CN115518104A - Method for processing fructus aurantii - Google Patents
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Abstract
The invention discloses a method for processing bitter orange, belonging to the field of traditional Chinese medicines. The method comprises the following steps: s1, taking fructus aurantii raw medicinal materials, removing impurities, cleaning, and soaking in water for 4-5 hours; s2, placing the soaked fructus aurantii for 60-96 hours at the temperature of 30-37 ℃ and the relative humidity of 70-90% for fermentation; s3, cleaning the fermented fructus aurantii, steaming for 5-7 hours, stewing for 10-15 hours, slicing and drying. The processing technology of the bitter orange is easy to operate, the production is not influenced by the environment and seasonal climate, the production period is controllable, and the production time in autumn and winter can be greatly shortened; the appearance character of the obtained product meets the processing standard requirement, the similarity of the fingerprint is more than 0.95, and the quality of the sample has better consistency; compared with unprocessed fructus Aurantii, the composition has stronger effect of promoting the small intestine of mice, and can be used for treating functional dyspepsia and other diseases caused by insufficient gastrointestinal motility.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicines, and particularly relates to a method for processing fructus aurantii.
Background
Fructus Aurantii is dry immature fruit of Citrus aurantium L of Rutaceae and its cultivar, and has effects of regulating qi-flowing, relieving epigastric distention, activating stagnancy and relieving flatulence. After being processed, the medicine can relieve the drastic nature of the medicine and achieve the purposes of 'enhancing the effect and reducing dryness'. The bran stir-frying method is the most commonly used method for processing fructus Aurantii, and is recorded in 2020 edition "Chinese pharmacopoeia". The common processing method of fructus aurantii in regions such as Guangdong hong Kong and Macao is a processing method of fermenting and steaming. The method is recorded in 1984 edition of Guangdong province Chinese medicinal decoction pieces processing Specification. The processing method can enhance the effects of relieving epigastric distention, activating stagnancy, regulating qi-flowing and relieving flatulence, and can also achieve the purpose of better relieving dryness.
The existing processing technology of fermenting and steaming the bitter orange has certain disadvantages. For example, the production operation depends on the experience technology of workers, and the key technological parameters are not clear, such as uncertain soaking time and mastery by experience; in addition, fermentation under natural conditions is greatly influenced by temperature and humidity changes of production environments in different seasons, and the production process is longer in dry and cold climates in winter; in summer with high temperature and high humidity, the fermentation process is difficult to control, namely, the phenomenon of drug decay is easy to occur; in addition, the quality of the processed products is also greatly different.
Therefore, the development of a processing method of fructus aurantii with high production efficiency, stable quality and low cost is urgently needed.
Reference:
[1] guangdong province health parlor, guangdong province Chinese medicine processing specification [ M ]. 1984.
[2] Zhanglianjian, li Wei and Liang Zhi Tao, etc. the change of components and the optimization of the process before and after the fermentation and the processing of the bitter orange, the Chinese pharmacy 2017,28 (07) 971-974.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide a method for processing fructus aurantii.
The purpose of the invention is realized by the following technical scheme:
a processing method of fructus aurantii comprises the following steps:
s1, taking fructus aurantii raw medicinal materials, removing impurities, cleaning, and soaking in water for 4-5 hours;
s2, placing the soaked fructus aurantii for 60-96 hours at the temperature of 30-37 ℃ and the relative humidity of 70-90%, and fermenting;
and S3, cleaning the fermented bitter orange, steaming for 5-7 hours, stewing for 10-15 hours, slicing and drying.
Preferably, the temperature in step S2 is 32-37 ℃.
More preferably, the temperature in step S2 is 35-37 ℃.
Preferably, the steaming in step S3 is atmospheric steaming.
Preferably, the steaming time in step S3 is 6 hours.
Preferably, the smoldering time described in step S3 is 12 hours.
Preferably, the temperature of the drying in step S3 is 70-80 ℃.
Compared with the prior art, the invention has the following beneficial effects:
(1) The processing technology of the bitter orange is easy to operate, the production is not influenced by the environment and seasonal climate, the production period is controllable, and the production time in autumn and winter can be greatly shortened.
(2) The processing technology of the fructus aurantii has short fermentation time, and the required fermentation effect can be achieved within 2.5 days at the fastest speed.
(3) The product obtained by the bitter orange processing technology meets the processing standard requirements on appearance characters, the similarity of fingerprint spectra is more than 0.95, and the quality of samples has better consistency.
(4) Compared with the raw bitter orange, the product obtained by the processing technology of the bitter orange has stronger effect of promoting the small intestine of a mouse, and can be used for treating diseases such as functional dyspepsia caused by insufficient gastrointestinal motility.
