CN111643560A - Preparation method and application of pyrrole derivative compound in noni ferment - Google Patents
Preparation method and application of pyrrole derivative compound in noni ferment Download PDFInfo
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Abstract
The invention discloses a preparation method of a pyrrole derivative compound in noni ferment, belonging to the technical field of phytochemistry and comprising the following steps: (1) cleaning and freezing treatment, (2) pressure swing treatment, (3) juicing and fermenting treatment, (4) extracting treatment and (5) chromatography treatment. The pyrrole derivative compound prepared by the invention has good prevention and treatment effects on three-high symptoms, can be used for preparing medicines for treating the three-high symptoms, and has good medical value and economic benefit.
Description
Technical Field
The invention belongs to the technical field of phytochemistry, and particularly relates to a preparation method and application of a pyrrole derivative compound in noni ferment.
Background
Noni (Morinda citrifolia Linn), a tropical evergreen perennial broadleaf shrub or small arbor originating from islands of the south pacific ocean, is a plant of the Morinda genus of the rubiaceae family. Noni fruit is called 'super fruit', is rich in 227 nutrient components, comprises 13 vitamins (A, B, C, E and the like), 16 minerals (potassium, sodium, zinc, calcium, iron, magnesium, phosphorus, copper, selenium and the like), 8 trace elements, more than 20 amino acids including 9 amino acids essential to human body (the total content of the amino acids is the first content in 16 tropical fruits, and the super combination containing the most amino acids in plants) and a plurality of substances with antioxidant effect. Also contains medicinal components such as belladonna, various compounds, various green onion wakening compounds, various alkaloids, sitosterol, various flavone glycosides, alizarin, rutin and the like, and has various effects of resisting aging, bacteria, viruses, tumors and parasites, relieving pain, reducing blood pressure, diminishing inflammation, improving immunity and the like. In 6 months and 1 day 2010, noni pulp is approved as new resource food by No. 9 notice of Ministry of health of China, the safety of noni pulp is fully demonstrated, and no toxic or side effect is caused after long-term use.
Three highs are the general term for hyperlipidemia, hypertension and hyperglycemia. Hypertension refers to the condition that the blood pressure in the circulatory system is higher than normal, and generally refers to the increase of the blood pressure of systemic circulation arteries, and is a common clinical syndrome. Usually, the blood pressure is normally lower than 140/90 mmHg and higher than 140/90 mmHg. It is a "silent killer" because it occurs unknowingly. Hyperlipidemia refers to the condition of high blood cholesterol or triglyceride or low high-density lipoprotein cholesterol, and is called as dyslipidemia in modern medicine. It is a main factor causing atherosclerosis and is a risk factor for occurrence and development of cardiovascular and cerebrovascular diseases. It is known as "invisible killer" because it has a hidden onset of disease and mostly has no clinical symptoms. Hyperglycemia is a condition in which the glucose level in the blood of the body is higher than normal. Is an independent pathological change in a body, the pathological change part is in blood, and the pathological change is blood sugar metabolic disorder. The clinical manifestations of hyperglycemia can have dominant symptoms, such as thirst, much drinking water, much urine and emaciation; may be an implicit symptom without significant subjective discomfort.
With the development of society and the increase of working and living pressure of people, the number of people with three highs is continuously increased, and noni has proved to have good effects of reducing three highs, but effective components for preventing and treating three highs are not well separated and prepared in the prior art.
Disclosure of Invention
The invention aims to provide a preparation method and application of a pyrrole derivative compound in noni ferment.
The technical purpose of the invention is realized by the following technical scheme:
a preparation method of a pyrrole derivative compound in noni ferment comprises the following steps:
(1) cleaning and freezing treatment:
cleaning fresh noni fruits, draining surface water, putting the noni fruits into a freezing box, freezing for 2-2.5 hours, and taking out for later use;
(2) pressure transformation treatment:
putting the noni fruits processed in the step (1) into a pressure swing tank, then performing pressure swing processing on the pressure swing tank, and taking out the noni fruits for later use after the pressure swing processing is completed;
(3) juicing and fermenting treatment:
juicing the noni fruits treated in the step (2), blending the obtained slurry and hydrogen peroxide, putting the mixture into a fermentation tank for sealed fermentation treatment, and taking out the mixture to obtain fermentation liquor for later use;
(4) and (3) extraction treatment:
extracting the fermentation liquor obtained in the step (3), and simultaneously carrying out ultrasonic treatment to obtain an extraction liquid for later use;
(5) chromatography treatment:
and (4) concentrating the extract obtained in the step (4), and then carrying out chromatography treatment on the extract to obtain the product.
