阅读说明：本技术 一种从华山参中分离纯化东莨菪碱单体的方法 (Method for separating and purifying scopolamine monomer from radix physochlainae ) 是由 于喜洋 刘东滨 刘千驰 于 2021-08-28 设计创作，主要内容包括：本发明提供一种从华山参中分离纯化东莨菪碱单体的方法,涉及中药成分提取技术领域。该方法包括以下步骤：向华山参中加入乙醚和体积浓度为70％-80％的乙醇,经过超声提取和离心,取上清液,将上清液抽真空得到第一残渣,向第一残渣加入氢氧化钠溶液,再加入氯仿进行萃取,得到氯仿萃取溶液,将氯仿萃取溶液蒸干得到第二残渣；向第二残渣中加入盐酸溶液,然后加入D151大孔吸附树脂吸附5-10h,吸附后,将D151大孔吸附树脂用洗脱液洗脱6-12h。通过该方法分离得到的东莨菪碱纯度更高。(The invention provides a method for separating and purifying scopolamine monomers from radix physochlainae, and relates to the technical field of extraction of traditional Chinese medicine components. The method comprises the following steps: adding diethyl ether and ethanol with the volume concentration of 70-80% into the physochlaina root, performing ultrasonic extraction and centrifugation, taking supernate, vacuumizing the supernate to obtain first residue, adding a sodium hydroxide solution into the first residue, adding chloroform for extraction to obtain a chloroform extraction solution, and evaporating the chloroform extraction solution to dryness to obtain second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorption resin for adsorption for 5-10h, and eluting the D151 macroporous adsorption resin with eluent for 6-12h after adsorption. The scopolamine separated by the method has higher purity.)

1. A method for separating and purifying scopolamine monomers from radix physochlainae is characterized by comprising the following steps:

adding diethyl ether and ethanol with the volume concentration of 70-80% into the physochlaina root, performing ultrasonic extraction and centrifugation, taking supernate, vacuumizing the supernate to obtain first residue, adding a sodium hydroxide solution into the first residue, adding chloroform for extraction to obtain a chloroform extraction solution, and evaporating the chloroform extraction solution to dryness to obtain second residue; and adding a hydrochloric acid solution into the second residue, then adding D151 macroporous adsorption resin for adsorption for 5-10h, and eluting the D151 macroporous adsorption resin with an eluent for 6-12h after adsorption.

2. The method according to claim 1, wherein the volume ratio of the diethyl ether to the ethanol is 10: (1-5).

3. The method according to claim 2, wherein the total mass of the diethyl ether and the ethanol is 10 to 20 times the mass of the radix physochlainae.

4. The method according to claim 1, wherein the ultrasonic extraction is performed for 2 times, the 1 st ultrasonic extraction is performed for 30-40min, the first filtrate and the filter residue are separated, the ethanol is added to the filter residue to perform the 2 nd ultrasonic extraction, the 2 nd ultrasonic extraction is performed for 20-30min, the second filtrate is separated, and the first filtrate and the second filtrate are mixed and then centrifuged.

5. The method as claimed in claim 4, wherein the centrifugation time is 30-60min and the centrifugation speed is 500-800 r/s.

6. The method of claim 1, wherein the supernatant is vacuumed at 55-65 ℃ for 50-100 min.

7. The method of claim 1, further comprising adding 0.5-1 mol/L sulfuric acid to the first residue before adding the sodium hydroxide solution to the first residue, followed by extraction with dichloromethane for 30-90min, and discarding the dichloromethane layer.

8. The method as claimed in claim 1, wherein the chloroform is used for 3 times, each time for 25-35min, and the chloroform is used in an amount of 10-20mL, 5-10mL, and 5-8mL in sequence.

9. The method according to claim 1, wherein the eluent is a hydrochloric acid solution with a mass concentration of 0.1% -0.5%.

10. The method as claimed in claim 1, further comprising centrifuging the solution obtained after elution at 1000-1500r/s for 10-20 min.

Technical Field

The invention relates to the technical field of extraction of traditional Chinese medicine components, and particularly relates to a method for separating and purifying scopolamine monomers from radix physochlainae.