Drawings
FIG. 1 is a liquid phase fingerprint of different fructus Aurantii samples.
FIG. 2 is a photograph of a fermentation reject; wherein, A is a fermentation unqualified product with black hypha, B is a fermentation unqualified product with green hypha, C is a fermentation unqualified product for producing yellow bacterial plaque, and D is a fermentation unqualified product with rotten medicinal material.
FIG. 3 is a photograph of fructus Aurantii at various stages of processing; wherein A is fructus Aurantii raw material, B is fermentation intermediate (slightly white mycelium), C is fermentation intermediate (all over white mycelium), and D is steamed intermediate.
FIG. 4 is a photograph of the processed fructus Aurantii product.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Unless defined otherwise, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
The instruments and reagents referred to in the following examples:
high performance liquid chromatograph, shimadzu LC-20A, inertsil ODS-3 high performance liquid chromatography column (250X 4.6mm,5 μm);
a constant temperature air-blast drying oven, purchased from Shanghai sperm macro experimental facilities, inc.;
a constant temperature and humidity chamber, LHS-150HC-II, available from Shanghai-Hengchun scientific instruments, inc.;
crude fructus Aurantii was purchased from Setaria Suffruticosa Seisakusho processing factory in Foshan city, and identified as dry immature fruit of Citrus aurantium L. And its cultivar of Rutaceae.
Example 1 determination of fermentation Process
(1) Screening of soaking time before fermentation of fructus aurantii
Weighing the same batch of crude fructus Aurantii, cleaning, soaking in water for 2h, 4h, 5h, 6h, 8h, and 9h respectively, weighing after soaking is stopped, and calculating water absorption capacity of fructus Aurantii; water absorption capacity of fructus aurantii = (weight of fructus aurantii before soaking/weight of fructus aurantii after soaking) × 100%;
and (3) putting the soaked fructus aurantii into a container with air and water permeability, placing the container in a constant-temperature and constant-humidity box, respectively observing the conditions of fermentation for 72h and 96h under the conditions that the temperature is 32 ℃ and the humidity is 70%, and screening the soaking time by taking the standard as the fermentation end point according to the property and the texture (no decay and deterioration) of the medicinal materials and the growth condition (more than 7.5 minutes) of hypha. Wherein, if the texture of the medicinal materials is soft, if the rotten condition exceeds 10%, the medicinal materials are judged to be rotten and go bad, and the scoring standard of the hypha growth condition is shown in a table 1.
The result shows that the soaking time is 1 to 3 hours, the water absorption is less than 60 percent, and the fermentation speed of the medicinal materials is lower; after the soaking time exceeds 6 hours, the water absorption is over 85 percent, and the medicinal materials are easy to soften, rot and deteriorate. See table 3 for details. Therefore, the soaking time is preferably 4 to 5 hours.
TABLE 1 evaluation Table for hypha growth
TABLE 2 soaking time and water absorption of fructus Aurantii
TABLE 3 influence of soaking time before fermentation of Citrus aurantium fruit on fermentation quality
Note: "-" is that the character and the texture meet the requirements; "+" indicates the appearance of rotting and deterioration.
(2) Screening of fermentation temperature and fermentation time of fructus aurantii
Weighing the same raw bitter orange, cleaning, adding water, soaking for 4h, placing in a container with air and water permeability, observing the fermentation conditions at different fermentation temperatures of 27 ℃, 30 ℃, 32 ℃, 35 ℃ and 37 ℃ and different fermentation times in a constant temperature and humidity box under the conditions of 70% relative humidity and 90% relative humidity, and screening the fermentation temperature and time by taking the standard as the fermentation end point according to the property and the texture (no putrefaction and deterioration) of the medicinal material and the growth condition (more than 7.5 minutes) of hypha.
As a result, it was found that the higher the fermentation temperature, the shorter the time required for fermentation. When the fermentation temperature is lower than 30 ℃, the fermentation time is longer, and the rotting phenomenon of the medicinal materials begins to appear, and the results are shown in table 4. Therefore, the fermentation temperature is preferably determined to be 30-37 ℃. During fermentation of fructus Aurantii, hyphae appear after 24h, hyphae grow rapidly after 48h, and fermentation requirements can be met at high temperature and relative humidity for 60 h. However, after the fermentation time exceeds 96h, black hypha is fully distributed on the surface of the medicinal material, and the fermentation time is continuously prolonged, so that the medicinal material is rotten or has yellow or green and other variegated bacterial plaques. The results are shown in Table 5. Therefore, the fermentation time is preferably 60-96 h.
