CN105267274A - Atractylenolide extract having antiparkinsonian effect, preparation method and application thereof - Google Patents

Abstract

Atractylodes lactone extract with anti-Parkinson effect, preparation method and application, said Atractylodes lactone extract contains Atractylodes lactone I with a weight percentage greater than 5%, Atractylodes II with a weight percentage greater than 10% and Atractylodes lactone The weight percent of III is greater than 20%, and the purity of the total lactones is greater than 35% by weight. The application of the atractylodes lactone extract with anti-Parkinson effect as the preparation of anti-Parkinson drugs. The Atractylodes macrolide extract, preparation method and application provided by the present invention have an anti-Parkinson effect. By adding an oxidizing agent to the Atractylodes macrocephala powder, the atractylodes ketones in the Atractylodes volatile oil are oxidized into Atractylodes lactone compounds, thereby increasing the Atractylodes atractylodes The content of the ester; zinc particles are added to the extraction solution to protect the generated compounds such as atractylodes lactone from being further oxidized and degraded. The preparation method is simple and practical, easy to operate, more suitable for industrial production, and has remarkable advancement and practical value.

Description

Atractylodes lactone extract with anti-parkinson effect, preparation method and application

technical field

The invention relates to the technical field of traditional Chinese medicine, in particular to an extract of atractylodes lactone with anti-parkinson effect, a preparation method and application.

Background technique

Parkinson's disease (PD) is a central nervous system degenerative disease characterized by progressive death of dopaminergic neurons in the substantia nigra of the midbrain. At present, the main treatment method is western medicine, but it only improves the symptoms, but cannot control its progress, and the curative effect is weakened and the toxicity increases after long-term application. And Chinese medicine treatment has advantages such as little toxic and side effects, strong synergistic effect and more development prospects.

Atractylodes macrocephala is the dried rhizome of Atractylodesmacrocephala Koidz., a plant of the Compositae family. It has the effects of invigorating the spleen and replenishing qi, drying dampness and promoting diuresis, antiperspirant, and preventing miscarriage. ". Atractylodes lactones play an important role in anti-inflammation and anti-tumor (KATSUYAENDO, TAKASHITAGUCHI, FUMIKOTAGUCHI, etc.AntiinflammationPrinciplesofAtractylodesRhizomes[J].Chem.Pharm.Bull.,1979,27(12):2954-2958, Shen Guoqing , He Falin, Li Fengxin, etc.; Atractylodes Rhizoma Atractylodes Rhizome volatile oil chemical constituents and experimental research on anti-tumor [J], Journal of Beijing University of Traditional Chinese Medicine, 2009,32(6):413-415.), which is the main substance of Atractylodes Rhizoma Atractylodes Rhizoma Atractylodes Rhizoma Atractylodes Rhizome. The main component of the volatile oil in Atractylodes macrocephala is atractylodes ketone. Studies have shown that atractylodes atractylodes has no anti-Parkinson effect, and it can be partially converted into atractylodes lactone under light conditions, but it takes a long time and is easy to volatilize. Therefore, we explored the preparation method of Atractylodes lactone extract and found that it has a good anti-Parkinson effect.

Contents of the invention

In view of the above problems, the present invention provides Atractylodes lactone extract with anti-Parkinson effect, preparation method and application, which effectively overcomes the disadvantage of low yield of Atractylodes lactone obtained in the prior art, significantly reduces production cost, improves extraction efficiency, It is more suitable for industrial production.

In order to achieve the above-mentioned purpose of the present invention, the present invention provides a kind of Atractylodes lactone extract that has anti-Parkinson effect, and described Atractylodes lactone extract contains the weight percentage of Atractylodes lactone I greater than 5%, the weight percentage of Atractylodes lactone II The percentage is greater than 10% and the weight percentage of Atractylodes lactone III is greater than 20%, and the purity of the total lactones is greater than 35% by weight.

The present invention also provides a preparation method of atractylodes atractylodes extract with anti-Parkinson effect, comprising the following steps.

Step 1, Atractylodes macrocephala medicinal material or decoction pieces are pulverized.

Step 2, oxidizing atractylone in Atractylodes rhizome powder with an oxidizing agent.

Step 3, drying the oxidized Atractylodes macrocephala powder.

Step 4, adding 1 to 5% of the dried Atractylodes rhizome powder mass of zinc granules, and then extracting the Atractylodes rhizome powder at least once to obtain a total lactone extract of Atractylodes rhizome.

Step 5, the extract is concentrated to obtain a concentrate.

Step 6, the concentrated solution is dried to obtain Atractylodes lactone extract.

The application of the atractylodes lactone extract with anti-Parkinson effect as the preparation of anti-Parkinson drugs.

In the step 1, the Atractylodes macrocephala medicinal material or decoction pieces are pulverized into 10-60 mesh powder.

Oxidant is ozone and hydrogen peroxide in the described step 2.

The oxidation conditions of the hydrogen peroxide are: (1) the concentration of hydrogen peroxide: 10-30% by volume; (2) the amount of hydrogen peroxide: the mass ratio is 3:1-1:1; (3) soaking time: 1-8h ; (4) Reaction temperature: 30-80°C; (5) Oxidation time: 2-24h.

The drying temperature in step 3 is 30-100°C.

The drying in the step 6 can adopt vacuum drying, spray drying and freeze drying.

The extraction of the lactone in the step 4 can adopt solvent reflux extraction method, ultrasonic extraction method, room temperature extraction method, percolation method; the solvent used for extraction is methanol, ethanol, acetone and the like.

Specifically, taking ethanol as an example, the extraction conditions are described as follows.

(1) Reflux extraction method: add 60-95% ethanol (volume percentage) solution to the oxidized and dried Atractylodes macrocephala powder in an amount of 5-12 times the mass of the medicinal material, and add zinc particles in an amount of 1-5% of the mass of the medicinal material. Reflux extraction for 1-5 hours, extract twice, combine extracts, filter, concentrate and dry to obtain high-content Atractylodes lactone extract.

(2) Ultrasonic extraction method: add 60-95% ethanol (volume percentage) solution to the oxidized and dried Atractylodes macrocephala powder in an amount of 5-12 times the mass of the medicinal material, and add zinc particles in an amount of 1-5% of the mass of the medicinal material. Ultrasonic extraction for 10-50 minutes, filtering, and evaporating to dryness to obtain Atractylodes lactone extract.

(3) Percolation method: use 60-95% ethanol to percolate the oxidized and dried Atractylodes macrocephala powder, the percolation volume is 10-20 times the mass of Atractylodes macrocephala powder, concentrate and dry to obtain Atractylodes macrolide extract.

Compared with the prior art, the present invention has beneficial effects.

The present invention provides Atractylodes macrolide extract with anti-Parkinson effect, preparation method and application, by adding hydrogen peroxide or ozone to the powder of Atractylodes macrocephala as an oxidizing agent to oxidize the main component atractylodes ketone in Atractylodes volatile oil into Atractylodes lactones compounds, thereby increasing the content of Atractylodes lactone; further in the extraction process, zinc particles are added to the extraction solution as a reducing agent to protect the generated compounds such as Atractylodes lactone from being further oxidized and degraded, effectively increasing the content of Atractylodes lactone extraction rate. The preparation method uses a common and safe oxidant to oxidize atractylone in the medicinal material, and after the oxidant reaction is completed, the residue is oxygen and water, which is environmentally friendly and pollution-free; moreover, the preparation method is simple and practical, easy to operate, and is more suitable for industrial production. Significant advancement and practical value. In addition, the present invention discovers for the first time that atractylodes lactone has an anti-Parkinson effect, and can be used alone or in combination with other drugs to prepare anti-Parkinson drugs and further treat Parkinson's disease.

Description of drawings

Figure 1 is the fingerprint of Atractylodes macrocephala before transformation.