Further, the temperature in the freezer is controlled to be-18 to-22 ℃ during the freezing treatment in the step (1).
Further, the specific operation of the pressure transformation process in the step (2) is: after the noni fruits are put into a pressure-changing tank, firstly increasing the pressure in the pressure-changing tank to 2.6-2.8 MPa, maintaining the pressure for 10-12 min under the pressure, and then quickly unloading the pressure-changing tank to normal pressure within 30 s; the temperature in the pressure swing tank is maintained at no more than 0 ℃ throughout this period.
Further, the hydrogen peroxide in the step (3) is food grade hydrogen peroxide, and the volume concentration of the hydrogen peroxide is 5-5.5%; the adding amount of the hydrogen peroxide is 15-20% of the total mass of the slurry.
Further, in the fermentation treatment in the step (3), composite zymocyte accounting for 1.5-2.0% of the total mass of the serous fluid is added into the fermentation tank, then the temperature in the fermentation tank is controlled to be 33-36 ℃ all the time, and the fermentation treatment is carried out in a sealing manner for 25-35 days; the composite zymocyte is composed of lactobacillus and yeast correspondingly according to the weight ratio of 2: 1.
Further, the extraction treatment in the step (4) adopts ethyl acetate for extraction, and the volume consumption of the ethyl acetate in the extraction treatment is 5-6 times that of the fermentation liquor.
Further, the frequency of the ultrasonic wave used in the ultrasonic treatment in the step (4) is 600-700 kHz.
Further, the chromatography treatment in the step (5) adopts gel column chromatography, and the control conditions are as follows: gel is SephadexLH-20, and a chloroform-methanol mixed solvent with the chloroform volume percentage of 15-30% is used as an eluent.
An application of pyrrole derivative compounds in noni ferment in preparing medicines for reducing high blood pressure, high blood lipid and high blood sugar is disclosed.
Compared with the prior art, the invention has the following advantages:
the invention provides a method for preparing and extracting pyrrole derivative compounds from noni enzyme, wherein noni is cleaned and subjected to pressure swing treatment, the fiber tissue structure of noni is fully loosened, a foundation is provided for the seepage of effective components, a channel among substances is communicated, then the noni enzyme is prepared by fermentation treatment, and a proper amount of food grade hydrogen peroxide component is added during the fermentation process, so that the generation and the residue of methanol in the fermentation process can be reduced, the generation and the extraction of the pyrrole derivative compounds can be influenced by the increase of the content of the methanol, and finally, the extraction and the chromatography treatment are carried out, so that the effective extraction of the pyrrole derivative compounds is realized; the method has high extraction efficiency and large extraction amount, and is easy to popularize and apply. The pyrrole derivative compound prepared by the invention has good prevention and treatment effects on three-high symptoms, can be used for preparing medicines for treating the three-high symptoms, and has good medical value and economic benefit.
Detailed Description
The present invention is further illustrated by the following specific examples, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Example 1
A preparation method of a pyrrole derivative compound in noni ferment comprises the following steps:
(1) cleaning and freezing treatment:
cleaning fresh noni fruits, draining surface water, putting into a freezing box, freezing for 2h, and taking out for later use; the temperature in the freezing box is controlled to be-18 ℃ during the freezing treatment;
(2) pressure transformation treatment:
putting the noni fruits processed in the step (1) into a pressure swing tank, then performing pressure swing processing on the pressure swing tank, and taking out the noni fruits for later use after the pressure swing processing is completed; the specific operation of the transformation treatment is as follows: after the noni fruits are put into a pressure swing tank, the pressure in the pressure swing tank is increased to 2.6MPa, the pressure is maintained for 10min under the pressure, and then the pressure swing tank is quickly unloaded to the normal pressure within 30 s; the temperature in the pressure swing tank is always kept not higher than 0 ℃ during the period;
(3) juicing and fermenting treatment:
juicing the noni fruits treated in the step (2), blending the obtained slurry and hydrogen peroxide, putting the mixture into a fermentation tank for sealed fermentation treatment, and taking out the mixture to obtain fermentation liquor for later use; the hydrogen peroxide is food grade hydrogen peroxide, and the volume concentration of the hydrogen peroxide is 5 percent; the adding amount of the hydrogen peroxide is 15% of the total mass of the slurry; the fermentation treatment is to add composite zymocyte with the total mass of 1.5 percent of the serous fluid into the fermentation tank, then control the temperature in the fermentation tank to be 33 ℃ all the time, and carry out sealed fermentation treatment for 25 days; the composite zymocyte is composed of lactobacillus and microzyme according to the weight ratio of 2: 1;
(4) and (3) extraction treatment:
extracting the fermentation liquor obtained in the step (3), and simultaneously carrying out ultrasonic treatment to obtain an extraction liquid for later use; the extraction treatment adopts ethyl acetate for extraction, and the volume consumption of the ethyl acetate is 5 times of that of the fermentation liquor during the extraction treatment; the frequency of the ultrasonic wave used in the ultrasonic treatment is 600 kHz;
(5) chromatography treatment:
concentrating the extract obtained in the step (4), and then carrying out chromatography treatment on the extract to obtain the extract; the chromatography treatment adopts gel column chromatography, and the control conditions are as follows: gel is SephadexLH-20, and a chloroform-methanol mixed solvent with the chloroform volume percent of 15 percent is used as an eluent.