TABLE 4 influence of different fermentation temperatures of Citrus aurantium fruit on fermentation quality
TABLE 5 influence of different fermentation times of Citrus aurantium fruit on fermentation quality
(3) Screening of fermentation relative humidity of fructus aurantii
Weighing the same raw bitter orange, cleaning, soaking in water for 4h, placing in a container with air and water permeability, observing fermentation conditions under the conditions of different relative humidities of 50%, 70%, 80%, 85%, 90% and 95% in a constant temperature and humidity box at 32 ℃, and screening the fermentation temperature according to the character and texture (no putrefaction and deterioration) of the medicinal material and the growth condition of hypha (more than 7.5 minutes) by taking the standard as the fermentation end point.
The results are shown in Table 6, the fermentation time is too long at a relative humidity of 50%; when the relative humidity is 95%, the medicinal materials are rotten and deteriorated in less than 48 hours, and yellow or green bacterial plaques are generated. Therefore, the relative humidity of fermentation is preferably 70-90%.
TABLE 6 influence of different relative humidities on fermentation quality of Citrus aurantium
FIG. 2 is a photograph of a partially fermented reject. Wherein, A is a fermentation unqualified product with black hypha, B is a fermentation unqualified product with green hypha, C is a fermentation unqualified product for producing yellow bacterial plaque, and D is a fermentation unqualified product with rotten medicinal materials.
The tests show that the fermentation speed and the fermentation temperature are in positive correlation with the relative humidity during fermentation. When the fermentation temperature is lower than 30 ℃ and the relative humidity is lower than 70%, the fermentation time is long, the fermentation degree is not uniform, and the sample is rotten. If the relative humidity is high during fermentation, and if the relative humidity exceeds 95%, the fermentation time is short, but the sample is easy to rot or variegated bacterial plaque is easy to appear.
Therefore, the optimized fermentation process is as follows: taking fructus Aurantii raw medicinal materials, removing impurities, cleaning, and soaking in water for 4-5 hr; and (3) placing the soaked fructus aurantii in a constant-temperature constant-humidity incubator, and fermenting for 60-96 hours at the temperature of 30-37 ℃ and the relative humidity of 70-90%.
Example 2 processing of fructus Aurantii
Taking fructus Aurantii raw medicinal materials, removing impurities, cleaning, and soaking in water for 4 hr; placing the soaked fructus Aurantii in a constant temperature and humidity incubator at 35 deg.C and 85% relative humidity for 72 hr; cleaning the fermented fructus aurantii, putting the fructus aurantii into a steaming container, steaming for 6 hours, sealing for 12 hours, slicing, drying, and recording as a fructus aurantii sample S1.
Taking fructus Aurantii raw medicinal materials, removing impurities, cleaning, and soaking in water for 5 hr; placing the soaked fructus Aurantii in a constant temperature and humidity incubator at 30 deg.C and relative humidity of 80% for 96 hr; cleaning the fermented fructus aurantii, putting the fructus aurantii into a steaming container, steaming for 6 hours, sealing for 12 hours, slicing, and drying to obtain a sample S2 for preparing the fructus aurantii.
Taking fructus Aurantii raw medicinal materials, removing impurities, cleaning, and soaking in water for 4 hr; placing the soaked fructus Aurantii in a constant temperature and humidity incubator at 37 deg.C and relative humidity of 90% for 60 hr; cleaning the fermented fructus aurantii, putting the fructus aurantii into a steaming container, steaming for 6 hours, sealing for 12 hours, slicing, drying, and recording as a fructus aurantii sample S3.
Taking fructus Aurantii raw medicinal materials, removing impurities, cleaning, and soaking in water for 5 hr; placing the soaked fructus Aurantii in a constant temperature and humidity incubator at 32 deg.C and relative humidity of 70% for 96 hr; and cleaning the fermented fructus aurantii, putting the fructus aurantii into a steaming container, steaming for 6 hours, sealing for 12 hours, slicing, drying, and recording as a fructus aurantii sample S4.
FIG. 3 is a photograph of fructus Aurantii at different stages of processing. Wherein A is fructus Aurantii raw material, B is fermentation intermediate (slightly white hypha), C is fermentation intermediate (all over white hypha), and D is steamed intermediate. Fig. 4 is a photograph of the processed product of fructus Aurantii.
Example 3 evaluation of quality stability of processed product of bitter orange
Establishing fructus Aurantii liquid phase fingerprint by high performance liquid chromatography, and evaluating quality stability according to fingerprint similarity of different samples
3.1 control solutions
The appropriate amount of naringin, neohesperidin and hesperidin as reference substances are precisely weighed, and methanol is added to prepare a mixed reference substance solution containing 80ug of naringin, 80 mu g of hesperidin and 80 mu g of neohesperidin per 1 mL.
