CN106474169B - Celery seed extract, preparation and preparation method thereof - Google Patents
Celery seed extract, preparation and preparation method thereof Download PDFInfo
- Publication number
- CN106474169B CN106474169B CN201610177185.2A CN201610177185A CN106474169B CN 106474169 B CN106474169 B CN 106474169B CN 201610177185 A CN201610177185 A CN 201610177185A CN 106474169 B CN106474169 B CN 106474169B
- Authority
- CN
- China
- Prior art keywords
- extract
- celery seed
- extraction
- volume
- seed extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/4816—Wall or shell material
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/4841—Filling excipients; Inactive ingredients
- A61K9/4866—Organic macromolecular compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
The invention relates to a celery seed extract, a preparation and a preparation method thereof. Taking dry celery seeds, and carrying out reflux extraction for 1-2 times by using an ethanol water solution with 2-4 times of volume amount and volume fraction of 95%; collecting extractive solution, and concentrating under reduced pressure to obtain soft extract; suspending the thick extract in water with 0.5-1 times of the volume of the raw materials, extracting for 2-4 times with the extraction reagent, and combining organic phases after extraction to obtain a crude extract; weighing silica gel with 35-55 times of crude extract mass, activating at 130 deg.C for 1h, and loading into chromatographic column; taking the crude extract, and loading the crude extract at the concentration of 0.2-0.5 g/ml; gradient eluting with petroleum ether-dichloromethane-ethyl acetate, collecting eluate, and concentrating under reduced pressure to obtain semen Apii Graveolentis extract. The preparation method has the advantages of simple preparation process, less extraction times and high yield, and the obtained celery seed extract has a better effect of reducing serum uric acid.
Description
Technical Field
The invention belongs to the field of pharmaceutical preparations, and particularly relates to a celery seed extract, a preparation and a preparation method thereof.
Background
Celery seeds are seeds of Apium graveolens L.of an Umbelliferae plant, have a century history in Australia for treating arthralgia, are regarded as a traditional folk prescription, and have multiple effects of reducing uric acid, blood pressure, blood fat and the like, so that the effect of reducing uric acid is most remarkable. Most researchers in China carry out related research and report on extracts of roots, stems and leaves of celery and celery seed essential oil, but the related research on the celery seed extracts is less involved; chinese patent CN100569241C discloses a celery seed ethyl acetate extract, the preparation method is as follows: 10kg of dried celery seeds are subjected to percolation extraction by 15-20 times of 75-95% (V/V) ethanol or heating extraction by 4-5 times of 75-95% (V/V) ethanol for 3-4 times, and the heating is performed for 1-1.5 hours each time; concentrating the extractive solution at 55 deg.C under reduced pressure until ethanol is almost absent to obtain concentrated solution; adding 1-2 times of water into the concentrated solution to dilute and suspend, extracting for 2-3 times by using petroleum ether with a boiling range of 60-90 ℃ and one third volume of the diluted solution, separating out a petroleum ether layer, and recovering the petroleum ether to obtain a petroleum ether extract; extracting the water layer with ethyl acetate of one third volume for 4 times, separating ethyl acetate layer, recovering ethyl acetate to obtain semen Apii Graveolentis ethyl acetate extract, and application of the extract in preparing medicine for preventing and treating gout, antiinflammatory agent or health food. The existing extraction method of the celery seed extract has the defects of complex extraction steps, low crude drug rate, basically no refining method, poor repeatability and the like, and is difficult to industrially produce.
Disclosure of Invention
The invention is characterized in that the celery seed extract and the preparation method thereof are provided, the obtained celery seed extract has better pharmaceutical performance and can effectively reduce uric acid; the preparation method has simple process, high yield and good reproducibility.
In order to achieve the purpose, the specific technical scheme of the invention is as follows:
a celery seed extract is prepared by the following steps:
a. pulverizing dried celery seeds, adding 95% ethanol water solution, reflux extracting for 1-2 times, wherein the ethanol water solution is added for each time to obtain 2-4 times volume of dried celery seeds, and the reflux time is 0.5-1.5 h; collecting and combining the extracting solutions, and concentrating under reduced pressure until no alcohol exists to obtain a thick extract;
b. suspending the thick extract obtained in the step a in water with the volume 0.5-1 time of that of the celery seed, extracting for 2-4 times by using an extraction reagent, adding the extraction reagent 0.2-0.4 times of the thick extract each time, merging organic phases after extraction is finished, drying, and concentrating under reduced pressure to obtain the celery seed extract;
wherein the extraction reagent is selected from: one or more of ethyl acetate, dichloromethane, petroleum ether and n-hexane.