An application of pyrrole derivative compounds in noni ferment in preparing medicines for reducing blood lipid is disclosed.
Example 2
A preparation method of a pyrrole derivative compound in noni ferment comprises the following steps:
(1) cleaning and freezing treatment:
cleaning fresh noni fruits, draining surface water, putting into a freezing box, freezing for 2.3h, and taking out for later use; the temperature in the freezing box is controlled to be-20 ℃ during the freezing treatment;
(2) pressure transformation treatment:
putting the noni fruits processed in the step (1) into a pressure swing tank, then performing pressure swing processing on the pressure swing tank, and taking out the noni fruits for later use after the pressure swing processing is completed; the specific operation of the transformation treatment is as follows: after the noni fruits are put into a pressure swing tank, the pressure in the pressure swing tank is increased to 2.7MPa, the pressure is maintained for 11min under the pressure, and then the pressure swing tank is quickly unloaded to the normal pressure within 30 s; the temperature in the pressure swing tank is always kept not higher than 0 ℃ during the period;
(3) juicing and fermenting treatment:
juicing the noni fruits treated in the step (2), blending the obtained slurry and hydrogen peroxide, putting the mixture into a fermentation tank for sealed fermentation treatment, and taking out the mixture to obtain fermentation liquor for later use; the hydrogen peroxide is food grade hydrogen peroxide, and the volume concentration of the hydrogen peroxide is 5.3 percent; the adding amount of the hydrogen peroxide is 18 percent of the total mass of the slurry; the fermentation treatment is to add composite zymocyte with the total mass of 1.7 percent of the serous fluid into the fermentation tank, then control the temperature in the fermentation tank to be 35 ℃ all the time, and carry out sealed fermentation treatment for 30 days; the composite zymocyte is composed of lactobacillus and microzyme according to the weight ratio of 2: 1;
(4) and (3) extraction treatment:
extracting the fermentation liquor obtained in the step (3), and simultaneously carrying out ultrasonic treatment to obtain an extraction liquid for later use; the extraction treatment adopts ethyl acetate for extraction, and the volume consumption of the ethyl acetate is 5.6 times of that of the fermentation liquor during the extraction treatment; the frequency of the ultrasonic wave used in the ultrasonic treatment is 680 kHz;
(5) chromatography treatment:
concentrating the extract obtained in the step (4), and then carrying out chromatography treatment on the extract to obtain the extract; the chromatography treatment adopts gel column chromatography, and the control conditions are as follows: gel is SephadexLH-20, and a chloroform-methanol mixed solvent with the chloroform volume percentage of 20 percent is used as an eluent.
An application of pyrrole derivative compounds in noni ferment in preparing antihypertensive drugs.
Example 3
A preparation method of a pyrrole derivative compound in noni ferment comprises the following steps:
(1) cleaning and freezing treatment:
cleaning fresh noni fruits, draining surface water, putting into a freezing box, freezing for 2.5h, and taking out for later use; the temperature in the freezing box is controlled to be-22 ℃ during the freezing treatment;
(2) pressure transformation treatment:
putting the noni fruits processed in the step (1) into a pressure swing tank, then performing pressure swing processing on the pressure swing tank, and taking out the noni fruits for later use after the pressure swing processing is completed; the specific operation of the transformation treatment is as follows: after the noni fruits are put into a pressure swing tank, the pressure in the pressure swing tank is increased to 2.8MPa, the pressure is maintained for 12min under the pressure, and then the pressure swing tank is quickly unloaded to the normal pressure within 30 s; the temperature in the pressure swing tank is always kept not higher than 0 ℃ during the period;
(3) juicing and fermenting treatment:
juicing the noni fruits treated in the step (2), blending the obtained slurry and hydrogen peroxide, putting the mixture into a fermentation tank for sealed fermentation treatment, and taking out the mixture to obtain fermentation liquor for later use; the hydrogen peroxide is food grade hydrogen peroxide, and the volume concentration of the hydrogen peroxide is 5.5 percent; the adding amount of the hydrogen peroxide is 20 percent of the total mass of the slurry; the fermentation treatment is to add composite zymocyte with the total mass of 2.0 percent of the serous fluid into the fermentation tank, then control the temperature in the fermentation tank to be 36 ℃ all the time, and carry out sealed fermentation treatment for 35 days; the composite zymocyte is composed of lactobacillus and microzyme according to the weight ratio of 2: 1;
(4) and (3) extraction treatment:
extracting the fermentation liquor obtained in the step (3), and simultaneously carrying out ultrasonic treatment to obtain an extraction liquid for later use; the extraction treatment adopts ethyl acetate for extraction, and the volume consumption of the ethyl acetate is 6 times of that of the fermentation liquor during the extraction treatment; the frequency of the ultrasonic wave used in the ultrasonic treatment is 700 kHz;
(5) chromatography treatment:
concentrating the extract obtained in the step (4), and then carrying out chromatography treatment on the extract to obtain the extract; the chromatography treatment adopts gel column chromatography, and the control conditions are as follows: gel is SephadexLH-20, and a chloroform-methanol mixed solvent with the chloroform volume percentage of 30 percent is used as an eluent.
An application of pyrrole derivative compounds in noni ferment in preparing hypoglycemic drugs.
Test example 1
To compare the effects of the present invention, the compound of example 1 was subjected to a blood lipid lowering test, specifically: healthy male SD rats were randomly divided into a normal control group, a model control group, a positive control group, and example 1 group, with 15 rats per group. The normal control group was fed with normal diet, and the other groups were fed with high-fat diet. Simvastatin of 20mg/kg and pyrrole derivative of 10mg/kg were administered to the positive control group and the group of example 1 daily for 8 weeks. After 8 weeks, blood is taken from tail vein to determine the blood lipid level of the serum of each group of rats, and the detection result is shown in the following table 1:
TABLE 1
| TC | TG | LDL | |
| Normal control group | 1.85±0.42 | 0.55±0.16 | 0.88±0.38 |
| Model control group | 11.85±0.46 | 1.40±0.20 | 4.52±0.45 |
| Positive control group | 1.98±0.43 | 0.92±0.17 | 0.85±0.34 |
| EXAMPLE 1 group | 2.05±0.44 | 0.68±0.16 | 0.96±0.40 |
As can be seen from the above table 1, the pyrrole derivative compound prepared by the method of the invention has a good cure effect on hyperglycemia.
Test example two
To compare the effects of the present invention, the compound of example 2 above was subjected to a hypotensive test, specifically: SD male rats 40 were normally kept for 1 week before the experiment, randomly divided into a blank group, a model group, a positive control group, and an example 2 group, and blood pressure of each rat was measured. Except for the blank group, the other rats used the surgical method to construct the renal hypertension model rats. After operation, 4 groups of rats were gavaged with physiological saline 0.2ml/100g, captopril 2.5mg/kg, pyrrole derivative compound 1mg/kg, 1 time/d, and administered continuously for 4 weeks. The blood pressure of each group of rats is measured before modeling, after modeling and after treatment respectively, and the detection results are shown in the following table 2:
TABLE 2
| Before molding | After molding | After treatment | |
| Blank group | 108.56±13.12 | 109.01±13.52 | 109.72±13.66 |
| Model set | 106.82±12.88 | 147.84±15.26 | 149.27±14.29 |
| Positive control group | 107.58±13.42 | 146.02±14.18 | 115.33±12.47 |
| EXAMPLE 2 group | 108.81±13.55 | 145.98±13.44 | 110.27±12.80 |
As can be seen from the above table 2, the pyrrole derivative compound prepared by the method of the invention has a good curative effect on hypertension.
Test example three
To compare the effects of the present invention, the compound of example 3 was subjected to a hypoglycemic test, specifically: 30 Kunming mice are randomly extracted as a blank group, the abdominal cavities of the other mice are injected with Streptozotocin (STZ), the equivalent buffer solution is injected into the abdominal cavities of the mice in the blank group, and a diabetic mouse model is established by taking the fasting blood glucose (FPC) of the mice as an index. The mice in the model are randomly divided into a model control group and an example 3 group, and each group contains 10 mice. Example 3 mice in group were gavaged daily with 8mg/kg of the pyrrole derivative compound, and the blank group and the model control group were given equal volumes of physiological saline, respectively, for 4 weeks continuously, and finally 1 fasting blood glucose was measured. The results of the measurements are shown in Table 3 below:
TABLE 3
As can be seen from the above table 3, the pyrrole derivative compound prepared by the method of the invention has a good cure effect on hyperglycemia.
Claims (9)
1. The preparation method of the pyrrole derivative compound in noni ferment is characterized by comprising the following steps:
(1) cleaning and freezing treatment:
cleaning fresh noni fruits, draining surface water, putting the noni fruits into a freezing box, freezing for 2-2.5 hours, and taking out for later use;
(2) pressure transformation treatment:
putting the noni fruits processed in the step (1) into a pressure swing tank, then performing pressure swing processing on the pressure swing tank, and taking out the noni fruits for later use after the pressure swing processing is completed;
(3) juicing and fermenting treatment:
juicing the noni fruits treated in the step (2), blending the obtained slurry and hydrogen peroxide, putting the mixture into a fermentation tank for sealed fermentation treatment, and taking out the mixture to obtain fermentation liquor for later use;
(4) and (3) extraction treatment:
extracting the fermentation liquor obtained in the step (3), and simultaneously carrying out ultrasonic treatment to obtain an extraction liquid for later use;
(5) chromatography treatment:
and (4) concentrating the extract obtained in the step (4), and then carrying out chromatography treatment on the extract to obtain the product.
2. The method for preparing a pyrrole derivative compound in noni ferment according to claim 1, wherein the temperature in the freezing chamber is controlled to be-18 to-22 ℃ during the freezing process in step (1).
3. The method for preparing pyrrole derivative compounds in noni ferment according to claim 1, wherein the specific operation of the pressure swing treatment in step (2) is: after the noni fruits are put into a pressure-changing tank, firstly increasing the pressure in the pressure-changing tank to 2.6-2.8 MPa, maintaining the pressure for 10-12 min under the pressure, and then quickly unloading the pressure-changing tank to normal pressure within 30 s; the temperature in the pressure swing tank is maintained at no more than 0 ℃ throughout this period.
4. The method according to claim 1, wherein the hydrogen peroxide in step (3) is food grade hydrogen peroxide, and the volume concentration of the hydrogen peroxide is 5-5.5%; the adding amount of the hydrogen peroxide is 15-20% of the total mass of the slurry.
5. The method for preparing a pyrrole derivative compound in noni ferment according to claim 1, wherein the fermentation treatment in step (3) is performed by adding composite fermentation tubes in an amount of 1.5-2.0% by mass of the total slurry into a fermentation tank, controlling the temperature in the fermentation tank to be 33-36 ℃ all the time, and performing sealed fermentation for 25-35 days; the composite zymocyte is composed of lactobacillus and yeast correspondingly according to the weight ratio of 2: 1.
6. The method for preparing pyrrole derivative compounds in noni ferment according to claim 1, wherein the extraction treatment in step (4) is performed with ethyl acetate, and the volume usage of ethyl acetate in the extraction treatment is 5-6 times of that of the fermentation broth.
7. The method for preparing pyrrole derivative compounds in noni ferment according to claim 1, wherein the ultrasonic frequency used in the ultrasonic treatment in the step (4) is 600-700 kHz.
8. The method for preparing pyrrole derivative compounds in noni ferment according to claim 1, wherein the chromatography in step (5) is performed by gel column chromatography under the following specific control conditions: gel is SephadexLH-20, and a chloroform-methanol mixed solvent with the chloroform volume percentage of 15-30% is used as an eluent.
9. An application of pyrrole derivative compounds in noni ferment in preparing medicines for reducing high blood pressure, high blood lipid and high blood sugar is disclosed.
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Application publication date: 20200911 |