Extraction and purification process of macleaya cordata alkaloid
阅读说明：本技术 一种博落回生物碱的提取和纯化工艺 (Extraction and purification process of macleaya cordata alkaloid ) 是由 谈满良 罗炼辉 李小冬 赵会平 于 2020-06-02 设计创作，主要内容包括：本发明提供了一种博落回生物碱的提取和纯化工艺,包括以下步骤：取博落回粉末,加入石油醚,水浴回流；取药渣,加入酸乙醇,静置,超声提取,抽滤,滤液浓缩；干燥成粗博落回生物碱粉末；用蒸馏水分别配制聚乙二醇溶液和硫酸铵溶液；将聚乙二醇溶液、硫酸铵溶液、蒸馏水、粗博落回生物碱粉末混合；置于摇床中振摇,充分混合均匀,离心；用胶头滴管吸取上下相溶液,测量紫外吸光度,并以血根碱、白屈菜红碱、博落回碱、小檗碱的标准溶液作参比；分别测定上、下相溶液中各生物碱的含量,计算萃取率。本发明方法简单,萃取率高,血根碱、白屈菜红碱、博落回碱、小檗碱的萃取率最高分别为99.74%、99.44%、99.65%、99.49%。(The invention provides a macleaya cordata alkaloid extraction and purification process, which comprises the following steps: adding petroleum ether into herba Macleayae Cordatae powder, and refluxing in water bath; collecting residue, adding acid ethanol, standing, ultrasonic extracting, vacuum filtering, and concentrating the filtrate; drying to obtain crude macleaya cordata alkaloid powder; preparing a polyethylene glycol solution and an ammonium sulfate solution by using distilled water respectively; mixing polyethylene glycol solution, ammonium sulfate solution, distilled water, and coarse Macleaya cordata alkaloid powder; shaking in shaking table, mixing, and centrifuging; sucking the upper and lower phase solutions with rubber dropper, measuring ultraviolet absorbance, and using standard solution of sanguinarine, chelerythrine, macleaya cordata alkali, and berberine as reference; and respectively measuring the content of each alkaloid in the upper phase solution and the lower phase solution, and calculating the extraction rate. The method is simple, the extraction rate is high, and the extraction rates of sanguinarine, chelerythrine, macleaya cordata alkali and berberine are respectively 99.74%, 99.44%, 99.65% and 99.49%.)
1. A technology for extracting and purifying macleaya cordata alkaloid is characterized in that: the method comprises the following steps:
step 1: adding petroleum ether into macleaya cordata powder according to the mass ratio of 1:10-15, and refluxing for 2-3h in water bath at 60 ℃;
step 2: taking the residue, adding 1% acid ethanol, standing for 30min, performing ultrasonic extraction for 30-50min, vacuum filtering, and concentrating the filtrate to 25mL, wherein the ratio of the residue to the 1% acid ethanol is 1: 25-35;
and step 3: drying to obtain crude macleaya cordata alkaloid powder;
and 4, step 4: preparing a polyethylene glycol solution with the mass fraction of 40% and an ammonium sulfate solution with the mass fraction of 30% by using distilled water respectively;
and 5: mixing polyethylene glycol solution, ammonium sulfate solution, distilled water, and coarse Macleaya cordata alkaloid powder;
step 6: placing in a shaking table, shaking for 2-3h at the speed of 200-;
and 7: sucking the upper and lower phase solutions with rubber dropper, measuring ultraviolet absorbance, and using standard solution of sanguinarine, chelerythrine, macleaya cordata alkali, and berberine as reference;
and 8: respectively measuring the content of sanguinarine, chelerythrine, macleaya cordata alkali and berberine in the upper phase solution and the lower phase solution, and calculating the extraction rate.
2. The process of claim 1 for the extraction and purification of macleaya cordata alkaloids, wherein: the extraction temperature in the step 2 is 40-60 ℃, and the ultrasonic power is 120-160W.
3. The process of claim 1 for the extraction and purification of macleaya cordata alkaloids, wherein: and in the step 4, the polyethylene glycol PEG is PEG1000, PEG2000 or PEG 6000.
4. The process of claim 1 for the extraction and purification of macleaya cordata alkaloids, wherein: and in the step 5, the liquid-solid ratio of the polyethylene glycol solution to the ammonium sulfate solution to the distilled water to the crude macleaya cordata alkaloid powder is 4:3:7: 1.
5. The process of claim 1 for the extraction and purification of macleaya cordata alkaloids, wherein: the calculation formula of the content measured in the step 8 is as follows: r = Vt/VbIn the formula: r is a phase ratio, Vt、VbThe volume of the upper phase and the lower phase is mL respectively; k = Ct/CbIn the formula: k is inA distribution coefficient; ct，CbThe mass concentrations of the upper phase alkaloid and the lower phase alkaloid are mg/L respectively; extraction ratio (Y)/% =1/[1+1/(RK)]。
Technical Field
The invention relates to the field of medicinal chemistry, in particular to a macleaya cordata alkaloid extraction and purification process.