3.2 test article solution
1g of coarse powder (which can completely pass through a second sieve, but is mixed with powder which can pass through a fourth sieve and is not more than 40 percent, namely the powder is precisely weighed, the mixture is placed in a conical bottle with a plug, 90mL of methanol is precisely added, the weight is weighed, the mixture is heated and refluxed for 1.5h at 70 ℃, the mixture is cooled, the weight is weighed again, the weight loss is complemented by methanol, the mixture is shaken up, and the filtrate is filtered to obtain the coarse powder.
3.3 chromatographic conditions
Column Syncronis C18 column (250 mm. Times.4.6 mm,5 μm); mobile phase methanol (a) -0.1% phosphoric acid (B), gradient elution (0-10min, 5% -6% a, 10-20min,6% -15% a, 20-75min,15% -27% a, 75-120min, 27% -65% a). The flow rate is 1mL/min, the column temperature is 25 ℃, the detection wavelength is 283nm, and the injection volume is 0.5 muL.
3.4 methodological considerations
And (3) precision test: taking the same sample solution, continuously injecting samples for 6 times according to the chromatographic condition under the item of 3.3, wherein the sample injection amount is 10uL, recording a chromatogram, taking a naringin peak as a reference peak, calculating the relative retention time of each chromatogram peak and the RSD value of the relative peak area, and if the RSD of the relative peak area of the main chromatogram peak is less than 3%, indicating that the precision of the instrument is good.
And (3) repeatability test: taking 6 parts of the same sample coarse powder, preparing a test solution according to the method under the item '3.3', determining under the chromatographic condition under the item '3.3', recording a chromatogram, taking a naringin peak as a reference peak, calculating the relative retention time of each chromatogram peak and the RSD value of the relative peak area, and if the RSD of the relative peak area of the main chromatogram peak is less than 3%, indicating that the method has good repeatability.
And (3) stability test: and taking the same test solution, respectively injecting samples at 0h,4h,8h,12h, 1111h, 20h and 24h after preparation, determining according to chromatographic conditions under the item of 3.3, recording a chromatogram, calculating the relative retention time of each chromatographic peak and the RSD value of the relative peak area by taking a naringin peak as a reference peak, and if the RSD of the relative peak area of a main chromatographic peak is less than 3%, indicating that the stability of the test solution in 24h is good.
3.5 Generation of liquid phase fingerprint and similarity evaluation
3.5.1 creation of fingerprint
Taking a proper amount of 4 samples prepared in the embodiment 2, preparing a sample solution according to the item 3.2, injecting samples according to the item 3.3, introducing a sample chromatogram into a traditional Chinese medicine chromatogram fingerprint similarity evaluation system 2012, performing chromatogram peak matching, and calculating the similarity. The results are shown in FIG. 1 and Table 7. The result shows that the similarity is more than 0.95, which shows that the similarity is better and the quality is stable.
TABLE 7 calculation of fingerprint similarity of different samples of fructus Aurantii
Example 4 evaluation of drug efficacy of processed product of Citrus aurantium
4.1 reagent and animal
The mosapride citrate dispersible tablet is purchased from Dookong pharmaceutical industry group, limited, batch number: 170801; SPF-grade Kunming (KM) mice, half male and female, with body weights (20 + -2 g), were provided by the Experimental animals center of Guangzhou university of traditional Chinese medicine.
4.2 methods and results
4.2.1 preparation of Water decoction
Taking 30g of raw fructus aurantii and prepared fructus aurantii (sample 1) respectively, adding 300mL of water, soaking for 30min, decocting for 30min, filtering with gauze, adding 240mL of water into residues, decocting for 20min, filtering with gauze, combining filtrates, concentrating to 30mL, preparing into liquid medicine with concentration of 1g/mL, and refrigerating and storing in a refrigerator.
4.2.2 preparation of solutions for Positive control and blank control
Mosapride citrate is used as a positive control drug, a reference (Chenfuxin, hou's book, liwenbin, luyuan Long, tang Jianhua. Mosapride is used for optimal dose screening [ J ]. Chinese veterinary journal, 2019,55 (12): 127-129) of a mouse test and a pre-test result determine that the administration dose of the mouse is 10mg/kg of body weight, and a certain amount of Mosapride citrate dispersible tablets are prepared into an aqueous solution with the concentration of 0.5mg/mL for later use. Distilled water was taken as a blank control solution.