Background

Scopolamine is a tropane-type alkaloid present in plants of the Solanaceae family. Scopolamine is an alkaloid with the strongest pharmacological action in belladonna, is a viscous syrup-like liquid at normal temperature, and is bitter and spicy in taste; is easily soluble in ethanol, chloroform, acetone and hot water, is slightly soluble in benzene and petroleum ether, and can form crystalline salt with various inorganic acids or organic acids. Scopolamine can be used for blocking parasympathetic nerve, and can also be used as central nervous system inhibitor, its hydrobromide is generally used clinically, and can be used for anesthesia, analgesia, cough relieving, asthma relieving, motion sickness relieving, and Parkinson's disease stiffness and tremor controlling.

At present, the extraction of scopolamine from radix codonopsis lanceolatae is one of the sources of scopolamine, but the extraction purity of scopolamine is not high by the existing extraction method. Therefore, in order to obtain scopolamine with higher purity, it is necessary to provide a new method for separating scopolamine monomers from radix physochlainae.

Disclosure of Invention

The invention aims to provide a method for separating and purifying scopolamine monomers from radix physochlainae, and the scopolamine separated by the method has higher purity.

The technical problem to be solved by the invention is realized by adopting the following technical scheme.

The invention provides a method for separating and purifying scopolamine monomers from radix physochlainae, which comprises the following steps:

adding diethyl ether and ethanol with the volume concentration of 70-80% into the physochlaina root, performing ultrasonic extraction and centrifugation, taking supernate, vacuumizing the supernate to obtain first residue, adding a sodium hydroxide solution into the first residue, adding chloroform for extraction to obtain a chloroform extraction solution, and evaporating the chloroform extraction solution to dryness to obtain second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorption resin for adsorption for 5-10h, and eluting the D151 macroporous adsorption resin with eluent for 6-12h after adsorption.

The method for separating and purifying the scopolamine monomer from the radix physochlainae in the embodiment of the invention at least has the following beneficial effects: according to the method, firstly, the Huashan mountain is mixed with the ether and the ethanol, the dispersion speed of the Huashan mountain ginseng in the solution is accelerated through ultrasonic extraction and the oscillation action generated by ultrasonic waves, the ether and the ethanol are easier to permeate into the Huashan mountain ginseng through continuous oscillation of the ultrasonic waves, and the mixing of the Huashan mountain with the ether and the ethanol is accelerated. The scopolamine and the hyoscyamine in the radix physochlainae can be extracted by ethanol with the volume concentration of 70-80%, namely the scopolamine and the hyoscyamine are dissolved in ethanol and ether. The first residue is obtained by centrifuging and vacuumizing, and other substances dissolved in ethanol or diethyl ether are mixed in the first residue, so that the first residue needs to be separated and purified for the second time. Adding a sodium hydroxide solution into the first residue, mainly for adjusting the pH, when the pH is alkalescent, extracting by using chloroform, and avoiding influencing the extraction effect of the chloroform in an acidic environment, specifically, salifying the alkaloid, wherein the alkaloid is easily dissolved in water after salifying, and thus the alkaloid is more easily extracted when extracting by using chloroform. The chloroform extract solution was evaporated to dryness, primarily to remove the chloroform, to obtain a second residue, a crude scopolamine extract. The crude extract of scopolamine contains other scopolamine, needs to be separated and purified again, and is adsorbed by D151 macroporous resin, and the D151 macroporous resin has good adsorption rate and elution rate to scopolamine and no adsorption to other scopolamine, so the scopolamine with good purity can be obtained after adsorption and elution.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail below with reference to specific examples.

A method for separating and purifying scopolamine monomers from radix Physochlainae comprises the following steps:

adding diethyl ether and ethanol with volume concentration of 70-80% into radix physochlainae, performing ultrasonic extraction for 2 times, performing ultrasonic extraction for 1 st time for 30-40min, separating and taking first filtrate and filter residue, adding ethanol into the filter residue to perform ultrasonic extraction for 2 times, performing ultrasonic extraction for 2 nd time for 20-30min, separating and taking second filtrate, mixing the first filtrate and the second filtrate, centrifuging for 30-60min at a centrifuging speed of 800r/s, taking supernatant after centrifuging, vacuumizing the supernatant at 55-65 ℃ for 50-100min to obtain first residue, adding sodium hydroxide solution into the first residue, adding chloroform for extraction, performing chloroform extraction for 3 times, performing extraction for 25-35min each time, mixing the extract solutions obtained each time to obtain chloroform extraction solution, and evaporating the chloroform extraction solution to obtain second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorption resin for adsorption for 5-10h, eluting the D151 macroporous adsorption resin with eluent for 6-12h, and centrifuging the solution obtained after elution at 1500r/s for 10-20min to obtain the scopolamine finished product.

Scopolamine is a tertiary amine antimuscarinic drug that generally acts like atropine, both centrally and peripherally, but it acts more strongly than atropine in inhibiting salivation, generally slowing rather than accelerating heart rate, especially when used in small doses. It has different central action from atropine, and scopolamine can inhibit cerebral cortex, produce lethargy and amnesia, and has stronger inhibiting effect on eye smooth muscle and gland secretion than atropine. The scopolamine has similar peripheral effect to atropine, has stronger inhibition effect on secretion of salivary glands, bronchus and sweat glands than atropine, has quicker effect on mydriasis of eyes and paralysis regulation than atropine, has quicker disappearance of the effect, has weaker effect on heart, intestinal canal and bronchial smooth muscle, and also has the effects of relieving vasospasm and improving microcirculation. The traditional Chinese medicine composition has obvious inhibition effect on the center at the general treatment dose, can generate hypnosis at a larger dose, can generate central excitation symptoms such as agitation, uneasiness, hallucination or delirium and the like at a large dose, but can quickly enter a sleeping state, and has more obvious cortical inhibition at a larger dose (intravenous injection of 0.08 mg/kg).

Radix Physochlainae Infangularis Kuang is dried root of Physochlaina Infangularis Kuang of Solanaceae, has effects of warming lung, eliminating phlegm, relieving asthma and cough, tranquilizing mind and relieving convulsion, and contains various alkaloids. Alkaloids are nitrogen-containing basic organic compounds existing in nature, and have alkali-like properties, so the alkaloids are also called pseudo-alkalis in the past. Most of the Chinese herbal medicines have complex ring structures, nitrogen is mostly contained in the ring, and the Chinese herbal medicines have remarkable biological activity, are one of important effective components in the Chinese herbal medicines and also have optical activity. The alkaloid has a cyclic structure, is difficult to dissolve in water, can form a salt with acid, has certain optical activity and absorption spectrum, mostly has bitter taste, is colorless crystals, and is a liquid in a small amount. Alkaloids, which are synthesized from different amino acids or their direct derivatives, are one of the secondary metabolites and have toxic or strong physiological effects on living organisms. At normal pressure, most alkaloids are nonvolatile, and can be directly heated to melt first and then decomposed, or melt and simultaneously decompose, and only under high vacuum can sublimation be caused by heating.

The macroporous adsorption resin is a polymer adsorption resin which does not contain exchange groups and has a macroporous structure, has a good macroporous network structure and a larger specific surface area, and is widely favored in separation and purification of natural medicine active ingredients due to the excellent characteristics of good adsorbability, rapid adsorption, easy desorption and the like. The D151 macroporous adsorption resin has high adsorption rate and elution rate on scopolamine, and has good separation and purification effects on scopolamine, so that the D151 macroporous adsorption resin is used for separating and purifying the scopolamine, and the scopolamine with higher purity can be obtained.

In this embodiment, since the alkaloid is insoluble in water and soluble in organic solvents, ethyl ether and ethanol are used as solvents for radix physochlainae. The ether and the ethanol are selected as solvents, and the biocompatibility of the solution can be improved after the ether and the ethanol are mixed, so that the alkaloid is more easily dissolved in the solution, and the scopolamine is prevented from being trapped by substances insoluble in organic solvents in the radix physochlainae, so that the scopolamine in the radix physochlainae cannot be completely dissolved in the ether and the ethanol, and the purity of the scopolamine obtained by separation and purification is reduced. In detail, ethanol with the volume concentration of 70-80% is selected, and scopolamine and hyoscyamine in the radix physochlainae can be extracted by the ethanol with proper concentration, namely the scopolamine and the hyoscyamine are dissolved in the ethanol and the ether, so that the subsequent extraction of the scopolamine is simpler. Alternatively, the volume ratio of diethyl ether to ethanol is 10: (1-5), the appropriate proportion of the ethyl ether and the ethanol can ensure that the biocompatibility effect of the solution is better, and further ensure that the scopolamine in the radix physochlainae can be completely separated and extracted. Wherein the total mass of the ether and the ethanol is 10-20 times of the mass of the radix physochlainae, so that the condition that the amount of the ether and the ethanol is too small to completely extract the scopolamine in the radix physochlainae can be avoided.

In the embodiment, the radix physochlainae is subjected to ultrasonic extraction for 2 times, the diffusion speed of the radix physochlainae in the solution is accelerated by the oscillation action generated by the ultrasonic waves, and the diethyl ether and the ethanol are easier to permeate into the radix physochlainae by the continuous oscillation of the ultrasonic waves, so that the radix physochlainae is accelerated to participate in the mixing of the diethyl ether and the ethanol. Extracting for 30-40min at 1 st time, because the content of the extract in the solution is high during the 1 st extraction, the extract can be extracted completely in the extraction time, and the extract which is not completely extracted may exist in the filter residue after the 1 st extraction, so that the 2 nd extraction is needed. The extraction time of the second extraction is 20-30min, and because the content of the extracted substances in the filter residue is low, the extraction can not only extract residual substances, but also save time and avoid overlong extraction time.

Optionally, before adding the sodium hydroxide solution into the first residue, adding 0.5mol/L-1mol/L sulfuric acid into the first residue, extracting with dichloromethane for 30-90min, and discarding dichloromethane layer. Sulfuric acid can remove water-soluble impurities from the first residue, and methylene chloride can remove fat-soluble impurities from the first residue. The sulfuric acid with the concentration of 0.5-1 mol/L is selected for impurity removal, the dissolution degree of strong acid is high, water-soluble impurities in the first residue can be thoroughly removed, the impurity content is reduced, and further the water-soluble impurities in the first residue can be completely removed. Extracting with dichloromethane for 30-90min, wherein dichloromethane can better infiltrate into the first residue within the extraction time, so as to ensure that dichloromethane can remove fat-soluble impurities on the surface inside the first residue, and further improve the content of the extracted substances of the first residue.

Optionally, when performing the 3-time chloroform extraction, the amount of chloroform is 10-20mL, 5-10mL, and 5-8mL in sequence. 10-20mL of chloroform was used for the 1 st extraction, since the scopolamine content in the solution was high, more chloroform was required to extract it. 5-10mL of chloroform is adopted for extraction for the 2 nd and the 3 rd time, the scopolamine content is lower at the moment, the scopolamine can be completely extracted by using 5-8mL of chloroform, and the chloroform waste caused by excessive chloroform dosage is avoided.

In this embodiment, the supernatant is vacuumized at 55-65 ℃ to obtain the first residue, and the first residue is vacuumized at the temperature, so that the phenomenon that the form of scopolamine in the first residue is changed or the crystal form of scopolamine is damaged due to overhigh temperature can be avoided, the purity of the obtained scopolamine is reduced, and ethanol and ether can be evaporated at the temperature to obtain a crude scopolamine extract. Vacuumizing for 50-100min to evaporate the ethanol and ether from the first residue and reduce the influence of ethanol and ether on the subsequent operation.

In detail, the eluent is a hydrochloric acid solution with the mass concentration of 0.1% -0.5%, the hydrochloric acid solution is selected to elute the D151 macroporous adsorption resin, the hydrochloric acid reacts with the scopolamine to obtain scopolamine hydrochloride, the scopolamine hydrochloride is easily dissolved in water, so that the scopolamine on the D151 macroporous adsorption resin can be eluted, in addition, the reaction speed of the hydrochloric acid and the scopolamine is higher, so that the elution speed of the scopolamine is higher, and the elution rate is higher. Hydrochloric acid with the mass concentration of 0.1% -0.5% is selected for elution, the ion concentration in the solution is higher at the concentration, and when scopolamine is eluted, the ions react with scopolamine more quickly, so that the elution speed can be improved.

The features and properties of the present invention are described in further detail below with reference to examples.

Example 1

A method for separating and purifying scopolamine monomers from radix Physochlainae comprises the following steps:

adding diethyl ether and 70% ethanol by volume into radix physochlainae, performing ultrasonic extraction for 2 times, performing ultrasonic extraction for 1 st time for 30min, separating to obtain a first filtrate and a filter residue, adding ethanol into the filter residue to perform ultrasonic extraction for 2 times, performing ultrasonic extraction for 2 nd time for 20min, separating to obtain a second filtrate, mixing the first filtrate and the second filtrate, centrifuging for 30min at a centrifuging speed of 500r/s, centrifuging to obtain a supernatant, vacuumizing the supernatant at 55 ℃ for 50min to obtain a first residue, adding a sodium hydroxide solution into the first residue, adding chloroform for extraction, performing chloroform extraction for 3 times, performing chloroform extraction for 25min each time, mixing the obtained extract liquids to obtain a chloroform extraction solution, and evaporating the chloroform extraction solution to obtain a second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorption resin, adsorbing for 5h, eluting D151 macroporous adsorption resin with eluent for 6h, and centrifuging the solution obtained after elution at 1000r/s for 10min to obtain scopolamine finished product.

In this example, the volume ratio of ethyl ether to ethanol is 10: 1, the total mass of the ether and the ethanol is 10 times of that of the physochlaina segetum, the mass concentration of the hydrochloric acid solution is 0.1 percent, and the dosage of the chloroform is 10mL, 5mL and 5mL in sequence when the chloroform is extracted for 3 times.

Example 2

A method for separating and purifying scopolamine monomers from radix Physochlainae comprises the following steps:

adding diethyl ether and 80% ethanol by volume into radix physochlainae, performing ultrasonic extraction for 2 times, performing ultrasonic extraction for 40min for the 1 st time, separating to obtain a first filtrate and filter residue, adding ethanol into the filter residue to perform ultrasonic extraction for the 2 nd time, performing ultrasonic extraction for the 2 nd time for 30min, separating to obtain a second filtrate, mixing the first filtrate and the second filtrate, centrifuging for 60min at a centrifuging speed of 800r/s, centrifuging to obtain a supernatant, vacuumizing the supernatant at 65 ℃ for 100min to obtain a first residue, adding a sodium hydroxide solution into the first residue, adding chloroform for extraction, performing chloroform extraction for 3 times, performing chloroform extraction for 35min each time, mixing the obtained extract liquids to obtain a chloroform extraction solution, and evaporating the chloroform extraction solution to obtain a second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorbent resin, adsorbing for 10h, eluting D151 macroporous adsorbent resin with eluent for 12h, and centrifuging the solution at 1500r/s for 20min to obtain scopolamine.

In this example, the volume ratio of ethyl ether to ethanol is 10: 5, the total mass of the ether and the ethanol is 20 times of that of the physochlaina hance, the mass concentration of the hydrochloric acid solution is 0.5 percent, and the dosage of the chloroform during 3-time chloroform extraction is 20mL, 10mL and 8mL in sequence.

Example 3

A method for separating and purifying scopolamine monomers from radix Physochlainae comprises the following steps:

adding diethyl ether and 75% ethanol by volume into radix physochlainae, performing ultrasonic extraction for 2 times, performing ultrasonic extraction for 35min for 1 st time, separating to obtain a first filtrate and a filter residue, adding ethanol into the filter residue to perform ultrasonic extraction for 2 nd time, performing ultrasonic extraction for 25min for 2 nd time, separating to obtain a second filtrate, mixing the first filtrate and the second filtrate, centrifuging for 40min at the centrifuging speed of 700r/s, centrifuging to obtain a supernatant, vacuumizing the supernatant at 60 ℃ for 70min to obtain a first residue, adding a sodium hydroxide solution into the first residue, adding chloroform for extraction, performing chloroform extraction for 3 times, performing chloroform extraction for 30min each time, mixing the obtained extract liquids to obtain a chloroform extraction solution, and evaporating the chloroform extraction solution to obtain a second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorbent resin, adsorbing for 6h, eluting D151 macroporous adsorbent resin with eluent for 8h, and centrifuging the solution obtained after elution at 1100r/s for 15min to obtain scopolamine finished product.

In this example, the volume ratio of ethyl ether to ethanol is 10: 3, the total mass of the ether and the ethanol is 15 times of that of the physochlaina hance, the mass concentration of the hydrochloric acid solution is 0.2 percent, and the dosage of the chloroform during 3-time chloroform extraction is 15mL, 8mL and 6mL in sequence.

Example 4

A method for separating and purifying scopolamine monomers from radix Physochlainae comprises the following steps:

adding diethyl ether and 72% ethanol by volume into radix physochlainae, performing ultrasonic extraction for 2 times, performing ultrasonic extraction for 32min for the 1 st time, separating to obtain a first filtrate and filter residue, adding ethanol into the filter residue to perform ultrasonic extraction for the 2 nd time, performing ultrasonic extraction for the 2 nd time for 22min, separating to obtain a second filtrate, mixing the first filtrate and the second filtrate, centrifuging for 50min at a centrifuging speed of 600r/s, centrifuging to obtain a supernatant, vacuumizing the supernatant at 58 ℃ for 80min to obtain a first residue, adding a sodium hydroxide solution into the first residue, adding chloroform for extraction, performing chloroform extraction for 3 times, extracting for 27min each time, mixing the obtained extract liquids to obtain a chloroform extraction solution, and evaporating the chloroform extraction solution to obtain a second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorbent resin, adsorbing for 8h, eluting D151 macroporous adsorbent resin with eluent for 10h, and centrifuging the solution obtained after elution at 1200r/s for 12min to obtain scopolamine finished product.

In this example, the volume ratio of ethyl ether to ethanol is 10: 4, the total mass of the ether and the ethanol is 12 times of that of the physochlaina hance, the mass concentration of the hydrochloric acid solution is 0.3 percent, and the dosage of the chloroform during 3-time chloroform extraction is 16mL, 7mL and 7mL in sequence.

Example 5

A method for separating and purifying scopolamine monomers from radix Physochlainae comprises the following steps:

adding diethyl ether and 78% ethanol, ultrasonic extracting for 2 times, ultrasonic extracting for 1 st time for 37min, separating to obtain first filtrate and residue, adding ethanol into the residue for 2 nd time ultrasonic extracting, ultrasonic extracting for 2 nd time for 27min, separating to obtain second filtrate, mixing the first filtrate and the second filtrate, centrifuging for 45min at a speed of 650r/s, centrifuging to obtain supernatant, vacuumizing the supernatant at 62 deg.C for 75min to obtain first residue, adding 0.5mol/L sulfuric acid into the first residue, extracting with dichloromethane for 40min, discarding dichloromethane layer, adding sodium hydroxide solution, adding chloroform, extracting with chloroform for 3 times (32 min each time), mixing the extractive solutions to obtain chloroform extractive solution, and evaporating the chloroform extractive solution to obtain second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorbent resin, adsorbing for 7.5h, eluting D151 macroporous adsorbent resin with eluent for 9h, and centrifuging the solution obtained after elution at 1300r/s for 17min to obtain scopolamine finished product.

In this example, the volume ratio of ethyl ether to ethanol is 10: 2, the total mass of the ether and the ethanol is 17 times of that of the physochlaina hance, the mass concentration of the hydrochloric acid solution is 0.4 percent, and the dosage of the chloroform during 3-time chloroform extraction is 13mL, 7.5mL and 6.5mL in sequence.

Example 6

A method for separating and purifying scopolamine monomers from radix Physochlainae comprises the following steps:

adding diethyl ether and 75% ethanol by volume into radix Physochlainae, ultrasonically extracting for 30min, centrifuging for 45min at 650r/s, collecting supernatant, vacuumizing the supernatant at 60 deg.C for 65min to obtain first residue, adding sodium hydroxide solution into the first residue, adding chloroform, extracting for 30min to obtain chloroform extract solution, and evaporating the chloroform extract solution to obtain second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorption resin for adsorption for 6.5h, and eluting the D151 macroporous adsorption resin with eluent for 8.5h after adsorption.

In this example, the volume ratio of ethyl ether to ethanol is 10: 2.5, the total mass of the ether and the ethanol is 13 times of that of the physochlaina hance, the mass concentration of the hydrochloric acid solution is 0.15 percent, and the dosage of the chloroform during 3-time chloroform extraction is 14mL, 8.5mL and 5.5mL in sequence.

Example 7

A method for separating and purifying scopolamine monomers from radix Physochlainae comprises the following steps:

adding diethyl ether and 76% ethanol into radix Physochlainae, ultrasonically extracting for 35min, centrifuging at 750r/s for 55min, collecting supernatant, vacuumizing the supernatant at 63 deg.C for 55min to obtain first residue, adding sodium hydroxide solution into the first residue, adding chloroform, extracting for 30min to obtain chloroform extract solution, and evaporating the chloroform extract solution to obtain second residue; adding hydrochloric acid solution into the second residue, adding D151 macroporous adsorption resin for adsorption for 8.5h, and eluting the D151 macroporous adsorption resin with eluent for 11h after adsorption.

In this example, the volume ratio of ethyl ether to ethanol is 10: 3.5, the total mass of the ether and the ethanol is 18 times of that of the physochlaina root, the mass concentration of the hydrochloric acid solution is 0.35 percent, and the dosage of the chloroform during 3-time chloroform extraction is 18mL, 9.5mL and 6.8mL in sequence.

Test results

Taking the scopolamine samples in the examples 1 to 7 to perform high performance liquid chromatography analysis, obtaining chromatographic peaks of the scopolamine samples in each example, comparing the chromatographic peaks with a standard substance, substituting the area of the chromatographic peaks obtained in each example into a formula of a standard curve to calculate the content, and obtaining the purity of the scopolamine in the samples through content calculation, wherein the specific data are as follows: (in the test, the standard was calibrated)

TABLE 1 statistical table of results

Examples	1	2	3	4	5	6	7
								Purity of	70％	65％	85％	81％	88％	60％	58％

As is clear from Table 1, the purity of scopolamine obtained in example 5 was the best. Example 5 compared with examples 1-4, the operation procedure was substantially the same, the operation conditions were different, and the addition of sulfuric acid to the first residue, extraction with dichloromethane, and removal of the dichloromethane layer allowed further removal of impurities in the alkaloid, thereby improving the purity of the obtained scopolamine, and it was found that the purity of the obtained scopolamine was the best under the operation conditions of example 5. Example 5 compared with examples 6-7, in example 5, the number of times of ultrasonic extraction and chloroform extraction was increased, and the number of times of treatment of the first residue was also increased, and the treatment of the first residue enabled the purity of scopolamine to be improved, and the number of times of ultrasonic extraction and chloroform extraction was increased, and multiple extractions allowed further improvement of the separation effect of scopolamine in the solution, and further improved the purity of the obtained scopolamine. Therefore, the scopolamine with higher purity can be separated and extracted from the physochlaina.

In summary, in the method for separating and purifying scopolamine monomers from radix physochlainae according to the embodiments of the present invention, radix physochlainae is first mixed and dissolved in ethyl ether and ethanol, and then subjected to ultrasonic extraction and the oscillation action generated by ultrasonic waves, so that the diffusion speed of radix physochlainae in a solution is increased, and the ethyl ether and the ethanol are more easily permeated into the radix physochlainae through the continuous oscillation of the ultrasonic waves, thereby increasing the mixing of the ethyl ether and the ethanol by the radix physochlainae. The first residue is obtained by centrifuging and vacuumizing, and other substances dissolved in ethanol or diethyl ether are mixed in the first residue, so that the first residue needs to be separated and purified for the second time. Adding a sodium hydroxide solution into the first residue, mainly for adjusting the pH, when the pH is alkalescent, extracting by using chloroform, and avoiding influencing the extraction effect of the chloroform in an acidic environment, specifically, salifying the alkaloid, wherein the alkaloid is easily dissolved in water after salifying, and thus the alkaloid is more easily extracted when extracting by using chloroform. The chloroform extract solution was evaporated to dryness, primarily to remove the chloroform, to obtain a second residue, a crude scopolamine extract. The crude extract of scopolamine contains other scopolamine, needs to be separated and purified again, and is adsorbed by D151 macroporous resin, and the D151 macroporous resin has good adsorption rate and elution rate to scopolamine and no adsorption to other scopolamine, so the scopolamine with good purity can be obtained after adsorption and elution.

The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.