CN111388534A - Olive leaf extract and preparation and application thereof - Google Patents
Olive leaf extract and preparation and application thereof Download PDFInfo
- Publication number
- CN111388534A CN111388534A CN202010353478.8A CN202010353478A CN111388534A CN 111388534 A CN111388534 A CN 111388534A CN 202010353478 A CN202010353478 A CN 202010353478A CN 111388534 A CN111388534 A CN 111388534A
- Authority
- CN
- China
- Prior art keywords
- leaf extract
- olive leaf
- olive
- solution
- dose
- 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.)
- Pending
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)
- A61K36/63—Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
-
- 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
- A61K2236/39—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
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- 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
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- 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 discloses a preparation method of olive leaf extract, which comprises the steps of adding dried olive leaves into an ethanol water solution, heating, refluxing and extracting, recovering a solvent from an extracting solution, placing, cooling, filtering to obtain a concentrated extracting solution, adsorbing effective components on a macroporous resin column, washing the resin column with water to remove impurities after adsorption is finished, desorbing with an ethanol water solution, collecting a desorption solution, recovering the solvent from the desorption solution, and drying to obtain the olive leaf extract, wherein the contents of the effective components in the olive leaf extract prepared by the invention are 40-65% of picroside, 3-7% of ligustrum lucidum ait glycoside, 2-8% of luteolin-7-O- β -D glucoside and 0.1-1% of hydroxytyrosol.
Description
Technical Field
The invention relates to an olive leaf extract with hypoglycemic activity and a preparation method thereof, belonging to the field of separation of functional components of natural products.
Background
Olive (Olea europaea L.) is evergreen tree of the genus Olea of the family meliaceae, is a world-known woody oil tree species, has a history of over four thousand years of cultivation in mediterranean regions, and is formally introduced in our country from the 60 th century, and is currently widely cultivated in our country in gansu, shanxi, sichuan, Yunnan, etc. a large number of epidemiological evidences indicate that the incidence of cardiovascular diseases, diabetes, breast cancer, and colon cancer of people in mediterranean regions is lower than that of other people in the world, mainly due to the fact that olive oil is rich in unsaturated fatty acids and polyphenolic compounds, modern phytology studies indicate that the olive leaves contain abundant iridoid terpenoids, flavonoids, polyphenols, trisubes, and the like, and the pharmacological effects of the olive leaves are arrhythmia resistant, antifungal, antihypercholesterolemic, blood pressure lowering, anti-inflammatory, antioxidant, diuretic, spasmolytic, vasodilatory, and the like, and the olive leaf decoction has long-medicinal effect on rat blood sugar, and the primary diabetic mouse blood sugar level is unknown, and the olive leaf extract has a high-blood pressure-reducing medicinal effect on rat (generally-treated rat), and the rat) and the rat with high blood sugar level of olive leaf extract of olive oil.
Disclosure of Invention
The invention aims to provide a preparation method of an olive leaf extract.
The invention also aims to research the active ingredients and the hypoglycemic activity of the olive leaf extract, so that the olive leaf extract can be used as the active ingredients for preparing the medicines for treating the hypoglycemic diabetes.
Preparation method of olive leaf extract
The preparation method of the olive leaf extract comprises the following process steps:
(1) adding dried olive leaves into an ethanol aqueous solution, heating and refluxing (at the temperature of 50-100 ℃) for 1-3 times, mixing extracting solutions, recovering a solvent, standing, cooling, and filtering to obtain a concentrated extracting solution, wherein the mass percent of the ethanol aqueous solution is 50-90%, the dried olive leaves are added into the ethanol aqueous solution according to the material-liquid ratio of 1: 5-1: 15 g/L, the refluxing extraction time is 30-120 minutes each time, and the concentration of the extracted solution after the solvent is recovered is equivalent to 0.05 g/m L-0.5 g/m L of a raw material.
(2) Adsorbing effective components on the concentrated extracting solution obtained in the step (1) through a macroporous resin column, wherein the used macroporous resin is L SA type macroporous resin, the sample loading flow rate of the concentrated extracting solution is 4 BV/h-15 BV/h, after adsorption, washing the resin column with water to remove impurities, desorbing with 10-70% ethanol water solution by mass percent, and collecting the desorption solution, wherein the desorption flow rate is 4 BV/h-15 BV/h.
