阅读说明：本技术 一种芹菜素的半合成方法 (Semi-synthesis method of apigenin ) 是由 张琦 郭涛 孙绿华 于 2020-06-19 设计创作，主要内容包括：一种芹菜素的半合成方法,包括：以二甲基亚砜为溶剂,将柚皮素与碘在100-120℃温度下进行反应；向反应液中加入保险粉或保原粉以及体积分数25%-50%的乙醇水溶液,在碱性环境下,于70-90℃温度下反应；将反应液趁热过滤,收集滤液,调节温度为30-40℃、pH值为6-7,加入体积分数50%-80%的乙醇水溶液,待晶体析出后过滤,收集滤饼,即得芹菜素产品。该方法反应条件温和、易于控制、步骤简单、操作方便、耗时短、成本低、收率高,且对环境无污染。(A semi-synthesis method of apigenin comprises the following steps: reacting naringenin and iodine at the temperature of 100-120 ℃ by using dimethyl sulfoxide as a solvent; adding sodium hydrosulfite or preservative powder and 25-50% of ethanol aqueous solution by volume fraction into the reaction solution, and reacting at 70-90 ℃ in an alkaline environment; filtering the reaction solution while the reaction solution is hot, collecting filtrate, adjusting the temperature to be 30-40 ℃ and the pH value to be 6-7, adding 50-80% ethanol water solution by volume fraction, filtering after crystals are separated out, collecting filter cakes, and obtaining the apigenin product. The method has the advantages of mild reaction conditions, easy control, simple steps, convenient operation, short time consumption, low cost, high yield and no pollution to the environment.)

1. A semi-synthesis method of apigenin is characterized by comprising the following steps:

1) reacting naringenin and iodine at the temperature of 100-120 ℃ by using dimethyl sulfoxide as a solvent;

2) after the reaction in the step 1) is finished, adding sodium hydrosulfite or preservative powder and 25-50% of ethanol aqueous solution by volume fraction into the reaction solution, and reacting at the temperature of 70-90 ℃ in an alkaline environment;

3) after the reaction in the step 2) is finished, filtering the reaction solution while the reaction solution is hot, and collecting filtrate;

4) adjusting the temperature of the filtrate obtained in the step 3) to be 30-40 ℃ and the pH value to be 6-7, adding 50-80% by volume of ethanol aqueous solution, and waiting for crystal precipitation;

5) and (4) after the crystal precipitation in the step 4) is finished, filtering, and collecting a filter cake to obtain the apigenin product.

2. The semi-synthesis method of apigenin according to claim 1, characterized in that: the temperature of the step 1) is 105-110 ℃.

3. The semi-synthesis method of apigenin according to claim 1, characterized in that: in the step 1), the dosage of the dimethyl sulfoxide is 5-10ml/g of the naringenin.

4. The semi-synthesis method of apigenin according to claim 1, characterized in that: in the step 1), the addition amount of the iodine is 0.05-0.1 time of the weight of the naringenin feeding material.

5. The semi-synthesis method of apigenin according to claim 1, characterized in that: in the step 2), the pH value of the alkaline environment is 8-11.

6. The semi-synthesis method of apigenin according to claim 1, characterized in that: the reaction temperature in step 2) was 85 ℃.

7. The semi-synthesis method of apigenin according to claim 1, characterized in that: in the step 2), the addition amount of the sodium hydrosulfite or the preservative powder is 10-20% of the weight of the naringenin feeding material.

8. The semi-synthesis method of apigenin according to claim 1, characterized in that: in the step 3), the temperature of the reaction solution during filtration is 50 ℃ or higher.

9. The semi-synthesis method of apigenin according to claim 1, characterized in that: in the step 4), the adding mode of the ethanol water solution is dripping at the speed of 70-140 ml/min.

Technical Field

The invention relates to the technical field of synthesis of pharmaceutical and chemical products, in particular to a semi-synthesis method for preparing apigenin.

Background

Apigenin, Apigenin and Apigenin, wherein the English name is Apigenin, the chemical name is 4, 5, 7-trihydroxyflavone, and the CAS number is 520-36-5, and the Apigenin is a flavonoid compound. It is widely distributed in nature, and mainly exists in vegetables, fruits, beans, tea leaves, and Scrophulariaceae plant such as Citrus reticulata Blanco, Polygonum cuspidatum of Polygonaceae, Liliaceae plant such as Veratrum nigrum and Pinaceae plant in warm and hot zone in the form of plant yellow pigment, wherein the content of celery is the highest. The pure apigenin is yellow powder, odorless, tasteless, hardly soluble in water, slightly soluble in hot alcohol, soluble in dilute alkali solution, and natural antioxidant.

Modern researches find that apigenin has various pharmacological effects, such as anti-tumor, anti-arteriosclerosis, antithrombotic, antiviral, antibacterial and anti-inflammatory, diuretic, tranquilizing, nerve soothing, blood pressure lowering, and Parkinson's disease resistance, wherein the anti-tumor effect is the most prominent, and the apigenin has inhibition effects on breast cancer, gastric cancer, prostate cancer, liver cancer, ovarian cancer and the like, so that the apigenin has high medicinal value.

In the traditional method for industrially producing apigenin, the traditional Chinese medicine extraction technology is adopted to extract and enrich the apigenin in natural plants, but due to the low content of the apigenin in the natural plants and the limitation of the traditional Chinese medicine extraction technology, the method has the defects of low extraction rate, high energy consumption and high cost, and is difficult to meet the increasing market demand. Therefore, a chemical synthesis method is commonly adopted at present, for example, chinese patent application CN1793137A discloses a semi-synthesis method of apigenin, which specifically uses naringenin as a raw material to react with iodine in a solvent of 1, 4-dioxane at a temperature of 50-130 ℃, and oxidative dehydrogenation is performed to generate a crude apigenin product, and the crude product is crystallized and recrystallized or purified by reflux to obtain refined apigenin. However, this method has the following disadvantages: 1. 1, 4-dioxane with high toxicity and high price is used as a solvent, so that environmental pollution is easily caused and the production cost is high; 2. the crude apigenin generated by oxidative dehydrogenation is obtained by concentration, water addition, standing and suction filtration, the crude product needs to be dissolved and crystallized by alkaline solution, and then recrystallized or refluxed by solvent, the steps are complex, and the time consumption is long; 3. the yield of the method is only 60-70%, and the method is relatively low and increases the production cost.

Technical problem

The invention aims to solve the technical problems of complex process, long time consumption, low yield, high cost and environmental pollution of the existing apigenin semi-synthesis method in the background art.

Technical solution

In order to achieve the above object, the present invention provides a semi-synthesis method of apigenin, comprising the steps of:

1) reacting naringenin and iodine at the temperature of 100-120 ℃ by using dimethyl sulfoxide as a solvent;

2) after the reaction in the step 1) is finished, adding sodium hydrosulfite or preservative powder and 25-50% of ethanol aqueous solution by volume fraction into the reaction solution, and reacting at the temperature of 70-90 ℃ in an alkaline environment;

3) after the reaction in the step 2) is finished, filtering the reaction solution while the reaction solution is hot, and collecting filtrate;

4) adjusting the temperature of the filtrate obtained in the step 3) to be 30-40 ℃ and the pH value to be 6-7, adding 50-80% by volume of ethanol aqueous solution, and waiting for crystal precipitation;

5) and (4) after the crystal precipitation in the step 4) is finished, filtering, and collecting a filter cake to obtain the apigenin product.

In the semi-synthesis method provided by the invention, the chemical name of the sodium hydrosulfite is sodium hydrosulfite and sodium hydrosulfite with the chemical formula of Na₂S₂O₄The product is a white sand-shaped crystal or light yellow powder chemical product, has a melting point of 300 ℃ (decomposition), an ignition temperature of 250 ℃, is insoluble in ethanol, is soluble in sodium hydroxide solution, and has strong reducibility, and the product can react strongly with water and burn. The so-called preservative powder is a conventional substitute for sodium hydrosulfite, and is generally higher than sodium hydrosulfite in reducing property and stability.

Preferably, in step 1) of the semi-synthesis method provided by the invention, the reaction temperature is 105-110 ℃, and the conversion rate of raw materials is the highest and the reaction speed is the fastest at the temperature.

From the viewpoint of maximizing resource utilization and saving cost, the amount of the dimethyl sulfoxide used in the step 1) of the semi-synthesis method provided by the invention is preferably 5-10ml/g naringenin.

From the viewpoint of maximizing resource utilization and saving cost, preferably, in step 1) of the semi-synthesis method provided by the present invention, the amount of iodine added is 0.05 to 0.1 times of the weight of naringenin fed.

In step 2) of the semi-synthesis method provided by the invention, the 25-50% volume fraction of ethanol aqueous solution has the functions of quenching the previous reaction on one hand and promoting the sodium hydrosulfite or preservative powder to participate in the next reaction on the other hand.

In step 2) of the semi-synthesis method provided by the invention, an alkaline environment is an important guarantee for ensuring that the reaction can be smoothly carried out, and preferably, the pH value of the alkaline environment is 8-11, so that the sodium hydrosulfite/preservative powder can lose efficacy when the pH value is lower than the range, and the by-product can be increased when the pH value is higher than the range.

Preferably, in step 2) of the semi-synthesis method provided by the present invention, the alkaline environment is achieved by adding an alkaline solution to the reaction solution, wherein the alkaline solution includes, but is not limited to, sodium hydroxide solution, potassium hydroxide solution, lithium hydroxide solution, sodium bicarbonate solution and sodium carbonate solution.

Preferably, in step 2) of the semi-synthesis method provided by the present invention, the reaction temperature is 85 ℃, and the optimal reaction effect can be achieved at the temperature.

Preferably, in the step 2) of the semi-synthesis method provided by the invention, the addition amount of the sodium hydrosulfite or the preservative powder is 10-20% of the weight of the naringenin feed.

Preferably, in step 3) of the semi-synthesis method provided by the present invention, the temperature of the reaction solution during filtration is ensured to be 50 ℃ or higher, and filtration is difficult if the temperature is too low.

More preferably, in step 3) of the semi-synthesis method provided by the present invention, the temperature of the reaction solution during filtration is ensured to be 50-60 ℃.

Preferably, in step 4) of the above semi-synthesis method provided by the present invention, the pH of the filtrate obtained in step 3) is adjusted by adding glacial acetic acid or phosphoric acid.

Preferably, in step 4) of the semi-synthesis method provided by the invention, the ethanol aqueous solution is added dropwise at a speed of 70-140ml/min, so that apigenin crystals are rapidly separated out, and the separated crystals are large particles uniformly distributed, thereby facilitating subsequent filtration and separation.

The filtration in the semi-synthesis method provided by the invention refers to a process of separating solid and liquid in a solution by a physical method, and common filtration methods are all applicable to the method provided by the invention, and include normal pressure filtration, reduced pressure filtration, centrifugal filtration and the like.

Advantageous effects

Compared with the semi-synthesis method of apigenin disclosed in the Chinese patent application CN1793137A (hereinafter referred to as a comparison document), the method provided by the invention has the following advantages:

1. the high-content apigenin is synthesized by adopting a one-pot method, the raw materials in the whole process are cheap and easy to obtain, the reaction condition is mild, the control is easy, the steps are simple, the operation is convenient, and the consumed time is short;

2. dimethyl sulfoxide is used as a reaction solvent, so that the method is environment-friendly, economic and environment-friendly, meets the requirement of waste liquid recycling, achieves the aim of zero discharge of three wastes, and is more suitable for industrial mass production;

3. after the chemical synthesis reaction is finished, related impurities, byproducts and the like in the reaction liquid can be effectively removed by innovatively adding sodium hydrosulfite or preservative powder into the reaction liquid and carrying out high-temperature reaction in an alkaline environment, and then the apigenin refined product with the purity equivalent to that in the comparison document can be obtained only by one-time crystallization (recrystallization and three-time crystallization are not needed), the yield is up to more than 90%, and the yield is improved by more than 20% -30% compared with that in 60% -70% of the comparison document.

Modes for carrying out the invention

The present invention will be described in further detail with reference to specific examples, which are illustrative of the present invention and are not to be construed as being limited thereto.

The raw materials and reagents used in the following examples were all commercially available except for those specifically mentioned. Sodium hydrosulfite (sodium hydrosulfite) used in the following examples was purchased from Shanghai Aladdin Biotech Co., Ltd; the preservative powder was purchased from kumquat science ltd, guangzhou city.

Example 1

Adding 10g naringenin and 50ml dimethyl sulfoxide into a 250ml three-necked bottle, stirring, dissolving and clarifying; adding 1g elemental iodine, heating to 100 deg.C for reaction, and stopping the reaction by monitoring naringenin residue content of less than 10% by HPLC (HPLC detection method, Asahi C18 chromatographic column, mobile phase is 50% acetonitrile water solution). After the reaction is finished, cooling to 70 ℃, adding 10ml of 50% ethanol water solution, adjusting the pH to 9 by 10mol/L sodium hydroxide solution, adding 2g of sodium hydrosulfite, and reacting for 2 hours; filtering while the solution is hot, and washing a filter cake by using 5ml of dimethyl sulfoxide; mixing the filtrates, cooling to 30 ℃, adding 5ml of 6mol/L hydrochloric acid to adjust the pH of the reaction solution to 6-7, dropwise adding 150ml of 50% ethanol aqueous solution at the speed of 70ml/min, precipitating out a large amount of product, monitoring the apigenin precipitation amount by HPLC to be more than 90%, filtering to obtain a filter cake which is an apigenin refined product, and performing vacuum drying at 50 ℃ to obtain 9.1g of red-brown apigenin dried product powder; the product weight yield was 91.00%. Loss on drying was 2.33%; HPLC purity 97.62%; the apigenin obtained by the detection is subjected to content determination by taking the apigenin standard substance (marked content is 98%) of the Zhongzhong institute as a control, and the content is detected to be 95.23%.

Example 2

Adding 10g naringenin and 70ml dimethyl sulfoxide into a 250ml three-necked bottle, stirring, dissolving and clarifying; 0.5g of elemental iodine was added, the reaction was warmed to 120 ℃ and stopped by monitoring the naringenin residue content of the reaction raw material by HPLC to less than 10% (HPLC detection method: Asahi C18 column, mobile phase: 50% acetonitrile aqueous solution). After the reaction is finished, cooling to 85 ℃, adding 10ml of 25% ethanol water solution, adjusting the pH to 8 by 10mol/L potassium hydroxide solution, adding 1g of sodium hydrosulfite, and reacting for 2 hours; filtering while the solution is hot, and washing a filter cake by using 5ml of dimethyl sulfoxide; mixing the filtrates, cooling to 35 deg.C, adding 5ml glacial acetic acid to adjust pH to 6-7, dripping 150ml 80% ethanol water solution at 120ml/min speed to precipitate a large amount of apigenin, monitoring apigenin precipitation amount by HPLC to be greater than 90%, filtering to obtain refined apigenin cake, and vacuum drying at 50 deg.C to obtain light yellow apigenin dried product 9.52 g; the weight yield was 95.2%. Loss on drying was 2.13%; HPLC purity 98.31%; the apigenin obtained by the previous step is subjected to content measurement by taking the apigenin standard (marked content is 98%) of the Zhongzhong institute as a control, and the content is 96.84%.

Example 3

Adding 1kg of naringenin and 10L of dimethyl sulfoxide into a 50L glass reaction kettle, stirring, dissolving and clarifying; adding 50g of elemental iodine, heating to 105 deg.C for reaction, and stopping the reaction by monitoring naringenin residue content of less than 10% by HPLC (HPLC detection method: Asahi C18 column, mobile phase is 50% acetonitrile water solution). After the reaction is finished, cooling to 85 ℃, adding 1L of 40% ethanol aqueous solution, adjusting the pH to 11 by 10mol/L lithium hydroxide solution, adding 100g of sodium hydrosulfite, and reacting for 2 hours; filtering while the solution is hot, and washing a filter cake by using 500ml of dimethyl sulfoxide; mixing the filtrates, cooling to 40 deg.C, adding 500ml glacial acetic acid to adjust pH to 6-7, adding dropwise 15L 70% ethanol water solution at 140ml/min to precipitate a large amount of apigenin, monitoring apigenin precipitation amount by HPLC to be greater than 90%, filtering to obtain filter cake (refined apigenin product), and vacuum drying at 50 deg.C to obtain pale yellow apigenin dried product 923 g; the weight yield was 92.30%. Loss on drying of 1.53%; HPLC purity 98.46%; the apigenin obtained by the previous step is subjected to content measurement by taking the apigenin standard substance (marked content is 98%) of the Zhongzhong institute as a control, and the content is detected to be 97.64%.

Example 4

Adding 1kg of naringenin and 5L of dimethyl sulfoxide into a 50L glass reaction kettle, stirring, dissolving and clarifying; adding 50g of elemental iodine, heating to 110 deg.C for reaction, and stopping the reaction by monitoring naringenin residue content of less than 10% by HPLC (HPLC detection method: Asahi C18 column, mobile phase is 50% acetonitrile water solution). After the reaction is finished, cooling to 90 ℃, adding 1L of 30% ethanol water solution, adjusting the pH to 10 by 10mol/L sodium carbonate solution, adding 200g of preservative powder, and reacting for 1 h; filtering while the solution is hot, and washing a filter cake by using 500ml of dimethyl sulfoxide; mixing filtrates, cooling to 40 deg.C, adding 500ml sulfuric acid to adjust pH of reaction solution to 6-7, dripping 15L 50% ethanol water solution at 90ml/min to precipitate a large amount of apigenin, monitoring by HPLC to obtain apigenin solution with a precipitation amount greater than 90%, filtering, and pulping filter cake with 15L 60% ethanol water solution at 70 deg.C for 10 hr; filtering to obtain refined apigenin filter cake, and vacuum drying at 50 deg.C for 30 hr to obtain pale yellow apigenin dried product 937 g; the weight yield was 93.70%. Loss on drying is 1.79%; HPLC purity 98.69%; the apigenin obtained by the previous step is subjected to content measurement by taking the apigenin standard substance (marked content is 98%) of the Zhongzhong institute as a reference, and the content is detected to be 97.42%.