CN101088553B - Effective Parts of Alkaloids and Their Applications - Google Patents

Abstract

The present invention relates to extraction process and medicinal application of sessile stemona alkaloid. Sessile stemona alkaloid is extracted through the following steps: 1. extracting with alcohol solution or acid aqua; and 2. purifying with cation exchange resin. The sessile stemona alkaloid product has the sessile stemona alkaloid content over 50 %. The extracting process uses alcohol, no toxic organic solvent and reusable cation exchange resin, and has high safety and low cost. The medicine composition with the sessile stemona alkaloid may be used in treating skin diseases and diseasesof respiratory tract, and in preventing and controlling plant diseases and pests.

Description

Effective parts of alkaloids and their application

technical field

The present invention relates to an effective part of a traditional Chinese medicine and the application of the pharmaceutical preparation thereof, in particular to an effective part of the alkaloids of the plant and a pharmaceutical preparation containing the effective part of the alkaloids of the plant and an application thereof. the

technical background

Checking the domestic and foreign literature reports on Ceprea, it is found that there are few extraction methods for the total alkaloids of Ceprea. Lin Wenhan {[1] Lin Wenhan, Ye Yang, Xu Rensheng. A Study on the Structure of New Alkaloids in Centipeda Antatus. Organic Chemistry, 1991, 11: 500-3; [2] Lin Wenhan, Xu Rensheng. Zhong Qiongxin. Chemical research on the new alkaloids of Phyllostachys chinensis. Acta Chimica Sinica, 1991, 49: 927-31. [3] Lin Wenhan, Xu Rensheng, Zhong Qiongxin. Chemical research on the alkaloids of Phyllostachys chinensis II. Structural studies of bases. Acta Chemie Sinica, 1991, 49: 1034-37.} and others reported several extraction methods in their above articles. the

Method 1: take 600g of crude drug roots, cut into slices, and cold percolate with 95% industrial alcohol for one week, concentrate the percolation solution under reduced pressure, acidify with 3% HCI, filter, wash the filtrate twice with dichloroethane, and alkalize with ammonia water to pH11, extracted with chloroform, and concentrated under reduced pressure to obtain 2.0 g of crude total alkali;

Method 2: Take 2.5kg of the roots of A. chinensis produced in Hainan Island, grind them into powder, and extract them with chloroform under reflux in a Soxhlet extractor. After concentration, the extract is acidified with 4% dilute hydrochloric acid. After filtration, the filtrate is alkalized with ammonia water. , extracted with chloroform, and concentrated under reduced pressure to obtain 8.2g of crude total alkali;

Method 3: Take 2.5kg of the roots of B. chinensis produced in Hainan Island, grind them into powder, and cold percolate with 95% industrial alcohol. The concentrated ammonia water was basified to pH-10, extracted with chloroform, and concentrated under reduced pressure to obtain 11 g of crude total alkali. the

Ge Lin et al. reported in their U.S. patent US2003229071 that the extraction process of the total alkaloids of the plant was as follows: the plant was crushed, extracted with 95% ethanol under reflux for 2 hours, poured out the extract and placed at 10°C overnight, filtered, and the filtrate was recovered under reduced pressure to The thick paste was acidified with 4% hydrochloric acid, centrifuged at 5°C at 3000 rpm for 40 minutes, the supernatant was basified with ammonia water to pH9, then extracted with diethyl ether and chloroform respectively, the organic phases were combined, recovered under reduced pressure, and dried to obtain Extract of total alkaloids. It can be seen from the above reports that it is found that a large number of organic solvents such as chloroform are used in the current extraction process, and the extraction amount is only 0.33% to 0.44%, which is not conducive to large-scale production. the

Contents of the invention

The purpose of the present invention is to provide a high-purity, effective part of the alkaloids of bacifera with good curative effect. the

Another object of the present invention is to provide the application of the effective fractions of the alkaloids of Phetaceae. the

Technical scheme of the present invention is as follows:

A kind of effective part of alkaloids of basilica, which is extracted by the following method, including two steps:

(1) Extraction with ethanol or acid water: grind the medicinal material of Herba Cinnamomi into a coarse powder, and use 70%-90% ethanol to reflux or extract by percolation or extract with acid water with a pH value of 1-5;