(3) Recovering solvent from the desorption solution, and drying to obtain common olive leaf extract. Wherein the drying is one or more of reduced pressure drying, spray drying, freeze drying or microwave drying.
FIG. 1 is a high performance liquid chromatogram of the olive leaf extract prepared by the invention, wherein 1 is oleuropein, 2 is ligustrin, 3 is luteolin-7-O- β -D-glucoside, 4 is hydroxytyrosol, the chromatographic detection conditions are SinoChrom ODS-BP chromatographic column (4.6 mm × 250 mm, 5 mu m) or other equivalent chromatographic columns, the mobile phases A is 0.2% of acetic acid water and B is acetonitrile, the gradient elution is 0-25 min, 18% -31% of B, the flow rate is 1.0m L/min, and the sample amount is 20 mu L.
High performance liquid chromatography detection results show that the olive leaf extract prepared by the method comprises 40-65% of oleuropein, 3-7% of ligustrin, 2-8% of luteolin-7-O- β -D glucoside and 0.1-1% of hydroxytyrosol.
Second, the hypoglycemic activity test of the olive leaf extract
1.2 diabetic rat hypoglycemic Activity test
The test selects 120 male SD rats with the weight of 180-:
1.1 Effect on fasting plasma glucose
After 1 time of administration of the olive leaf extract, the blood sugar of rats with high, medium and low doses is remarkably reduced, and compared with a model group, the blood sugar of the rats is remarkably different, and the blood sugar of the rats is basically stabilized at the same level after continuous administration for 4 weeks. The olive leaf extract shows a certain dose-effect relationship on the inhibition of blood sugar by each group. The blood sugar reducing effect of the olive leaf extract at the dose of 500mg/kg after 1 week of administration is equivalent to that of the positive medicament, and the blood sugar reducing effect of the olive leaf extract at the dose of 250mg/kg after 4 weeks of administration is equivalent to that of the positive medicament (see table 1).
1.2 Effect on glucose tolerance in rats
500. 250 and 125mg/kg of olive leaf extract can remarkably reduce the blood sugar of rats after being given glucose for 120 min, remarkably reduce the AUC area, and remarkably improve the abnormal glucose tolerance of rats with type 2 diabetes, and the olive leaf extract with 500mg/kg dose has lower effect than a positive drug (see table 2).
1.3 Effect on rat insulin tolerance
Compared with the model group, after the injection of insulin, the blood sugar level of the olive leaf extract is reduced in the rats of the 500mg/kg and 250mg/kg dose groups, and the area under the curve is obviously reduced, which shows that the olive leaf extract can obviously improve the insulin tolerance abnormality of the type 2 diabetic rats and increase the sensitivity of the type 2 diabetic rats to exogenous insulin, and the effect of the olive leaf extract of the 500mg/kg dose is slightly lower than that of a positive drug (see table 3).
1.4 Effect on serum GSP in type 2 diabetes model rats
The high, medium and low dose groups of the olive leaf extract can obviously reduce the serum GSP content of rats, which indicates that the blood sugar of each dose group is in a stable state and has small fluctuation after continuous administration for 5 weeks, and the olive leaf extract with the doses of 500, 250 and 125mg/kg has the effect equivalent to a positive drug (see table 4).
1.5 Effect on serum blood lipids in rats
Compared with the model group, the olive leaf extract high, medium and low dose groups can obviously reduce the serum TG and TCHO contents, and the high and medium dose groups can obviously reduce the serum NEFA content, compared with the positive drug group, the olive leaf extract high and low dose groups can obviously reduce the serum TG content, the low dose group can obviously reduce the serum L DC-C content, and the high dose group can obviously increase the serum HD L-C content, which indicates that the olive leaf extract can regulate the lipid metabolism disorder of diabetic rats, and the capability of regulating blood fat is better than that of the positive drug (see Table 5).
1.6 Effect on serum insulin content and insulin sensitivity index of type 2 diabetes model rat
The medium, low and low dose groups of the olive leaf extract can obviously reduce the content of insulin, and the high, medium and low dose groups can obviously improve the insulin sensitivity index, which indicates that the medicament can obviously reduce the blood sugar of a model rat, increase the insulin sensitivity index and relieve the insulin resistance, and the effect of the dose group in the olive leaf extract for increasing the insulin sensitivity index is equivalent to that of a positive medicament (see table 6).
