wine is 15 to 20 degrees.

6. The method for preparing leeches with wine and vinegar according to claim 4, characterized in that The vinegar in the wine-vinegar mixed solution is rice vinegar, and the degree of the rice vinegar is 3.5 degrees to 9 degrees.

7. The method for preparing leeches with wine and vinegar according to claim 1, characterized in that The soaking time is 30 minutes to 150 minutes.

8. The method for preparing leeches with wine and vinegar according to claim 1, characterized in that The power of the medicine frying machine is 400W-800W, and the frying time is 20min-30min.

9. The method for preparing leeches with wine and vinegar according to claim 1, characterized in that The shaking process is as follows: shaking at room temperature for 5 hours to 7 hours, with a shaking frequency of 100 times / min.

10. The method for preparing leeches with wine and vinegar according to claim 1, characterized in that The sieving is through a No. 3 sieve.

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