Preferably, the preparation method further comprises the following refining method:
c. dissolving the crude extract in ethyl acetate at a concentration of 0.2-0.5g/ml, and purifying with silica gel column chromatography;
d. gradient elution: eluting with petroleum ether at an elution flow rate of 7-10ml/min for 8-10 column volumes, and removing fraction; eluting 10-12 column volumes by using a mixed solution of petroleum ether and dichloromethane in a volume ratio of 7:1, and collecting fractions; eluting 3-5 column volumes with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 9:4:1, and removing fraction; eluting with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 5:7:3 for 15-20 column volumes, collecting fractions, mixing the collected fractions, and concentrating under reduced pressure to obtain refined herba Apii Graveolentis seed extract.
The celery seed extract provided by the invention has a very good effect on reducing uric acid, and is beneficial to treatment of hyperuricemia and protection of human kidney.
On the other hand, the preparation method of the celery seed extract comprises the following specific steps:
a. pulverizing dried celery seeds, adding 95% ethanol water solution, reflux extracting for 1-2 times, wherein the ethanol water solution is added for each time to obtain 2-4 times volume of dried celery seeds, and the reflux time is 0.5-1.5 h; collecting and combining the extracting solutions, and concentrating under reduced pressure until no alcohol exists to obtain a thick extract;
b. suspending the thick extract obtained in the step a in water with the volume of 0.5-1 time of that of the dry celery seeds, extracting for 2-4 times by using an extraction reagent, adding the extraction reagent in an amount of 0.2-0.4 times of the volume of the thick extract each time, merging organic phases after the extraction is finished, drying, and concentrating under reduced pressure to obtain a crude extract;
wherein the extraction reagent is selected from: one or more of Ethyl Acetate (EA), Dichloromethane (DCM), Petroleum Ether (PE) or n-hexane (Hex); preferably a mixed solution of ethyl acetate and n-hexane in a mass ratio of 1:1-1:4, more preferably a mass ratio of ethyl acetate to n-hexane of 1: 2;
c. dissolving the crude extract in ethyl acetate at a concentration of 0.2-0.5g/ml, and purifying with silica gel column chromatography;
d. gradient elution: the elution flow is 7-10ml/min, preferably 8 ml/min; eluting 8-10 column volumes with petroleum ether, and removing fraction; eluting 10-12 column volumes by using a mixed solution of petroleum ether and dichloromethane in a volume ratio of 7:1, and collecting fractions; eluting 3-5 column volumes with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 9:4:1, and removing fraction; eluting with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 5:7:3 for 15-20 column volumes, collecting fractions, mixing the collected fractions, and concentrating under reduced pressure to obtain the semen Apii Graveolentis extract.
Furthermore, in order to improve the yield of the product, ultrasonic wave assistance is arranged in the reflux extraction step, and the frequency of the ultrasonic wave is 60-80 KHz.
The preparation method of the celery seed extract provided by the invention is summarized by a large number of experiments, reasonable overall planning and comparison, and has the advantages of simple process, less extraction times, high drug yield, mild conditions and the like; the purification and refining steps are reasonable, the purification yield is high, the active ingredients contained in the celery seed extract obtained through the purification steps are improved by 12-15% compared with the active ingredients in the prior art, and the method can be suitable for industrial production.
On the other hand, the invention also provides a soft capsule prepared from the celery seed extract and pharmaceutically acceptable auxiliary materials, the soft capsule comprises a soft capsule filler and a capsule wall material, and the soft capsule filler comprises the following raw materials in parts by weight: 10-25 parts of celery seed extract, 20-35 parts of diluent, 3-6 parts of disintegrating agent and 1-2 parts of adhesive.
Preferably, the diluent is a mixture of N-dodecylamine xyloside and sesame oil in a weight ratio of 2: 3; the disintegrating agent is a mixture of crospovidone, sodium lauryl sulfate and lauroylsarcosine with the weight part ratio of 1:2: 2.
More preferably, the capsule wall material is mainly prepared from the following raw materials in parts by weight, namely 10 parts of gelatin, 1.2 parts of cellulose acetate phthalate, 0.8 part of pullulan, 10 parts of water, 2.8 parts of titanium dioxide and 1.9 parts of galactose.
According to the soft capsule provided by the invention, the auxiliary materials and the celery extract do not react, the bioavailability of the celery seed extract in the capsule can be improved, the curative effect of the capsule is fully exerted, and the content uniformity is high; by strictly screening the types and the technological parameters of the auxiliary materials, the capsule wall material prepared from the raw materials is selected to be matched with the celery seed extract, so that the problem that the medicine is unstable when encountering damp and heat is solved, the stability of the capsule is further ensured, and the stability of the celery seed extract is further increased.
In addition, the invention also provides application of the celery seed extract in preparing a medicament for reducing uric acid.
