CN114868919A - Composition for clearing liver and nourishing lung and processing method thereof - Google Patents
Composition for clearing liver and nourishing lung and processing method thereof Download PDFInfo
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Abstract
The invention discloses a composition for clearing liver and nourishing lung and a processing method thereof. The composition mainly comprises silybum marianum seed oil, naringin extract, loquat leaf extract, loquat powder, tangerine powder, phospholipid, mono-diglycerol fatty acid ester and gelatin skin. The processing method of the composition mainly utilizes the immobilized alpha-glucosyltransferase with higher enzyme activity, larger specific surface area and nano-scale structure to ensure that the alpha-glycosylated naringin extract has solubilization, improves the solubility and bioavailability of the composition for clearing liver and nourishing lung, promotes the absorption of the composition in vivo and has excellent effects of clearing liver and nourishing lung.
Description
Technical Field
The invention relates to the technical field of health food processing, in particular to a composition for clearing liver and nourishing lung and a processing method thereof.
Background
In the current generation of the 21 st century, haze, water pollution and automobile exhaust are ubiquitous, insomnia, fatigue stress, frequent social benefits, alcoholism and smoking gradually become the daily lives of young people, and the young people also drink drinks and overeat too much, so that the modern daily life is taken as the most important 'transfer station' of a human body, the most main detoxifying organs of the human body, and the liver is increasingly burdened. The lung is also the most delicate organ of the human body, is easily damaged by internal and external factors, haze and smoking, and oil smoke and excessive sadness generated by cooking can directly influence the lung health, and is the defense line of the human body which is most vulnerable to being out of guard. Therefore, due to factors such as unhealthy diet, irregular life, overlarge pressure, overstrain work, environmental pollution and the like, more and more subhealthy people with weak liver and lung are caused, and related diseases are easily caused.
CN103392886B discloses a health tea for clearing liver and nourishing lung, which comprises the following main components: the traditional Chinese medicine composition comprises the following components in parts by weight: 20-30 parts of leatherleaf mahonia, 2-6 parts of uncaria, 1-3 parts of cicada slough, 2-6 parts of lumbricus, 3-9 parts of houttuynia cordata, 3-9 parts of honeysuckle stem, 3-9 parts of loquat leaf and 2-6 parts of chrysanthemum. The preparation method comprises the steps of weighing the raw materials according to the weight ratio, placing the raw materials into a crusher, stirring and crushing the raw materials, sieving the raw materials by a 6-mesh sieve, and packaging the raw materials by aluminum foil paper or preparing the raw materials into tea, wherein each bag contains 20g of the raw materials. The invention has the following remarkable effects: the traditional Chinese medicine composition has the effects of tonifying liver and kidney, nourishing qi and blood, clearing heat, expelling toxin, expelling wind-damp and removing rubbish in lung, achieves the functions of clearing liver and nourishing lung and recovering respiratory system, and has the advantages of simple treatment, low treatment cost, total effective rate of more than 90% and the like.
CN105477333A discloses a tea for nourishing lung and clearing liver for treating keratitis and a preparation method thereof, the tea for nourishing lung and clearing liver for treating keratitis and the preparation method thereof comprise honeysuckle, cassia seed, radix rehmanniae, adenophora stricta, bletilla striata, white paeony root, gentiana scabra bunge and scutellaria baicalensis tea.
In order to clear liver and nourish lung and keep body healthy, many people start a large amount of traditional Chinese medicine food, however, toxins produced by some Chinese herbal medicines need secondary metabolism of liver, and the burden of liver is increased. Therefore, there is a need to develop a composition for clearing liver and nourishing lung.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention provides a composition for clearing liver and nourishing lung, mainly comprising silybum marianum seed oil, naringin extract, loquat leaf extract, loquat powder, tangerine peel powder, phospholipid, mono-diglycerol fatty acid ester and gelatin skin.
Silybum marianum, alternative name: milk thistle, rat tendon, silymarin, compositae, silybum one year or biennial compositae herbs. Silybum marianum is a herbal medicine, which has been used for thousands of years to protect the liver, and the Silybum marianum seed oil is squeezed from Silybum marianum seeds, which are used as both medicine and food. The oil content in the seeds of the silybum marianum is about 28 percent, wherein the content of unsaturated fatty acids such as oleic acid, linoleic acid and the like is as high as 80 percent, the silybum marianum oil has high nutritional and medicinal values, is also rich in various vitamins and mineral substances, particularly contains effective components such as silymarin which is a flavonoid substance of the silybum marianum and the like, and is special healthy edible oil with health care effect.
Naringin extract, also called naringin and naringin, is a natural dihydroflavonoids compound, which is a light yellow dihydroflavonoids compound extracted from immature or near mature dry pericarp of Citrus paradisi and grapefruit of Rutaceae. Because the A ring and the B ring are not conjugated at all, the naringin has strong ultraviolet absorption peak at 282nm, so that the naringin shows various biological activities and pharmacological actions. Has antiinflammatory, antiviral, anticancer, antimutagenic, antiallergic, antiulcer, analgesic, blood pressure lowering, blood cholesterol reducing, thrombosis reducing, local microcirculation improving and nutrition supply improving effects, and can be used for preventing and treating cardiovascular disease and cerebrovascular disease. Naringin has abundant sources in nature and extremely wide pharmacological actions, and is widely applied to the aspects of medicine, food science, chemistry and the like.
The folium Eriobotryae extract is leaf of Ligustrum lucidum ait of Oleaceae or leaf of Eriobotrya japonica of Rosaceae, and contains effective components such as oleanolic acid, saponin, caffeic acid, amygdalin and ursolic acid. In loquat leavesOleanolic acid has effects of protecting liver, removing toxic substance, and treating CCl 4 The damage of acute and chronic hepatitis caused by the hepatitis virus has obvious protective effect, obviously reduces the activity of glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase, and can prevent experimental cirrhosis. The saponin has effects of inducing bronchial mucosa secretion, and eliminating phlegm by oral administration. Caffeic acid has effects of relieving cough and eliminating phlegm. Amygdalin is decomposed into hydrocyanic acid and benzaldehyde by gastric acid or amygdalin, and after absorption, cytochrome oxidase is inhibited, the oxidation metabolism of common carotid artery body is inhibited to reflectively deepen breath, so that the respiratory center is calmed, and the respiratory movement tends to be quiet, thereby achieving the effect of relieving cough and asthma.
The loquat powder is a special food obtained by drying fresh loquat fruits and grinding the fresh loquat fruits into powder, retains most of the nutrition in the loquat fruits, has a much longer shelf life than the loquat fruits, and is easy to carry and eat. The loquat powder contains various mineral substances and vitamins, can eliminate inflammation of trachea and lung, and has the important effects of moistening lung and relieving cough.
The exocarpium Citri Tangerinae powder is prepared from dried outer pericarp of Rutaceae plant, fructus Citri Tangerinae and its cultivar, by collecting in autumn and winter after fruit ripening, cutting off exocarpium, sun drying or drying in shade, and making into powder for use. The main components of the tangerine peel powder comprise citric acid, geraniol, aromatic alcohol, naringin, neohesperidin, and citrin, so that the tangerine peel powder has the effects of dispelling cold, eliminating dampness, promoting qi circulation, eliminating phlegm, and the like.
Caramel color, also known as caramel color, is a complex reddish-brown or blackish-brown mixture of carbohydrates (e.g., maltose, sucrose, molasses, invert sugar, lactose, maltose syrup, and hydrolysates of starch, etc.) dehydrated, decomposed, and polymerized at high temperatures, some of which are colloidal aggregates, which are more widely used semi-natural food colorants.
The composition mainly comprises 470-475 parts by weight of silybum marianum seed oil, 28-32 parts by weight of naringin extract, 115-125 parts by weight of loquat leaf extract, 4.5-5.5 parts by weight of loquat powder, 28-32 parts by weight of tangerine peel powder, 6.5-7.5 parts by weight of phospholipid, 33-38 parts by weight of mono-diglycerol fatty acid ester and 900-1000 parts by weight of gelatin skin.
Preferably, the main components of the rubber comprise gelatin, glycerin, water, trehalose, erythritol, mogroside, titanium dioxide and caramel color.
Further preferably, the main components of the rubber comprise 290-310 parts by weight of gelatin, 110-130 parts by weight of glycerol, 290-310 parts by weight of water, 18-22 parts by weight of trehalose, 4.5-5.5 parts by weight of erythritol, 0.001-0.002 part by weight of mogroside, 2.7-3.3 parts by weight of titanium dioxide and 8-10 parts by weight of caramel color.
Because natural biological macromolecules such as the naringin extract, the silybum marianum seed oil, the loquat leaf extract, the loquat powder, the tangerine peel powder and the like have poor solubility in gastrointestinal tract liquid, the absorption is poor when the oral administration is carried out, and the bioavailability is low, so that the improvement of the dissolution rate of the natural biological macromolecules has important significance for improving the oral administration absorption. CN110205351A discloses a preparation method of glycosylated naringenin, which is characterized by comprising the following specific steps: the method comprises the steps of putting raw materials and glycosyl donors into a reaction buffer solution, uniformly mixing the feed liquid, and then adding glycosidase and glycosyl transferase into a reaction system, wherein the raw materials are naringin, naringenin extracts or converted substances and derivatives thereof. The glycosylated naringenin prepared by the method has the advantages that 1-9 glycosyl groups are added on the flavone parent body, so that the solubility of the naringenin is greatly improved, the problem of poor solubility of the naringenin is solved, the problem of decomposition and absorption of the naringenin in a human body is solved, and the bioavailability of the naringenin is greatly improved. In addition, the inventor finds that when the alpha-glycosylated naringin extract is synthesized by adding one glucose unit in the naringin extract, the solubility and the bioavailability of the alpha-glycosylated naringin extract are improved, and the alpha-glycosylated naringin extract is also beneficial to the improvement of the solubility and the bioavailability of other natural biological macromolecules such as silybum marianum seed oil, loquat leaf extract, loquat powder, tangerine powder and the like.
Preferably, the naringin extract is an alpha-glycosylated naringin extract, and the preparation method comprises the following steps: mixing naringin extract and glycosyl donor in buffer solution, adding glycosyltransferase for reaction, heating to inactivate enzyme, and cooling to obtain alpha-glycosylated naringin extract.
Specifically, the naringin extract is an alpha-glycosylated naringin extract, and the preparation method comprises the following steps: adding 1-2 parts by weight of naringin extract and 1-3 parts by weight of glycosyl donor into 20-200 parts by weight of buffer solution, mixing, stirring at 200-300 rpm for 5-20 min, adding 0.3-1.5 parts by weight of glycosyl transferase at 35-50 ℃ for reaction for 12-24 h, heating to 70-90 ℃, and cooling to 20-30 ℃ to obtain the alpha-glycosylated naringin extract.
More preferably, the glycosyl donor is one of glucose, xylan, sucrose, uridine diphosphate glucose, xylose galactose, starch, cyclodextrin or maltodextrin.
Further preferably, the buffer solution is 0.05-1 wt% of phosphate buffer solution, and the pH value is 4.5-6.
Further preferably, the glycosyltransferase is one, two or more of rhamnosyl transferase, cyclomaltodextrin glucanotransferase, rhizopus glucoamylase, alpha-glucosyl transferase and xylosyltransferase.