Figure 2 is the fingerprint of Atractylodes macrocephala after transformation.

detailed description

The present invention will be further described in detail below in conjunction with specific examples.

Example 1.

This example provides an extract of Atractylodes macrolide with anti-Parkinson effect. The extract contains Atractylodes atractylodes I in a weight percentage of 7.5%, Atractylodes atractylodes II in a weight percentage of 12.3% and Atractylodes atractylodes The weight percent of ester III is 25.7%, and the purity of total lactones is 45.5% by weight.

This embodiment also provides a preparation method of Atractylodes lactone extract with anti-Parkinson effect, including the following steps.

Step 1. Take 50 g of Atractylodes macrocephala medicinal material, pulverize it, and pass through a 10-mesh sieve to obtain Atractylodes macrocephala powder.

Step 2. Use hydrogen peroxide to oxidize atractylone in Atractylodes macrocephala powder: use 50 mL of hydrogen peroxide with a volume percentage of 20%, infiltrate for 4 hours, and oxidize at 50° C. for 8 hours.

Step 3. After the oxidation of the Atractylodes macrocephala powder is completed, it is dried at 40° C. for 12 hours.

Step 4. Put the dried Atractylodes macrocephala powder in a 1L round bottom flask, add 500ml of 95% ethanol, and then add 1g of zinc particles, reflux for extraction for 1 hour, and filter to obtain the total lactone extract of Atractylodes macrocephala. When the Atractylodes macrocephala powder is extracted, in order to prevent the generated Atractylodes macrolide from being further oxidized, zinc particles need to be added as a protective agent; the amount of the zinc particles added is 1-5% of the mass of the dry Atractylodes macrocephala powder.

Step 5. The extract of total lactones of Atractylodes macrocephala is concentrated under reduced pressure or normal pressure to obtain a concentrated solution.

Step 6, the concentrated solution is freeze-dried to obtain Atractylodes lactone extract. The yield of the obtained atractylodes lactone extract was 10.1% (weight ratio).

The obtained atractylodes atractylodes extract can be used alone or in combination with other drugs as a raw material drug and used in anti-Parkinson medicaments.

In order to further verify the beneficial effects of the present invention, the following experimental cases are provided.

1. The content of Atractylodes lactone was determined by HPLC external standard method.

1. HPLC chromatographic conditions: Agilentzorbax C18 (250mm×4.6mm, 5μm) column, diode array detector (DAD), gradient elution with acetonitrile-water as mobile phase, the gradient elution table is shown in Table 1, flow rate: 1.0ml /min; injection volume: 20 μL; column temperature: 25°C. Table 2 shows the change schedule of Atractylodes macrophage oxidation wavelength. The fingerprint of Atractylodes macrocephala before transformation is shown in Figure 1, and the fingerprint of Atractylodes macrocephala after transformation is shown in Figure 2, where A is the peak of Atractylodes lactone III, B is the peak of Atractylodes lactone I, and C is the peak of Atractylodes lactone Ⅱ is the peak, D is the peak of atractylone.

Table 1: Gradient elution schedule for Atractylodes macrophage oxidation process.

time Acetonitrile (%) water 10～15min 10～40 90～60 15～30min 40～60 60～40 30～40min 60～100 40～0 40～50min 100 0 50～60min 100～10 0～90

Table 2: Program table of wavelength change of Atractylodes macrophage oxidation.

time wavelength (nm) 0～37min 220 37～40min 280 40～60min 220

2. Anti-Parkinsonian experiment of Atractylodes lactone in vitro.

1. Experimental materials: dopaminergic human neuroblastoma cell line (SH-SY5Y cells, purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences); 10% inactivated calf serum (purchased from GIBCO, batch number: 8166872); penicillin 100U mL ^{- 1.} Streptomycin 100 μg mL ^-1 (purchased from HyClone Company, lot number: J120721); DMEMF-121:1 culture medium (purchased from HyClone Company, lot number: NXA0559); MPP ⁺ (purchased from Sigma Company, lot number: 100M4713V ); the arctiin and aglycon used were obtained from our laboratory.

2. Cell culture: Culture SH-SY5Y cells in DMEMF-121:1 medium containing 10% inactivated calf serum, penicillin 100U·mL ^-1 , streptomycin 100μg·mL ^-1 , placed at 37°C, 5% CO ₂ incubator. Replace the culture medium every other day, culture until the monolayer cells are about 80% confluent, subculture, and use the cells in the logarithmic growth phase for experiments.

3. Modeling and administration methods: adjust the SH-SY5Y cell density to about 5×10 ⁴ /mL, inoculate 100 μL/well in a 96-well plate, and add arctiin and burdock at different concentrations (1 μM, 5 μM and 10 μM) after 16 hours Aglycone, set 3 duplicate wells for each concentration, and set equal amount of control group (without treatment reagent) and model group (only added MPP ⁺ ), after 24h, the control group was added with complete medium, model group and blank group Add 1 mM MPP ⁺ (1-methyl-4-phenylpyridinium ion) diluted with culture medium to each well.

4. Detection of cell viability by tetramethylazolium salt (MTT ⁺ ) method: after 48 hours, add 5 mg/mL MTT ⁺ 10 μL to each well of cells in different groups, culture for 4 hours, suck out the medium in the well, add 100 μL of DMSO to each well, After the crystals were fully dissolved, the microplate reader detected the absorbance value (OD value) of each well at 490nm (reference wavelength 620nm), and calculated the cell survival rate. Cell survival rate%=(OD value of experimental group-OD value of blank group/OD value of control group-OD value of blank group)×100%.

5. Statistical processing Data are expressed as mean ± standard deviation, and statistical processing is performed by one-way ANOVA.

6. Results.

The activity test results of Atractylodes lactone are shown in Table 3, which shows that the anti-parkinsonism activity of Atractylodes lactone extract is stronger than that of monomer Atractylodes lactone, and the activity of Atractylodes lactone II is stronger.

Table 3: Protective effect of atractylodes lactone on MPP ⁺ -induced cell injury (mean ± standard deviation, n = 3).

Survival rate (%) Survival rate (%) Survival rate (%) Survival rate (%) 1μM 10μM 100μM 200μM blank control 100.00±2.38 100.00±2.38 100.00±2.38 100.00±2.38 Model 51.32±0.88 ^## 51.32±0.88 ^## 51.32±0.88 ^## 51.32±0.88 ^## Atractyloid Ⅰ 52.23±2.17 54.96±2.28 ^* 59.62±0.22 ^** 61.39±0.82 ^** Atractyloid Ⅱ 51.46±1.62 54.15±2.16 64.99±0.66 ^** 73.09±1.88 ^** Atractyloid Ⅲ 51.99±1.51 51.41±1.8 52.04±1.58 55.59±0.58 ^* Atractylodes lactone extract 51.46±1.62 56.13±2.08* 70.09±0.65 ^** 83.10±1.78 ^**

Note: ^## P＜0.01 compared with the blank control group; ^* P＜0.05 compared with the model group; ^** P＜0.01 compared with the model group.

3. Animal experiments on the anti-Parkinson effect of Atractylodes macrolide extract.

1. Experimental materials: C57BL/6 mice were purchased from the Animal Experiment Center of Dalian Medical University. The Atractylodes lactone extract used was self-made according to Example 1. The positive drug selegiline was produced by Shenzhen Zhonglian Pharmaceutical Co., Ltd.

2. Animal grouping: 60 C57BL/6 mice were randomly divided into six groups: blank group, model group, positive control group (selegiline group) and low, medium and high doses of Atractylodes lactone extract group (referred to as low, medium and high dose extract groups), 10 rats in each group.

3. Modeling and administration methods: Low, medium and high doses of extracts were administered at 20 mg/kg, 40 mg/kg and 80 mg/kg respectively, once a day for 13 consecutive days. The dose of selegiline group was 15mg/kg. On the 9th day after administration, MPTP (1-methyl-4-phenyl-1,2,3,6-tetrafluoroethylene) was subcutaneously injected into the model group, low-, medium-, and high-dose extract groups, and Hydropyridine (1-methyl4-phenyltetrahydropyridine, purchased from Sigma, batch number: 1001054803) 25 mg/kg, once a day, for 5 consecutive days. The blank group was injected with the same amount of normal saline intraperitoneally instead of MPTP, once a day. At the same time, animal behavior studies were performed every day. Experimental examination.

4. Behavioral examination.

(1) Suspension test (Tractiontest): The purpose is to detect the coordination of limb movement in C57BL/6 mice. The test mice were suspended on a horizontal wire with a diameter of about 0.2 mm. The timing starts when the mouse grabs the wire with its front paws and ends when the mouse falls. 6 points for more than 25s; 5 points for 20-25s; 4 points for 15-20s; 3 points for 10-15s; 2 points for 5-10s; 1 point for 0-5s. Finally, the scores are calculated and analyzed statistically. The higher the score, the firmer the mouse grasped the wire; the better the coordination of the limb movement of the mouse, otherwise, the more severe the tremor and muscle rigidity of the mouse. The results are shown in Table 4.

Table 4: Hanging experiment scoring results (mean ± standard deviation).

dose Day 9 day 10 day 11 day 12 day 13 blank group -- 7.00±0.00 6.45±1.22 6.00±1.46 5.97±1.24 5.05±1.70 model group -- 6.32±1.35 ^△ 5.12±1.03 ^△ 3.21±1.42 ^△△ 2.84±1.53 ^△△ 2.03±1.03 ^△△ positive group 15 6.77±0.73 6.05±1.45 ^▲ 5.79±1.43 ^▲ 5.32±1.24 ^▲▲ 4.33±1.29 ^▲▲ low dose group 20 5.92±0.11 4.98±1.22 3.40±1.92 3.80±1.41 ^▲ 3.00±1.82 ^▲ Middle dose group 40 6.190±0.09 5.12±1.09 5.22±1.31 ^▲▲ 5.64±1.55 ^▲▲ 4.20±0.87 ^▲▲ high dose group 80 6.30±0.73 4.78±1.26 3.71±1.20 ^▲ 4.26±1.29 ^▲▲ 3.52±1.13 ^▲▲

Note: △ is different from the blank group (p<0.05); △△ is significantly different from the blank group (p<0.01); ▲ is different from the model group (p<0.05), ▲▲ is different from the model Compared with positive control group, there was significant difference (p<0.01); ■Compared with positive control group, there was no difference (p>0.05).

(2) Swimming test (Swimming test): The purpose is to detect the coordination of limb movement in C57BL/6 mice. Put the tested mice into a 20cm×30cm×20cm water tank, the water temperature is 22°C-25°C, and record the floating time of the mice within 1min. The scoring criteria are as follows: 6 points for floating time within 10s within 1min; 5 points for floating within 20s; 4 points for within 30s; 3 points for within 40s; 2 points for floating within 50s; 2 points for exceeding 50s. 1 point. Finally, the scores are calculated and analyzed statistically. The higher the score, the longer the swimming time of the mouse; the better the coordination of the limb movement of the mouse, otherwise, the severe tremor and muscle rigidity of the mouse. The results are shown in Table 5.

Table 5: Swimming test score results (mean ± standard deviation).

dose Day 9 day 10 day 11 day 12 day 13 blank group -- 6.00±0.00 5.80±0.05 5.67±0.32 5.98±0.60 6.00±0.39 model group -- 5.71±0.51 4.32±0.62 4.55±1.30 ^△ 4.31±1.23 ^△△ 3.81±0.90 ^△△ positive group 15 6.00±0.00 5.62±0.54 ^▲ 5.85±0.71 ^▲▲ 5.02±0.61 ^▲ 5.15±0.84 ^▲▲ low dose group 20 6.00±0.45 4.71±0.55 5.04±0.65 4.90±1.01 ^▲▲ 4.75±0.81 ^▲ Middle dose group 80 5.85±0.45 5.45±0.72 ^▲ 5.68±0.33 ^▲▲ 5.35±0.43 ^▲▲ 5.24±0.45 ^▲▲ high dose group 40 6.00±0.03 5.04±0.65 ^▲ 5.16±0.43 ^▲ 5.11±0.70 ^▲▲ 5.23±0.73 ^▲▲

Note: △ is different from the blank group (p<0.05); △△ is significantly different from the blank group (p<0.01); ▲ is different from the model group (p<0.05); ▲▲ is different from the model Compared with positive control group, there was significant difference (p<0.01); ■Compared with positive control group, there was no difference (p>0.05).

The results of behavioral experiments showed that the medium-dose extract group could enhance the suspension index and swimming index of the mice, and had the effect of significantly improving the behavior of the MPTP model C57BL/6 mice, which was equivalent to the effect of the positive control drug selegiline group .

4. Study on the mechanism of action of the extract against Parkinson's disease.

1. Experimental materials: dopaminergic human neuroblastoma cell line (SH-SY5Y cells, purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences); DMEMF-121:1 culture medium (purchased from HyClone); MPP ⁺ (purchased from Sigma, batch number : 100M4713V); the arctigenin used is self-made. β-actin antibody (purchased from Zhongshan Jinqiao, lot number 111212); Caspase-3 (activated) antibody (purchased from Beyotim); Bcl-2 antibody (purchased from abcam company, lot number: GR73243-1); Bax antibody (purchased from From abcam company, batch number: GR45349-3).

2. Cell culture: SH-SY5Y cells were cultured in DMEMF-121:1 medium containing 10% inactivated fetal bovine serum, penicillin 100U·mL ^-1 and streptomycin 100μg·mL ^-1 , placed at 37°C, 5% CO ₂ incubator. Replace the culture medium every other day, culture until the monolayer cells are about 80% confluent, subculture, and use the cells in the logarithmic growth phase for experiments.

3. Modeling and administration methods: adjust the density of SH-SY5Y cells to about 10 ⁵ /mL, passage them in culture flasks, culture them until the cells are about 80% confluent, and then add extracts of different concentrations (1μM, 5μM and 10μM) , and set up a control group (without treatment reagent) and a model group (only added with MPP ⁺ ). After 24 hours, the control group was added with complete medium and the model group was added with 1mM MPP ⁺ in each bottle.

4. Western blotting was used to detect the level of related apoptotic proteins in cells.

(1) The level of pro-apoptotic protein caspase-3.

The whole protein of cells in each group was extracted (the kit was purchased from KGI), and the protein was quantified by the BCA method (the kit was purchased from Beyotim). After the samples were processed, 10 μL of each sample was loaded into electrophoresis. The membrane was transferred by semi-dry method, blocked with 5% skimmed milk powder, β-actin antibody and Caspase-3 (activated) antibody were mixed with TBST at a volume ratio of 1:1000, and left overnight at 4°C. Wash away the above antibodies, respectively, with horseradish peroxidase-labeled goat anti-mouse IgG (H+L) (volume ratio 1:2000) and horseradish peroxidase-labeled goat anti-rabbit IgG (H+L) (purchased (from Beyotim Company) (volume ratio 1:1000) was incubated at room temperature for 2 hours, after the above antibodies were washed away, ECL chemiluminescence (purchased from Beyotim Company), developed, fixed and then scanned and analyzed, the experiment was repeated 3 times.

(2) The levels of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax.

The protein sample processing steps were the same as above. After blocking, β-actin antibody, Bcl-2 antibody and Bax antibody were mixed with TBST at a volume ratio of 1:1000, 1:500 and 1:1000, respectively, and left overnight at 4°C. Wash off the above antibodies, select the corresponding horseradish peroxidase-labeled goat anti-rabbit IgG (H+L) (1:1000, volume ratio) and horseradish peroxidase-labeled goat anti-mouse IgG (H+L) (1:2000, volume ratio) incubate at room temperature for 2 hours, wash away goat anti-mouse IgG (H+L), perform ECL chemiluminescence, develop, and fix for scanning and analysis, and repeat the experiment 3 times.

5. The results are shown in Table 6.

The expression of Bcl-2, Bax and Caspase-3 proteins in the control group and the model group had statistically significant differences (p<0.05), and the SH-SY5Y cells treated with arctigenin could reduce the ratio of Bax/Bcl-2, Decrease the expression of Caspase-3, indicating that arctigenin may up-regulate Bax/Bcl-2 protein and down-regulate Caspase-3 protein, and the degree of regulation is concentration-dependent with drugs.

Table 6: Effects of extracts on expression of apoptosis-related proteins in SH-SY5Y cells after MPP ⁺ induction.

Bax/Bcl-2 Caspase-3/β-actin control group 100±0.00 100±0.00 model group 1512.20±0.05 ^△ 161.85±0.02 ^△ 1mM extract 133.84±0.07 ^*** 135.61±0.04 ^* 5mM extract 123.32±0.05 ^*** 130.23±0.03 ^** 10mM extract 115.31±0.06 ^*** 117.52±0.03 ^***

Note: △Compared with the control group, there is a difference (p<0.05); *Compared with the model group, there is a difference (p<0.05); **Compared with the model group, there is a significant difference (p<0.005); ***With Compared with the model group, there was a significant difference (p<0.001).

In summary, it can be seen that different concentrations of Atractylodes lactone extracts have anti-Parkinson activity, and in vivo experiments have also confirmed that Atractylodes lactone is more suitable for long-term use in Parkinson's patients due to its low toxicity and side effects.

Example 2.

This example provides an extract of Atractylodes macrolide with anti-Parkinson effect. The extract contains Atractylodes atractylodes I at a weight percentage of 8.3%, Atractylolide II at a weight percentage of 14.2% and Atractylodes atractylodes The weight percent of ester III is 27.2%, and the purity of total lactones is 49.7% by weight.

This embodiment also provides a preparation method of Atractylodes lactone extract with anti-Parkinson effect, including the following steps.

Step 1. Take 50 g of Atractylodes macrocephala medicinal material, pulverize it, and pass through a 20-mesh sieve to obtain Atractylodes macrocephala powder.

Step 2. Use hydrogen peroxide to oxidize atractylone in Atractylodes macrocephala powder: use 50 mL of hydrogen peroxide with a volume percentage of 30%, infiltrate for 3 hours, and oxidize at 40° C. for 6 hours.

Step 3. After the oxidation of the Atractylodes macrocephala powder is completed, it is dried at 40° C. for 12 hours.

Step 4. Put the dried Atractylodes macrocephala powder in a 1L round bottom flask, add 500ml of 95% ethanol, and then add 3g of zinc particles, reflux for extraction for 2 hours, and filter to obtain the total lactone extract of Atractylodes macrocephala.

Step 5. The extract of total lactones of Atractylodes macrocephala is concentrated under normal pressure to obtain a concentrated solution.

Step 6, the concentrated solution is dried under reduced pressure to obtain the extract of atractylodes lactone.

Example 3.

This example provides an extract of Atractylodes macrolide with anti-Parkinson effect. The extract contains Atractylodes atractylodes I in a weight percentage of 8.7%, Atractylodes atractylodes II in a weight percentage of 11.1% and Atractylodes atractylodes The weight percent of ester III is 28.7%, and the purity of total lactones is 48.5% by weight.

This embodiment also provides a preparation method of Atractylodes lactone extract with anti-Parkinson effect, including the following steps.

Step 1. Take 50 g of Atractylodes macrocephala medicinal material, pulverize it, and pass through a 20-mesh sieve to obtain Atractylodes macrocephala powder.

Step 2. Oxidation of atractylone in Atractylodes macrocephala powder with hydrogen peroxide: use 100 mL of hydrogen peroxide with a volume percentage of 30%, infiltrate for 6 hours, and oxidize at 50° C. for 4 hours.

Step 3. After the oxidation of the Atractylodes macrocephala powder is completed, it is dried at 40° C. for 12 hours.

Step 4. Put the dried Atractylodes macrocephala powder in a 1L round bottom flask, add 700ml of 95% ethanol, and then add 3g of zinc particles, reflux for extraction for 2 hours, and filter to obtain the total lactone extract of Atractylodes macrocephala.

Step 5. The extract of total lactones of Atractylodes macrocephala is concentrated under normal pressure to obtain a concentrated solution.

Step 6, the concentrated solution is dried under reduced pressure to obtain the extract of atractylodes lactone.

Claims (9)

1. there is the atractylodes lactone extract of anti-Parkinson effect, it is characterized in that, the percentage by weight that described atractylodes lactone extract contains atractylenolide Ⅰ is greater than 5%, the percentage by weight of atractylenolideⅡ be greater than 10% and the percentage by weight of atractylenolideⅢ be greater than 20%, the purity of total lactones is weight percentage and is greater than 35%.

2. there is the preparation method of the atractylodes lactone extract of anti-Parkinson effect, it is characterized in that, comprise the following steps:

Step 1, Rhizoma Atractylodis Macrocephalae or decoction pieces are pulverized;

Step 2, with oxidant, atractylone in Rhizoma Atractylodis Macrocephalae powder to be oxidized;

Step 3, to oxidation after Rhizoma Atractylodis Macrocephalae powder carry out bake drying;

Step 4, add the zinc granule of dry gained Rhizoma Atractylodis Macrocephalae opaque amount 1 ~ 5%, then Rhizoma Atractylodis Macrocephalae powder is extracted at least one times, obtain Rhizoma Atractylodis Macrocephalae total lactones extracting solution;

Step 5, described extracting solution liquid obtain concentrated solution through concentrated;

Step 6, described concentrated solution drying obtain atractylodes lactone extract.

3. have the atractylodes lactone extract of anti-Parkinson effect as claimed in claim 2, it is characterized in that, in described step 1, Rhizoma Atractylodis Macrocephalae or decoction pieces are ground into 10 ~ 60 order powder.

4. have the atractylodes lactone extract of anti-Parkinson effect as claimed in claim 2, it is characterized in that, in described step 2, oxidant is ozone and hydrogen peroxide.

5. have the atractylodes lactone extract of anti-Parkinson effect as claimed in claim 4, it is characterized in that, the oxidizing condition of described hydrogen peroxide is: the concentration of (1) hydrogen peroxide: 10 ~ 30%; (2) consumption of hydrogen peroxide: mass ratio is 3:1 ~ 1:1; (3) infiltrating time: 1 ~ 8h; (4) reaction temperature: 30 ~ 80 DEG C; (5) oxidization time: 2 ~ 24h.

6. have the atractylodes lactone extract of anti-Parkinson effect as claimed in claim 5, it is characterized in that, in described step 3, bake out temperature is 30 ~ 100 degree.

7. have the atractylodes lactone extract of anti-Parkinson effect as claimed in claim 6, it is characterized in that, in described step 6, drying can adopt drying under reduced pressure, spraying dry, lyophilization.

8. have the atractylodes lactone extract of anti-Parkinson effect as claimed in claim 7, it is characterized in that, in described step 4, the extraction of lactone can adopt solvent refluxing extraction method, ultrasonic extraction, room temperature extraction, percolation; Extracting solvent used is methanol, ethanol, acetone etc.

9. there is the atractylodes lactone extract of anti-Parkinson effect as the application preparing anti-Parkinson medicine.