Application of Rhizopus delemar in the preparation of traditional Chinese medicine Fructus Aurantii Immaturus slices

Abstract

The present invention relates to Rhizopus delavirdii Rhizopus delemar The application in the preparation method of the Chinese medicine Citrus aurantium decoction slices belongs to the field of drug preparation. The present invention also provides a preparation method of Citrus aurantium decoction slices. The preparation method of the present invention clarifies the fermentation strain, avoids the interference of miscellaneous bacteria in the air during the fermentation process, and ensures the safety of Citrus aurantium decoction slices. At the same time, the key process parameters of fermentation and steaming of Citrus aurantium decoction slices are optimized and determined to ensure the stability of the quality of Citrus aurantium decoction slices. Compared with the currently naturally fermented Citrus aurantium decoction slices, the Citrus aurantium decoction slices prepared by the method of the present invention have significantly increased contents of synephrine and flavonoids effective ingredients, no harmful components such as aflatoxin are detected, and the quality is better, which is suitable for large-scale production of Citrus aurantium decoction slices.

Description

Technical Field

[0001] The present invention relates to Rhizopus delavirdii Rhizopus delemar The application of the invention in the preparation method of the Chinese medicine Fructus Aurantii Immaturus decoction pieces belongs to the field of Chinese medicine preparation. Background Art

[0002] Citrus aurantium, a plant of the Rutaceae family Citrus aurantium L. and its cultivars or sweet orange Citrus sinensis The dried young fruits of Osbeck are collected from the fallen fruits in May and June, and the impurities are removed. The fruits are cut into two halves from the middle, and dried in the sun or at a low temperature. The smaller ones are directly dried in the sun or at a low temperature.

[0003] At present, the main processing methods of Zhishi are: 1. Remove impurities, wash, moisten, cut into thin slices, and dry. 2. Stir-fry Zhishi with bran: Take Zhishi slices and stir-fry them according to the bran stir-fry method (the method of fumigating the cleaned or cut medicine with wheat bran) until the color becomes darker. Yu Xiaoli, Research on the processing technology and quality standards of Zhishi, a master's thesis of Liaoning University of Traditional Chinese Medicine in 2010, determined the specific cutting technology of Jishi slices, optimized the best processing technology of stir-fried Jishi, and compared the effects of Jishi processing on gastrointestinal function before and after, and formulated the quality standards of stir-fried Jishi. Huang Yanqiu et al., Historical evolution and research progress of the processing technology of Citrus aurantium, Guangdong Chemical Industry, Vol. 50, No. 490, Issue 8, 2023. The processing methods of the past dynasties, changes in chemical composition before and after processing, pharmacological effects and processing technology were summarized. Due to the complex chemical composition of Citrus aurantium, there are certain differences in the chemical composition before and after processing. Most of the research is limited to laboratory research, and the process has not been scaled up, which cannot fully reflect the actual situation of scaled-up production. Wang Cheng, et al., Research Progress on the Processing of Citrus aurantium, Asia-Pacific Traditional Medicine, Volume 19, Issue 6, June 2023, disclosed the processing methods of Citrus aurantium in the past dynasties, including cleaning, cutting, stir-frying (stir-frying until yellow, stir-frying until charcoal), stir-frying with solid auxiliary materials (stir-frying with bran, soil, and flour), roasting (roasting with wine, vinegar, ginger, and honey), burning, steaming, roasting, boiling, soaking in swill, vinegar paste, etc. More than 20 methods; modern pharmacopoeias and local processing specifications basically follow the traditional processing methods of Citrus aurantium, mainly cleaning, cutting, and stir-frying with bran. The research on the processing technology of Citrus aurantium mainly focuses on stir-frying with bran, softening and cutting, and water roasting. After processing, the volatile oil content of Citrus aurantium decreases, the types of components also change, and the content of components such as hesperidin, naringin, and synephrine also decreases.

[0004] There are literature reports that fermentation is used to process Zhishi, such as Xia Quan et al., "Exploring the characteristics of Lingnan processing from the perspective of Lingnan medicine", Chinese Medicinal Materials, Vol. 40, No. 6, June 2017, reported that the processing method of Zhishi is steaming after fermentation, the purpose of which is to alleviate the pungent and dry medicinal properties. The fermentation and steaming process of Zhishi slices is a characteristic processing technology in the Lingnan region. Its standard process flow is included in the Guangdong Provincial Traditional Chinese Medicine Processing Specifications (1984 Edition) under Zhishi, and its processing method is stipulated as follows: Zhishi, remove impurities, wash, soak until 70% thorough, take out, ferment for 3-4 days, wash, steam for 4-6 hours, simmer for 1 night, until the inside is purple-brown, take out, slice, and dry. According to the standard records, the process is relatively rough, and the details of the fermentation, steaming and other processes are not clear. The processing time, heat and degree are mostly determined by the experience of the processing workers. There is a lack of standardized and standardized processes in the processing process, and the quality lacks stability and controllability. At the same time, due to the presence of many harmful bacteria in the air, in the actual fermentation process of Citrus aurantium, it is easy to be accompanied by the growth of miscellaneous bacteria, which will produce a large number of harmful toxins (such as aflatoxin) after growth, affecting the safety of the clinical application of fermented and steamed Citrus aurantium slices. All these are not conducive to the promotion and large-scale processing of Lingnan-style Citrus aurantium slices. Summary of the invention

[0005] Aiming at the problems existing in the fermentation and steaming process of Citrus aurantium decoction pieces, the present invention optimizes a strain of Rhizopus delle that is suitable for fermentation of Citrus aurantium decoction pieces, and determines the preparation process of fermentation and steaming of Citrus aurantium decoction pieces by using the strain of Rhizopus delle through process parameter screening experiments. The shortcomings of the existing processing specifications are solved, which is conducive to ensuring the quality stability and safety of the Citrus aurantium decoction pieces with Lingnan characteristics, and is conducive to the large-scale production of Citrus aurantium decoction pieces.

[0006] The invention provides Rhizopus delavayi Rhizopus delemar Application of the traditional Chinese medicine Zhishi decoction pieces in the processing method, the Dale Rhizopus Rhizopus delemar The registration number with the China Industrial Microbiology Culture Collection Center is CICC40308.

[0007] The present invention also provides a method for preparing Fructus Aurantii Immaturus, which comprises using Rhizopus delavayi Rhizopus delemar The fermentation process of CICC40308 includes the following steps:

[0008] a. Take the sterilized Zhishi medicinal material and moisten it with water;

[0009] b. Take Rhizopus delbrueckii, activate it, and prepare a strain activation solution. The amount of bacteria in the activation solution is (2.0±2.0×10%)×10 8 CFU / mL, dilute with water and spray evenly into the medicinal materials;

[0010] c. Fermentation: temperature 20-30℃, relative humidity 70-90%, fermentation time 3-5d;

[0011] d. Steaming: Steam the fermented medicinal materials in step c for 4-6 hours;

[0012] e.Muffled:

[0013] f. Slice and dry for 3.5-8 hours.

[0014] Further preferably, the sterilization method described in step a is ultraviolet sterilization, and the sterilization time is 30 minutes; the moistening time is 5 hours; the amount of water added is 60% w / w of the weight of Citrus aurantium; the dosage of the strain activation liquid, water and Citrus aurantium described in step b is: 10 ml of strain activation liquid and 40 ml of water per kg of Citrus aurantium; the fermentation conditions described in step c are temperature 25°C, relative humidity 90%, and fermentation time 4 days; the moistening time described in step e is 5 hours; the drying temperature described in step f is: 60°C, and the drying time is 4 hours.

[0015] The present invention also provides a Fructus Aurantii Immaturus decoction piece prepared by the Fructus Aurantii Immaturus processing method, wherein the content of synephrine (mg / g) is not less than 3.0 mg / g; and the content of naringin, naringin, hesperidin and neohesperidin (mg / g) is not less than 160 mg / g.

[0016] Further preferably, the synephrine content (mg / g) is not less than 4.0 mg / g.

[0017] The beneficial effects of the present invention are:

[0018] The processing method of the present invention clarifies the fermentation strain, avoids the interference of miscellaneous bacteria in the air during the fermentation process, and ensures the safety of the Citrus aurantium slices. At the same time, the key process parameters of fermentation and steaming of Citrus aurantium slices are optimized and determined, ensuring the stability of the quality of the Citrus aurantium slices. Compared with the Citrus aurantium slices currently fermented naturally, the Citrus aurantium slices prepared by the method of the present invention have significantly increased contents of synephrine and flavonoids, no harmful components such as aflatoxin are detected, and the quality is better, which is suitable for large-scale production of Citrus aurantium slices. DETAILED DESCRIPTION

[0019] Example 1 Process for preparing fermented Citrus aurantium slices

[0020] Take the Citrus aurantium, sort by size, wash with sterile purified water, sterilize under ultraviolet light for 30 min, add 0.6 times the amount of sterile purified water, mix well, cover and soak for 5 h until there is no white core inside, and place in a sterilized and clean incubator. Rhizopus delemar (Strain number CICC40308) was activated using Sabouraud liquid medium (peptone 10g, glucose 40g, chloramphenicol 0.1g, distilled water to 1000ml) at 25°C and shaker culture for 48h. The bacterial count after activation was (2.0±2.0×10%)×108 CFU / mL. Take 10ml of the above-mentioned bacterial activation solution, add 40ml of distilled water, shake well, spray into 1kg of clean Citrus aurantium, mix well, cover with wet gauze, and cover. Place in an incubator, culture at 25℃ and 90% relative humidity for 4 days, take out, clean the fermented Citrus aurantium, steam it in a steamer for 4-6 hours, steam it overnight (12 hours), take out, slice it, and dry it at 60℃.

[0021] The Rhizopus delbrueckii CICC40308 used in the present invention has a Latin name: Rhizopus delemar , which is a strain of China Center of Industrial Culture Collection (CICC, address: Building 6, No. 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing). The Dale Rhizopus strain CICC40308 described in the present invention can be purchased through domestic and foreign commercial channels before the filing date of the present invention, or can be purchased from China Center of Industrial Culture Collection. CICC has a special website at http: / / www.china-cicc.org. The public can directly order strains online at its website address:

[0022] http: / / m.china-cicc.org / search / ?classtype=0&keyword=40308.

[0023] Example 2 Screening experiment of strains for preparing fermented Citrus aurantium slices

[0024] Take the Zhishi medicinal material, sort it by size, wash it with sterile purified water, sterilize it under ultraviolet light for 30 minutes, add 1 times the amount of sterile purified water, mix it well, cover it and soak it thoroughly until there is no white core inside, and place it in 3 disinfected and clean culture vessels. One of them is naturally fermented; the other is taken from the activation liquid of the Dale Rhizopus strain [the bacterial count is (2.0±2.0×10%)×10 8 CFU / mL], add 4 times the amount of sterile purified water, mix well, and spray evenly into the medicinal materials at a rate of 100g:5ml; take 1 portion of 0.4% of the weight of the immature bitter orange powder (spore content 10 billion / g, manufacturer: Jining Xiangyuan Biotechnology Co., Ltd.), add 20 times the sterile purified water, and spray evenly into the medicinal materials. The above three samples of immature bitter orange are all covered with wet gauze and covered. Place in an incubator and culture for 4 days at a temperature of 25°C and a relative humidity of 90%. Wash the fermented immature bitter orange, steam it in a steamer for 4-6 hours, simmer overnight (12 hours), take out the slices, and dry at 60°C.

[0025] Evaluation indicators:

[0026] 1. Appearance evaluation: Describe the properties of the obtained medicinal pieces according to whether there are any bacteria growing during the fermentation process, and grade them according to the grades. The scoring criteria are shown in Table 1.

[0027] Table 1 Scoring table for the fermentation process of Citrus aurantium

[0028]

[0029] 2. Determination of synephrine content: Determined by high performance liquid chromatography method (General Rule 0512) of Part IV of the 2020 edition of the Chinese Pharmacopoeia.

[0030] Chromatographic conditions: Column Agilent Eclipse plus C 18 (4.6×250mm, 5μm); the mobile phase is methanol-potassium dihydrogen phosphate solution (take 0.6g of potassium dihydrogen phosphate, 1.0g of sodium dodecyl sulfate, 1ml of glacial vinegar, dissolve in water and dilute to 1000ml) (50:50) as the mobile phase; the detection wavelength is 275nm; the flow rate is 1 mL / min; the column temperature is 30 ℃; the theoretical plate number calculated according to the synephrine peak should be not less than 2000.

[0031] Preparation of reference solution: Take an appropriate amount of synephrine reference, weigh accurately, and add water to make a solution containing 30 μg per 1 ml.

[0032] Preparation method of test sample: Take about 0.5g of the powder of this product, weigh it accurately, put it in a stoppered conical flask, add 25ml of methanol accurately, treat it with ultrasound for 30 minutes, filter it after cooling, take 5ml of the filtrate, dilute it with water and make up to 20ml, shake it well, and you have it.

[0033] Determination method: Accurately pipette 10μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0034] 3. Determination of flavonoid components (naringin, naringin, hesperidin, neohesperidin)

[0035] Chromatographic conditions: chromatographic column Agilent Eclipse plus C18 (4.6×250mm, 5μm); mobile phase acetonitrile (A)-0.1% formic acid aqueous solution (B); gradient elution conditions: 0-10 min, 10%-19%A; 10-25 min, 19%A; 25-27 min, 19%-30%A; 27-50 min, 30%-53%A; 50-51 min, 53%-75%A; 51-56 min, 75%A; 56-57 min, 75%-10%A; 57-62 min, 10%A. Detection wavelength is 330nm; volume flow rate is 1 mL / min; column temperature is 35℃; theoretical plate number calculated based on synephrine peak should be no less than 2000.

[0036] Preparation of reference solution: Take appropriate amount of naringin, naringin, hesperidin, and neohesperidin reference substances, weigh accurately, and use methanol to prepare mixed reference solutions with mass concentrations of 100 μg / mL (naringin, naringin) and 200 μg / mL (hesperidin, neohesperidin), respectively.

[0037] Preparation method of test sample: Take 0.5g of Citrus aurantium powder, weigh it accurately, put it in a stoppered conical flask, add 25ml of methanol accurately, weigh it, ultrasonicate it for 30min, let it cool, weigh it again, make up the lost weight with methanol, shake it well, filter it, accurately measure 5ml of the filtrate, transfer it to a 25ml volumetric flask, add methanol to the scale, shake it well, and obtain it.

[0038] Determination method: Accurately pipette 10μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0039] 4. Aflatoxin test shall be carried out in accordance with the second method of aflatoxin determination (liquid chromatography-tandem mass spectrometry) in Part IV, 2351, Determination of Mycotoxins in the 2020 edition of the Chinese Pharmacopoeia.

[0040] The measurement results are shown in Table 2.

[0041] Table 2 Screening experiment of strains for preparing fermented Citrus aurantium slices

[0042] Bacteria Fermentation process scoring Synephrine content (mg / g) Naringin, naringin, hesperidin, neohesperidin (mg / g) Aflatoxin (ppb) Natural fermentation 70 3.49 180.22 12 Rhizopus daelensis 95 4.53 200.15 Not detected Mucor 85 3.58 185.74 Not detected

[0043] In addition, the 2020 edition of the Chinese Pharmacopoeia stipulates that the content of synephrine in the medicinal materials and slices of Citrus aurantium is not less than 3 mg / g (0.3%), and there is no limit on the content of flavonoids. As shown in Table 2, the slices obtained by fermentation with Rhizopus delavayi are superior to those obtained by other fermentation methods in all indicators.

[0044] Example 3 Screening experiment of fermented Citrus aurantium slice preparation process

[0045] 1. Research on the lubrication process

[0046] 1.1. Water absorption test

[0047] Take 50g of clean Citrus aurantium, weigh it, add 3 times the amount of water, leave it for 4 hours, filter out the water, weigh the Citrus aurantium after absorbing water, and calculate the water absorption. The measurement results are shown in Table 3.

[0048] Table 3 Water absorption test of Citrus aurantium

[0049]

[0050] According to experimental results, the water absorption capacity of Citrus aurantium is about 60%, so 60% of the weight of water can be added to moisten it.

[0051] 1.2 Run-in time test

[0052] Take 50g of clean Citrus aurantium, weigh it, wash it, add 60% of the weight of Citrus aurantium (30g), observe the degree of moistening every 60 minutes, until there is no white core inside and it is not hard when pinched by hand. Determine the appropriate moistening time. The test results are shown in Table 4.

[0053] Table 4 Experiment on the time of processing of Citrus aurantium

[0054] time Degree of lubrication 60min The water has not been completely absorbed, and it still feels hard when squeezed, and it has not been moistened to meet the requirements. 120min The water has not been completely absorbed, and it still feels hard when squeezed, and it has not been moistened to meet the requirements. 180min The water is basically absorbed, and there is a white core when cut open 240min The water is absorbed, but there is still a little white heart 300min Water is absorbed cleanly, no white core, easier to cut

[0055] According to the experimental results, it is determined that the soaking time of Citrus aurantium is about 5 hours, until all the water is absorbed.

[0056] 2. Fermentation process research

[0057] 2.1 Fermentation process conditions

[0058] In the preparation process of fermented Citrus aurantium, fermentation temperature, fermentation humidity and fermentation time are the main influencing factors. Therefore, in the study of Citrus aurantium fermentation process, appearance, synephrine content and flavonoid content (naringin, naringin, hesperidin, neohesperidin) were used as evaluation indicators, and fermentation temperature, fermentation humidity and fermentation time were selected as investigation factors for orthogonal research. The process parameter design is shown in Table 5, and the process condition design table is shown in Table 6.

[0059] Table 5 Level table of factors in the fermentation process of Citrus aurantium

[0060]

[0061] Table 6 Fermentation process conditions of Citrus aurantium

[0062]

[0063] 2.2 Fermentation process test methods

[0064] All processing tools need to be sterilized by boiling water (15-20 minutes). Take 9 portions of Zhishi medicinal materials, grade them by size, 50g each, wash them with sterile purified water, sterilize them under ultraviolet light for 30 minutes, add 30g of sterile purified water and mix well, cover and moisten for 5 hours until there is no white core inside, place them in a sterilized and clean incubator, cover them with wet gauze, and cover them. Then place the incubator in an incubator and ferment according to the conditions in Table 5 (check the humidity of the gauze during the fermentation process, and spray distilled water appropriately to keep it moist). After the fermentation is completed, clean the Zhishi, steam it in a steamer for 5 hours, and simmer it overnight (12 hours). Take out the slices and dry them at 60℃.

[0065] 2.3 Fermentation process evaluation indicators and determination methods

[0066] In order to evaluate the fermentation process of Fructus Aurantii Immaturus objectively and comprehensively, three evaluation indicators were selected, including appearance, synephrine content, and flavonoids content (naringin, naringin, hesperidin, and neohesperidin), and the comprehensive score was calculated according to the weights.

[0067] (1) Appearance: Describe the properties of the pieces according to whether bacteria grow during the fermentation process, and grade them according to the test method, see Example 2. It accounts for 40% of the comprehensive score of the fermentation process evaluation of Citrus aurantium.

[0068] (2) Synephrine content: the determination method is shown in Example 2. It accounts for 30% of the comprehensive score of the fermentation process evaluation of Citrus aurantium.

[0069] (3) Flavonoid content (naringin, naringin, hesperidin, neohesperidin): the determination method is shown in Example 2. It accounts for 30% of the comprehensive score of the fermentation process evaluation of Citrus aurantium.

[0070] (4) Comprehensive score: calculated according to the following formula.

[0071]

[0072] 2.4 Fermentation process results: The test results are shown in Table 7.

[0073] Table 7 Results of the fermentation process of Citrus aurantium

[0074]

[0075] Based on the test results, the optimal process parameters for fermentation of Citrus aurantium were determined to be A2B3C2, that is, fermentation was performed at 25°C and 90% relative humidity for 4 days (the humidity of the gauze was checked during the fermentation process, and distilled water could be sprayed appropriately to keep it moist).

[0076] 3. Steaming time study

[0077] According to the determined optimal fermentation process, a batch of fermented Citrus aurantium (100g) was prepared and divided into 5 portions, each of 20g, and steamed for 0 hours, 3 hours, 4 hours, 5 hours, and 6 hours respectively, and then steamed overnight (12 hours) until the inside turned purple-brown. The slices were taken out and dried in an oven at 60℃ for 4 hours for later use.

[0078] Determination index: The properties of the decoction pieces were described by observation; the determination method of synephrine content and flavonoid content (naringin, naringin, hesperidin, and neohesperidin) was the same as that of Implementation Case 2. The determination results are shown in Table 8.