Glycosyltransferase catalysis is widely applied to the fields of food, biology, chemistry and the like due to good selectivity and environmental friendliness, and becomes an effective tool for green and sustainable catalysis methods. However, glycosyltransferases have poor stability of operation, difficulty in isolation and low reproducibility, and thus have been hindered from being used in industry. These problems can be solved by immobilizing glycosyltransferase, which improves biocatalytic ability by immobilizing glycosyltransferase on a solid support, minimizes product contamination, gives the possibility of reuse of biocatalysts, improves the cost efficiency of catalytic conversion process and improves the safety of material handling. Both graphene and graphene oxide can bind different amphiphiles, such as peptides, polysaccharides and fatty acids, exhibiting different binding affinities depending on the type of graphene derivative and biomolecule. The inventor finds that the glycosyl transferase immobilized by using graphene derivatives such as single-layer graphene oxide, graphene oxide nano colloid and polycarboxylated graphene nano sheet can influence the biochemical characteristics of the enzyme, increase the performance of the enzyme in a non-aqueous solvent and enhance the stability of the enzyme.
Further preferably, the naringin extract is an alpha-glycosylated naringin extract, and the preparation method comprises the following steps: mixing naringin extract and glycosyl donor in buffer solution, adding glycosyltransferase for reaction, centrifuging, and collecting supernatant to obtain alpha-glycosylated naringin extract.
Specifically, the preparation method of the alpha-glycosylated naringin extract comprises the following steps: adding 1-2 parts by weight of naringin extract and 1-3 parts by weight of glycosyl donor into 20-200 parts by weight of buffer solution, mixing, stirring at 200-300 rpm for 5-20 min, adding 0.3-1.5 parts by weight of glycosyl transferase at 35-50 ℃ for reaction for 12-24 h, centrifuging at 1000-3000 rpm for 10-20 min, and taking supernatant to obtain the alpha-glycosylated naringin extract.
Preferably, the glycosyltransferase is an immobilized alpha-glucosyltransferase.
Further, the preparation method of the immobilized alpha-glucosyltransferase comprises the following steps:
adding X1 and graphene derivatives into a phosphate buffer solution for ultrasonic treatment to obtain a suspension;
x2, dissolving alpha-glucosyltransferase in phosphate buffer solution to obtain enzyme solution;
and X3, mixing the suspension obtained in the step X1 with the enzyme solution obtained in the step X2, and reacting, centrifuging and washing to obtain the immobilized alpha-glucosyltransferase.
Specifically, the preparation method of the immobilized alpha-glucosyltransferase comprises the following steps:
adding X1 and 0.6-0.7 part by weight of graphene derivative into 90-110 parts by weight of phosphate buffer solution for ultrasonic treatment to obtain a suspension;
x2, dissolving 0.1-0.6 weight part of alpha-glucosyltransferase in 90-110 weight parts of phosphate buffer solution to obtain enzyme solution;
and X3, mixing the suspension obtained in the step X1 with the enzyme solution obtained in the step X2, reacting at the temperature of 0-4 ℃ for 12-16 hours at the shaking speed of 200-300 rpm, taking the reaction supernatant, centrifuging at the rotating speed of 2000-3000 rpm for 10-20 min, washing the centrifugal precipitate with phosphate buffer solution for 3-5 times, and obtaining the immobilized alpha-glucosyltransferase.
Preferably, the phosphate buffer solutions in the steps X1, X2 and X3 are sodium phosphate buffer solutions with pH of 5.4-6.8 and concentration of 20-25 mmol/L.
Preferably, the ultrasonic treatment condition in the step X1 is ultrasonic treatment for 30-100 min at a frequency of 20-30 kHz and a power of 100-300W.
Preferably, the graphene derivative in step X1 is one of a single-layer graphene oxide, a graphene oxide nano colloid, and a polycarboxylated graphene nanoplatelet.
The invention also provides a processing method of the composition for clearing liver and nourishing lung, which comprises the following steps:
(1) accurately weighing each raw and auxiliary material according to the formula amount;
(2) heating trehalose, erythritol, mogroside, residual water and glycerol, adding gelatin, stirring, adding titanium dioxide and caramel color suspension, stirring, vacuumizing, sieving, and keeping the temperature to obtain a rubber sheet;
(3) heating and dissolving diglycerol fatty acid ester and part of silybum marianum seed oil, cooling, adding naringin extract, loquat leaf extract, loquat powder, tangerine peel powder, phospholipid and the rest of silybum marianum seed oil, stirring, and passing through a colloid mill and a sieve to obtain a content material;
(4) pressing the rubber sheet obtained in the step (2) and the content material obtained in the step (3) into soft capsules;
(5) shaping and drying the soft capsules obtained in the step (4) to obtain capsules;
(6) and (4) sorting out unqualified capsules from the capsules obtained in the step (5) to obtain the composition for clearing liver and nourishing lung.
Specifically, the processing method of the composition for clearing liver and nourishing lung comprises the following steps:
(1) accurately weighing each raw and auxiliary material according to the formula amount;
(2) mixing trehalose, erythritol, mogroside, 1/3-1/2 water and glycerol, heating to 60-80 ℃, adding gelatin, stirring to be completely dissolved, adding titanium dioxide and caramel color suspension, stirring for 10-20 min, vacuumizing to-0.06-0.10 MPa, sieving by a 100-200-mesh sieve, and keeping the temperature at 58-62 ℃ to obtain a rubber;
(3) heating and dissolving mono-diglycerol fatty acid ester and 1/3-1/2 silybum marianum seed oil to 50-60 ℃, cooling to 35-40 ℃, adding naringin extract, loquat leaf extract, loquat powder, tangerine powder, phospholipid and 1/2-2/3 silybum marianum seed oil, mixing and stirring for 15-30 min, passing through a colloid mill for 1-2 times, and passing through a 80-100-mesh sieve to obtain a content material;
(4) pressing the rubber sheet obtained in the step (2) and the content material obtained in the step (3) into soft capsules at the temperature of 18-26 ℃ and the relative humidity of less than or equal to 50%;
(5) shaping the soft capsule obtained in the step (4) for 1-4 hours at the temperature of 18-26 ℃ and the relative humidity of less than or equal to 50%, and drying for 16-24 hours at the temperature of 15-28 ℃ and the relative humidity of less than or equal to 50% to obtain a capsule;
(6) and (4) sorting out unqualified capsules with oil leakage, flat pills, deformity, bubbles, large and small heads, black spots, impurities in contents and the like from the capsules obtained in the step (5) to obtain the composition for clearing liver and nourishing lung.
Preferably, the preparation method of the titanium dioxide and caramel color suspension in the step (2) comprises the steps of mixing the titanium dioxide and the caramel color with water of 1/2-2/3, and grinding the mixture by a colloid mill to obtain the titanium dioxide and caramel color suspension.
Preferably, the capsule in the step (5) is in a 20# cylindrical shape, and the content is 0.7 g/capsule with a +/-7.5% difference.
The invention has the following beneficial effects:
the invention provides a composition for clearing liver and nourishing lung and a processing method thereof. The composition with the effects of clearing liver and nourishing lung is prepared from effective active substances such as silybum marianum seed oil, naringin extract, loquat leaf extract, loquat powder, tangerine peel powder and the like. According to the invention, the graphene oxide, graphene oxide nano colloid, polycarboxylated graphene nano sheet and other nanoscale graphene derivatives with strong adsorption capacity and large specific surface area are combined with alpha-glucosyltransferase, so that the graphene derivative immobilized alpha-glucosyltransferase has higher enzyme activity, larger specific surface area and a nanoscale structure, the contact surface of the alpha-glucosyltransferase and a substrate is increased, the formation of nanoscale alpha-glycosylated naringin extract molecular aggregates is facilitated, the alpha-glycosylated naringin extract has a solubilization function, the solubility and bioavailability of the composition for clearing liver and nourishing lung are improved, the absorption of the composition in vivo is promoted, and the composition has excellent effects of clearing liver and nourishing lung.
Detailed Description
Some raw material introductions in this application:
the Graphene oxide monolayer is Graphene oxide with the purity of 98%, the diameter of a lamella is 0.5-5 mu m, and the Graphene oxide monolayer is obtained by Shanghai-sourced leaf Biotech Co.
Graphene oxide nanocolloid, Graphene oxide anocolloids, product number: 795534 SIGMA corporation.
Polycarboxylated Graphene nanoplatelets, Graphene nanoplatelets polycarboxylated functionalized, product number: 806625 SIGMA corporation.
Example 1
A composition for clearing liver and nourishing lung, its preparation method comprises the following steps:
(1) weighing each raw and auxiliary material;
(2) mixing 20g of trehalose, 5g of erythritol, 0.001g of mogroside, 150g of water and 120g of glycerol, heating to 75 ℃, adding 300g of gelatin, stirring to be completely dissolved, adding a suspension of titanium dioxide and caramel color, stirring for 15min, vacuumizing to-0.06 MPa, sieving by a 100-mesh sieve, and keeping the temperature at 60 ℃ to obtain a rubber;
(3) heating 35g of mono-diglycerol fatty acid ester and 200g of silybum marianum seed oil to 55 ℃ for dissolving, cooling to 35 ℃, adding 30g of naringin extract, 120g of loquat leaf extract, 5g of loquat powder, 30g of tangerine peel powder, 7g of phospholipid and 273g of silybum marianum seed oil, mixing and stirring for 20min, sieving by a colloid mill for 2 times, and sieving by a 80-mesh sieve to obtain a content material;
(4) pressing the rubber sheet obtained in the step (2) and the content material obtained in the step (3) into 1000 soft capsules under the conditions that the temperature is 20 ℃ and the relative humidity is 50%;
(5) shaping the soft capsule obtained in the step (4) at the temperature of 20 ℃ and the relative humidity of 50% for 3h, and drying at the temperature of 20 ℃ and the relative humidity of 50% for 24h to obtain a capsule;
(6) and (4) sorting out unqualified capsules with oil leakage, flat pills, deformity, bubbles, large and small heads, black spots, impurities in contents and the like from the capsules obtained in the step (5) to obtain the composition for clearing liver and nourishing lung.
The preparation method of the titanium dioxide and caramel color suspension in the step (2) is to mix 3g of titanium dioxide, 9g of caramel color and 150g of water, and grind the mixture by a colloid mill to obtain the titanium dioxide and caramel color suspension.
The capsule in the step (5) is in a 20# cylindrical shape, and the difference of the filling amount of the content of 0.7 g/capsule is +/-7.5%.
Example 2
A composition for clearing liver and nourishing lung, its preparation method comprises the following steps:
(1) weighing each raw and auxiliary material;
(2) mixing 20g of trehalose, 5g of erythritol, 0.001g of mogroside, 150g of water and 120g of glycerol, heating to 75 ℃, adding 300g of gelatin, stirring to be completely dissolved, adding a suspension of titanium dioxide and caramel color, stirring for 15min, vacuumizing to-0.06 MPa, sieving by a 100-mesh sieve, and keeping the temperature at 60 ℃ to obtain a rubber;
(3) heating 35g of mono-diglycerol fatty acid ester and 200g of silybum marianum seed oil to 55 ℃ for dissolution, cooling to 35 ℃, adding 30g of alpha-glycosylated naringin extract, 120g of loquat leaf extract, 5g of loquat powder, 30g of tangerine peel powder, 7g of phospholipid and 273g of silybum marianum seed oil, mixing and stirring for 20min, sieving by a colloid mill for 2 times, and sieving by a 80-mesh sieve to obtain a content material;
(4) pressing the rubber sheet obtained in the step (2) and the content material obtained in the step (3) into 1000 soft capsules under the conditions that the temperature is 20 ℃ and the relative humidity is 50%;
(5) shaping the soft capsule obtained in the step (4) at the temperature of 20 ℃ and the relative humidity of 50% for 3h, and drying at the temperature of 20 ℃ and the relative humidity of 50% for 24h to obtain a capsule;
(6) and (4) sorting out unqualified capsules with oil leakage, flat pills, deformity, bubbles, large and small heads, black spots, impurities in contents and the like from the capsules obtained in the step (5) to obtain the composition for clearing liver and nourishing lung.