(2), cation exchange resin purification: pass the supernatant extracted in step (1) through a positive exchange resin, elute to remove water-soluble impurities, elute with ammonia water and ethanol solution, until bismuth potassium iodide reacts negatively, stop washing The eluate is recovered to dryness under reduced pressure, and pulverized to obtain the total alkaloid extract of the plant. The purity of the effective parts of the obtained basilica alkaloids is above 50%. the

In the above method, preferably, step (1) is extracted with 90% ethanol, without soaking, and extracted 3 times, each time for 1 hour; the volume ratio of ammonia water and ethanol eluent in step (2) is: 4: 1～ 1:4. the

The cation exchange resin used is a strongly acidic or weakly acidic cation exchange resin. Cation exchange resins can be regenerated repeatedly. the

The effective parts of the alkaloids above can be used to prepare medicines or cosmetics for treating diseases such as mites, athlete's foot, tinea corporis, rosacea, dermatitis, eczema, urticaria, body lice, head lice, trichomonas vaginitis, and the dosage form of medicines It can be ointment, gel, lotion, suppository medicine. It can also be used to prepare drugs for cough, bronchitis, pneumonia, tuberculosis and other respiratory diseases. Intestinal targeting preparations, oral liquid preparations, injections, powder injections and other pharmaceutically acceptable dosage forms. It can also be used to prepare medicines for treating human and animal parasitic diseases such as pinworms and ascariasis, and can be applied to prepare medicines for preventing and controlling various plant diseases and insect pests such as fruit trees, vegetables, flowers, and corns. the

The advantages of the present invention: the patent adopts the ethanol/acid water extraction-cation exchange resin purification technology that can be produced in a large scale to prepare and enrich the total alkaloids of the plant, and the content of the effective parts of the plant is more than 50%. The entire extraction and separation process does not use other toxic organic solvents except ethanol. The ion exchange resin can be used repeatedly, the industrialized production is safe, the cost is low, and the effective part of the prepared basil alkaloid has high purity. the

Detailed ways

Example 1

2000g of herbal medicines, pulverized, percolated and extracted with an aqueous acid solution with a pH of 1, and the supernatant was passed through a HZ004 type positive exchange resin, resin: medicinal materials (1:3), and firstly eluted with deionized water to remove water-soluble impurities. Until there is no polysaccharide reaction, elute with a solution of ammonia water and ethanol in a ratio of 2:1, until the reaction of bismuth potassium iodide becomes negative, stop the elution, and recover the eluate to dryness under reduced pressure, crush it, and obtain the total alkaloids of the plant. Material 35g. the

Example 2

100kg of herbal medicines, crushed and extracted by percolation with 90% ethanol. Ethanol was recovered under reduced pressure to obtain a thick extract. Heat and dissolve with 1% hydrochloric acid in a water bath, the best dosage is 1 ml of 1% hydrochloric acid per gram of medicinal material, centrifuge the acid water, take the supernatant, pass it through JK008 positive exchange resin (resin: medicinal material 1:4), and wash with deionized water first Remove water-soluble impurities until there is no polysaccharide reaction, elute with a solution of ammonia water and ethanol in a ratio of 1:1, stop the elution until the reaction of potassium bismuth iodide becomes negative, and recover the eluent to dryness under reduced pressure, crush to obtain 1.5kg of total alkaloid extract of 100 stems. the

Example 3

Baibu alkaloid active part ointment

Alkaloids 1g, stearic acid 15g, lanolin 2g, liquid paraffin 25ml, triethanolamine 2g, glycerin 5ml, distilled water 50ml. the

Take the alkaloids of the basilica and grind it evenly with a small amount of liquid paraffin, and set it aside. Put stearic acid, lanolin and liquid paraffin in a container, heat in a water bath until melted, and continue to heat to 70-80°C; take another triethanolamine, glycerin and distilled water, heat to 70-80°C, slowly pour in stearic acid, etc. The mixture is stirred in the same direction as it is added, and alkaloids are added until emulsified and coagulated. For external use, it is used to kill Demodex mites parasitic on the human body. the

Example 4

The effective parts of the alkaloids are scattered

According to 50-120g of the active parts of the alkaloids, 250g of lactose, 50g of mannitol, 50g of cross-linked polyvinylpyrrolidone (cross-linked PVP), an appropriate amount of 95% ethanol, and 8g of magnesium stearate are compressed into tablets. It can be used to treat respiratory diseases such as cough, bronchitis, pneumonia and tuberculosis. the

Example 5

Baibu alkaloid active parts aerosol

Add 100ml of dimethyl sulfoxide to 100-200g of the active part of the alkaloids, shake 1000ml of propylene glycol to dissolve, add water to 10,000, shake well, filter to obtain a clarified drug solution, and quantify the above drug solution in a sterile room Dispensed in an aerosol container, fix the valve on the container and press 7.0g of propellant F ₁₂ filtered through a microporous membrane into the bottle under a pressure of 784.6kPa (8kg/cm ² ), namely A solution-type aerosol is obtained. A total of 1000 bottles were produced. Can be used as a safe and effective insecticide.

The present invention is the result of the invention obtained by the inventor through repeated tests, including the extraction experimental research on the total alkaloids of the plant, the refining test of the effective parts and the pharmacodynamic experiment. The contents are as follows:

1. Extraction experiment of total alkaloids

1.1 Pre-test

The study of the extraction process found that the decoction pieces of Baibu (Zhejiang Yinzhou Pharmaceutical and Medicinal Materials Co., Ltd., vines, with a total alkaloid content of 1.32%) were directly extracted by heating and reflux, and the extraction rate of total alkaloids was low. The reason for the analysis may be due to The content of total alkaloids in medicinal materials is relatively low. When extracting directly from decoction pieces as raw materials, alkaloids are difficult to extract from plant cells, resulting in a low extraction rate. And the Baibu decoction pieces are crushed into coarse powder and then extracted. The extraction rate is obviously improved, the extraction rate is 76.3% when water is used as the solvent, and the extraction rate is 91.6% when 90% ethanol is used as the solvent. The test results are shown in Table 1. the

The preliminary test result of alkaloid extraction in table 1

Amount of medicinal materials (g) degree of crushing Extraction solvent extraction conditions Extraction rate (%) 50 Pieces 90% ethanol 12 times, 2 hours, 2 times 16 50 Semolina water 12 times, 2 hours, 2 times 76 50 Semolina 90% ethanol 12 times, 2 hours, 2 times 92

The extraction process of Baibu was determined as follows: the medicinal materials of Baibu were crushed into coarse powder, extracted with ethanol, and the extraction process conditions of ethanol were further optimized through orthogonal experiments. the

1.2 Orthogonal test

1.2.1 Factor level table

According to the preliminary test, the fixed extraction solvent is 8 times the amount, and the four influencing factors of ethanol concentration (A), soaking time (B), extraction time (C), and extraction times (D) are respectively optimized at three levels. , according to the L ₉ (3 ⁴ ) orthogonal table to carry out the experiment, and the arrangement of factor levels is shown in Table 2.

Table 2 Alcohol Extraction Orthogonal Experiment Factor Level Table

level Alcohol concentration (%)/A Soaking time (h)/B Extraction time (h)/C Extraction times/D 1 90 0 1 1 2 70 1 2 2 3 50 2 3 3

1.2.2 Orthogonal test and results

Take by weighing 27 parts of 50 grams (parallel 3 parts) of Ceprea (Zhejiang Yinzhou Pharmaceutical and Medicinal Materials Co., Ltd., vines, total alkaloid content 1.32%) coarse powder respectively, extract according to the conditions selected in Orthogonal Test Table 5, and combine the extraction liquid, filtered, and accurately pipette an appropriate amount of filtrate, evaporated to dryness, and dilute to 10ml with methanol. Precisely draw 100 μl of sample solution, put it in a separatory funnel, add 5.0ml of distilled water, add 2.0ml of buffer solution of 0.1% bromocresol green (pH5.0), 10.0ml of chloroform, shake for 2 minutes, and let it stand for 1 hour. Precisely pipette 5.0ml of chloroform solution, accurately add 1ml of absolute ethanol solution containing 0.01mol/l potassium hydroxide, shake well, put it in a colorimetric cell, and accompany it with a blank, measure the absorbance at 620nm with a spectrophotometer, and calculate the sample The content of total alkaloids in the test results are shown in Table 3. the

Table 3 Alcohol Extraction Orthogonal Test Result Table

1.2.3 Analysis of variance of orthogonal test results

The results of the variance analysis of the orthogonal test results are shown in Table 4. the

The results of analysis of variance of table 4 orthogonal test results

source of variance sum of squared deviations degrees of freedom Variance F value Significance A 3732.43 2 1866.22 3.59 P＜0.05 B 3558.66 2 1779.33 3.42 P＞0.05 C 32906.93 2 16453.47 31.61 P＜0.01 D 38287.60 2 19143.80 36.78 P＜0.01 Error E 9368.86 18 520.49

F _{0.05(2, 18)} = 3.55 F _{0.01(2, 18)} = 6.01

Intuitive analysis, the best process is A ₁ B ₃ C ₁ D ₃ , that is, 90% ethanol, soaking for 2 hours, extracting 3 times, 1 hour each time, where A (alcohol concentration), C (extraction time) and D (extraction time) Times) factor has a significant impact on the extraction rate. Factor B (soaking time) has no significant impact on the extraction rate, and different levels can be selected according to needs. In order to save time and cost, the best process to choose is 90% ethanol, no soaking, extraction 3 times, 1 hour each time.

1.2.4 Verification test of orthogonal test results

In order to further investigate the stability of the above-mentioned preferred process, according to the best conditions obtained, a verification test is carried out, and 3 parts of Herba Cinnamomi (Zhejiang Yinzhou Pharmaceutical and Medicinal Materials Co., Ltd.) are extracted in parallel, and each part of 600g is used to determine the content of total alkaloids of Cinnamomum japonicus. The results are shown in Table 5. the

Table 5 Verification test result table of orthogonal test results

2. Refining test of effective fractions of total alkaloids

According to the test results, after drying under reduced pressure for the ethanol extract of Papaya, the yield of ointment is 32%, which is unfavorable for the preparation of the preparation. On the premise of ensuring the transfer rate of total alkaloids of bacuroni, in order to reduce the yield of ointment, the total alkaloids of baicalensis is refined by ion exchange resin method. the

2.1 Investigation of ion exchange resin type and maximum adsorption capacity

Method: Weigh 1kg of the bacurbita coarse powder, extract according to the optimal method of the orthogonal test, extract the solution under reduced pressure and recover the solvent to a thick extract, fully dissolve it with about 1000ml of 1% hydrochloric acid, centrifuge the acid-water solution, and take the supernatant, Pass through different types of strongly acidic cation exchange resins until the effluent reacts positive with bismuth potassium iodide, stop loading the sample, and elute with deionized water until the Molish reaction is negative (no polysaccharide), stop elution, and measure For the amount of total alkaloids in the sample solution, effluent and washing solution, the maximum adsorption capacity per unit of resin is calculated according to the following formula:

Maximum adsorption capacity = (m _loading - m _outflow - m _washing ) / m _resin

Resin: 4 types of cation ion exchange resins, HZ004, HD008, JK008, and HZ0016 produced by Huazhen Technology Trading Company of East China University of Science and Technology, and SK ₁ B cation exchange resin produced by Mitsubishi Chemical Corporation.

Results: Weigh the appropriate amount of each of the above resins (10 g as dry resin), put them into a chromatographic column with R=1.8 cm, operate according to the above method, and calculate the maximum adsorption capacity. The results are shown in Table 6. the

Table 6 The maximum adsorption capacity investigation result table of unit resin

Resin model Moisture content (%) The amount of total alkaloids adsorbed by 1g dry resin (g) HZ004 68.5 0.3175 HD008 57.5 0.1832 JK008 57.1 0.3052 HZ0016 28.2 0.0196 SK ₁ B 46.0 0.1086

Results: From Table 6, it can be known that the HZ004 strongly acidic cation exchange resin has the largest adsorption capacity for the total alkaloids of Phetaceae, so the HZ004 strongly acidic cation exchange resin was initially selected. the

2.2Optimization of conditions for refining total alkaloids with HZ004 type strongly acidic cation exchange resin

Method: Take 100g of HZ004 type strongly acidic cation exchange resin (about 31g in terms of resin), put it into a chromatographic column, and load 600g of the extract of medicinal materials. The flow rate of the sample solution is 1BV/h. After the sample is loaded, Elute with deionized water until the Molish reaction is negative, stop the elution, change to the eluent listed in Table 1.9 and continue the elution until the eluent bismuth iodide potassium reaction becomes negative, measure the total The content of alkaloids is shown in Table 7. the

Table 7HZ-4 type strongly acidic cation exchange resin refining condition optimization of total alkaloids

Resin amount Total alkali content in sample solution Eluent (V _ammonia : V _ethanol ) Eluent dosage Anointing rate Total alkali content in dry paste total alkali content in dry paste Elution rate 100g 6.77g 2:1 280ml 1.19% 51.5% 3.68g 54.32% 100g 6.77g 1:1 240ml 1.17% 58.6% 4.12g 60.79% 100g 6.77g 1:2 380ml 1.38% 42.2% 3.50g 51.63%

Results: V _{ammonia water} : V _ethanol = 1:1 was selected as the eluent for elution.

2.3 Verification experiment of refining total alkaloids with HZ004 strong acidic cation exchange resin

Method: Take 200g of HZ004 type strongly acidic cation exchange resin (62g as dry resin), put it into a chromatographic column, and load 1200g of the extract of medicinal materials. The flow rate of the sample solution is 1BV/h. After the sample is loaded, use Elute with deionized water until the Molish reaction becomes negative, stop the elution, change V _ammonia water: V _ethanol = 1:1 as the eluent for elution, until the eluent bismuth potassium iodide reacts negative, measure the concentration of the eluent in the eluent For the content of total alkaloids, the relative elution rate was calculated according to the following formula.

Relative relative elution rate (%)=(m _elution /m _loading )×100%, the results are shown in Table 8.

Table 8HZ004 type strong acidic cation exchange resin refining conditions verification of total alkaloids

Summary: The average alcohol extraction rate of Baibu medicinal materials is 32%, the total alkaloid content in the dry extract is 3.5%, and the total alkaloid transfer rate is 85%. After alcohol extraction and refining by strong acidic cation exchange resin, the average extract rate is reduced to 1.18%, the total alkaloid content in the extract is 55.8%, and the total alkaloid transfer rate is 49.8%. It shows that most of the index components can be retained by refining with strong acidic cation exchange resin, while the yield of extract is greatly reduced, the dosage is reduced, and the formulation is facilitated. the

2.4 Investigation on the usage times and regeneration times of HZ004 strong acidic cation exchange resin

When the resin is used for a certain period of time and times, the resin will be polluted and the adsorption capacity will be reduced. At this time, it must be regenerated before it can be used again. the

The resin regeneration method is: ① 1N hydrochloric acid elution: elution with 1N hydrochloric acid, the dosage is 2 to 3 times the volume of the resin, and washed with water until the pH is 5. The flow rate is 2 to 3 times the resin volume per hour. ②5% sodium chloride elution: use 5% sodium chloride 5 times the volume of the resin for elution. The flow rate is 2 to 3 times the resin volume per hour. ③ Elution with 1N sodium hydroxide: elution with 1N sodium hydroxide, the dosage is 2 to 3 times the volume of the resin, and washed with water until the pH is 9. The flow rate is 2 to 3 times the resin volume per hour. ④ 1N hydrochloric acid elution: elution with 1N hydrochloric acid, the dosage is 2 to 3 times the volume of the resin, and washed with water until the pH is 6. The flow rate is 2 to 3 times the resin volume per hour. the

Refer to the manufacturer's instructions, after using HZ-4 strong acidic cation exchange resin, rinse the column with distilled water at pH < 8, rinse with 1N hydrochloric acid twice the volume of the resin to convert to H type, rinse the column with distilled water at pH > 6 and continue to use it. It is generally believed that when the resin adsorption capacity drops by 30%, the resin should be regenerated according to the above-mentioned regeneration method. The results of the investigation are shown in Table 9. the

Table 9 Investigation of the usage times and regeneration times of HZ-4 type strongly acidic cation exchange resin

usage count Adsorption capacity of total alkaloids (mg/g dry resin) Accounted for the initial percentage (%) 1 0.3175 100 5 0.3023 95.2 10 0.2715 85.5 14 0.2273 71.6 15 0.2108 66.4 Regenerate 1 time 1 0.3026 95.3 5 0.2388 75.2 6 0.2184 68.8

The experimental results show that: HZ004 strong acidic cation exchange resin needs to be regenerated after being used for 15 times, and it cannot be reused after being used for 5 times. the

2.5 pilot test

Carry out four batches of pilot scale setting out according to the refined refining process of determination, measure the content of total alkali in each step process, total transfer rate, the result is shown in Table 10. the

Table 10 Preparation process pilot test results

batch number 050525 050526 050527 050408 Variety To Ye Baibu To Ye Baibu vines vines Feeding amount (kg) 15 15 15 225 TA content of medicinal materials (%) 1.58 1.58 0.81 0.79 The total amount of medicinal material TA (g) 236.4 236.4 121.5 1777.5 Amount of extract (g) 260 269 130 4400 Extract yield (%) 1.73 1.79 0.87 ? TA content in extract (%) 50.2 50.8 42.4 ? TA transfer rate in extraction and refining process (%) 55.21 57.81 45.37 ?

The results show that the pilot test process conditions are stable. the

3. Pharmacodynamic experiments on the effective parts of the total alkaloids of Baibu

3.1 Anti-mite test on the effective parts of Baibu alkaloids

Use the improved scraping and pressure method to take human Demodex mites as much as possible onto the glass slides, and observe and count them under a microscope. Each sample contains more than 20 active and active Demodex mites. After the mites are successfully collected, add them to the glass slides immediately Add 3 drops of different concentrations of the total alkaloid solution of Phetopaceae, and spread it evenly to ensure full contact between the liquid medicine and the worm body. The same amount of liquid paraffin and 2% metronidazole aqueous solution were added dropwise to the blank group and the control group respectively. Wait for a while after dropping the medicinal solution, put the slides under a microscope to observe, calculate the number of live mites in each group and mark the location of each insect (to facilitate observation during the experiment). If the room temperature is lower than 25°C during the experiment, put the specimen in an incubator with a temperature of 25-28°C and a relative humidity of 70%-80% to keep warm, take it out every 1-2 hours and observe it under a microscope to record the number of mites activity until all mites on the slide are dead. The criteria for judging the death of mites are as follows: ① those with shrunken or transparent mites; The results are shown in Table 11. the

Table 11 Baibu alkaloid mite removal test

It can be seen from the table that there is no significant difference between the alkaloids of 10mg/ml and metronidazole at the concentration of 10mg/ml, better than metronidazole at the concentration of 20mg/ml, and weaker than metronidazole at the concentration of 5mg/ml. the

3.2 Experimental study on the antitussive and expectorant effects of the effective parts of alkaloids

3.2.1 Ammonia induced cough test

Intragastric administration of the corresponding medicinal solution once, 1 hour after the administration, the mice were respectively placed in a self-made cough-inducing device (300ml wide-mouth bottle with a cover), so that the mice received ammonia water (quantitatively draw 50ul of ammonia water on a filter paper strip) (above) Stimulate for a predetermined time, take out the mouse immediately, and observe the number of coughs of the mouse within 1 min. The coughing sound is recorded and amplified by a microphone, and analyzed in the computer with WaveCN1.90 software. If there are more than 3 typical coughing movements (abdominal muscle contraction or chest shrinkage, and mouth opening at the same time, it is a cough, sometimes there may be a coughing sound) within 1 minute. , counted as "with cough", otherwise counted as "no cough". According to the principle of statistical "up and down method/sequential method" to find half of the number ( _EDT50 ), change the time for each mouse to receive stimulation (the logarithmic difference between two adjacent moments is fixed in an experiment as 0.1) to observe whether the cough response is "with cough" or "without cough". If the previous mouse had a cough reaction, the next mouse would be stimulated with ammonia gas at a lower level; on the contrary, if there was no cough, the next mouse would be stimulated with ammonia gas at a higher level; Only animals were not tested, and the opposite response of the previous mouse was counted. The half cough-induced time (EDT50) and the relative value R of the treatment group and the control group were calculated respectively by weighting method.

EDT ₅₀ ＝log ^-1 ∑r _i x _i /∑n _i

n _i is the number of test animals in each stimulation time period (whether coughing or not), x _i is the logarithm of each stimulation time period, and i is the difference between adjacent stimulus logarithms.

R=(EDT ₅₀ of administration group/EDT ₅₀ of control group)×100%

If R>130%, it is considered that the drug has antitussive effect; if R>150%, it is considered that the drug has obvious antitussive effect. the

3.2.2 Grouping and administration

Sixty Kunming mice, half male and half male, were randomly divided into 5 groups, that is, the normal saline group, the extracts of Phyllostachys chinensis, Phyllostachys erecta and Phytophthora vines, and the codeine phosphate group. The route of administration is all intragastric administration, and the administration volume is 0.01ml/g. The dosage of codeine phosphate is 0.030g/kg, the dosage converted by crude drug is 20g/kg, and the dosage converted by extract is 0.408g respectively /kg, 0.200g/kg, 0.240g/kg. the

3.1.3 Test results of antitussive effect

Antitussive experiments were carried out on three kinds of Bubu Bu extracts, and the results are shown in Table 12. the

Table 12 Comparison of antitussive experiments of three kinds of Bubu extracts

group Dose (g/kg) EDT ₅₀ (s) R normal saline / 15.5 100% codeine 0.030 33.9 219% Herba Leaf Extract 0.408 28.2 182% Centipeda erectus extract 0.200 25.1 162% vine root extract 0.240 29.1 188%

It can be seen from Table 12 that the three kinds of Bubu extracts all have obvious antitussive effect. the

See Table 13 for the results of the antitussive test on the effective parts of the alkaloids of the three kinds of Bubu Bu extracts. the

Table 13 Antitussive test results of the alkaloid part and non-alkaloid part of the three kinds of Bacillus chinensis extracts

3.3 Research on expectorant effect

Methods: The mice were fasted for 8 to 12 hours before administration, and 0.5 hours after intragastric administration, 0.5 mL of 0.5% phenol red solution was injected into the intraperitoneal cavity, and the mice were killed by cervical dislocation after 0.5 hours, and the dorsal position was fixed. Separate the trachea, insert a smoothed No. 7 needle into the trachea about 0.3cm below the throat, ligature and fix it with silk thread, draw 0.5mL of 5% sodium bicarbonate solution with a 1ml syringe, and lavage the respiratory tract back and forth for 3 times through the needle. Put it in a test tube and repeat it 2 more times. Combine the washing liquid and centrifuge at a high speed (9500r·min ^-1 ) for 5min. Carefully take the supernatant and measure the absorbance value at a wavelength of 560nm with a UV-visible spectrophotometer. Curve to calculate the amount of excreted phenol red. The results are shown in Table 14.

Table 14 Comparison of expectorant effects in each group (n=10, X±S) 

group Dose (mg/g) Phenol red content (ug/ml) normal saline / 1.667±0.1665 ammonium chloride 0.03 3.152±0.4359 # To Ye Baibu 0.204 3.147±0.8160 # upright hundred 0.20 2.136±0.4008 # * vines 0.24 2.185±0.4999 # *

#Compared with normal saline, there is a significant difference; *Compared with ammonium chloride, there is a significant difference. the

It can be seen from the table that the expectorant effect on the total alkaloids of C. chinensis, C. erectus and C. vines is significantly different from that of normal saline, and there is no significant difference between ammonium chloride and the total alkaloids of C. However, there were significant differences compared with the total alkaloids of Litonia erectus and Litonia vines. the

3.4 Liquid culture method to detect the anti-tuberculosis growth effect of the effective parts of alkaloids

Method: After milling, strain H37Rv was prepared in Middlebrook 7H9 medium (containing improved OADC nutritional supplement) to a concentration of 1mg/ml, and 0.1ml of bacterial liquid was added to 1.9ml of 7H9 medium containing different concentrations of drugs; the experiment was set as a blank Control group, solvent control group, positive control drug is rifampicin (final concentration 1ug/ml), every group is set up 2 parallel tubes; Mix the reaction tube, after cultivating for 15 days at 37 ℃, detect the effect of drug on the growth of Mycobacterium tuberculosis. The control group with a bacterial count of ++ or more was considered an effective experiment. The results are shown in Table 15. the

Table 15 liquid culture method to detect drug anti-tuberculosis growth efficacy result table

It was found by the liquid culture method that the alkaloids of Libidota alkaloids could resist the growth of Mycobacterium tuberculosis at 250ug/ml, which indicated that the alkaloids of Libidoceae alkaloids had the effect of anti-Mycobacterium tuberculosis growth in vitro, but the effect was weaker than that of rifampicin. the

3.5 Insecticidal test of effective parts of Baibu alkaloids

Carry out according to GB13917. 2-92 (i.e. closed cylinder method); test insects (20 freshwater Culex mosquitoes, non-blood-sucking female adult mosquitoes on the 2nd to 3d after eclosion; 30 houseflies, adults on the 4th to 5th day after eclosion, half male and half male) were anesthetized and placed in the cylinder. After the test insects returned to normal, the liquid was sprayed into the cylinder at the dose of 0.5g/tube and 1.0g/tube, and the round hole was plugged with a rubber stopper immediately. Stop, take out the baffle after 1 minute, and record the number of test insects knocked down in each period of time in time. After 20 minutes, move all the test insects in the cylinder to a clean cage and raise them normally with 5% glucose water cotton balls, and observe The number of dead insects in 24 hours was calculated to calculate KT50 and mortality. The experiment was repeated 3 times, and a blank control group was set at the same time. The results are shown in Table 16. the

Table 16 Baibu alkaloid insecticidal test

It can be seen from the table that the alkaloids of Phetopus have killing effects on freshwater Culex mosquitoes and houseflies. the

3.6 Calculation and comparison of pharmacokinetic parameters of different extracts in rats and mice

Table 17 shows the comparison of pharmacokinetic parameters between the active parts of the alkaloids of the vine and the protobarkine in rats and mice. the

Table 17 Comparison of pharmacokinetic parameters in mice and mice

It can be seen from Table 4.22 that whether it is rats or mice, the bioavailability of the effective part of the alkaloids administered by intragastric administration is higher than that of the original monobasine monomer. It is suggested that the effective parts of crotonine alkaloids may contain substances that promote the absorption of crocetin or components with a structure similar to crocetin, which can saturate the enzymes that metabolize crocetin during absorption and reduce the first over effect. the

Claims (10)

1. an effective part of alkaloids of bacifera, characterized in that: the effective parts of alkaloids of bacifera are extracted by the following method, comprising two steps: (1), using ethanol or acid water extraction: crush the medicinal material of Herba Cinnamomi into coarse powder, and extract with 70%-90% ethanol or acid water with a pH value of 1-5; (2), cation exchange resin purification: pass the supernatant extracted in step (1) through a positive exchange resin, elute to remove water-soluble impurities, elute with ammonia water and ethanol solution, until bismuth potassium iodide reacts negatively, stop washing The eluate is recovered to dryness under reduced pressure, and crushed to obtain the total alkaloid extract of the plant, the content of the active parts of the plant being more than 50%. 2. The active part of the plant alkaloid according to claim 1, characterized in that: in the step (1), 70%-90% ethanol is used for extraction by heating to reflux or percolation. 3. The effective part of the alkaloids according to claim 2, characterized in that: 90% ethanol extraction is adopted in the (1) step, and the extraction is 3 times, 1 hour each time. 4. The active part of the alkaloids of A. mellifera according to claim 1, characterized in that: the volume ratio of ammonia water and ethanol eluent in the step (2) is: 4:1-1:4. 5 . The active part of the alkaloids according to claim 1 , characterized in that: the cation exchange resin used in the (2) step is a strongly acidic or weakly acidic cation exchange resin. 6 . 6 . The active part of the alkaloids according to claim 5 , characterized in that: the cation exchange resin is HZ004 or JK008. 7. the effective parts of the alkaloids described in claims 1-6 are used in the preparation of medicines for the treatment of mites, athlete's foot, tinea corporis, rosacea, dermatitis, eczema, urticaria, body lice, head lice or trichomonal vaginitis Or the application in cosmetics, the pharmaceutical dosage form is ointment, gel, lotion or suppository. 8. the application of the effective part of the alkaloids described in claim 1-6 in the preparation of the medicine for the treatment of cough, bronchitis, pneumonia or tuberculosis, the dosage form of the medicine is tablet, soft capsule, microcapsule, granule Drugs, pills, powders, sustained-release preparations, controlled-release preparations, gastrointestinal targeted preparations, oral liquid preparations, injections, powder injections and other pharmaceutically acceptable dosage forms. 9. The application of the effective part of the alkaloids described in claims 1-6 in the preparation of medicines for treating pinworms, ascariasis or animal parasitic diseases. 10. The application of the effective parts of the alkaloids of the plant according to claims 1-6 in the preparation of medicines for the prevention and treatment of fruit trees, vegetables, flowers or corn plant diseases and insect pests.