Background
Macleaya cordata, named as Bizhao, Sanqian, Bolerui, mountain phoenix tree, etc. is a whole herb with root of Macleaya cordata of Papaveraceae, widely used as a medicine, widely distributed in southern provinces of Yangtze river basin, especially in clouds, honours and Sichuan, and is widely grown in wild slopes of barren mountains, is visible everywhere, and can be artificially planted. Macleaya cordata is pungent, bitter, warm, and toxic, and has the functions of expelling wind and removing toxicity, dispelling stasis and swelling, and killing and expelling parasites. It is used by folk to treat superficial body diseases such as carbuncle, furuncle, malignant ulcer, scald, intractable tinea, rheumatic arthralgia, traumatic injury, scabies, etc., and is not used internally because of its strong toxicity. Modern medical research shows that the effective medicinal component of macleaya cordata is alkaloid, and various kinds of sanguinarine, chelerythrine, macleaya cordata alkali, protopine, berberine and the like are extracted and separated simultaneously, and are called total alkaloids. The macleaya cordata total alkaloids have the effects of resisting bacteria and inflammation, inhibiting various pathogenic bacteria, resisting viruses, clearing away heat and toxic materials, enhancing the immune function and resisting various tumors, and are widely used for treating various infectious diseases of people and livestock; the pesticide can kill pests and bacteria, is mainly used as a raw material of various bioactive medicines, veterinary medicines and biological pesticides for controlling plant diseases and insect pests of vegetables, fruits, cotton and grain crops, and is a pure natural green biological 'three-medicine' product.
In order to better utilize macleaya cordata alkaloid and maximize the application and the effect of the macleaya cordata alkaloid, the macleaya cordata alkaloid must be purified, so that scientific basis is provided for most effectively developing and utilizing the macleaya cordata total alkaloid and exerting the maximum economic benefit of macleaya cordata in production.
Disclosure of Invention
The technical problem to be solved is as follows: the invention aims to provide a macleaya cordata alkaloid extraction and purification process, which has the advantages of simple method and high extraction rate, and the extraction rates of sanguinarine, chelerythrine, macleaya cordata alkali and berberine are respectively 99.74%, 99.44%, 99.65% and 99.49% at the highest.
The technical scheme is as follows: a process for extracting and purifying macleaya cordata alkaloid comprises the following steps:
step 1: adding petroleum ether into macleaya cordata powder according to the mass ratio of 1:10-15, and refluxing for 2-3h in water bath at 60 ℃;
step 2: taking the residue, adding 1% acid ethanol, standing for 30min, performing ultrasonic extraction for 30-50min, vacuum filtering, and concentrating the filtrate to 25mL, wherein the ratio of the residue to the 1% acid ethanol is 1: 25-35;
and step 3: drying to obtain crude macleaya cordata alkaloid powder;
and 4, step 4: preparing a polyethylene glycol solution with the mass fraction of 40% and an ammonium sulfate solution with the mass fraction of 30% by using distilled water respectively;
and 5: mixing polyethylene glycol solution, ammonium sulfate solution, distilled water, and coarse Macleaya cordata alkaloid powder;
step 6: placing in a shaking table, shaking for 2-3h at the speed of 200-;
and 7: sucking the upper and lower phase solutions with rubber dropper, measuring ultraviolet absorbance, and using standard solution of sanguinarine, chelerythrine, macleaya cordata alkali, and berberine as reference;
and 8: respectively measuring the content of sanguinarine, chelerythrine, macleaya cordata alkali and berberine in the upper phase solution and the lower phase solution, and calculating the extraction rate.
Further preferably, the extraction temperature in the step 2 is 40-60 ℃, and the ultrasonic power is 120-160W.
Further preferably, the polyethylene glycol PEG in step 4 is PEG1000, PEG2000, or PEG 6000.
Further preferably, the liquid-solid ratio of the polyethylene glycol solution, the ammonium sulfate solution, the distilled water and the crude macleaya cordata alkaloid powder in the step 5 is 4:3:7: 1.
Further preferably, the calculation formula for determining the content in step 8 is: r = Vt/VbIn the formula: r is a phase ratio, Vt、VbThe volume of the upper phase and the lower phase is mL respectively; k = Ct/CbIn the formula: k is a distribution coefficient; ct，CbThe mass concentrations of the upper phase alkaloid and the lower phase alkaloid are mg/L respectively; extraction ratio (Y)/% =1/[1+1/(RK)]。
Has the advantages that:
1. the extraction and purification method adopted by the invention is a liquid-liquid extraction method, and has the advantages of small loss of active ingredients, few separation steps, mild operation conditions, easy amplification and no organic solvent residue.
2. The method of the invention obtains the best separation and purification process, and has high extraction rate, and the extraction rates of sanguinarine, chelerythrine, macleaya cordata alkali and berberine are respectively 99.74%, 99.44%, 99.65% and 99.49%.
Detailed Description