The type 2 diabetes rat model experiment shows that: after 1 time of administration of the olive leaf extract, the blood sugar of rats in 500, 250 and 125mg/kg dose groups is remarkably reduced, and compared with a model group, the blood sugar of the rats is remarkably different, and the blood sugar of the rats is basically stabilized at the same level after continuous administration for 4 weeks. The olive leaf extract shows a certain dose-effect relationship on the inhibition of blood sugar by each group. The blood sugar reducing effect of the olive leaf extract with the dose of 500mg/kg after 1 week of administration is equivalent to that of the positive drug, and the blood sugar reducing effect of the olive leaf extract with the dose of 250mg/kg after 4 weeks of administration is equivalent to that of the positive drug; the olive leaf extract can obviously improve the glucose tolerance of rats, improve the insulin tolerance of rats, enhance the sensitivity of rats to insulin, and reduce the content of glycated serum protein and triglyceride and free fatty acid, which indicates that the olive leaf extract can regulate the lipid metabolism disorder of diabetic rats, has better capability of regulating blood fat than a positive drug, has equivalent hypoglycemic effect to the positive drug, and has slightly weaker capability of improving the glucose tolerance and the insulin tolerance than the positive drug.
2. Effect on spontaneous type 2 diabetic db/db mice
Measuring fasting blood glucose after model animal fasting for 10h, and selecting fasting blood glucose value>13.0 mmol/L db/db mice 66 are divided into 6 groups according to blood sugar level and body weight, each group comprises 11 mice, 6 males, and 5 females, and are respectively model group, small, low, medium, and high dose groups (corresponding to dose of 125, 250, 500, 1000 mg/kg) of Olea europaea leaf extract, positive drug control group (dose of 250 mg/kg), and C576 males and 5 females of the B L/6J mice are taken as a normal control group, 11 mice are taken as a normal control group, the groups are subjected to intragastric administration for 1 time every day according to the dose, the continuous administration is carried out for 5 weeks, and the normal control group and the model group are subjected to intragastric administration by deionized water every day.
2.1 Effect on fasting plasma glucose in db/db mice
After 1 week of administration, the olive leaf extract with 1000mg/kg dose has significant hypoglycemic effect, and after 2 weeks of administration, the olive leaf extract with 500mg/kg dose begins to have significant hypoglycemic effect, and the hypoglycemic effect lasts until the end of the test. As can be seen from the data in Table 7, the blood sugar reducing effect of the olive leaf extract on fasting blood sugar of a model mouse is gradually strengthened along with the increase of the administration days, and the blood sugar reducing effect 2 hours after the administration shows an obvious dose-effect relationship; comparison with the positive drug the hypoglycemic effect of the 1000mg/kg dose of olive leaf extract on fasting plasma glucose was comparable to the 250mg/kg dose of the positive drug at 5 weeks of administration (see table 7).
2.2 Effect on serum Glycated Serum Proteins (GSPs) of db/db mice
The high, medium, low and small dose groups of the olive leaf extract can obviously reduce the serum GSP content of rats, which indicates that the blood sugar of each dose group is in a stable state and has small fluctuation after continuous administration for 5 weeks, and the olive leaf extract with the dose of 1000mg/kg has the function equivalent to a positive drug (see table 8).
Spontaneous type 2 diabetes animal db/db mouse model experiments show that: after 1 week of administration of the olive leaf extract, the extract at a dose of 1000mg/kg has a significant hypoglycemic effect, and after 2 weeks of administration, the olive leaf extract at a dose of 500mg/kg starts to have a significant hypoglycemic effect, and thereafter, the hypoglycemic effect is continued until the end of the test. The data show that the blood sugar reducing effect of the olive leaf extract on fasting blood sugar of a model mouse is gradually strengthened along with the increase of administration days, and the blood sugar reducing effect 2 hours after the administration shows an obvious dose-effect relationship; the hypoglycemic effect of the olive leaf extract on fasting plasma glucose at the dose of 1000mg/kg compared with the positive drug was comparable to that of the positive drug at the dose of 250mg/kg at 5 weeks of administration.