The celery seed extract has a very good effect on reducing uric acid, and is beneficial to treatment of hyperuricemia and protection of human kidney; in addition, the preparation method of the celery seed extract provided by the invention is summarized by a large number of experiments, reasonable overall planning and comparison, has the advantages of simple process, less extraction times, high drug yield, mild conditions and the like, and provides a better choice for industrial production of the celery seed extract; the soft capsule is prepared from the celery seed extract and the auxiliary materials, so that the carrying and using convenience of a patient is improved, the auxiliary material formula is optimized, the bioavailability of the celery seed extract is improved, and the stability of the celery seed extract and the soft capsule is enhanced; the storage condition of the capsule becomes milder.
Drawings
FIG. 1 is a graph of the effect of reflux time on yield;
FIG. 2 is a graph showing the effect of column chromatography temperature on column chromatography yield.
Detailed Description
Example 1
A celery seed extract is prepared by the following steps:
a. taking dry celery seeds, crushing, adding 2 times of ethanol water solution with volume fraction of 95% and performing reflux extraction for 1.5 h; collecting extractive solution, concentrating under reduced pressure until no alcohol exists to obtain thick extract;
b. suspending the thick extract obtained in the step a in water with the volume of 1 time of that of the raw material, extracting for 2 times by using a mixed solution of petroleum ether and ethyl acetate with the volume ratio of 3:1, adding an extraction reagent with the volume of 0.2 time of that of the raw material each time, merging organic phases after extraction is finished, drying, and concentrating under reduced pressure to obtain a celery seed extract;
example 2
A celery seed extract is prepared by the following steps:
a. taking dry celery seeds, crushing, adding ethanol water solution with volume fraction of 95%, refluxing for 2 times, wherein the volume of the ethanol water solution added each time is 2.5 times, and the refluxing time is 0.5h, collecting and combining extracting solutions, and concentrating under reduced pressure until no alcohol exists to obtain a thick extract;
b. suspending the thick extract obtained in the step a in water with the volume 0.5 times that of the raw material, extracting for 4 times by using ethyl acetate or dichloromethane, adding an extracting agent with the volume 0.4 times that of the raw material each time, combining organic phases after extraction is finished, drying, and concentrating under reduced pressure to obtain a crude extract;
c. dissolving the crude extract in ethyl acetate at a concentration of 0.2g/ml, and purifying with silica gel column;
d. gradient elution: eluting with petroleum ether at an elution flow rate of 7ml/min for 10 column volumes, and removing fractions; eluting 12 column volumes by using mixed solution of petroleum ether and dichloromethane in a volume ratio of 7:1, and collecting fractions; eluting 3 column volumes with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 9:4:1, and removing fraction; eluting with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 5:7:3 for 15 column volumes, collecting fractions, mixing the collected fractions, and concentrating under reduced pressure to obtain semen Apii Graveolentis extract.
Example 3
A preparation method for preparing a celery seed extract comprises the following steps:
a. extraction: taking dry celery seeds, crushing, adding an ethanol water solution with 4 times volume of volume and volume fraction of 95% for reflux extraction for 1h, and simultaneously carrying out ultrasonic assistance; collecting extractive solution, concentrating under reduced pressure until no alcohol exists to obtain thick extract;
b. and (3) extraction: suspending the thick extract obtained in the step a in water with the volume 0.9 times that of the raw material, extracting for 3 times by using a mixed solution of ethyl acetate and n-hexane with the mass ratio of 1:2, adding an extraction reagent in an amount of 0.3 times that of the raw material each time, merging organic phases after extraction is finished, drying, and concentrating under reduced pressure to obtain the celery seed extract;
c. dissolving the crude extract in ethyl acetate at a concentration of 0.5g/ml, and purifying with silica gel column;
d. gradient elution: eluting with petroleum ether at flow rate of 10ml/min for 8 column volumes, and removing fraction; eluting 10 column volumes by using mixed solution of petroleum ether and dichloromethane in a volume ratio of 7:1, and collecting fractions; eluting 5 column volumes by using a mixed solution of petroleum ether, dichloromethane and ethyl acetate in a volume ratio of 9:4:1, and removing fractions; eluting 20 column volumes with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 5:7:3, collecting fractions, mixing the collected fractions, and concentrating under reduced pressure to obtain semen Apii Graveolentis extract.
Example 4
A preparation method for preparing a celery seed extract comprises the following steps:
a. extraction: taking dry celery seeds, crushing, adding an ethanol water solution with 4 times volume of volume and volume fraction of 95% for reflux extraction for 1h, and simultaneously carrying out ultrasonic assistance; collecting extractive solution, concentrating under reduced pressure until no alcohol exists to obtain thick extract;
b. and (3) extraction: suspending the thick extract obtained in the step a in water with the volume 0.9 times that of the raw material, extracting for 3 times by using a mixed solution of ethyl acetate and n-hexane with the mass ratio of 1:2, adding an extraction reagent in an amount of 0.3 times that of the raw material each time, merging organic phases after extraction is finished, drying, and concentrating under reduced pressure to obtain the celery seed extract;
c. dissolving the crude extract in ethyl acetate at a concentration of 0.5g/ml, and purifying with silica gel column;
d. gradient elution: eluting with petroleum ether at an elution flow rate of 8ml/min for 9 column volumes, and removing fractions; eluting 11 column volumes by using a mixed solution of petroleum ether and dichloromethane in a volume ratio of 7:1, and collecting fractions; eluting 4 column volumes with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 9:4:1, and removing fraction; eluting 17 column volumes with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 5:7:3, collecting fractions, mixing the collected fractions, and concentrating under reduced pressure to obtain semen Apii Graveolentis extract.
EXAMPLE 5 Soft Capsule
The soft capsule filling material:
capsule material:
the preparation method comprises the following steps:
1) conventionally mixing gelatin, water, galactose and cellulose acetate phthalate, heating and melting to obtain a glue solution, adding titanium dioxide and pullulan, uniformly mixing, grinding by a colloid mill, and pressing into a rubber sheet for later use;
2) uniformly mixing the celery seed extract prepared in the example 2, N-dodecylamine xyloside, sesame oil and a mixture of two-thirds of crospovidone, sodium lauryl sulfate and lauroylsarcosine, and then crushing into micro powder for later use; dissolving povidone in 75% ethanol water solution by volume fraction to obtain solution, dissolving the micropowder in the solution, mixing, making into wet pill, mixing the wet pill with the rest mixture of crospovidone, sodium lauryl sulfate and lauroyl sarcosine, and drying to obtain pellet;
3) and (3) pressing the pellets and the rubber obtained in the step 1) to obtain the soft capsule.
EXAMPLE 6 Soft Capsule
The soft capsule filling material:
capsule material:
the preparation method comprises the following steps: the preparation was carried out as described in example 4.
Example 7 Soft capsules
The soft capsule filling material:
capsule material:
the preparation method comprises the following steps: the preparation was carried out as described in example 4.
Comparative example 1 Soft Capsule
The soft capsule filling material:
capsule material:
the preparation method comprises the following steps:
1) conventionally mixing gelatin, water and galactose, heating and melting to obtain a glue solution, adding titanium dioxide, uniformly mixing, passing through a colloid mill, and pressing to obtain a rubber sheet for later use;
2) uniformly mixing the celery seed extract prepared in the example 2, N-dodecylamine xyloside, sesame oil and a mixture of two-thirds of crospovidone, sodium lauryl sulfate and lauroylsarcosine, and then crushing into micro powder for later use; dissolving povidone in 75% ethanol water solution by volume fraction to obtain solution, dissolving the micropowder in the solution, mixing, making into wet pill, mixing the wet pill with the rest mixture of crospovidone, sodium lauryl sulfate and lauroyl sarcosine, and drying to obtain pellet;
3) and (3) pressing the pellets and the rubber obtained in the step 1) to obtain the soft capsule.
Comparative example 2 Soft Capsule
The soft capsule filling material:
capsule material:
the preparation method comprises the following steps:
1) conventionally mixing gelatin, water and glycerol, heating and melting to obtain a glue solution, adding titanium dioxide, uniformly mixing, passing through a colloid mill, and pressing to obtain a rubber sheet for later use;
2) uniformly mixing the celery seed extract prepared in the example 2, sorbitol and alginic acid, crushing into micro powder, dissolving methylcellulose in 75% volume fraction ethanol water solution to form a solution, dissolving the micro powder in the solution, uniformly mixing, preparing into wet pills, uniformly mixing the wet pills with sodium carboxymethyl cellulose, and drying to obtain micro pills;
3) and (3) pressing the pellets and the rubber obtained in the step 1) to obtain the soft capsule.
1 method of experiment
(1) Influence of celery seed extract on serum uric acid of normal mice: 72 ICR mice, SPF grade, male and female halves, weight 18.2-22.9 g, randomly divided into 6 groups according to sex and weight, which are respectively a blank control group, an allopurinol group (39mg/kg), a celery seed ethyl acetate extract disclosed in Chinese patent CN100569241 as a positive control group (3.3g celery seed/kg), a low, medium and high dose group (3.3, 9.9, 29.7g celery seed/kg) of the celery seed extract, and each group comprises 12 animals. The corresponding dose of the liquid medicine is administrated to each group of mice by gastric lavage with 20mL/kg, and the equal volume of peanut oil is administrated to the blank control group by gastric lavage for 1 time/day for 14 consecutive days. Fasting is not forbidden for 12h before the last administration, blood is collected from orbital venous plexus of each group of mice 1h after the last administration, and serum uric acid level is detected.
(2) The influence of the celery seed extract on the mouse hyperuricemia model caused by oteracil potassium salt is as follows: 70 ICR mice, SPF grade, male, 24.1-27.9 g weight, randomly divided into 7 groups according to weight, which are respectively a normal control group, a model control group, an allopurinol group (39mg/kg), a celery seed ethyl acetate extract disclosed in Chinese patent CN100569241 as a positive control group (9.9g/kg), a celery seed extract low, medium and high dose group (3.3, 9.9, 29.7g/kg), and 10 animals in each group. The corresponding dose of liquid medicine is administrated to each group of mice by gastric lavage of 20mL/kg, the equal volume of distilled water is administrated to the normal control group by gastric lavage, the equal volume of peanut oil is administrated to the model control group by gastric lavage for 1 time/day for 7 consecutive days. Before the last administration, fasting is carried out for 12 hours without water prohibition, 1 hour before the administration, except for a normal control group, the intraperitoneal injection of 300mg/kg of oteracil potassium salt is carried out on the other groups of mice, and blood is collected from orbital venous plexus before model building and 1 hour after the last administration of the mice, and the serum uric acid level is detected, and the results are shown in table 1.
(3) Effect of celery seed extract on the model of hyperuricemia in rats caused by adenine and ethambutol: 70 SD rats with SPF grade and male body weight of 198.4-228.4 g are randomly divided into a normal control group, a model control group, an allopurinol group (27mg/kg) and a celery seed ethyl acetate extract disclosed in Chinese patent CN100569241 as a positive control group (5.1g/kg), wherein the celery seed extract is low, medium and high dose groups (1.7, 5.1 and 15.3g/kg), and each group comprises 10 animals. The rats in each group are subjected to intragastric administration according to 10mL/kg of liquid medicine with corresponding dose, the normal control group is subjected to intragastric administration of equal volume of distilled water, and the model control group is subjected to intragastric administration of equal volume of peanut oil for 1 time/day for 17 consecutive days. Except for the normal control group, the rats in the other groups are administered with adenine (300mg/kg) and ethambutol (250mg/kg) in the morning on the 10 th day, the rat hyperuricemia model is duplicated, and the corresponding liquid medicine is administered at the 10mL/kg intragastric administration in the afternoon. The rats were fasted for 12h before the last administration and 1h after the last administration, blood was collected from orbital venous plexus of each group, and serum uric acid levels were measured, and the results are shown in table 2.
2 results of the experiment
TABLE 1 Effect of celery seed extract on Potassium Oxonate induced hyperuricemia model in mice
Note: comparison with Positive control group Pa＜0.05，PbLess than 0.01, P compared with allopurinol groupc＜0.05.
TABLE 2 Effect of celery seed extract on the model of hyperuricemia in rats induced by adenine and ethambutol
Note: comparison with Positive control group Pa＜0.05，PbLess than 0.01, P compared with allopurinol groupc＜0.05.
(1) Influence of celery seed extract on serum uric acid of normal mice: mice were gavaged with different doses of celery seed extract (3.3, 9.9, 29.7g/kg) for 14 consecutive days, wherein low doses of celery seed extract significantly reduced serum uric acid levels in normal mice.
(2) The influence of the celery seed extract on the mouse hyperuricemia model caused by oteracil potassium salt is as follows: continuously administering for 7 days, and making the mouse into a hyperuricemia model by injecting potassium oxonate into the abdominal cavity, wherein the serum UA of the mouse is obviously increased after the model is made; the celery seed extract has low, medium and high doses (3.3, 9.9 and 29.7g/kg), can obviously reduce the serum UA level of a mouse after molding, can obviously reduce the rising degree of the mouse serum UA after molding, has difference compared with a positive control group in a low dose group, has obvious difference compared with a high dose group and a positive control group, and has better effect compared with the existing allopurinol tablets in a high dose group.
(3) Effect of celery seed extract on the model of hyperuricemia in rats caused by adenine and ethambutol: the rat replicates hyperuricemia model by continuously intragastrically administering adenine and ethambutol for 7 days, the serum UA of the rat is obviously increased after model building, the low, medium and high doses (1.7, 5.1 and 15.3g/kg) of the celery seed extract can obviously reduce the serum urea nitrogen BUN level, serum creatine CRE level and serum UA level of the model rat, and the differences are obvious compared with a positive control group.
The test results show that the celery seed extract has obvious effects of preventing and treating hyperuricemia and protecting the kidney. In the experimental process, no death occurs when the mice are administrated, other organs are not abnormal, and the toxicity is slight.
20kg of celery seeds are taken and extracted by the method described in the embodiment 3, under the condition that other experimental conditions are not changed, single factors such as extraction reagents, reflux time or column chromatography temperature are respectively changed, the change of the extraction yield is inspected, and the experimental results are shown in a table 3, a figure 1 and a figure 2.
TABLE 3 Effect of different extraction reagents on yield
From the above experimental results, EA/Hex is the best extraction reagent, crude drug yield can reach 9.2%, pure EA as extraction reagent has higher crude product yield but more impurities, and total yield after purification is slightly worse than that of EA/Hex as extraction reagent.
TABLE 4 influence of different ratios of ethyl acetate and n-hexane on the yield
From the above experimental results, it is known that the yield gradually increases as the n-hexane content decreases, and when n-hexane/ethyl acetate is 1:2, the yield reaches the highest, and the n-hexane content further increases, and the yield decreases.
As can be seen from FIG. 1, the yield is low after 0.5h of reflux, and the yield does not increase with the time after 1h of reflux, and the reflux time of 1h can be selected as the optimal reflux time in view of the cost-saving principle; as can be seen from FIG. 2, in the extraction method of the present invention, the column-passing purification yield is high, and under the condition of ensuring that other conditions are not changed, the column-passing temperature is independently changed, and it is observed that the column-passing yield increases with the increase of temperature, which is optimal when the temperature reaches 40 ℃, and the yield decreases after the temperature further increases to 50-55 ℃, and the adsorption of silica gel to the product increases.
Comprehensively, the preparation method of the celery seed extract has the advantages of less extraction times, mild extraction conditions, simple method, high purification efficiency and the like, greatly improves the crude drug extraction yield of natural drugs, can reach 9.3 percent to the maximum, and provides an effective way for industrial production.
3.1 accelerated test
Taking the soft capsules described in the embodiment 5, the embodiment 7, the comparison embodiment 1 and the comparison embodiment 2, placing the soft capsules for 6 months under the constant temperature of 40 ℃ +/-2 ℃ and the constant humidity condition that the relative humidity is 75% +/-5%, sampling once at the end of 1 month, 2 months, 3 months and 6 months in the test period, respectively detecting the properties of the capsules and the labeled amount (%) containing the apigenin according to the regulation in Chinese pharmacopoeia, and the accelerated test result is shown in a table 5;
TABLE 5 accelerated test results of inventive and control samples
As can be seen from the table, in example 7, after being placed in a high-temperature and high-humidity environment for 6 months, the soft capsule of the invention has no cracks and fissures, and the marked amount of the apigenin-containing soft capsule has no obvious change; example 7 is different from example 5 only in the weight ratio of the capsule wall material, the stability of example 5 is slightly poor, and the marked amount of apigenin is reduced by about 6%; example 7 is different from the comparative example in the capsule material, and the celery seed extract soft capsule described in the comparative example has a rough surface and a marked amount containing apigenin is significantly reduced after being placed in a high-temperature and high-humidity environment for 2-3 months; therefore, the soft capsule of the invention has obviously improved stability.
3.2 Long term test
Taking the soft capsules described in the embodiment 5, the embodiment 7, the comparison embodiment 1 and the comparison embodiment 2, placing the soft capsules at the temperature of 25 ℃ +/-2 ℃ and the relative humidity of 60% +/-10% for 24 months, sampling every 3 months, sampling at 0 month, 3 months, 6 months, 9 months, 12 months, 18 months and 24 months respectively, detecting the properties of the capsules and the marking amount (%) containing apigenin, and the long-term test result is shown in table 6;
TABLE 6 Long-term test results for inventive samples and controls
The table shows that the soft capsule of the invention is placed for a long time, the properties of the soft capsule meet the standard, and the quantity of the apigenin is not obviously changed; in contrast, after the soft capsule of the comparative example is placed for 9-12 months for a long time, spots appear on the surface of the capsule and the fracture phenomenon occurs; the amount of the danapigenin is obviously reduced; the soft capsule of the invention is stable in property after long-term storage compared with the control soft capsule.
Claims (9)
1. The celery seed extract is characterized in that the preparation method of the celery seed extract comprises the following steps:
a. crushing dried celery seeds, adding an ethanol water solution with the volume fraction of 95% for reflux extraction for 1-2 times, wherein the ethanol water solution is added for each time to obtain 2-4 times of volume of the dried celery seeds, and the reflux time is 1 h; collecting and combining the extracting solutions, and concentrating under reduced pressure until no alcohol exists to obtain a thick extract;
b. suspending the thick extract obtained in the step a in water with the volume 0.5-1 time of that of the celery seed, extracting for 2-4 times by using an extraction reagent, adding the extraction reagent 0.2-0.4 times of the thick extract each time, merging organic phases after extraction is finished, drying, and concentrating under reduced pressure to obtain the celery seed extract;
wherein the extraction reagent is a mixed solution of ethyl acetate and n-hexane in a mass ratio of 1:2-1: 4.
2. The celery seed extract according to claim 1, wherein the preparation method further comprises the following purification method:
c. dissolving the celery seed extract obtained in the step b in ethyl acetate at the concentration of 0.2-0.5g/ml, and loading on a silica gel chromatographic column;
d. gradient elution: eluting with petroleum ether at an elution flow rate of 7-10ml/min for 8-10 column volumes, and removing fraction; eluting 10-12 column volumes by using a mixed solution of petroleum ether and dichloromethane in a volume ratio of 7:1, and collecting fractions; eluting 3-5 column volumes with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 9:4:1, and removing fraction; eluting with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 5:7:3 for 15-20 column volumes, collecting fractions, mixing the collected fractions, and concentrating under reduced pressure to obtain refined semen Apii Graveolentis extract.
3. The preparation method of the celery seed extract is characterized by comprising the following steps:
a. crushing dried celery seeds, adding an ethanol water solution with the volume fraction of 95% for reflux extraction for 1-2 times, wherein the ethanol water solution is added for each time to obtain 2-4 times of volume of the dried celery seeds, and the reflux time is 1 h; collecting and combining the extracting solutions, and concentrating under reduced pressure until no alcohol exists to obtain a thick extract;
b. suspending the thick extract obtained in the step a in water with the volume of 0.5-1 time of that of the dry celery seeds, extracting for 2-4 times by using an extraction reagent, adding the extraction reagent in an amount of 0.2-0.4 times of the volume of the thick extract each time, merging organic phases after the extraction is finished, drying, and concentrating under reduced pressure to obtain a crude extract; wherein the extraction reagent is a mixed solution of ethyl acetate and n-hexane in a mass ratio of 1:2-1: 4;
c. dissolving the crude extract in ethyl acetate at a concentration of 0.2-0.5g/ml, and purifying with silica gel column chromatography;
d. gradient elution: eluting with petroleum ether at an elution flow rate of 7-10ml/min for 8-10 column volumes, and removing fraction; eluting 10-12 column volumes by using a mixed solution of petroleum ether and dichloromethane in a volume ratio of 7:1, and collecting fractions; eluting 3-5 column volumes with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 9:4:1, and removing fraction; eluting with mixed solution of petroleum ether, dichloromethane and ethyl acetate at volume ratio of 5:7:3 for 15-20 column volumes, collecting fractions, mixing the collected fractions, and concentrating under reduced pressure to obtain the semen Apii Graveolentis extract.
4. The method of claim 3, wherein the elution flow rate is 8 ml/min.
5. The method of any one of claims 3-4, wherein the reflux extraction step is assisted by ultrasound at a frequency of 60-80 KHz.
6. The soft capsule prepared from the celery seed extract and pharmaceutically acceptable auxiliary materials according to claim 1, wherein the soft capsule comprises a soft capsule filling material and a capsule wall material, and the soft capsule filling material comprises the following raw materials in parts by weight: 10-25 parts of celery seed extract, 20-35 parts of diluent, 3-6 parts of disintegrating agent and 1-2 parts of adhesive.
7. The soft capsule of claim 6, wherein the diluent is a mixture of N-dodecylamine xyloside and sesame oil in a weight ratio of 2: 3; the disintegrating agent is a mixture of crospovidone, sodium lauryl sulfate and lauroylsarcosine with the weight part ratio of 1:2: 2.
8. The soft capsule of claim 7, wherein the capsule wall material is prepared from the following raw materials, by weight, 10 parts of gelatin, 1.2 parts of cellulose acetate phthalate, 0.8 parts of pullulan, 10 parts of water, 2.8 parts of titanium dioxide and 1.9 parts of galactose.
9. Use of the celery seed extract according to claim 1 for the manufacture of a medicament for lowering uric acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610177185.2A CN106474169B (en) | 2016-03-25 | 2016-03-25 | Celery seed extract, preparation and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610177185.2A CN106474169B (en) | 2016-03-25 | 2016-03-25 | Celery seed extract, preparation and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106474169A CN106474169A (en) | 2017-03-08 |
| CN106474169B true CN106474169B (en) | 2022-04-12 |
Family
ID=58238720
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610177185.2A Active CN106474169B (en) | 2016-03-25 | 2016-03-25 | Celery seed extract, preparation and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106474169B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107998369A (en) * | 2017-12-13 | 2018-05-08 | 国药肽谷有限公司 | A kind of preparation method of the oligomeric peptides products of ox bone collagen albumen |
| CN107997184A (en) * | 2017-12-13 | 2018-05-08 | 国药肽谷有限公司 | The preparation method of one seed oyster oligopeptide product |
| CN108606124A (en) * | 2018-05-03 | 2018-10-02 | 桂林漓峰医药用品有限责任公司 | A kind of celery seed fat decreasing tea |
| CN108434184A (en) * | 2018-06-07 | 2018-08-24 | 安徽易秦生物科技有限公司 | A kind of plant extracts oral tablet and preparation method thereof reducing uric acid |
| CN110585252A (en) * | 2019-08-27 | 2019-12-20 | 杨凌萃健生物工程技术有限公司 | Celery seed extract and preparation method thereof |
| CN112716989B (en) * | 2020-10-26 | 2022-06-28 | 山东大学 | Celery seed extract and preparation method and application thereof |
| CN113893269A (en) * | 2021-10-05 | 2022-01-07 | 浙江省农业科学院 | Preparation and purification method of celery seed extracting solution with blood pressure lowering effect |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003099303A1 (en) * | 2002-05-23 | 2003-12-04 | Sheffield, Hallam University | Anti-helicobacter activity of celery seed extract |
| CN101007015A (en) * | 2007-01-17 | 2007-08-01 | 北京天川军威医药技术开发有限公司 | Application of flavonoids of celery seed and coumarins in preparation of drug for preventing and treating gout |
| CN102008525A (en) * | 2010-12-09 | 2011-04-13 | 北京人福军威医药技术开发有限公司 | Pellet of celery seed extract and preparation method thereof |
| CN102697772A (en) * | 2012-05-08 | 2012-10-03 | 中国人民解放军第二军医大学 | Pharmaceutical composition for treating gout and application thereof |
-
2016
- 2016-03-25 CN CN201610177185.2A patent/CN106474169B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003099303A1 (en) * | 2002-05-23 | 2003-12-04 | Sheffield, Hallam University | Anti-helicobacter activity of celery seed extract |
| CN101007015A (en) * | 2007-01-17 | 2007-08-01 | 北京天川军威医药技术开发有限公司 | Application of flavonoids of celery seed and coumarins in preparation of drug for preventing and treating gout |
| CN102008525A (en) * | 2010-12-09 | 2011-04-13 | 北京人福军威医药技术开发有限公司 | Pellet of celery seed extract and preparation method thereof |
| CN102697772A (en) * | 2012-05-08 | 2012-10-03 | 中国人民解放军第二军医大学 | Pharmaceutical composition for treating gout and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| Phytopathogenic Fungal Inhibitors from Celery Seeds;Tao Liu et al.;《Natural Product Communications》;20121231;第7卷(第7期);第890页左栏第2段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106474169A (en) | 2017-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106474169B (en) | Celery seed extract, preparation and preparation method thereof | |
| CN103432193A (en) | Microwave-assisted aqueous two-phase extraction and separation method of kudzu root total flavones | |
| CN112716989A (en) | Celery seed extract and preparation method and application thereof | |
| WO2020093510A1 (en) | Separation and purification method for polysaccharide in ganoderma lucidum spores | |
| CN102134268B (en) | Method for preparing panax japonicus saponin IVa and application of panax japonicus saponin IVa in preparing a medicament for protecting liver and lowering transaminase | |
| CN101480422A (en) | Tibetan oriental wormwood extract as well as preparation method and use thereof | |
| CN102321129A (en) | Stilbene glycoside injection and preparation process thereof | |
| CN107033045B (en) | A kind of preparation method of high-purity natural garlic 4,5,9-trithiadodeca-1,6,11-triene 9-oxide | |
| CN106822331A (en) | To lock application of the lichee bark extract based on shape polymer polyphenol in treatment antihyperuricemic disease drug or health products are prepared | |
| CN103830374B (en) | The application in hyperuricemia clearly of three leaf glycolipids | |
| CN110960569A (en) | Phyllanthus emblica extract and preparation method and application thereof | |
| CN104592322A (en) | Method for extracting and separating curculigoside from curculigo gaertn plants | |
| CN104983767A (en) | Method for preparing cynanchum wallichii wight total saponins | |
| CN101683396A (en) | Extraction method of tropaeolum total lavonoids | |
| CN108392506A (en) | The extracting method and pharmaceutical composition of a kind of wilsonii activity extract and its application | |
| CN101618069B (en) | Capsule preparation for treating bruise as well as bleeding and easing pain and preparation method and use thereof | |
| CN106822071B (en) | Chinese medicinal effective component for treating coronary heart disease and hyperlipidemia, its preparation method and method for separating effective component from the same | |
| CN107485615A (en) | Purposes of the Ligustrum robust glycosides C and combinations thereof in treatment hyperlipidemia and slimming medicine is prepared | |
| CN103800418A (en) | Composition for promoting blood circulation and stopping pain, capsule preparation technology and application thereof | |
| CN103585208A (en) | Preparation method of high-quality andrographolide component | |
| CN102614222A (en) | Sea cucumber extract rich in triterpene sapogenin and preparation method and application thereof | |
| CN110025659B (en) | Traditional Chinese medicine active site with anti-HBV effect, preparation method and application | |
| CN109467581B (en) | Method for extracting flavonoid glycoside from lotus leaves and pharmaceutical preparation containing flavonoid glycoside from lotus leaves | |
| CN110882246B (en) | Extraction method and application of coptis alkaloid with different biological activities | |
| CN105968150A (en) | Preparation method for 7-O-ethylmorroniside |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |