US 12,636,333 B1 · Herbal composition for supporting restoration of digestive system function and method for producing the same

Abstract

A herbal composition for supporting restoration of digestive system function comprising the following components: a Leucaena leucocephala extract, an eleutherin extract component from Eleutherine bulbosa, an Andrographis paniculata extract, a Panax vietnamensis extract, a Pouzolzia zeylanica extract, a first herbal mixture extract, a second herbal mixture extract, a sugar, a salt, a honey, a royal jelly, a vegetable oil, and a plant powder. Method for producing the composition comprising: (i) preparing materials; (ii) creating a base mixture; (iii) creating a first temporary mixture; (iv) creating a second temporary mixture; (v) creating a third temporary mixture; (vi) creating a fermented mixture; and (vii) packaging the fermented mixture to obtain the herbal composition for supporting restoration of digestive system function.

A method for producing a herbal composition for supporting restoration of digestive system function, comprising the following steps (i) to (vii):

(i) preparing materials comprising:
(a) a Leucaena leucocephala extract component having a predetermined first percentage (%) by weight, prepared by mixing a first extract, a second extract and a third extract at a weight ratio of 1:1:2;
wherein the first extract is prepared by performing steps (a1) to (a4):
(a1) preparing material:
collecting Leucaena leucocephala roots, washing, cutting into pieces, drying to a moisture content of 10%-12%, grinding, and sieving through a 1-2 mm mesh to obtain a first powder;
(a2) extracting the first powder with 60% ethanol at a ratio of 1:30 w/v at a temperature of 50° C.-60° C., in combination with ultrasonic treatment at a frequency of 30 KHz for 30 minutes to obtain a first temporary extract;
(a3) filtering the first temporary extract to remove residues to obtain a filtered first temporary extract;
and
(a4) concentrating the filtered first temporary extract at a temperature of 65° C.-70° C. until a ratio of the first powder to the first extract reaches (1.5-2):1 w/w to obtain the first extract;
wherein the second extract is prepared by performing steps (a1′) to (a4′):
(a1′) preparing material:
collecting Leucaena leucocephala stems, washing, cutting into pieces, drying to a moisture content of 10%-12%, grinding, and sieving through a 1-2 mm mesh to obtain a second powder;
(a2′) extracting the second powder with 70% ethanol at a ratio of 1:30 w/v at a temperature of 50° C.-60° C., in combination with ultrasonic treatment at a frequency of 25 kHz for 25 minutes to obtain a second temporary extract;
(a3′) filtering the second temporary extract to remove residues to obtain a filtered second temporary extract;
and
(a4′) concentrating the filtered second temporary extract at a temperature of 65° C.-70° C. until a ratio of the second powder to the second extract reaches (1.5-2):1 w/w to obtain the second extract;
wherein the third extract is prepared by performing steps (a1″) to (a4″):
(a1″) preparing material:
collecting Leucaena leucocephala seeds, washing, drying to a moisture content of 10%-12%, grinding, and sieving through a 1-2 mm mesh to obtain a third powder;
(a2″) extracting the third powder with 70% ethanol at a ratio of 1:30 w/v at a temperature of 50° C.-60° C., in combination with ultrasonic treatment at a frequency of 20 KHz for 20 minutes to obtain a third temporary extract;
(a3″) filtering the third temporary extract to remove residues to obtain a filtered third temporary extract;
and
(a4″) concentrating the filtered third temporary extract at a temperature of 65° C.-70° C. until a ratio of the third powder to the third extract reaches (1.5-2):1 w/w to obtain the third extract;
(b) an eleutherin extract component from Eleutherine bulbosa having a predetermined second percentage (%) by weight, prepared by performing steps (b1) to (b6):
(b1) preparing material:
collecting Eleutherine bulbosa tubers, washing, drying to a moisture content of 10%-12%, grinding, and sieving through a 1-2 mm mesh to obtain an Eleutherine bulbosa powder;
(b2) extracting the Eleutherine bulbosa powder with 96% ethanol at a ratio of 1:25 w/v at a temperature of 50° C.-60° C., in combination with ultrasonic treatment at a frequency of 20 KHz for 20 minutes, and filtering to obtain an Eleutherine bulbosa temporary extract;
(b3) concentrating the Eleutherine bulbosa temporary extract by rotary evaporation at a temperature of 65° C.-70° C. to obtain an extract residue;
(b4) crystallizing the extract residue at 30° C.-35° C. for 48-72 hours, filtering and washing the crystals with 96% ethanol to obtain a crystalline residue;
(b5) treating the crystalline residue with 5% NaOH solution at 50° C.-60° C. for 10-15 minutes, then recrystallizing the material at 30° C.-35° C. and filtering the crystals from the NaOH solution to obtain alkali-treated crystals;
and
(b6) dissolving the alkali-treated crystals in 96% ethanol at 30° C.-35° C. to obtain a solution, cooling the solution to 5° C.-10° C. for 2-3 hours, filtering the crystals from ethanol and drying the crystals at 40° C.-50° C. for 90-120 minutes to obtain the eleutherin extract component;
(c) an Andrographis paniculata extract component having a predetermined third percentage (%) by weight, prepared by performing steps (c1) to (c4):
(c1) preparing material:
collecting Andrographis paniculata leaves, washing, drying to a moisture content of 10%-12%, grinding, and sieving through a 1-2 mm mesh to obtain an Andrographis paniculata leaf powder;
(c2) extracting the Andrographis paniculata leaf powder with 70% ethanol at a ratio of 1:20 w/v at a temperature of 50° C.-60° C., in combination with ultrasonic treatment at a frequency of 25 kHz for 25 minutes, and filtering to remove residues to obtain an Andrographis paniculata extract;
(c3) concentrating the Andrographis paniculata extract by rotary evaporation at a temperature of 65° C.-70° C. to obtain an Andrographis paniculata extract residue;
and
(c4) mixing the Andrographis paniculata extract residue with water at a ratio of 1:1 w/w, then performing liquid-liquid extraction with ethyl acetate (EtOAc) in three repetitions, allowing phase separation and separating the ethyl acetate phase, combining the three ethyl acetate phases, and concentrating at a temperature of 50° C.-60° C. to remove the solvent, to obtain the Andrographis paniculata extract component;
(d) a Panax vietnamensis extract component having a predetermined fourth percentage (%) by weight, prepared by mixing a Panax vietnamensis root extract with a Panax vietnamensis stem and leaf extract at a ratio of 2:1 w/w;
wherein the Panax vietnamensis root extract is prepared by performing steps (d1) to (d4):
(d1) preparing material:
collecting Panax vietnamensis roots of 3-5 years old, washing, cutting into pieces, drying to a moisture content of 10%-12%, grinding, and sieving through a 1-2 mm mesh to obtain a Panax vietnamensis root powder;
(d2) mixing the Panax vietnamensis root powder with water at a ratio of 1:10 w/w, adding cellulase to achieve an activity of 280-300 U/g in the mixture and adding pectinase to achieve an activity of 220-250 U/g in the mixture, then incubating at a temperature of 45° C.-50° C. with agitation at 120-150 rpm for 1.5-2 hours, followed by ultrasonic-assisted extraction at a frequency of 25 kHz with agitation at 80-100 rpm at 60° C.-65° C. for 25-30 minutes, and centrifuging at 800 rpm for 20 minutes to obtain a solid residue from the root material and a first Panax vietnamensis root extract;
(d3) extracting the solid residue from the root material with 60% ethanol at a ratio of 1:20 w/v at a temperature of 50° C.-60° C., in combination with ultrasonic treatment at a frequency of 30 kHz for 30 minutes, and filtering to obtain a second Panax vietnamensis root extract;
and
(d4) combining the first Panax vietnamensis root extract and the second Panax vietnamensis root extract, concentrating the combined root extract at a temperature of 65° C.-70° C. until a ratio of the Panax vietnamensis root powder to the combined root extract reaches (1.5-2):1 w/w to obtain the Panax vietnamensis root extract;
wherein the Panax vietnamensis stem and leaf extract is prepared by performing steps (d1′) to (d4′):
(d1′) preparing material:
collecting Panax vietnamensis stems and leaves, washing, cutting into pieces, drying to a moisture content of 10%-12%, grinding, and sieving through a 1-2 mm mesh to obtain a Panax vietnamensis stem and leaf powder;
(d2′) mixing the Panax vietnamensis stem and leaf powder with water at a ratio of 1:10 w/w, adding cellulase to achieve an activity of 220-250 U/g in the mixture, then incubating at a temperature of 45° C.-50° C. with agitation at 100-120 rpm for 1.5-2 hours, followed by ultrasonic-assisted extraction at a frequency of 20 KHz with agitation at 80-100 rpm at 60° C.-65° C. for 20-25 minutes, and centrifuging at 800 rpm for 20 minutes to obtain a solid residue from the stem and leaf material and a first Panax vietnamensis stem and leaf extract;
(d3′) extracting the solid residue from the stem and leaf material with 70% ethanol at a ratio of 1:20 w/v at a temperature of 50° C.-60° C., in combination with ultrasonic treatment at a frequency of 20 kHz for 20 minutes, and filtering to obtain a second Panax vietnamensis stem and leaf extract;
and
(d4′) combining the first Panax vietnamensis stem and leaf extract and the second Panax vietnamensis stem and leaf extract, concentrating the combined stem and leaf extract at a temperature of 65° C.-70° C. until a ratio of the Panax vietnamensis stem and leaf powder to the combined stem and leaf extract reaches (1.5-2):1 w/w to obtain the Panax vietnamensis stem and leaf extract;
(e) a Pouzolzia zeylanica extract component having a predetermined fifth percentage (%) by weight, prepared by performing steps (e1) to (e4):
(e1) preparing material:
collecting Pouzolzia zeylanica stems and leaves, washing, cutting into pieces, drying to a moisture content of 10%-12%, grinding, and sieving through a 1-2 mm mesh to obtain a Pouzolzia zeylanica powder;
(e2) mixing the Pouzolzia zeylanica powder with water at a weight ratio of 1:10 w/w, adding cellulase to achieve an activity of 280-300 U/g in the mixture, then incubating at a temperature of 45° C.-50° C. with agitation at 100-120 rpm for 1.5-2 hours, followed by ultrasonic-assisted extraction at a frequency of 25 kHz with agitation at 80-100 rpm at 60° C.-65° C. for 20-25 minutes, and centrifuging at 800 rpm for 20 minutes to obtain a solid residue from the material and a first Pouzolzia zeylanica extract;
(e3) extracting the solid residue from the material with 70% ethanol at a ratio of 1:20 w/v at a temperature of 50° C.-60° C., in combination with ultrasonic treatment at a frequency of 25 kHz for 30 minutes, and filtering to obtain a second Pouzolzia zeylanica extract;
and
(e4) combining the first Pouzolzia zeylanica extract and the second Pouzolzia zeylanica extract, concentrating the combined extract at a temperature of 65° C.-70° C. until a ratio of the Pouzolzia zeylanica powder to the combined extract reaches (1.5-2):1 w/w to obtain the Pouzolzia zeylanica extract component;
(f) a first herbal mixture extract component having a predetermined sixth percentage (%) by weight, prepared by performing steps (f1) to (f4):
(f1) preparing a first herbal mixture powder by mixing six herbals at a weight ratio of 2 parts of Curcuma zedoaria root, 2 parts of Curcuma longa root, 2 parts of Zingiber officinale root, 2 parts of Alpinia galanga root, 1 part of Platycodon grandiflorus root and 1 part of Citrus aurantium fruit, each herbal having a moisture content of 10%-12%, and grinding and sieving the blended materials through a 1-2 mm mesh to obtain a first herbal mixture powder;
(f2) mixing the first herbal mixture powder with water at a ratio of 1:(15-20) w/w, adding cellulase to achieve an activity of 650-700 U/g in the mixture and adding pectinase to achieve an activity of 450-500 U/g in the mixture, then incubating at a temperature of 40° C.-45° C. with agitation at 100-150 rpm for 2-3 hours, followed by ultrasonic-assisted extraction at a frequency of 30 KHz with agitation at 80-100 rpm at 65° C.-70° C. for 30 minutes, and centrifuging at 800 rpm for 20 minutes to obtain a solid residue from the first herbal mixture and a first extract of the first herbal mixture;
(f3) extracting the solid residue from the first herbal mixture with 70% ethanol at a ratio of 1:(10-15) w/v at a temperature of 55° C.-60° C., in combination with ultrasonic treatment at a frequency of 25 kHz with agitation at 80-100 rpm for 20 minutes, followed by centrifuging at 800 rpm for 20 minutes and filtering to obtain a second extract of the first herbal mixture;
and
(f4) combining the first extract of the first herbal mixture and the second extract of the first herbal mixture, concentrating the combined first herbal mixture extract at a temperature of 65° C.-70° C. until a ratio of the first herbal mixture powder to the combined first herbal mixture extract reaches (1.5-2):1 w/w to obtain the first herbal mixture extract component;
(g) a second herbal mixture extract component having a predetermined seventh percentage (%) by weight, prepared by performing steps (g1) to (g4):
(g1) preparing a second herbal mixture by mixing five herbals at a weight ratio of 3 parts of Plumbago zeylanica stem, 3 parts of Cleistocalyx operculatus leaf, 2 parts of Ampelopsis cantoniensis leaf, 1 part of whole plant of Lycopodiella cernua, and 1 part of Ficus carica fruit, each herbal material having a moisture content of 10%-12%, and grinding and sieving the blended materials through a 1-2 mm mesh to obtain a second herbal mixture powder;
(g2) mixing the second herbal mixture powder with water at a weight ratio of 1:(20-25) w/w, adding cellulase to achieve an activity of 550-600 U/g in the mixture and adding pectinase to achieve an activity of 350-400 U/g in the mixture, incubating at a temperature of 40° C.-45° C. with agitation at 100-150 rpm for 2-3 hours, followed by ultrasonic-assisted extraction at a frequency of 30 kHz with agitation at 80-100 rpm at 65° C.-70° C. for 30 minutes, and centrifuging at 800 rpm for 20 minutes to obtain a solid residue from the second herbal mixture and a first extract of the second herbal mixture;
(g3) extracting the solid residue from the second herbal mixture with 70% ethanol at a ratio of 1:(10-15) w/v at a temperature of 55° C.-60° C., in combination with ultrasonic treatment at a frequency of 20 KHz with agitation at 80-100 rpm for 20 minutes, followed by centrifuging at 800 rpm for 20 minutes and removing the solid residue to obtain a second extract of the second herbal mixture;
and
(g4) combining the first extract of the second herbal mixture and the second extract of the second herbal mixture, concentrating the combined second herbal mixture extract at a temperature of 65° C.-70° C. until a ratio of the second herbal mixture powder to the combined second herbal mixture extract reaches (1.5-2):1 w/w to obtain the second herbal mixture extract component;
(h) a sugar component having a predetermined eighth percentage (%) by weight;
wherein the sugar component is selected from the group consisting of glucose, fructose, galactose, lactose, maltose, sucrose, and combinations thereof;
(i) a salt component having a predetermined ninth percentage (%) by weight;
wherein the salt component is selected from the group consisting of sea salt, himalayan pink salt, kosher salt, and combinations thereof;
(j) a honey component having a predetermined tenth percentage (%) by weight;
(k) a royal jelly component having a predetermined eleventh percentage (%) by weight;
(l) a vegetable oil component having a predetermined twelfth percentage (%) by weight;
wherein the vegetable oil component is selected from the group consisting of rice bran oil, sunflower oil, olive oil, and combinations thereof;
and
(m) a plant powder component having a predetermined thirteenth percentage (%) by weight;
wherein the plant powder component is selected from the group consisting of rice flour, wheat flour, cereal flour, and combinations thereof;
wherein the cereal flour is obtained by mixing five legume seed powders at a weight ratio of 1 part of Vigna unguiculata seed powder, 1 part of Glycine max seed powder, 1 part of Vigna radiata seed powder, 1 part of Vigna angularis seed powder, and 1 part of Phaseolus vulgaris seed powder;
(ii) heating the honey component at 65° C.-70° C., adding the royal jelly component, the sugar component and the salt component, then stirring at 500-600 rpm for 45 minutes to obtain a base mixture;
(iii) mixing the plant powder component with the base mixture at step (ii) and stirring at 400-500 rpm for 30 minutes to obtain a first temporary mixture;
(iv) mixing the components (a) to (g) in order with the first temporary mixture at step (iii) to obtain a second temporary mixture, ensuring homogeneous mixing after each addition;
(v) mixing the vegetable oil component with the second temporary mixture at step (iv) and stirring at 400-500 rpm for 30 minutes to obtain a third temporary mixture;
(vi) creating a fermented mixture by performing steps (i′) to (iii′):
(i′) preparing ingredients comprising a substrate mixture and a fermentation microbial mixture;
wherein the substrate mixture is obtained by sterilizing the third temporary mixture at step (v) by autoclaving at 121° C. for 15 minutes;
wherein the fermentation microbial mixture is obtained by mixing a first microbial biomass, a second microbial biomass, a third microbial biomass and a fourth microbial biomass at a weight ratio of 1:1:1:1;
wherein
the first microbial biomass is obtained by culturing Lactobacillus plantarum ATCC 14917 on a propagation medium for 24 hours at 37° C., then centrifuging at 4° C. at 6000 rpm for 10 minutes, removing the supernatant and collecting the microbial pellet to obtain the first microbial biomass;
the second microbial biomass is obtained by culturing Lactobacillus fermentum ATCC 23271 on a propagation medium for 24 hours at 37° C., then centrifuging at 4° C. at 6000 rpm for 10 minutes, removing the supernatant and collecting the microbial pellet to obtain the second microbial biomass;
the third microbial biomass is obtained by culturing Lactobacillus rhamnosus ATCC 53103 on a propagation medium for 24 hours at 37° C., then centrifuging at 4° C. at 6000 rpm for 10 minutes, removing the supernatant and collecting the microbial pellet to obtain the third microbial biomass;
and
the fourth microbial biomass is obtained by culturing Lactobacillus paracasei ATCC 25598 on a propagation medium for 24 hours at 37° C., then centrifuging at 4° C. at 6000 rpm for 10 minutes, removing the supernatant and collecting the microbial pellet to obtain the fourth microbial biomass;
wherein the propagation medium comprises glucose 20 g/L, peptone 10 g/L, yeast extract 5 g/L, CaCO3 5 g/L, K2HPO4 2 g/L, MgSO4 0.6 g/L, and MnSO4 0.3 g/L;
(ii′) mixing the fermentation microbial mixture with the substrate mixture at a ratio of (1-2):100 w/w, and mixing until homogeneous to obtain a fermentation substrate mixture;
and
(iii′) fermenting the fermentation substrate mixture at 30° C.-35° C. for 40-48 hours to obtain the fermented mixture;
wherein the total viable cell density in the fermented mixture is 1×107 to 1×109 CFU/g;
and
(vii) packaging the fermented mixture at step (vi) to obtain the herbal composition for supporting restoration of digestive system function.

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Herbal composition for supporting restoration of digestive system function and method for producing the same

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Herbal composition for supporting restoration of digestive system function and method for producing the same

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Abstract

Claims18