3. Effect on spontaneous type 2 diabetic KK-Ay mice
Measuring fasting blood glucose of model animal after fasting for 4 hr, and selecting fasting blood glucose value>13.0 mmol/L KK-Ay miceThe blood sugar and the body weight of 60 olive leaves are divided into 6 groups at random according to the layering of the blood sugar and the body weight, wherein each group comprises 10 olive leaves, 4 male olive leaves and 6 female olive leaves, and the groups respectively comprise a model group, a low-dose group, a medium-dose group and a high-dose group (corresponding to the doses of 250mg/kg, 500mg/kg and 1000 mg/kg) of olive leaves and a positive drug group (the dose of 250 mg/kg). C574 males and 6 females of the B L/6J mice are taken as a normal control group, 10 mice are taken as a normal control group, the groups are subjected to intragastric administration for 1 time every day according to the dose, the continuous administration is carried out for 6 weeks, and the normal control group and the model group are subjected to intragastric administration by deionized water every day.
3.1 Effect on 2h blood glucose after administration to KK-Ay mice
After 1 time of administration, the olive leaf extract with 1000mg/kg dose has obvious hypoglycemic effect 2h after administration, the olive leaf extract with 500mg/kg dose and 1000mg/kg dose has obvious hypoglycemic effect 1 week after administration, the olive leaf extract with 250mg/kg dose starts to have obvious hypoglycemic effect 2 weeks after administration, and the hypoglycemic effect lasts until the end of the test. As can be seen from the data in Table 9, the hypoglycemic effect of the olive leaf extract on fasting blood glucose of the model mice is gradually enhanced along with the increase of the administration days, and the hypoglycemic effect 2 hours after the administration shows an obvious dose-effect relationship; comparison with the positive drug the hypoglycemic effect of the 1000mg/kg dose of extract on fasting plasma glucose was comparable to the 250mg/kg dose of the positive drug at 3 weeks of administration (see table 9).
3.2 Effect on blood glucose before administration to KK-Ay mice (i.e. 24h blood glucose after each administration)
After 1 week of administration, the blood sugar of 24 hours after the medicine is prepared at 500mg/kg and 1000mg/kg doses, and the blood sugar is obviously reduced at each detection time point in the whole test period, which shows that the blood sugar reducing effect of the olive leaf extract can be continued to 24 hours after the administration, and compared with the positive medicine, the blood sugar reducing effect of the olive leaf extract at the 1000mg/kg dose for 24 hours is stronger than that of the positive medicine. The 250mg/kg dose group showed hypoglycemic effect starting at 2 weeks of administration (see Table 10).
3.3 Effect on blood glucose at different time points 2 weeks after administration to KK-Ay mice
After 2 weeks of administration, the blood glucose levels of the mice were substantially lowest in each administration group at 2h after administration, indicating that monitoring of blood glucose 2h after administration may show the optimal effect of the drug. Meanwhile, the hypoglycemic effect of each administration group was significant at each time point after administration, and the hypoglycemic effect of the olive leaf extract was significant for the 1000mg/kg and 250mg/kg dose groups for a period of 24 hours after administration (see table 11).
4. Effect on adrenergic diabetic rats
SD male rats with blood sugar values of 4.5-6.0 (mmol/L) are selected for experiments, the SD male rats are divided into groups according to the blood sugar values in a layered random mode, the SD male rats are subjected to intragastric administration for 3 weeks according to the dose shown in table 12, the control group and the model group are given distilled water with the same volume, 1 drop of blood is pricked by a tail vein needle of the rat before the last administration (fasting for 4 hours), blood sugar at a point of 0 time is measured by a glucometer after the administration, 30 minutes after the administration, blood sugar at 90 minutes, 135 minutes and 180 minutes after the molding is measured by the method for the remaining rat sc adrenaline hydrochloride (1 mg/m L) injection liquid 0.2 m L/kg. except the normal control group, and the result is subjected to statistical treatment, and the result (shown in table 12) shows that the blood sugar at each time point after the molding of the olive leaf extract is high, medium, low and small dose animals are remarkably lower than the model group, and the olive leaf extract has the remarkable hypoglycemic effect on the adrenaline induced hyperglycemia.
The results of rat model experiments on adrenergic diabetes show that: the blood sugar of animals with high, medium, low and small dosages of olive leaves extract at each time point after model building is obviously lower than that of model groups, which shows that the olive leaves extract also has obvious blood sugar reducing effect on rats with adrenaline induced hyperglycemia.