[0079] Table 8 Test results of fermented Citrus aurantium pieces at different steaming times

[0080] Steaming time (h) Appearance Synephrine content (mg / g) Naringin, naringin, hesperidin, neohesperidin (mg / g) 0 This product is a half-moon-shaped flake, yellowish brown, with a fragrant smell. 4.37 190.04 3 This product is a half-moon-shaped flake, purple-brown, with a fragrant smell. 4.43 194.21 4 This product is a half-moon-shaped flake, purple-brown, with a fragrant smell. 4.58 199.36 5 This product is a half-moon-shaped flake, purple-brown, with a fragrant smell. 4.62 200.78 6 This product is a half-moon-shaped flake, purple-brown, with a fragrant smell. 4.49 198.20

[0081] The results showed that the quality indicators of fermented Citrus aurantium slices steamed for 4 to 6 hours were better.

[0082] 4. Drying method research

[0083] A batch of fermented Citrus aurantium (100 g) was prepared according to the determined optimal fermentation process and steaming process, taken out and cut into thin slices, divided into 4 portions, and placed in an oven for drying (40°C, 50°C, 60°C, 70°C) respectively.

[0084] Determination indicators:

[0085] (1) Drying time: The drying time required when the moisture content of the medicinal material is less than 10%.

[0086] (2) Synephrine content: same as in Implementation Case 2.

[0087] (3) Contents of naringin, naringin, hesperidin and neohesperidin: same as in Implementation Case 2.

[0088] The measurement results are shown in Table 9.

[0089] Table 9 Determination results of fermented Citrus aurantium pieces obtained at different drying temperatures

[0090] Drying temperature (℃) Drying time (h) Synephrine content (mg / g) Naringin, naringin, hesperidin, neohesperidin (mg / g) 40 8 4.49 200.04 50 5.5 4.43 200.31 60 4 4.58 199.74 70 3.5 4.20 190.48

[0091] The results showed that the contents of active ingredients in the fermented Citrus aurantium slices obtained at 40, 50 and 60°C were higher, and the drying time required at 60°C was shorter. After comprehensive consideration, the drying temperature was determined to be 60°C.

[0092] Example 4: Experimental verification of the preparation process of fermented Citrus aurantium slices

[0093] According to the optimal process conditions determined in Example 3, three batches of fermented Citrus aurantium pieces were trial-produced, each batch of 200 g. The appearance of the pieces, synephrine content, flavonoid content (naringin, naringin, hesperidin, neohesperidin), aflatoxin limit and other indicators were measured to comprehensively evaluate the repeatability of the process. The measurement results are shown in Table 10.

[0094] Table 10 Results of the experimental verification of the preparation process of fermented Citrus aurantium slices

[0095]

[0096] In summary, compared with the currently naturally fermented Citrus aurantium decoction slices, the Citrus aurantium decoction slices prepared by the method of the present invention have significantly improved contents of synephrine and flavonoids, and no harmful components such as aflatoxin are detected, so the Citrus aurantium decoction slices are suitable for large-scale production.

Claims

1. A method for processing Fructus Aurantii Immaturus, characterized in that: Citrus aurantium is a kind of Rhizopus delemar ) for fermentation processing, comprising the following steps: a. Take the sterilized Zhishi medicinal material and moisten it with water; b. Take Rhizopus deltoides and activate it to prepare a strain activation solution. The amount of bacteria in the activation solution is (2.0±2.0×10%)×10 8 CFU / mL, dilute with water and spray evenly into the medicinal materials; use 10ml of bacterial activation solution per kg of Citrus aurantium, dilute with 40ml of water; c. Fermentation: temperature 25°C, relative humidity 90%, fermentation time 4 days; d. Steaming: Steam the fermented herbs in step c for 4-6 hours; e.Muffled: f. Slice and dry for 3.5-8h; The Rhizopus delbrueckii ( Rhizopus delemar ) was purchased from China Industrial Microbiological Culture Collection Center, No.: CICC40308.

2. The method for processing Fructus Aurantii Immaturus according to claim 1, characterized in that: The sterilization method described in step a is ultraviolet sterilization, and the sterilization time is 30 minutes; the soaking time is 5 hours; and the amount of water added is 60% of the weight of the immature bitter orange.

3. The method for processing Fructus Aurantii Immaturus according to claim 1, characterized in that: The moistening time described in step e is 5 hours.

4. The method for processing Fructus Aurantii Immaturus according to claim 1, characterized in that: The drying temperature in step f is 60°C and the drying time is 4 hours.

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