4.2.3 preparation of a nutritional semi-solid paste
Dissolving 5g of sodium carboxymethylcellulose (CMC-Na), 8g of milk powder, 5g of sugar, 4g of starch and 5g of active carbon in 250mL of water, uniformly stirring to prepare black semisolid paste, and refrigerating in a refrigerator until the temperature is recovered to the room temperature when the paste is used.
4.2.4 semi-solid paste Propulsion experiment
Taking 32 KM mice, weighing 18-22g, half each female and half, adaptively feeding for 3 days, randomly dividing the KM mice into 4 groups, 8 groups each, namely a blank group, a positive medicine group, a raw Zhi shell group and a prepared Zhi shell group. The positive medicine group and each administration group are subjected to intragastric administration according to the dose of 0.2mL/10g, corresponding liquid medicine is administered to the blank group, distilled water with the corresponding volume is administered for 4 days continuously, the medicine is taken once a day, fasting is carried out for 14 hours before the last administration, 0.6 mL/half nutritional semisolid paste is administered after the last administration for 1 hour, the patient is killed after taking off the neck after 5 minutes, the abdominal cavity is immediately cut open, the cardia and the pylorus of the stomach are ligated, the patient quickly leaves the stomach and intestine from the cardia to the cecum part, the separated stomach and intestine is taken out to be placed on an experiment table, and the small intestine is naturally laid on white paper without being dragged. The length of the pylorus to the cecum and the distance from the pylorus to the front of the semi-solid paste were measured, and the small intestine propulsion rate was calculated according to the following formula:
small intestine propulsion (%) = [ distance from pylorus to indicator front edge (cm)/pylorus to full length of ileocecal portion (cm) ] + 100%.
4.2.5 statistical processing methods
The experimental data were processed using SPSS 22.0 statistical software toExpressed, a one-way analysis of variance was used for the test.
4.2.6 Effect of raw and processed fructus Aurantii on the intestinal motility of mice
The results are shown in Table 8. The results in the table show that the mean values of the small intestine propulsion rates of the raw fructus aurantii and the prepared fructus aurantii are both obviously greater than those of the blank group, and the prepared fructus aurantii has better effect on the small intestine propulsion of mice than the raw fructus aurantii.
TABLE 8 influence of raw and processed fructus Aurantii on the intestinal motility of normal mice
Note: * P <0.05 (bilateral level) compared to blank; * P <0.01 (bilateral levels) compared to blank.
And (4) conclusion: fructus Aurantii has the effects of regulating qi, relieving epigastric distention, activating stagnancy and relieving flatulence, and is clinically used for treating gastrointestinal diseases such as abdominal distension and abdominal pain. Experiments show that the processed fructus aurantii has stronger effect of promoting the small intestine of a mouse compared with the unprocessed fructus aurantii. The indication shows that the traditional Chinese medicine composition has better curative effect on diseases such as functional dyspepsia caused by insufficient gastrointestinal motility.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such modifications are intended to be included in the scope of the present invention.
Claims (7)
1. A processing method of fructus aurantii is characterized by comprising the following steps: the method comprises the following steps:
s1, taking fructus aurantii raw medicinal materials, removing impurities, cleaning, and soaking in water for 4-5 hours;
s2, placing the soaked fructus aurantii for 60-96 hours at the temperature of 30-37 ℃ and the relative humidity of 70-90% for fermentation;
and S3, cleaning the fermented bitter orange, steaming for 5-7 hours, stewing for 10-15 hours, slicing and drying.
2. The processing method of fructus aurantii according to claim 1, wherein:
the temperature in step S2 is 32-37 ℃.
3. The processing method of fructus aurantii as claimed in claim 1, wherein:
the temperature in step S2 is 35-37 ℃.
4. The processing method of bitter orange according to any one of claims 1 to 3, wherein:
the steaming in the step S3 is normal-pressure steaming.
5. The processing method of fructus aurantii according to any one of claims 1 to 3, wherein:
the steaming time in step S3 is 6 hours.
6. The processing method of bitter orange according to any one of claims 1 to 3, wherein:
the smoldering time described in step S3 was 12 hours.
7. The processing method of bitter orange according to any one of claims 1 to 3, wherein:
the temperature of the drying in step S3 is 70-80 ℃.
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CN101612234A (en) * | 2009-07-15 | 2009-12-30 | 江西樟树天齐堂中药饮片有限公司 | A kind of method of Chinese medicine processing |
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夏荃等: "岭南特色炮制工艺对枳壳挥发油成分的影响", 中国实验方剂学杂志 * |
张栋健等: "枳壳发酵炮制前后的成分变化及工艺优化", 中国药房 * |
张金莲等: "多指标正交法优选樟帮枳壳饮片炮制工艺", 中成药 * |
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