In conclusion, the olive leaf extract is prepared by taking olive leaves as a raw material, heating, refluxing and extracting, concentrating an extracting solution, adsorbing effective components by adopting L SA macroporous adsorption resin, desorbing by using an ethanol water solution, recovering a solvent from a desorbing solution, and drying to obtain the olive leaf extract containing 40-65% of oleuropein, 3-7% of ligustrum lucidum glycoside, 2-8% of luteolin-7-O- β -D glucoside and 0.1-1% of hydroxytyrosol, wherein the olive leaf extract has a very obvious hypoglycemic effect on 4 different type 2 diabetes animal models and has an obvious time-effect and dose-effect relationship through activity tests of 4 different diabetes animal models such as a type 2 diabetes rat, a spontaneous type 2 diabetes db/db mouse, a spontaneous type 2 diabetes KK-AY mouse and an epinephrine diabetes rat.
Drawings
FIG. 1 is a high performance liquid chromatogram of the olive leaf extract prepared by the present invention.
Detailed Description
The preparation of the olive leaf extract and the content of the effective ingredients of the present invention will be further described by the following specific examples.
Example 1
(1) Weighing 5 kg of dried olive leaves, respectively extracting twice with 70% ethanol water solution under heating and refluxing (at 70 ℃), adding 60L extraction solvent (material-liquid ratio of 1: 12) for the first time, adding 50L extraction solvent (material-liquid ratio of 1: 10) for the second time, extracting for 120 min each time, combining the two extraction solutions, recovering solvent under reduced pressure, standing, and filtering to obtain concentrated extract 25L (equivalent to 0.2 g/m L of raw material);
(2) adsorbing effective components on an L SA macroporous resin column, wherein the height ratio of the diameter of the macroporous resin column is 1:10, the adsorption flow rate is 8 BV/h (BV is the volume of a resin column bed), washing the macroporous resin column by using 5BV of deionized water to remove part of impurities with weak adsorption after adsorption is finished, desorbing by using 60% ethanol aqueous solution, using the desorption solution at 6BV, and collecting the desorption solution, wherein the desorption flow rate is 15 BV/h;
(3) and concentrating the collected desorption solution under reduced pressure to recover the solvent, and then putting the recovered desorption solution into a vacuum drying oven to be dried to obtain 421 g of a dried product, namely the olive leaf extract.
The obtained olive leaf extract contains oleuropein 52%, ligustrin 4.6%, luteolin-7-O- β -D-glucoside 6.9%, and hydroxytyrosol 0.2% by high performance liquid chromatography.
Example 2
(1) Weighing 100 g of dried olive leaves, respectively heating and refluxing with 60% ethanol water solution twice, adding 1500 m L extraction solvent (material-liquid ratio is 1: 15) for the first time, adding 1200 m L extraction solvent (material-liquid ratio is 1: 12) for the second time, extracting for 90min each time, combining the two extraction solutions, recovering solvent under reduced pressure, standing and filtering to obtain 1000m L (equivalent to 0.1 g/m L) of raw material;
(2) adsorbing effective components on an L SA macroporous resin column, wherein the height ratio of the diameter of the macroporous resin column is 1:10, the adsorption flow rate is 5BV/h (BV is the volume of a resin column bed), washing the macroporous resin column by using 5BV deionized water to remove part of impurities with weak adsorption after adsorption is finished, desorbing by using 70% ethanol water solution, wherein the consumption of the desorption solution is 6BV, and the desorption flow rate is 10BV/h, and collecting the desorption solution;
(3) and concentrating the collected desorption solution under reduced pressure to recover the solvent, and then putting the recovered desorption solution into a vacuum drying oven to be dried to obtain 9.1 g of a dried product, namely the olive leaf extract.
The obtained olive leaf extract contains oleuropein 45%, ligustrin 3.5%, luteolin-7-O- β -D-glucoside 5.7%, and hydroxytyrosol 0.6% by high performance liquid chromatography.
Example 3
(1) Weighing 1 kg of dried olive leaves, respectively heating and refluxing for three times by using 80% ethanol water solution, adding 10L extraction solvent (material-liquid ratio is 1: 10) for the first time, adding 10L extraction solvent (material-liquid ratio is 1: 10) for the second time, adding 10L extraction solvent (material-liquid ratio is 1: 10) for the third time, extracting for 60 min each time, combining three extraction solutions, recovering solvent under reduced pressure, standing and filtering to obtain a concentrated extraction solution 20L (equivalent to 0.05 g/m L of a raw material);
(2) adsorbing effective components on an L SA macroporous resin column of the obtained concentrated extracting solution, wherein the height ratio of the diameter of the macroporous resin column is 1:10, the adsorption flow rate is 12 BV/h (BV is the volume of a resin column bed), washing the macroporous resin column by using 5BV of deionized water to remove part of impurities with weak adsorption after adsorption is finished, desorbing by using 50% ethanol water solution, using the desorbing solution for 8 BV, and collecting the desorbing solution, wherein the desorption flow rate is 6 BV/h;
(3) and concentrating the collected desorption solution under reduced pressure to recover the solvent, and then putting the recovered desorption solution into a vacuum drying oven to be dried to obtain 67.3 g of a dried product, namely the olive leaf extract.
The obtained olive leaf extract contains oleuropein 47%, ligustrin 5.1%, luteolin-7-O- β -D-glucoside 2.8%, and hydroxytyrosol 0.1% by high performance liquid chromatography.
Claims (10)
1. A preparation method of an olive leaf extract comprises the following process steps:
(1) adding dried olive leaves into an ethanol water solution, heating, refluxing and extracting for 1-3 times, combining extracting solutions, recovering a solvent, standing, cooling and filtering to obtain a concentrated extracting solution;
(2) adsorbing effective components of the concentrated extract obtained in the step (1) by using a macroporous resin column; after adsorption, washing the resin column with water to remove impurities, then desorbing with an ethanol aqueous solution, and collecting a desorption solution;
(3) recovering solvent from the desorption solution, and drying to obtain common olive leaf extract.
2. The preparation method of the olive leaf extract as claimed in claim 1, wherein in the step (1), the mass percent of the ethanol aqueous solution is 50% -90%, and the dried olive leaves are added into the ethanol aqueous solution according to the feed-liquid ratio of 1: 5-1: 15 g/L.
3. The method for preparing olive leaf extract as claimed in claim 1, wherein the olive leaf extract comprises: in the step (1), the reflux extraction temperature is 50-100 ℃, and the reflux extraction time is 30-120 minutes each time.
4. The method for producing an olive leaf extract according to claim 1, wherein in the step (1), the concentration of the solvent in the extract after the solvent is recovered is 0.05 g/m L-0.5 g/m L.
5. The method for preparing the olive leaf extract as claimed in claim 1, wherein the macroporous resin used in the step (2) is L SA type macroporous resin.
6. The method for preparing olive leaf extract as claimed in claim 1, wherein the olive leaf extract comprises: in the step (2), the sample feeding flow rate of the concentrated extracting solution is 4 BV/h-15 BV/h.
7. The method for preparing olive leaf extract as claimed in claim 1, wherein the olive leaf extract comprises: in the step (2), the mass percent of the ethanol water solution for macroporous resin column desorption is 10-70%, and the desorption flow rate is 4 BV/h-15 BV/h.
8. The method for preparing olive leaf extract as claimed in claim 1, wherein the olive leaf extract comprises: in the step (3), the drying is performed in one or more modes of reduced pressure drying, spray drying, freeze drying or microwave drying.
9. The olive leaf extract prepared by the method as claimed in claim 1, wherein the contents of the effective components are 40% -65% of oleuropein, 3% -7% of ligustrin, 2% -8% of luteolin-7-O- β -D-glucoside and 0.1% -1% of hydroxytyrosol.
10. The olive leaf extract as claimed in claim 1, which is used as an active ingredient for the preparation of hypoglycemic agents.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010353478.8A CN111388534A (en) | 2020-04-29 | 2020-04-29 | Olive leaf extract and preparation and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010353478.8A CN111388534A (en) | 2020-04-29 | 2020-04-29 | Olive leaf extract and preparation and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111388534A true CN111388534A (en) | 2020-07-10 |
Family
ID=71418417
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010353478.8A Pending CN111388534A (en) | 2020-04-29 | 2020-04-29 | Olive leaf extract and preparation and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111388534A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113854432A (en) * | 2021-09-26 | 2021-12-31 | 中国林业科学研究院林产化学工业研究所 | Olive leaf polyphenol extract feed additive rich in oleuropein and capable of improving quality of broiler chickens and preparation method of olive leaf polyphenol extract feed additive |
| CN114469820A (en) * | 2022-03-17 | 2022-05-13 | 中国科学院兰州化学物理研究所 | Olive leaf extract and application thereof in preparation of skin whitening cosmetics |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102451235A (en) * | 2010-10-27 | 2012-05-16 | 深圳劲创生物技术有限公司 | Preparation method of olive leaf extract |
| CN103830333A (en) * | 2012-11-20 | 2014-06-04 | 中国科学院兰州化学物理研究所 | Method for extracting anti-diabetic effective part of olive leaf by adopting macro-porous resin adsorption |
-
2020
- 2020-04-29 CN CN202010353478.8A patent/CN111388534A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102451235A (en) * | 2010-10-27 | 2012-05-16 | 深圳劲创生物技术有限公司 | Preparation method of olive leaf extract |
| CN103830333A (en) * | 2012-11-20 | 2014-06-04 | 中国科学院兰州化学物理研究所 | Method for extracting anti-diabetic effective part of olive leaf by adopting macro-porous resin adsorption |
Non-Patent Citations (1)
| Title |
|---|
| 刘天亮等: "橄榄苦苷的药理作用及其研究进展 ", 《西南国防医药》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113854432A (en) * | 2021-09-26 | 2021-12-31 | 中国林业科学研究院林产化学工业研究所 | Olive leaf polyphenol extract feed additive rich in oleuropein and capable of improving quality of broiler chickens and preparation method of olive leaf polyphenol extract feed additive |
| CN114469820A (en) * | 2022-03-17 | 2022-05-13 | 中国科学院兰州化学物理研究所 | Olive leaf extract and application thereof in preparation of skin whitening cosmetics |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112716989A (en) | Celery seed extract and preparation method and application thereof | |
| CN111388534A (en) | Olive leaf extract and preparation and application thereof | |
| CN109453237B (en) | Pharmaceutical application of jasminum grandiflorum extract | |
| CN102836188A (en) | Folium microcotis total flavone extract and preparation method and application thereof | |
| CN103550265A (en) | Extraction method of active ingredients of tuckahoe peels and tuckahoe peel extracts | |
| US20210187008A1 (en) | Use of acanthopanax trifoliatus polysaccharide atp1-1 in preparation of drugs for treatment of diabetes | |
| CN1203083C (en) | Rhizoma anemarrhenae extract and preparation process and use thereof | |
| CN109820959B (en) | Application of dendrobium officinale extract in preparation of medicines for preventing and treating dyslipidemia | |
| CN107349244B (en) | Extraction method of malonyl ginsenoside | |
| CN101618096A (en) | Madder extract and preparation method and application thereof | |
| CN109954024B (en) | A fructus Syzygii Aromaticae effective part, its preparation method and application in preparing medicine or food with blood sugar lowering effect | |
| CN110194758B (en) | Method for separating and purifying aristolochic acid compounds from caulis Aristolochiae Manshuriensis | |
| CN101322734B (en) | Codonopsis lanceolata total saponins with anti-inflammatory immunity function and preparation thereof | |
| CN113925948B (en) | Use of rhizoma Zingiberis recens total extract or its active component, pharmaceutical composition and preparation method | |
| CN101301374A (en) | Preparation of loquat leaf total flavones and functions thereof for reducing blood sugar and blood fat | |
| CN109966283B (en) | Application of degreased cinnamon polyphenol extract in preparation of product for preventing and treating diabetic nephropathy | |
| CN104257756B (en) | The application in preparing hypoglycemic medicine of a kind of Cortex Mori fatty oil | |
| CN109806287B (en) | General flavone glycoside of folium Microcoris paniculatae, and preparation method and application thereof | |
| CN102225095A (en) | Effective fraction of plantain seeds as well as preparation method and application thereof | |
| CN1475222A (en) | Vine leaf extract and its preparation process | |
| CN105106276A (en) | Tradescantia extract as well as preparation method and medical application of tradescantia extract | |
| CN116354912B (en) | Compound and application thereof in serving as or preparing ABCG2 agonist | |
| CN1322539A (en) | Diabetes-treating medicine preparing process with Chigenteng vane leaf | |
| CN116606330A (en) | Dendrobium nobile total alkaloids, extraction method thereof and application thereof in preparing medicament for treating diabetes | |
| CN101181336A (en) | Method for refining valid target of bitter gourd for dropping blood sugar |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |