Processing technology for removing aristolochic acid in medicinal materials

Abstract

The invention discloses a processing technology for removing aristolochic acid in medicinal materials, and relates to the technical field of traditional Chinese medicine processing, and the technical scheme is as follows: the processing technology comprises the following steps: S1, taking crude medicinal materials, and crushing the crude medicinal materials at room temperature to obtain medicinal material powder; s2, heating and processing the medicinal material powder prepared in the step S1 in a drying oven, taking out the medicinal material powder after processing is finished, and cooling to obtain a medicinal material processed product. According to the processing technology, the content of aristolochic acid in the medicinal materials can be better reduced, meanwhile, active ingredients of the medicinal materials are not damaged, and the curative effect of the traditional Chinese medicine is ensured. The medicinal material containing aristolochic acid is processed by heating, so that on one hand, the effect of reducing toxicity can be achieved, and on the other hand, other auxiliary materials are not added to influence the components of the traditional Chinese medicine. The processing period is short, the processing technology is simple and easy to operate, and industrial production can be achieved.

Description

Technical Field

[0001] The invention relates to the technical field of Chinese medicine processing, and more specifically, to a processing technology for removing aristolochic acid from medicinal materials. Background Art

[0002] Aristolochic acid substances are mainly found in Aristolochic acid and Asarum in the Aristolochiaceae family, and have pharmacological properties such as anti-inflammatory, antitussive, and antiplatelet aggregation. For centuries, aristolochic acid substances have been used as traditional Chinese medicine ingredients to treat various diseases. However, because aristolochic acid substances can produce toxic side effects such as hepatotoxicity, nephrotoxicity, carcinogenicity, and mutagenicity, it has become a key issue in health care. The reason why aristolochic acid substances produce toxicity is that aristolochic acid is metabolized into aristolochic acid lactam carbon cations in the human body, which then binds to DNA and damages kidney cells. Long-term low-dose or short-term high-dose use may lead to the occurrence of nephrotoxic reactions. There are many types of traditional Chinese medicines containing aristolochic acid substances, many of which have shown excellent pharmacological effects. Therefore, completely banning these traditional Chinese medicines will cause huge losses to the application of traditional Chinese medicine resources. In order to give full play to the efficacy of these traditional Chinese medicines, seeking a method to remove aristolochic acid from medicinal materials is of great significance for the application of traditional Chinese medicine resources.

[0003] Studies have shown that the toxicity of aristolochic acid substances can be reduced and controlled through reasonable compatibility and processing methods, so as to improve the clinical safety of aristolochic acid-containing medicinal materials. The carbon skeleton of aristolochic acid is the same, but the hydroxyl, methoxy and nitro groups are different in position and quantity. Its toxicity is related to nitro, methoxy and hydroxyl groups. If chemical reactions such as nitro reduction, demethylation and increase of hydroxyl groups can occur during the processing of medicinal materials, the toxicity of aristolochic acid-containing medicinal materials can be reduced.

[0004] The detoxification of Chinese medicine is one of the unique traditional processing methods of traditional Chinese medicine. By processing Chinese medicinal materials, they are subjected to a series of treatments and processes to reduce their toxicity or side effects. This process usually includes steps such as frying, roasting, and boiling to change the nature, taste, efficacy, and toxicity characteristics of the medicinal materials. The current process for processing medicinal materials containing aristolochic acid is mainly honey roasting, salt roasting, and alkaline water processing. The current process causes the loss of effective ingredients of medicinal materials containing aristolochic acid to varying degrees, and the resulting processed products also contain other ingredients, thereby affecting the purity of the Chinese medicine. At the same time, its processing process is cumbersome, the production cycle is long, and it is not suitable for industrial production. Therefore, the present invention provides a different processing process for removing aristolochic acid from medicinal materials, and adopts a heating processing method to treat a process for medicinal materials containing aristolochic acid. Summary of the invention

[0005] The purpose of the present invention is to provide a processing technology for removing aristolochic acid from medicinal materials. The heating processing method is used to treat medicinal materials containing aristolochic acid, which can greatly reduce the content of aristolochic acid in the medicinal materials. The processing cycle is short and the processing technology is simple and easy to operate.

[0006] The above technical objectives of the present invention are achieved through the following technical solutions: A processing process for removing aristolochic acid from medicinal materials, comprising the following steps:

[0007] S1: taking a raw medicinal material, and crushing the raw medicinal material at room temperature to obtain medicinal material powder;

[0008] S2: placing the medicinal material powder obtained in step S1 in an oven for heating and processing, taking it out after processing, and letting it cool to obtain the processed medicinal material product.

[0009] The present invention is further configured as follows: the raw medicinal material in step S1 is a medicinal material containing aristolochic acid, including but not limited to Rhizoma Aristolochiae, Asarum and / or Aristolochia lappa.

[0010] The present invention is further configured as follows: no substance is added as auxiliary material for co-processing.

[0011] The present invention is further configured as follows: in step S2, an oven is used for heating and preparation.

[0012] The present invention is further configured as follows: the processing temperature of step S2 is 250°C-350°C.

[0013] The present invention is further configured as follows: the processing temperature of step S2 is 300°C.

[0014] The present invention is further configured as follows: the preparation time of step S2 is at least 30 minutes.

[0015] The present invention is further configured as follows: in step S2, a tray made of stainless steel is used as a container for heating and processing the medicinal materials.

[0016] The present invention further provides a prepared medicinal material containing aristolochic acid, which is prepared according to the above-mentioned processing process for removing aristolochic acid from the medicinal material.

[0017] In summary, the present invention has the following beneficial effects:

[0018] 1. The processing technology provided by the present invention can better reduce the content of aristolochic acid in medicinal materials, while preventing the effective ingredients of the medicinal materials from being destroyed, thereby ensuring the efficacy of traditional Chinese medicine.

[0019] 2. The present invention processes the aristolochic acid-containing medicinal material by heating, which can achieve the effect of reducing toxicity on the one hand, and on the other hand, does not add other auxiliary materials to affect the components of the traditional Chinese medicine.

[0020] 3. The processing cycle of the invention is short, the processing technology is simple and easy to operate, and it can be industrialized. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 is a typical standard curve of aristolochic acid A in the embodiment of the present invention;

[0022] Figure 2 is a chromatogram of aristolochic acid A reference substance in an embodiment of the present invention;

[0023] Figure 3 is a chromatogram of aristolochic acid A in the Jiuyueshengsheng product in an embodiment of the present invention;

[0024] Figure 4 is a chromatogram of aristolochic acid A in the Jiuyuesheng processed product in an embodiment of the present invention;

[0025] Figure 5 is a chromatogram of aristolochic acid A in the raw Asarum product in the embodiment of the present invention;

[0026] Figure 6 is a chromatogram of aristolochic acid A in the processed Asarum product in the embodiment of the present invention;

[0027] Figure 7 is a chromatogram of aristolochic acid A in the raw product of Aristolochia sinensis in an embodiment of the present invention;

[0028] Figure 8 is a chromatogram of aristolochic acid A in the processed product of Aristolochia chinensis in an embodiment of the present invention;

[0029] Fig. 9 It is a temperature-time change line graph of aristolochic acid A in the aristolochic acid-containing medicinal material processed product according to an embodiment of the present invention;

[0030] Fig.10 It is a comparison chart of the content of aristolochic acid A after processing of aristolochic acid-containing medicinal materials with different grinding finenesses in the embodiments of the present invention. DETAILED DESCRIPTION

[0031] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific implementations, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0032] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing specific embodiments rather than for limiting the scope of protection of the present invention.

[0033] Embodiment: A processing technology for removing aristolochic acid from medicinal materials, such as Figure 1-Figure 10 As shown, the following steps are included:

[0034] S1: taking a raw medicinal material, and crushing the raw medicinal material at room temperature to obtain medicinal material powder;

[0035] S2: placing the medicinal material powder obtained in step S1 in an oven for heating and processing, taking it out after processing, and letting it cool to obtain the processed medicinal material product.

[0036] The raw medicinal materials are medicinal materials containing aristolochic acid, including but not limited to Rhizoma Aristolochiae, Asarum and / or Aristolochia sinensis. The above-mentioned processed medicinal materials containing aristolochic acid are processed without adding any substances as auxiliary materials. Instead of using a traditional wok for frying, an oven is used to achieve the purpose of heating and processing, and the heating is more uniform. When heating the above-mentioned processed medicinal materials containing aristolochic acid, a stainless steel tray is used as a container.

[0037] September Raw Preparation

[0038] Take 100g of raw Jiuyuesheng, put it in a grinder for grinding, then weigh 1.0g of Jiuyuesheng powder with an analytical balance and put it in a clean tray, use an oven to heat it at 300℃ for 30min, then take out the heated Jiuyuesheng powder and cool it to room temperature to obtain the Jiuyuesheng processed product. Detect the content of aristolochic acid A in the obtained Jiuyuesheng processed product.

[0039] Processing of Asarum

[0040] Take 100g of raw Asarum, put it in a grinder for grinding, then weigh 1.0g of Asarum powder on an analytical balance and put it in a clean tray, use an oven to heat it at 300℃ for 30min, then take out the heated Asarum powder and cool it to room temperature to obtain the processed Asarum. Detect the content of aristolochic acid A in the processed Asarum.

[0041] Preparation of Aucklandia

[0042] Take 100g of raw Aucklandia lappa and grind it in a grinder, then weigh 1.0g of Aucklandia lappa powder on an analytical balance and place it in a clean tray, heat it in an oven at 300°C for 30 minutes, then take out the heated Aucklandia lappa powder and cool it to room temperature to obtain the processed Aucklandia lappa. Detect the content of aristolochic acid A in the processed Aucklandia lappa.

[0043] Unprocessed raw September raw medicinal materials: Take 100g of raw September raw materials, put them into a grinder for grinding, put the ground September raw powder into a container and seal it for storage, and then test the content of aristolochic acid A in the raw September raw materials.

[0044] Unprocessed Asarum: 100 g of raw Asarum was taken and crushed in a grinder, and the crushed Asarum powder was placed in a sealed container for storage and standby use. The content of aristolochic acid A in the raw Asarum was detected.

[0045] Unprocessed Aristolochia sinensis medicinal material: 100 g of raw Aristolochia sinensis was taken and crushed in a grinder, and the crushed Aristolochia sinensis powder was placed in a sealed container for storage and standby use. The content of aristolochic acid A in the raw Aristolochia sinensis was detected.

[0046] The processed Rhizoma Aristolochiae, Asarum and Aucklandiae Radix are used as Examples 1-3, respectively, and the unprocessed Rhizoma Aristolochiae, Asarum and Aucklandiae Radix are used as Control Examples 1-3, respectively. The following is the experimental part of the processed medicinal materials containing aristolochic acid of the present invention. Determination of the content of aristolochic acid A in the medicinal materials containing aristolochic acid The aristolochic acid-containing medicinal materials and their processed products prepared in the examples and comparative examples of the present invention are determined by high-resolution liquid chromatography-mass spectrometry.

[0047] (1) Liquid phase conditions

[0048] Chromatographic column: Hypersil GOLD C18 column (150×2.1 mm, 1.9 μm, Thermo Fisher Scientific, Bellefonte, PA, USA);

[0049] Column temperature (℃): 40;

[0050] Injector temperature (°C): 10;

[0051] Flow rate (mL / min): 0.3;

[0052] Injection volume (μL): 3;

[0053] Mobile phase: 0.1% formic acid aqueous solution (A), methanol (B). Gradient elution conditions are as follows: 0 min, 5% B; 1-5 min, 5-10% B; 5-20 min, 10-40% B; 20-30 min, 40-56% B; 30-45 min, 56-68% B; 45-47 min, 68-90% B; 47-48 min, 90-100% B; 48-51 min, 100% B; 51-51.5 min, 100-5% B; 51.5-55 min, 5%.

[0054] (2) Mass spectrometry conditions

[0055] Ion detection mode: Full scan (data acquisition), FS-ddMS2 (secondary mass spectrometry acquisition);

[0056] Ionization method: electrospray ionization (ESI);

[0057] Ion polarity: positive ion mode;

[0058] Scanning range m / z: 100-800;

[0059] Collision energy (%): CE20, 30;

[0060] Electrospray voltage (kV): 3.5;

[0061] Sheath gas flow (units): 35;

[0062] Auxiliary gas flow (units): 15;

[0063] Capillary temperature (℃): 350;

[0064] Auxiliary gas heater temperature (℃): 350.

[0065] Preparation of aristolochic acid A reference solution: Accurately weigh 0.2 mg of aristolochic acid A reference substance, place it in a 1 mL centrifuge tube, add 1 mL of methanol to dissolve, and shake well to obtain 0.2 mg / mL aristolochic acid A stock solution. Dilute the stock solution with methanol to prepare a 500 ng / mL aristolochic acid A reference solution.

[0066] Preparation of the test solution of aristolochic acid-containing medicinal materials: Take 1.0 mg of raw powder of aristolochic acid-containing medicinal materials, place it in a 1 mL centrifuge tube, add 1 mL of 70% methanol, and ultrasonically treat (power 320 W, frequency 40 kHz) for 40 min. Aspirate the supernatant and centrifuge it at 14000 rpm for 10 min in a high-speed centrifuge. Take out the liquid after centrifugation and take the supernatant.

[0067] Preparation of processed medicinal material solution containing aristolochic acid: Take 1.0 mg of processed powder, place in 1 mL centrifuge tubes, add 1 mL of 70% methanol, and ultrasonically treat (power 320 W, frequency 40 kHz) for 40 min. Take out the supernatant and centrifuge at 14000 rpm for 10 min in a high-speed centrifuge. Take out the liquid after centrifugation and take the supernatant.

[0068] Determination method: Accurately pipette 100 μL of each of the solutions of aristolochic acid A reference substance, aristolochic acid-containing medicinal materials, and aristolochic acid-containing medicinal materials preparation test samples, and inject them into a high-resolution liquid spectrometer for sampling and detection.

[0069] Aristolochic acid A stock solution was diluted with pure water to prepare a series of reference solutions, which were: 4ng / mL, 10ng / mL, 40ng / mL, 100ng / mL, 400ng / mL and 1000ng / mL. The series of reference solutions were injected and analyzed, and the chromatograms were recorded. The standard curve was drawn with the concentration C of the reference solution as the horizontal axis (X) and the peak area as the vertical axis (Y). Figure 1 ), the linear regression equation is: Y = -1466459.019 + 552194.270X (R 2 =0.992). The peak area of ​​aristolochic acid A obtained after sampling the raw product and processed product of aristolochic acid-containing medicinal materials was substituted into the standard curve to calculate the content of aristolochic acid A therein.

[0070] Test results: The chromatograms of the raw aristolochic acid-containing medicinal materials and their processed products prepared in the examples and comparative examples of the present invention are as follows: Figure 3-8 shown; from Figure 2 It can be seen that the retention time of the aristolochic acid A reference substance is 35.66 min, and the peak areas of aristolochic acid A in the processed aristolochic acid-containing medicinal materials prepared in the embodiments of the present invention are smaller than the peak areas of aristolochic acid A in the comparative examples. The content of aristolochic acid A in the processed aristolochic acid-containing medicinal materials was calculated by the external standard method, and the results are shown in Table 1. Compared with the unprocessed medicinal materials, the content of aristolochic acid A in the processed aristolochic acid-containing medicinal materials prepared in the embodiments of the present invention is significantly reduced, indicating that the processing technology is reliable.

[0071] Table 1 Content of aristolochic acid A in medicinal materials containing aristolochic acid before and after processing

[0072]

[0073]

[0074] Investigation of processing conditions

[0075] By designing different processing temperatures (250°C, 270°C, 300°C) and different processing times (10min, 30min, 50min, 70min) (n=2), the optimal processing conditions for medicinal materials containing aristolochic acid were determined, such as Fig. 9 . After weighing several portions of aristolochic acid-containing medicinal materials, they were processed under the above different processing conditions, and the processed products were sampled and analyzed to compare the percentage of aristolochic acid A in the processed products under different processing conditions. The aristolochic acid-containing medicinal materials used in this study were September-born. The results showed that the aristolochic acid-containing medicinal materials had the best processing effect when the processing temperature was 300°C and the processing time was 30 minutes, and the aristolochic acid A content in the processed products was low at this time. After comprehensive consideration, the present invention finally chose to use the processing conditions of 300°C and 30 minutes to process the aristolochic acid-containing medicinal materials.

[0076] Grinding fineness inspection

[0077] Taking the drug sieves of the national standard of the 2020 edition of the "Chinese Pharmacopoeia" as a reference, the percentage of aristolochic acid A in the processed products of aristolochic acid-containing medicinal materials that passed through different sieve numbers (No. 2 sieve - 24 mesh, No. 3 sieve - 50 mesh, No. 4 sieve - 65 mesh and No. 5 sieve - 80 mesh) was investigated to determine the optimal grinding fineness for processing. The aristolochic acid-containing medicinal materials that passed the corresponding sieve number were processed at a processing temperature of 300°C and a processing time of 30 minutes, and the content of aristolochic acid A in the processed products was calculated respectively. The aristolochic acid-containing medicinal materials used in this investigation were September-grown. The results are as follows Fig.10 The comparison of the content of aristolochic acid A after processing of aristolochic acid-containing medicinal materials with different grinding finenesses is shown in the figure (the "-" in the horizontal axis indicates the processed aristolochic acid-containing medicinal materials that have not been screened by the standard pharmacopoeia sieve). From the figure, we can see that the aristolochic acid-containing medicinal materials with a grinding fineness of 65 mesh (passing the No. 4 sieve) have the lowest content of aristolochic acid A after processing. Therefore, the optimal grinding fineness for processing in the present invention is 65 mesh.

[0078] In summary, the present invention discloses for the first time the use of a heating processing process to treat aristolochic acid-containing medicinal materials. The aristolochic acid A content in the processed medicinal materials containing aristolochic acid obtained by the processing process provided by the present invention is low, and the processing process has the characteristics of reliable processing technology and good reproducibility, which fills the gap in the existing processing specifications that the processing technology of aristolochic acid-containing medicinal materials has not been included, and provides a guarantee for the clinical application of related medicinal materials in the later stage.

[0079] The present invention does not limit the source of the raw materials used. Unless otherwise specified, the raw materials used in the present invention are all common commercial products in the technical field. The aristolochic acid-containing medicinal material Jiuyuesheng used in the examples and comparative examples of the present invention is from Guizhou Wansheng Pharmaceutical Co., Ltd., and the aristolochic acid-containing medicinal materials Asarum and Aristolochia chinensis are both from Chengdu Ruifensi Biotechnology Co., Ltd.

[0080] The above technical features constitute the embodiments of the present invention, which have strong adaptability and implementation effect. Non-essential technical features can be added or reduced according to actual needs to meet the requirements of different situations.

[0081] This specific embodiment is merely an explanation of the present invention and is not a limitation of the present invention. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed. However, as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. A processing process for removing aristolochic acid from medicinal materials, characterized in that: The following steps are involved: S1: taking a raw medicinal material, and crushing the raw medicinal material at room temperature to obtain medicinal material powder; S2: placing the medicinal material powder obtained in step S1 in an oven for heating and processing, taking it out after processing, and letting it cool to obtain the processed medicinal material product.

2. The processing process for removing aristolochic acid from medicinal materials according to claim 1, characterized in that: The raw medicinal materials in step S1 are medicinal materials containing aristolochic acid, including but not limited to Rhizoma Aristolochiae, Asarum and / or Radix Aristolochiae.

3. The processing process for removing aristolochic acid from medicinal materials according to claim 1, characterized in that: No substance is added as auxiliary material during the preparation.

4. The processing process for removing aristolochic acid from medicinal materials according to claim 1, characterized in that: In step S2, an oven is used for heating and preparation.

5. The processing process for removing aristolochic acid from medicinal materials according to claim 1, characterized in that: The processing temperature of step S2 is 250°C-350°C.

6. The processing process for removing aristolochic acid from medicinal materials according to claim 5, characterized in that: The processing temperature of step S2 is 300°C.

7. The processing technology for removing aristolochic acid from medicinal materials according to claim 1, characterized in that: The preparation time of step S2 is at least 30 minutes.

8. The processing process for removing aristolochic acid from medicinal materials according to claim 1, characterized in that: In step S2, a stainless steel tray is used as a container to heat and process the medicinal materials.

9. A processed medicinal material containing aristolochic acid, characterized in that: A medicinal material processed product containing aristolochic acid prepared by the processing process according to any one of claims 1 to 8.

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