And (3) mixing 3g of titanium dioxide, 9g of caramel color and 150g of water, and grinding by a colloid mill to obtain the titanium dioxide and caramel color suspension.
The preparation method of the alpha-glycosylated naringin extract in the step (3) comprises the following steps: adding 1g naringin extract and 1.5g uridine diphosphate glucose into 50 weight parts of 1 wt% phosphate buffer solution with pH value of 6, mixing, stirring at 200rpm for 15min, adding 0.5g alpha-glucosyltransferase at 45 deg.C for reaction for 16h, heating to 80 deg.C, and cooling to 20 deg.C to obtain alpha-glycosylated naringin extract.
The capsule in the step (5) is in a 20# cylindrical shape, and the difference of the filling amount of the content of 0.7 g/capsule is +/-7.5%.
Example 3
A composition for clearing liver and nourishing lung, its preparation method comprises the following steps:
(1) weighing each raw and auxiliary material;
(2) mixing 20g of trehalose, 5g of erythritol, 0.001g of mogroside, 150g of water and 120g of glycerol, heating to 75 ℃, adding 300g of gelatin, stirring to be completely dissolved, adding a suspension of titanium dioxide and caramel color, stirring for 15min, vacuumizing to-0.06 MPa, sieving by a 100-mesh sieve, and keeping the temperature at 60 ℃ to obtain a rubber;
(3) heating 35g of mono-diglycerol fatty acid ester and 200g of silybum marianum seed oil to 55 ℃ for dissolution, cooling to 35 ℃, adding 30g of alpha-glycosylated naringin extract, 120g of loquat leaf extract, 5g of loquat powder, 30g of tangerine peel powder, 7g of phospholipid and 273g of silybum marianum seed oil, mixing and stirring for 20min, sieving by a colloid mill for 2 times, and sieving by a 80-mesh sieve to obtain a content material;
(4) pressing the rubber sheet obtained in the step (2) and the content material obtained in the step (3) into 1000 soft capsules under the conditions that the temperature is 20 ℃ and the relative humidity is 50%;
(5) shaping the soft capsule obtained in the step (4) at the temperature of 20 ℃ and the relative humidity of 50% for 3h, and drying at the temperature of 20 ℃ and the relative humidity of 50% for 24h to obtain a capsule;
(6) and (4) sorting out unqualified capsules with oil leakage, flat pills, deformity, bubbles, large and small heads, black spots, impurities in contents and the like from the capsules obtained in the step (5) to obtain the composition for clearing liver and nourishing lung.
The preparation method of the titanium dioxide and caramel color suspension in the step (2) is to mix 3g of titanium dioxide, 9g of caramel color and 150g of water, and grind the mixture by a colloid mill to obtain the titanium dioxide and caramel color suspension.
The capsule in the step (5) is in a 20# cylindrical shape, and the difference of the filling amount of the content of 0.7 g/capsule is +/-7.5%.
The preparation method of the alpha-glycosylated naringin extract in the step (3) comprises the following steps: adding 1g naringin extract and 1.5g uridine diphosphate glucose into 50 weight parts of 1 wt% phosphate buffer solution with pH value of 6, mixing, stirring at 200rpm for 15min, adding 0.5g alpha-glucosyltransferase at 45 deg.C for reaction for 16h, centrifuging at 1000rpm for 10min, and collecting supernatant to obtain alpha-glycosylated naringin extract.
The preparation method of the immobilized alpha-glucosyltransferase comprises the following steps:
adding X1 and 0.6g of monolayer graphene oxide into 100mL of phosphate buffer solution, and carrying out ultrasonic treatment for 60min at the frequency of 25kHz and the power of 100W to obtain a suspension;
x2, dissolving 0.6g of alpha-glucosyltransferase in 100mL of phosphate buffer solution to obtain an enzyme solution;
and X3, mixing the suspension obtained in the step X1 with the enzyme solution obtained in the step X2, reacting at 4 ℃ for 14 hours at a shaking speed of 300rpm, centrifuging the reaction supernatant at 2000rpm for 10 minutes, and washing the centrifugal precipitate for 3 times by using a phosphate buffer solution to obtain the immobilized alpha-glucosyltransferase.
The phosphate buffer described in steps X1, X2 and X3 was a 25mmol/L sodium phosphate buffer at pH 6.5.
Example 4
A composition for clearing liver and nourishing lung, its preparation method comprises the following steps:
(1) weighing each raw and auxiliary material;
(2) mixing 20g of trehalose, 5g of erythritol, 0.001g of mogroside, 150g of water and 120g of glycerol, heating to 75 ℃, adding 300g of gelatin, stirring to be completely dissolved, adding a suspension of titanium dioxide and caramel color, stirring for 15min, vacuumizing to-0.06 MPa, sieving by a 100-mesh sieve, and keeping the temperature at 60 ℃ to obtain a rubber;
(3) heating 35g of mono-diglycerol fatty acid ester and 200g of silybum marianum seed oil to 55 ℃ for dissolution, cooling to 35 ℃, adding 30g of alpha-glycosylated naringin extract, 120g of loquat leaf extract, 5g of loquat powder, 30g of tangerine peel powder, 7g of phospholipid and 273g of silybum marianum seed oil, mixing and stirring for 20min, sieving by a colloid mill for 2 times, and sieving by a 80-mesh sieve to obtain a content material;
(4) pressing the rubber sheet obtained in the step (2) and the content material obtained in the step (3) into 1000 soft capsules under the conditions that the temperature is 20 ℃ and the relative humidity is 50%;
(5) shaping the soft capsule obtained in the step (4) at the temperature of 20 ℃ and the relative humidity of 50% for 3h, and drying at the temperature of 20 ℃ and the relative humidity of 50% for 24h to obtain a capsule;
(6) and (4) sorting out unqualified capsules with oil leakage, flat pills, deformity, bubbles, large and small heads, black spots, impurities in contents and the like from the capsules obtained in the step (5) to obtain the composition for clearing liver and nourishing lung.
The preparation method of the titanium dioxide and caramel color suspension in the step (2) is to mix 3g of titanium dioxide, 9g of caramel color and 150g of water, and grind the mixture by a colloid mill to obtain the titanium dioxide and caramel color suspension.
The capsule in the step (5) is in a 20# cylindrical shape, and the difference of the filling amount of the content of 0.7 g/capsule is +/-7.5%.
The preparation method of the alpha-glycosylated naringin extract in the step (3) comprises the following steps: adding 1g naringin extract and 1.5g uridine diphosphate glucose into 50 weight parts of 1 wt% phosphate buffer solution with pH value of 6, mixing, stirring at 200rpm for 15min, adding 0.5g alpha-glucosyltransferase at 45 deg.C for reaction for 16h, and centrifuging at 1000rpm for 10min to obtain alpha-glycosylated naringin extract.
The preparation method of the immobilized alpha-glucosyltransferase comprises the following steps:
adding X1 and 0.6g of graphene oxide nano colloid into 100mL of phosphate buffer solution, and carrying out ultrasonic treatment for 60min at the frequency of 25kHz and the power of 100W to obtain a suspension;
x2, dissolving 0.6g of alpha-glucosyltransferase in 100mL of phosphate buffer solution to obtain an enzyme solution;
and X3, mixing the suspension obtained in the step X1 with the enzyme solution obtained in the step X2, reacting at 4 ℃ for 14 hours at a shaking speed of 300rpm, centrifuging the reaction supernatant at 2000rpm for 10 minutes, and washing the centrifugal precipitate for 3 times by using a phosphate buffer solution to obtain the immobilized alpha-glucosyltransferase.
The phosphate buffer described in steps X1, X2 and X3 was a 25mmol/L sodium phosphate buffer at pH 6.5.
Example 5
A composition for clearing liver and nourishing lung, its preparation method comprises the following steps:
(1) weighing each raw and auxiliary material;
(2) mixing 20g of trehalose, 5g of erythritol, 0.001g of mogroside, 150g of water and 120g of glycerol, heating to 75 ℃, adding 300g of gelatin, stirring to be completely dissolved, adding a suspension of titanium dioxide and caramel color, stirring for 15min, vacuumizing to-0.06 MPa, sieving by a 100-mesh sieve, and keeping the temperature at 60 ℃ to obtain a rubber;
(3) heating 35g of mono-diglycerol fatty acid ester and 200g of silybum marianum seed oil to 55 ℃ for dissolution, cooling to 35 ℃, adding 30g of alpha-glycosylated naringin extract, 120g of loquat leaf extract, 5g of loquat powder, 30g of tangerine peel powder, 7g of phospholipid and 273g of silybum marianum seed oil, mixing and stirring for 20min, sieving by a colloid mill for 2 times, and sieving by a 80-mesh sieve to obtain a content material;
(4) pressing the rubber sheet obtained in the step (2) and the content material obtained in the step (3) into 1000 soft capsules under the conditions that the temperature is 20 ℃ and the relative humidity is 50%;
(5) shaping the soft capsule obtained in the step (4) at the temperature of 20 ℃ and the relative humidity of 50% for 3h, and drying at the temperature of 20 ℃ and the relative humidity of 50% for 24h to obtain a capsule;
(6) and (4) sorting out unqualified capsules with oil leakage, flat pills, deformity, bubbles, large and small heads, black spots, impurities in contents and the like from the capsules obtained in the step (5) to obtain the composition for clearing liver and nourishing lung.
The preparation method of the titanium dioxide and caramel color suspension in the step (2) is to mix 3g of titanium dioxide, 9g of caramel color and 150g of water, and grind the mixture by a colloid mill to obtain the titanium dioxide and caramel color suspension.
The capsule in the step (5) is in a 20# cylindrical shape, and the difference of the filling amount of the content of 0.7 g/capsule is +/-7.5%.
The preparation method of the pomelo alpha-glycosylated naringin extract in the step (3) comprises the following steps: adding 1g naringin extract and 1.5g uridine diphosphate glucose into 50 weight parts of 1 wt% phosphate buffer solution with pH value of 6, mixing, stirring at 200rpm for 15min, adding 0.5g alpha-glucosyltransferase at 45 ℃ for reaction for 16h, and centrifuging at 1000rpm for 10min to obtain the alpha-glycosylated naringin extract.
The preparation method of the immobilized alpha-glucosyltransferase comprises the following steps:
adding X1 and 0.6g of polycarboxylated graphene nanosheet into 100mL of phosphate buffer solution, and carrying out ultrasonic treatment for 60min at the frequency of 25kHz and the power of 100W to obtain a suspension;
x2, dissolving 0.6g of alpha-glucosyltransferase in 100mL of phosphate buffer solution to obtain an enzyme solution;
and X3, mixing the suspension obtained in the step X1 with the enzyme solution obtained in the step X2, reacting at 4 ℃ for 14 hours at a shaking speed of 300rpm, centrifuging the reaction supernatant at 2000rpm for 10 minutes, and washing the centrifugal precipitate for 3 times by using a phosphate buffer solution to obtain the immobilized alpha-glucosyltransferase.
The phosphate buffer described in steps X1, X2 and X3 was a 25mmol/L sodium phosphate buffer at pH 6.5.
Test example 1
The immobilized α -glucosyltransferases of examples 3 to 5 were subjected to enzyme activity and substrate binding property measurement: 4-nitrobenzene-A-D-glucopyranoside is used as a substrate to determine the activity of the alpha-glucosyltransferase. The substrate solution consisted of 65. mu.L of 1.6 mmol/L4-nitrophenyl-A-D-glucopyranoside and 25mmol/L phosphate buffer pH 6.5. To start the enzymatic reaction, 10. mu.L of enzyme diluent containing 7.5mg/mL bovine serum albumin was added to the substrate solution on a 96F microplate, reacted at 50 ℃ for 15 minutes, and 75. mu.L of 0.25mol/L sodium carbonate was added to stop the reaction. The absorbance was measured spectrophotometrically at 405nm and the enzyme concentration was determined by Bradford assay. The percent binding of alpha-glucosyltransferase to different graphene derivatives was calculated as follows and the test results are shown in table 1.
Wherein the activity of the free enzyme is the activity value of the free enzyme before the immobilization reaction, and the unit is U/mg; the enzyme activity of the supernatant is the activity value of free enzyme in the supernatant recovered after the enzyme immobilization reaction, and the unit is U/mg.
TABLE 1 results of the measurement of immobilized alpha-glucosyltransferase Activity and binding Properties
Table 1 shows the percent binding of α -glucosyltransferase in different graphene derivatives, the amount of enzyme immobilized per mg of graphene derivative, and the relative activity retained after immobilization. As can be seen from table 1, due to the strong adsorption capacity and the extremely large specific surface area of the single-layer graphene oxide, graphene oxide nanocolloid, and polycarboxylated graphene nanoplatelet, the single-layer graphene oxide, graphene oxide nanocolloid, and polycarboxylated graphene nanoplatelet can be combined with α -glucosyltransferase, and the enzymatic activity thereof is improved. The single-layer graphene oxide and graphene oxide nano-colloids have high loading capacity, and the polycarboxylated graphene nanosheets have poor loading performance when being used as carriers of alpha-glucosyltransferase, which is probably because the polycarboxylated graphene nanosheets are easier to stack, and the adsorption of the alpha-glucosyltransferase is inhibited.
The relative activity of alpha-glucosyltransferase remaining in the graphene derivative after the immobilization reaction was determined with the enzyme activity of free alpha-glucosyltransferase as a reference. Due to different nano structures of the graphene derivatives, different aggregation states of the enzyme and the nano particles can be caused, so that the relative activity reserved after enzyme immobilization is influenced. The graphene oxide nano colloid and the polycarboxylic acid graphene nanosheet aggregate are more dispersed and smaller in size, and particularly the polycarboxylic acid graphene nanosheet enlarges the contact surface of the adsorbed alpha-glucosyltransferase and a substrate, so that the alpha-glycosylated naringin extract with small particle size is more favorably formed. In addition, the presence of polycarboxylic acid groups in the polycarboxygraphene nanoplatelets increases the spacing between stacked planar layers and the relatively low number of oxygen-containing functional groups, and is also responsible for increasing the immobilized alpha-glucosyltransferase activity of the polycarboxygraphene nanoplatelets at low adsorption levels.
Test example 2
And (3) measuring the solubility: the liver-clearing and lung-nourishing compositions prepared in examples 1 to 5 were added to distilled water and simulated fasted intestinal juice as dissolution media, and shaken in a water bath at a speed of 100rpm at 37 ℃. After 5, 15, 30, 60, 120 and 240min, 1mL of the suspension was taken out, a boron trifluoride methanol solution was added for methyl esterification, and methyl linoleate in the sample solution was detected by gas chromatography. A chromatographic column: 30 m.times.0.32 mm.times.0.5 μm of DB-FFAP, carrier gas: nitrogen gas; flow rate of carrier gas: 4 mL/min; sample inlet temperature: 260 ℃; the split ratio is as follows: 10: 1; a detector: a hydrogen flame ionization detector; detection of mouth temperature: 280 ℃; sample introduction amount: 1.0 uL; column temperature: the initial temperature is 140 ℃ and kept for 1min, then the temperature is raised to 160 ℃ at 20 ℃, and then raised to 220 ℃ at 4 ℃/min, and kept for 2 min. The test results are shown in tables 2 and 3.
Measurement of average particle diameter: the average particle size of the naringin extract or the α -glycosylated naringin extract prepared in examples 1 to 5 was measured by a dynamic light scattering method using a Naring-Nanotrac UPA-UT151 particle size analyzer, and the results are shown in table 2.
TABLE 2 results of sample solubility in distilled water
TABLE 3 results of measurement of solubility of samples in intestinal fluid simulating fasted state
As is clear from the results of the average particle size measurement in table 1, examples 3 to 5, compared to examples 1 and 2, had the effect of reducing the particle size of the naringin extract to several nanometers by α -glycosylation of the naringin extract by immobilized α -glucosyltransferase, forming a nano-aggregate structure, and further contributing to the improvement of the solubility of the naringin extract.
As can be seen from tables 1 and 2, in examples 2 to 5, compared to example 1, the solubility of the prepared composition for clearing liver and nourishing lung was improved in both distilled water and intestinal fluid simulating a fasting state by α -glycosylation of the naringin extract, which indicates that the α -glycosylated naringin extract has a solubilizing effect on the composition for clearing liver and nourishing lung and is more favorable for absorption of the composition for clearing liver and nourishing lung. This is probably because alpha-glucosyltransferase grafts the glycosyl donor onto the naringin extract, increasing 1 hydrophilic glycosyl on the hydrophobic flavone parent, and simultaneously making the alpha-glycosylated naringin extract form a nano-scale aggregation structure, thereby improving the solubility and bioavailability of the alpha-glycosylated naringin extract.
Compared with the embodiment 2, the composition for clearing liver and nourishing lung prepared by the immobilized alpha-glucosyltransferase in the embodiments 3 to 5 has better solubility, because the graphene derivative immobilized alpha-glucosyltransferase has higher enzyme activity, larger specific surface area and nano-scale structure, and particularly, the graphene oxide nano colloid and the polycarboxylated graphene nanosheet aggregate are more dispersed and smaller in size, so that the contact area of the adsorbed alpha-glucosyltransferase and the substrate is increased, and the formation of the nano-scale alpha-glycosylated naringin extract molecular aggregate is facilitated, so that the alpha-glycosylated naringin extract has a solubilizing effect, and the solubility and the bioavailability of the composition for clearing liver and nourishing lung are improved.
Test example 3
Liver-clearing effect test: 180 patients with liver function impairment and liver fire flaming based on the syndrome differentiation of traditional Chinese medicine are selected. Wherein 108 men and 72 women are aged 40-50 years old. Diagnostic criteria for impaired liver function: dull pain in the liver area with asthenia; the glutamic-pyruvic transaminase and the glutamic-acyltransferase enzyme are slightly (less than or equal to 3 times of normal) or moderately (more than 3 times of normal) increased; the course of the disease is more than half a year. Diagnosis standard of liver fire flaming up: headache or dizziness; sour and astringent eyes or red eyes; dry mouth and bitter taste; hypochondriac distending pain; constipation or yellow urine; dysphoria and irritability; those who have more than 3 of the above symptoms can be diagnosed as the syndrome of liver-fire flaming upward. The selected cases are divided into 6 groups according to a random principle, each group contains 30 people, wherein 5 groups respectively take the liver-clearing and lung-nourishing composition prepared in the embodiment 1-5, 2 granules are taken once a day, and the other group does not take any medicine. The treatment effect is evaluated after 30 days as a treatment course, the treatment effect standard is shown in table 4, and the test result is shown in table 5.
The lung nourishing effect test: 180 patients with lung heat according to the syndrome differentiation of the traditional Chinese medicine are selected. 113 of the males and 63 of the females, wherein the age is 40-50 years. Diagnostic criteria: the lung fails to disperse and descend, and symptoms such as fever, cough, chest pain, abundant and thick sputum, yellow color, fishy smell and the like appear, and the cough attacks repeatedly and is not cured for a long time. The selected cases are divided into 6 groups according to a random principle, each group contains 30 people, wherein 5 groups respectively take the liver-clearing and lung-nourishing composition prepared in the embodiment 1-5, 2 granules are taken once a day, and the other group does not take any medicine. The treatment effect is evaluated after 30 days as a treatment course, the treatment effect standard is shown in table 4, and the test result is shown in table 5.
TABLE 4 evaluation criteria for liver-clearing and lung-nourishing effects
TABLE 5 test results of liver-clearing and lung-nourishing effects
As can be seen from Table 5, the compositions prepared in examples 1 to 5 all had the effect of clearing liver and nourishing lung, compared with the blank group. Compared with example 1, the compositions prepared by the alpha-glycosylated naringin extract in examples 2 to 5 have better effects of clearing liver and nourishing lung, because the hydrophilicity of the naringin extract is improved by the alpha-glycosylation, and the solubility of the naringin extract is further improved. Compared with the embodiment 2, the compositions prepared by the immobilized alpha-glucosyltransferase in the embodiments 3 to 5 have better effects of clearing liver and nourishing lung. The graphene derivative immobilized alpha-glucosyltransferase has higher enzyme activity, larger specific surface area and nano-scale structure, and particularly, graphene oxide nano colloid and polycarboxylated graphene nanosheet aggregate are more dispersed and smaller in size, so that the contact surface of the adsorbed alpha-glucosyltransferase and a substrate is increased, and the formation of nano-scale alpha-glycosylated naringin extract molecular aggregate is facilitated, so that the alpha-glycosylated naringin extract has a solubilizing effect, the solubility and bioavailability of the composition for clearing liver and nourishing lung are improved, the absorption of the composition in vivo is promoted, and the better effect of clearing liver and nourishing lung is achieved.
Claims (5)
1. A composition for removing heat from the liver and nourishing the lung, comprising: silybum marianum seed oil, naringin extract, loquat leaf extract, loquat powder, tangerine peel powder, phospholipid, mono-diglycerol fatty acid ester and gelatin skin.
2. The composition of claim 1, comprising: 470-475 parts by weight of silybum marianum seed oil, 28-32 parts by weight of naringin extract, 115-125 parts by weight of loquat leaf extract, 4.5-5.5 parts by weight of loquat powder, 28-32 parts by weight of tangerine peel powder, 6.5-7.5 parts by weight of phospholipid, 33-38 parts by weight of mono-diglycerol fatty acid ester and 900-1000 parts by weight of rubber skin.
3. The liver-clearing and lung-nourishing composition of claim 1 or 2, wherein: the rubber comprises gelatin, glycerol, water, trehalose, erythritol, mogroside, titanium dioxide and caramel color.
4. A liver-clearing and lung-nourishing composition as claimed in claim 3, wherein: the rubber comprises 290-310 parts by weight of gelatin, 110-130 parts by weight of glycerol, 290-310 parts by weight of water, 18-22 parts by weight of trehalose, 4.5-5.5 parts by weight of erythritol, 0.001-0.002 part by weight of mogroside, 2.7-3.3 parts by weight of titanium dioxide and 8-10 parts by weight of caramel color.
5. The method for processing the composition for clearing liver and nourishing lung according to any one of claims 1 to 4, which is prepared by the following steps:
(1) accurately weighing each raw and auxiliary material according to the formula amount;
(2) heating trehalose, erythritol, mogroside, 1/3-1/2 water and glycerol, adding gelatin, stirring, adding a suspension of titanium dioxide and caramel color, stirring, vacuumizing, sieving, and preserving heat to obtain a rubber;
(3) heating and dissolving mono-diglycerol fatty acid ester and 1/3-1/2 silybum marianum seed oil, cooling, adding naringin extract, loquat leaf extract, loquat powder, tangerine powder, phospholipid and 1/2-2/3 silybum marianum seed oil, stirring, and passing through a colloid mill and a sieve to obtain a content material;
(4) pressing the rubber sheet obtained in the step (2) and the content material obtained in the step (3) into soft capsules;
(5) shaping and drying the soft capsules obtained in the step (4) to obtain capsules;
(6) and (5) sorting out unqualified capsules from the capsules obtained in the step (5) to obtain the composition for clearing liver and nourishing lung.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116531444A (en) * | 2023-07-06 | 2023-08-04 | 健码制药（广东）有限公司 | Cough-relieving lung-heat-clearing soft capsule and preparation process thereof |
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