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MX2008003885A - Compositions and methods comprising panax species - Google Patents

Compositions and methods comprising panax species

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Publication number
MX2008003885A
MX2008003885A MXMX/A/2008/003885A MX2008003885A MX2008003885A MX 2008003885 A MX2008003885 A MX 2008003885A MX 2008003885 A MX2008003885 A MX 2008003885A MX 2008003885 A MX2008003885 A MX 2008003885A
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MX
Mexico
Prior art keywords
ginsenoin
acid
extraction
composition according
ginsenoside
Prior art date
Application number
MXMX/A/2008/003885A
Other languages
Spanish (es)
Inventor
T Gow Robert
W Sypert George
Yan Xun
Li Dan
Original Assignee
Herbalscience Singapore Pte Ltd
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Application filed by Herbalscience Singapore Pte Ltd filed Critical Herbalscience Singapore Pte Ltd
Publication of MX2008003885A publication Critical patent/MX2008003885A/en

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Abstract

Compositions comprising elevated amounts of a ginsenosides, polyacetylenes, essential oil and/or polysaccharide are described, as are preferred extraction methods.

Description

COMPOSITIONS AND METHODS THAT INCLUDE PANAX SPECIES FIELD OF THE INVENTION Ginseng, the rhizome (root) of Panax ginseng (Asian ginseng, Korean ginseng) of the family Araliaceae, has been used in oriental medicine since ancient times as a stimulant, tonic, diuretic, and digestive aid. In Europe ginseng phytomedicines are marketed as self-acquired medicines and are taken to increase mental and physical performance, to provide resistance to fatigue and disease, and to alleviate exhaustion. Because of continuous harvests and use for thousands of years, the natural supply of P. ginseng roots has been exhausted for some time. Currently, almost all roots of P. ginseng are cultivated in China, Korea, and Japan.
BACKGROUND OF THE INVENTION Many congeners of ginseng are used in medicines. The root of P. quinquefoli us L. (American ginseng), which originally grew in North America, is now grown for export to the Asian market, where it is used medicinally for slightly different purposes than P. ginseng, P. notoginseng (also known as Sanchi ginseng) has been used as a special herb in Traditional Chinese Medicine (TCM) from ancestral times to the present. Other species such as, but not limited to P. japonicus, P. pseudo-ginseng, P. vietnamensis, Eleutherococcus senticosus (Siberian ginseng), and other species, subspecies, or varieties have also been used in Asian phytomedicine. The constituents of Panax rhizomes (ginseng roots) have been investigated since the beginning of the 20th century. Several of the classes of compounds have been isolated and some of the individual chemical constituents have been studied for their biological effects. Some of the classes of chemical compounds that are ubiquitous in the various roots of ginseng include the triterpene saponins, an essential oil in which are contained the chemicals known as polyacetylenes, polysaccharides, sesquiterpenes, peptidoglycans, compounds containing nitrogen, and others such as fatty acids, carbohydrates and phenolic compounds (2). The chemical constituents of ginsengs that are believed to contribute to their pharmacological effects have been investigated extensively since the 1950's. The main bioactive compounds based on these investigations are triterpene saponins (for example, ginsenosides), polyacetylenes, and polysaccharides (2-7). All Panax species contain a class of triterpene saponins collectively termed ginsenoids (or panaxosides). The ginsenosides contain 4 rigid trans-anular steroid skeletons, and the individual ginsenosides differ in the number, type, and location of their sugars, and the central structure of triterpene or steroid portion (3). The ginsenosides are called "ginsenosides Rx" where "x" corresponds to the Rf value sequence of the spots when they are analyzed by thin layer chromatography. Among the known ginsenosides are the R0, Rba ?, Rt, 2 Rc, R¿, Re, Rf Rgl, g2 f Rg3 Rg5 / Rhl i R ai R t Y 2 • L? S ginsenosides are further classified into groups based on the central structure of the steroid portion, and include those ginsenosides based on the 20 (S) protopanaxdiol core structures (collectively the Rb group), the 20 (S) protopanaxtriol core structures (collectively the Rg group), the octylol core structure, and the olene central structure. Specific ginsenosides in the Rb group include bi / Rb2 / C Rd and several other related compounds. Specific Ginsenosides in the Rg group include Rg, Re, Rf, Rg2 and various other related compounds. Figure 5 shows the chemical structures for Rhl, Rh2, Rg3, and Rgs (Gic = ß-D-glucopyranosyl-, G? CG? C = ß-D-glucopyranosyl- (l- »2) ß-D-glucopyranosyl- ). Compendiums of the pharmacological effects of extracts and preparations of P. ginseng (ginsenoside fractions) have been presented, polyacetylene and polysaccharide) (2-8). The preparation and definition of ginseng products is specified in several European pharmacopoeias. The Swiss Pharmacopoeia, Pharmacopoea Helvetica (Commission Suisse de Pharmacopée), requires a total content of ginsenosides, calculated as in relation to the abundance of the ginsenoside Rg ?, of not less than 2.0%. In accordance with the German Pharmacopoeia (Herbal Medicine -Expanded Commission E Mongraphs, Blumenthal, M. et al., Integrative Medicine Communications, 2000, pp. 170-177), the total content of gnosenosides will not be less than 1.5% using a method quantification spectrometer. In contrast, the 4th. edition of the European Pharmacopoeia (European Pharmacopoeia Commission, European Directorate for the Quality of Medicines - Council of Europe, 2001) requires the content of ginsenosides Rg? and Rbi is not less than 0.3% measured using High Performance Liquid Chromatography (HPLC) methods. However, consumers in the United States who purchase a ginseng product should carefully consider the source and the product. No federal agency imposes quality control on the ingredients of many products. In a study of 54 products called ginseng, it was found that 25% did not contain ginsenosides of all. And 60% contained only trace amounts (8-10).
The main physiological effects from the ingestion of ginseng that were documented in the ancient literature (2) include the following: general tonic, stimulation of the immunological function; beneficial effects on the cardiovascular system including a reduction in blood pressure, reductions in serum total cholesterol levels, low density lipoprotein cholesterol and triglycerides and an increase in serum high density lipoprotein cholesterol levels; stimulation of alcoholic dehydrogenase and oxidation of alcohol in the liver, depletion of blood sugar levels; stimulation of the pituitary adrenocorticoid system, anti-aging, and inhibition of tumor growth. Interestingly, Rgl is claimed to stimulate the central nervous system and improves the synthesis of proteins, DNA and RNA while Rb? it has tranquilizing effects and improves memory which can again be considered for the different biological effects associated with different Panax species (6). Experiments and recent clinical studies have demonstrated the following bioactive properties of the various chemical fractions and chemicals of the Panax species: potent antioxidant activity (Rg ?, Rb ?, extract) (11-17); cardiovascular protection (Rg ?, Rb ?, extract) (11-21); immunological and anti-viral improvement, anti-influenza (Rg ?, polysaccharides, extract) (22-34); cytoprotection (polysaccharides, extract) (17, 20, 35); neuroprotection and antidementia (ginsenosides, Rb ?, Rg2 Rg3 r extract) (36-40); inhibition of platelet agglutination (ginsenosides, R0, Rgl, Rg2, polyacetylenes, extract) (41-44); inhibition of the calcium channel (Rf) (45): anticancer (Rg3, R 2 r polyacetylenes, polysaccharides) (46-52); enti-inflammatory (polyacetylenes, extract) (53, 54); anti-cholesterol (extract) (55, 56); hypoglycemic and antidiabetes (polysaccharide, extract) (57-60); protection and therapy for lung diseases (extract) (34, 61); disease treatment and liver protection (extract) (62,63); improvement of the erectile capacity (extract) (64-67); memory and improved knowledge (Rbi, Rgi r polyacetylenes, extract) (37, 38, 68, 69); and chronic fatigue (extract) (70, 71). Currently, the available ginseng products are suspect with respect to their chemical composition not only with respect to the content of ginsenosides but also with respect to the crucial chemical constituents such as essential oils and polysaccharides. Methods are required to extract Panax and related species and Panax extraction compositions with improved bioactive profiles, such as, but not limited to, triterpene saponins (e.g., ginsenosides), essential oils (e.g., polyacetylenes), and fractions polysaccharides, which can be produced with standard and reliable quantities of these constituents of Panax beneficial bioactives medically and physiologically.
SUMMARY OF THE INVENTION In certain aspects the invention characterizes new compositions and pharmaceutical preparations thereof. In certain embodiments, the compositions comprise at least one ginsenoside in an amount greater than 10% by weight. The ginsenoside may comprise R0, Ri, R2 / Ra r Rb Rb2 r Rc r Rd r R / Rf Rgl f Rg2 1 Rg3 Rg5 1 Rhl and Rh2 / Rh4, Rs ?, RS2, Rs3 / or Rs. Certain compositions comprise at least one ginsenoside in an amount greater than 15, 20, 25, 30, 35, 40, 45 or 50% by weight. Additional embodiments characterize compositions comprising a polyacetylene in an amount greater than 1, 2, 4, 6, 8, or 10% by weight. The polyacetylene can comprise pananaxinol, panaxidol, panaxtriol, acetylpanaoxidol, chloropapaxidol, panaxidolchlorohydrin, panaxin, ginsenoin A, ginsenoin B, ginsenoin C, ginsenoin D, ginsenoin E, ginsenoin F, ginsenin G, ginsenoin H, ginsenin I, ginsenoin J, or ginsenoin K. In certain modalities, the polyacetylene comprises pananaxinol, panaxidol, panaxitriol, acetylpanaxidol, chloropapaxidol, panaxidolchlorohydrin, panaxin, ginsenoin A, ginsenoin B, ginsenoin C, ginsenoin D, ginsenoin E, ginsenoin F, ginsenoin G, ginsenoin H, ginsenoin I, ginsenoin J, or ginsenoin K. Additional moieties characterize compositions comprising a polysaccharide in an amount greater than 25, 30, 35, 40, 45, 50, 55, or 60% by weight. The polysaccharide may comprise glucose, arabinose, galactose, rhamnose, xylose or uronic acid. Still other embodiments characterize compositions comprising an essential oil selected from the group consisting of (+) spatulenol; (-) spatulenol; caffeine; hexadecanoic acid; (-) - caryophyllenium oxide; ethyl heptanoate; trans methyl ester, trans-octadeca-9, 12-dienoic; octadec-9-inoic acid methyl ester; phenylacetylene; ethylentiourea; linoleic acid; 4- methyl-pent-2-enoic acid; 2- methyl-4-nitroimidazole; 9, 12- octadecadienal; mevinfos; undec-10-inoic acid; falcarinol ((Z) -1,9-heptadecadien-4,6-diyl-3-ol); [IR- (1, 4β, 4aa, 6β, 8a)] - octahydro-4, 8a, 9, 9-tetramethyl-1, 6-methano-1 (2H) -naphthol; 4,6-diamino- 1, 3, 5-triazin-2 (1H) -one; 2, 2'-methyliminodiethanol; dihydrouracil; stearic acid; 4- nitrophenol; 3- nitrotoluene; 2,3-dihydroxypropyl palmitate; oleic acid; cinnamyl acetate; 7- octenoic acid; 1-methyl-5-nitro-1H-imidazole; 2-ethyl-2-methyloxirane; (9E, 12E) - octadeca- 9, 12-methyl-dienoate; sinalbin, stigmasta- 5, 22- dien- 3-4- ol; (3ß, 24S) - stigmast- 5- in- 3- ol; stigmast- 5-in-3 ß- ol; (3ß, 24E) - stigmast- 5- in- 3- ol; 4-methyl-1, 4-heptadiene; 9, 12- octadecadienal; 7, 8-epxyoctene; 4- nonino; 2-cyclopentene-1-undecanoic acid; 3-hydroxy-2-methyl-4-pyrone; pyrogallol; [laR- (laa, 7, 7aa, 7b)] - la, 2, 3, 5, 6, 7, 7a, 7b-octahydro-1, 1, 7, 7a-tetramethyl-1H-cyclopropa [a] naphthalene: [la R- (laa, 4aa, 7a, 7aβ, 7ba)] - decahydro-1, 1, 7-trimethyl-4-methylene-1H-cycloprop [e] azulene; caryophyllene; [IR- (1R *, 4Z, 9S *)] - 4, 11, 11-trimethyl-8-methylenebicyclo [7.2.0] undec-4-ene; 4- methyl-2-phenyl-2-pentenal; (Z) - 9, 17, octadecadienal; ethyleneidecycloheptane; octa-1, 7-diine; 3- (phenylmethyl) sidnon; diisopropyl adipate; 2-, 3- dihydroxypropyl palmitate; acid (3- linoleoyl glycerol) 9Z, 12Z, octadecadienoic; and 3-ethenyl-cyclooctene. Additional embodiments characterize compositions comprising at least one ginsenoside, polyacetylene, polysaccharide and / or essential oil of the specified minimum weights of hundreds of weights. In another embodiment, the compositions are formulated alone or in combination with other active ingredients in pharmaceutical and / or food formulations. The compositions of the present invention may be useful to provide certain physiological, psychological, and / or medicinal benefits after ingestion, including, but not limited to, antioxidant activity, protection and cardiovascular treatment, cytoprotection, nervous system protection, anti-inflammatory disease. -neurodegenerative (eg, Alzheimer's disease, Parkinson's disease, stroke), inhibition of platelet agglutination, anti-cholesterol, hypoglycemia and diabetes mellitus, anti-inflammatory, immune enhancement, anti-viral (eg, influenza), disease anti-pulmonary, protection and treatment of liver diseases, prophylaxis and therapy for cancer, improvement of male erectile function, improvement of memory and knowledge and relief of chronic fatigue syndromes. In another aspect, the present invention features methods of treating a mammal (e.g., a human) for a disease or disorder, comprising administering to the mammal in need therefor a therapeutically effective amount of any of the aforementioned compositions. In another aspect, the present invention concerns methods for extracting Panax compositions having a predetermined characteristic, such as, but not limited to, a high amount of a chemical constituent selected from the group consisting of: a triterpene saponin, polyacetylene, and / or polysaccharide. By "elevated" is meant an amount greater than the amount present in the native plant material or Panax extraction products of the prior art. In general, said methods comprise the extraction of compounds, such as triterpene saponins, polyacetylenes, and polysaccharides from extracts of native Panax plant materials or from native Panax plant materials using one or more extraction steps described herein. The various aspects and embodiments are further described in the detailed description, figures and claims below.
BRIEF DESCRIPTION OF THE FIGURES Figure 1, shows an exemplary method for the preparation of an essential oil fraction from vegetable raw material. Figure 2, exposes an exemplary method for the preparation of ginsenoside fractions. Figure 3 sets forth an exemplary method for the preparation of a purified ginsenoside fraction using a polymeric adsorbent. Figure 4 shows an exemplary method for the preparation of polysaccharide fractions. Figure 5, exposes the chemical structure of 20 (S) -ginsenoside Ri, 20 (S) -ginsenoside Rh2 / 20 (S) -ginsenoside Rg3, and ginsenoside Rg5.
DETAILED DESCRIPTION OF THE INVENTION Definitions It should be noted that, as used in this specification and the appended claims, the singular forms "a" (before consonant) a "one" (before a vowel), and " , "include plural referents unless the context clearly dictates otherwise. As used herein, the term "essential oil fraction" comprises compounds that are volatile, insoluble in water, and extractable using non-polar solvents. As used herein, the essential oil fraction further comprises polyacetylenes obtained from Panax and related species. The essential oil fraction may further comprise one or more compounds from sesquiterpenes, azulene, patchuleno, sesquiterpene alcohols, panasinsanol A, B panasinsanol, methoxypyrazine, beta-elmeno, dien panaxynoles or alquilpirazinas. Polyacetylenes fractions of essential oils can comprise one or more compounds of pananaxinol, panaxidiol, panaxytriol, acetilpanaxidol, panaxidolclorohidrina, panaxina, ginsenosina A, ginsenosina B, ginsenosina C, ginsenosina D, ginsenosina E, ginsenosina F, ginsenosina G, ginsenosina H , ginsenosine I, ginsenosine J, ginsenosine K, panaxacol, panaxidol, falcarinol or falcarintriol. As used herein, the term "raw material" refers to raw plant material, comprising whole vegetables alone, or in combination with one or more constituent parts of a plant comprising leaves, rhizomes, roots, including, but not limited to, main roots, tail roots, and fibrous roots, shoots, leaves, seeds, and flowers, wherein the vegetable or constituent parts may comprise material that is crude, dried, vaporized, heated, or otherwise subjected to physical processing to facilitate processing, which may additionally comprise material that is intact, crushed, shredded, ground or otherwise processed to affect the size and physical integrity of the plant material. As used herein, the term "fraction" means a composition that comprises a specific group of compounds characterized by certain chemical, physical, or chemical and physical properties. As used herein, the term "constituents of ginseng" will mean compounds found in each of the individual and related Panax species and will include all of the chemical compounds identified above as well as other compounds found in each of the Panax species. and related, including but not limited to essential oils, polyacetylenes, ginsenosides, and polysaccharides. As used herein, the term "ginsenoide fraction" comprises saponin triterpene obtained from Panax and related species, which further comprise compounds based on the core structure protopanaxadiol, the central structure protopanaxatriol, the central structure octilol, or the central structure oleanano, and related compounds. As used herein, the term "increased" or "elevated" a fraction amount, including but not limited to, moieties such as triterpene saponin, polyacetylene, and polysaccharide fractions, means the weight percent of the fraction, either in toto or a single constituent of the fraction, in a mixture or sample is increased as compared to the weight percent of the constituent or fraction in the native tissue or in the native vegetable. As used herein, the term "one or more compounds" means that at least one compound, such as panaxitriol (a polyacetylene essential oil), Rg? (a triterpene saponin ginsenoside), or ginsenan PA (a ginseng polysaccharide soluble in water) is planned, or that more than one compound, for example, panaxitriol and Rg? are planned As is known in the art, the term "compound" does not mean a single molecule, but multiple or moles of molecules. As is known in the art, the term "compound" means a specific chemical entity having different chemical and physical properties, while "compounds" refers to one or more chemical constituents. As used herein, the term "Panax" includes the Panax genera and related species, including, but not limited to, Eleutherococcus senticosus. Additionally, as used herein, Panax refers to the vegetable or plant material derived from the plant family Araliaceae, where the species include but are not limited to P. ginseng, P. quinqué folius, P. notoginseng, P. pseidoginseng, P. jamonicum, P. vietnamensis, and E. senticosus. The term also includes all clones, cultivars, variants, and mutants of Panax and related species. The term "Panax" can also be used here indistinctly with "ginseng" and means, these plants, clones, cultivars, variants and mutants. As used herein, the term "polysaccharide moiety" comprises compounds obtained or derivatives of Panax and related species. The fraction of the polysaccharide extract of the chemical constituents of the Panax species has been defined in the scientific literature as the fraction of water-soluble extraction insoluble in ethanol (26, 31, 63, 72-74). The polysaccharide fraction may comprise one or more ginsan and panaxan A to U compounds, including, but not limited to, the panaxan neutral polysaccharides A to E and the panaxan acid polysaccharides A to U. The polysaccharide moiety may further comprise polymers and oligomers which they comprise monomer units of glucose, arabinose, galactose, rhamnose, xylose, or uronic acid. As used herein, the term "rhizome" refers to the constituent part of Panax and related species comprising a horizontal root stem, which may be in part or whole, underground, additionally comprising the offshoots in the upper part and lower roots, including but not limited to roots, tail roots, and fibrous roots. Compositions The compositions of the present invention comprise fractions of Panax extract comprising high levels of ginsenosides which may additionally comprise a polyacetylene, a polysaccharide, or both.
The Panax species extraction products of the prior art contain variable native plant materials. For example, widely fluctuating amounts of ginsenosides only, if any are present. In contrast, the compositions herein comprise defined amounts of isolated and purified fractions of essential oils, ginsenosides and polysaccharides from one or more Panax species. These individual fraction compositions can be combined in specific proportions (profiles) to provide beneficial combination compositions and can provide extract products that are not found in commonly known extract products. For example, an essential oil fraction of a species can be combined with a ginsenoside fraction of the same or different species, and that combination composition may or may not be combined with a polysaccharide fraction thereof or different Panax species. Table 1 through 4 list the fractions of major beneficial bioactive chemical constituents found in the rhizome raw materials of the four major Panax species. Used to produce ginseng products.
Table 1. Fractions of chemical constituents of Rizoma * of Panax notoginseng *% of mass in dry weight = weight of mass of fraction / weight of mass of raw material). * USDA Agricultural Research Service. *** HerbalScience Laboratory. Table 2. Fractions of American Ginseng Chemical Constituents (Panax Quinquefolius L. Rhizome) Table 3. Fractions of White Ginseng Chemical Constituents (Panax ginseng C. Rhizome) Table 4. Fractions of Red Ginseng Chemical Constituents (Panax ginseng C. Rhizome) The Panax and related species compositions described herein have ginsenosides in amounts greater than those found in the native Panax plant material and related species or extract products of commonly available Panax species. Importantly, natural Panax and related species have low amounts of key ginsenoside ingredients (86-91% by weight at most). The commercially available products comprise ~ 3% by weight of ginsenosides which reflects the difficulty in isolated Panax ingredients that currently exist. The embodiments also comprise compositions wherein one or more of the fractions, including essential oils, ginsenosides, or polysaccharides, were found in a concentration that is higher than that found in plant material of native Panax species. The embodiments also comprise compositions wherein one or more of the fractions, including essential oils, ginsenosides, or polysaccharides, were found in a concentration that is lower than that found in native Panax species. Known amounts of four Panax species are shown in Tables 1-4. For example, compositions of the present invention comprise compositions wherein the concentration of essential oils is from 0.001 to 200 times the concentration of native Panax species, and / or compositions where the concentration of ginsenosides is from 0.001 to 100 times the concentration of native Panax species, and / or compositions where the concentration of polysaccharides is from 0.01 to 6 times the concentration of native Panax species. Compositions of the present invention comprise compositions wherein the concentration of essential oils is from 0.1 to 50 times the concentration of native Panax species, and / or compositions where 1 concentration of ginsenosides is from 0.1 to 50 times the concentration of native Panax species, and / or compositions where the concentration of polysaccharides is from 0.01 to 6 times the concentration of native Panax species. Compositions of Panax species for antioxidant activity and cardiovascular protection may have a concentration of the composition of increasing ginsenoside fraction, a concentration of the reduced essential oil fraction composition, and a concentration of the composition of increasing polysaccharide fraction, in% by weight , that found in the native plant material of native Panax species or conventional known extraction products. A composition of new Panax species for immune enhancement may have a composition of increasing ginsenoside fraction and a composition of polysaccharide fraction, and a concentration of the composition of reduced essential oil fraction, in% by weight, than that found in plant material of native Panax species or conventional known extraction products. Another example of a composition of new Panax species, for the treatment of Alzheimer's disease, dementia, and memory and knowledge improvement, comprises a composition having a concentration of the growing essential oil composition and a ginsenoside fraction composition. , and a reduced polysaccharide fraction composition that is found in plant material of native Panax species or known conventional extraction products. Additional embodiments comprise compositions comprising altered profiles (distribution of proportions) of the chemical constituents of Panax species in relation to that found in the native plant material or extract products of commonly available Panax species. For example, the fraction of essential oil can be increased or decreased in relation to the concentrations of ginsenoside and / or polysaccharide. In a similar way, the ginsenosides or polysaccharides can be increased or decreased in relation to the other constituent fractions of the extract to allow new compositions of chemical constituent profile for specific biological effects. Extraction Methods The initial material for extraction is plant material of one or more species of Panax, it is thought that P. notoginseng, P. ginseng, which can be either in the form commonly known as "white ginseng" or in the commonly known as "red ginseng", or P. quinquefoli us are the preferred initial materials. As used herein, "white ginseng" comprises the material derived from P. ginseng which has been dried in the open air or in a dryer after harvesting so that the color does not turn red to brown. As used herein, "red ginseng" comprises material derived from P. ginseng, which has been heated, typically by evaporation, and then dried in a manner so that the material turns red to brown. The material may be the aerial portion of the plant, which includes leaves, stems, or other parts of the plant, it is thought that the rhizome is the preferred initial material. The plant material of Panax species may undergo pre-extraction steps to convert the material into any particular form, and any form that is useful for extraction is contemplated in the present invention. Said pre-extraction steps include, but are not limited to, material that is minced, shredded, grated, crushed, pulverized, cut, or torn, and the initial material, prior to the pre-extraction stages, is fresh plant material. or dried. A pre-extraction step comprises crushing and / or pulverizing the rhizome plant material of Panax species into a fine powder. The initial material or material after the pre-extraction stages can be dried or have moisture added to it. Once the plant material of Panax species is in a form for extraction, extraction methods are contemplated in the present invention. The extraction methods of the present invention comprise processes described herein. In general, the methods of the present invention comprise, in part, methods in which the plant material of Panax species is extracted using supercritical carbon dioxide (SCC02) which is followed by one or more solvent extraction steps, such as as, but not limited to, water extraction processes, hydroalcoholic, by polymeric absorption. Additional methods contemplated by the present invention comprise the extraction of plant material from Panax species using other organic solvents, chemical refrigerants, compressible gases, sonification, liquid pressure extraction, high-speed countercurrent chromatography, molecular printing polymers and other methods known extraction. Said techniques are known to those skilled in the art. In one aspect, compositions of the present invention can be prepared by means of a method comprising the steps set forth schematically in Figures 1, 2, 3 and. Extraction of Panax by Supercritical Fluid Due to the hydrophobic nature of the essential oil, non-polar solvents, including but not limited to SCC02, hexane, petroleum ether and ethyl acetate can be used for this extraction process.
A generalized description of the extraction of the essential oil fraction from the rhizome of the Panax species using SSC02, is diagrammed in Figure 1. The raw material 10 is rhizome of crushed Panax species (8 to 20 meshes). Solvent 210 is pure C02. The raw material is loaded in a basket that is placed inside an extraction vessel by supercritical fluid (SFE) 20. After testing leaks and purge, the process comprises the flow of liquefied C02 from a storage container to through a cooler to the C02 pump. Then the C02 is compressed to the desired pressure, it flows through the raw material into the extraction vessel where the pressure and temperature are maintained at the desired levels. The pressures for extraction range from about 100 to about 800 bars, from about 200 to about 600 bars, from about 300 to about 400 bars, and the temperature ranges from about 50 ° C to about 120 ° C and from 60 ° C to about 100 ° C, and from about 80 ° C to about 90 ° C. The time for extraction ranges from about 30 minutes to about 2.5 hours, from about 1 hour to about 2 hours, to about 1.5 hours. The ratio of solvent to feed is typically 17 -18 to 1 for each of the extractions by SCC02. The purified and extracted essential oil fraction is then collected in a collection vessel, set aside and stored in a dark refrigerated at 5 ° C. The C02 is recycled. The residue of the raw material of Panax 40 species is also collected from the extraction vessel, sectioned and used for additional extractions of the chemical constituents in the rhizome of Panax species. Typically, the total yield of essential oil fractions of Panax species ranges from 0.2% to 0.05% by weight dry mass of the original raw material having an essentially 100% pure essential oil fraction chemical constituent composition. Purity is measured using HPLC and OC-MS analysis (see Examples 1-4). Ginsenoside Extraction Process In one aspect the present invention comprises the extraction and concentration of the trinipepenic ginsenosides or saponins. A generalized description of this extraction step is diagrammed in Figure 2. This ginsenoside extraction process is a 3-stage solvent percolation process. The raw material for this process for ginsenosides is the residue 40 or 45 after extraction of the essential oil fraction. The extraction solvent 220, 230, 240 is typically 63% ethanol in water. In this method, the residue of Panax species and the extraction solvent are loaded in a heated extraction vessel, 50.
It can be heated to less than 100 ° C, to approximately 90 ° C, 80 ° C, or to approximately 60-60 ° C. The extraction is carried out in 1 to 4 hours, for approximately 3 hours, for approximately 2 hours. The resulting fluid extract is filtered 60. The filtrate is collected as product 310, measured for volume and weight of dry mass contained in solids after evaporation of the solvent. The extraction residue material 70 was retained in the filter. The extraction can be repeated as many times as necessary or desired. It can be repeated two or more times, etc. For example, Figure 2 shows a three-stage process, where the second stage 80 and the third stage 110 use the same methods and conditions. The Examples are seen in Examples 5 to 8 and Tables 5 to 8, respectively. Although more than 30 individual ginsenoside compounds have been detected and characterized in the genus Panax, most of these exist only in trace amounts. The seven ginsenosides (Rg ?, Re, Rf, Rbi Rc, Rb2, d) representing more than 95% of the total ginsenoside content of the Panax species were measured in the course of the extraction process using the HPLC analysis that allows the calculation of% dry weight of total and individual ginsenosides contained in the extract products Tables 5 to 20). HPLC analysis was achieved using a Shimadzu SE0405003 HPLC with analytical reference standards obtained from Chromadex, KIT-00007226-005 (ginsenoside standards set) (see examples 22-25). As shown in Tables 5 through 12, a two-stage solvent percolation process can yield a total ginsenoside yield of at least 96% for each of the four Panax species used as a raw material studied while simultaneously increasing the purity of the ginsenosides in the fraction of ginsenoside extract at least 4 times. In the particular case of P. notogensing, the ginsenoside concentration of the raw material was 12.26% by weight of mass and the concentration of ginsenoside in the ginsenoside fraction from the extraction recombination of stage I and stage II was 50.5 % in dry weight, consequently a 5-fold increase in the concentration of ginsenoside. For P. quinquefoli us, the concentration of ginsenoside in the raw material was 2.32% by weight mass and the concentration in the extract fraction of ginsenoside percolated from stage 2 was 15.2%, consequently, an increase of 6 times in the concentration of ginsenoside. For white ginseng (P. ginseng), the concentration of ginsenoside in the raw material was 3.19 and the concentration in the fraction of the ginsenoside percolate extract from stage 2 was 8.9%, consequently 3 times the increase in the concentration of ginsenoside Finally, for ginseng Red (P. ginseng), the concentration of ginsenoside in the raw material was 1.84% and the concentration in the two fractions of ginsenoside extract percolated in stage 2 was 4.6%, therefore a total increase of 2.6 times the concentration of ginsenoside. In addition, the concentration or profile distribution of the individual ginsenosides in the extract fraction obtained from the two-stage solvent percolation method was well preserved in relation to the ginsenoside profile found in each of the original raw material materials. In conclusion, a two-stage solvent percolation process is a very efficient and cost-effective method for extracting the highly purified ginsenoside fraction from the plant material of Panax species. Purification of Ginsenosides by polymeric adsorption An extract of ginsenoside purified from raw material of Panax species can be obtained by contacting an aqueous extract of a ginseng raw material with a polymeric affinity adsorbent resin for solids to adsorb the ginsenosides contained in the aqueous extract on the adsorbent resin. The bound ginsenosides are then eluted by means of methods set forth herein. Before eluting the ginsenosides, the polymeric adsorbent resin with the ginsenosides adsorbed thereon can be separated from the remainder of the extract in any convenient way, preferably by passing the extract through a column containing the resin. A variety of polymeric adsorbent resins can be used to purify ginsenosides, including, but not limited to, Amberlite XAD-2 (Rohm &; Haas), Duolite S-30 (Diamond Alkai Co.), or ADS-8 (Nankai University, Tianjin, China). ADS-8 has high affinity for triterpene saponin ginsenosides. The ADS-8 resin beads, particle size 0.5 - 0.6 mm, are a polystyrene copolymer with an ester group. It is believed that polystyrene adsorbs chemical compounds by hydrophobic interactions between the highly hydrophobic surface of polystyrene and hydrophobic sites or sorbates. The ester groups are used for the adsorption of chemical products by means of hydrogen bonding interactions. These two interactions work together to achieve high selectivity for the binding of the triterpene saponin ginsenosides. Since the hydrophobic interaction is one of the activation forces in this separation, an aqueous solution free of alcohol is the solvent used to contain the chemical constituents to be adsorbed. An alcohol is then used as the de-adsorption agent. Although several eluents can be employed to recover the ginsenosides from the polymeric adsorbent resin, in one aspect of the present invention, the eluent comprises a low molecular weight alcohol, including, but not limited to, methanol, ethanol, or propanol In a second aspect, the eluent comprises a low molecular weight alcohol and water. In another aspect, the eluent comprises a low molecular weight alcohol in a mixture with another organic solvent. In a further aspect, the eluent comprises a low molecular weight alcohol, a second organic solvent, and water. The raw materials of Panax species may have undergone one or more preliminary processes including, but not limited to, processes for removal of essential oils or solvent percolation steps, shown in Figures 1 and 2, before contacting the extract containing the ginsenoside in aqueous solution with the polymeric adsorbent resin.
The use of polymeric adsorption resins as disclosed in the present invention resulted in highly purified ginsenoside extracts of Panax species that are free from other chemical constituents, which are normally present in the natural plant material or in commercially extracted products. available. For example, the processes set forth in the present invention can result in purified ginsenoside extracts containing total ginsenosides in excess of 90% by weight dry mass. The use of the methods set forth herein makes it possible to achieve a fraction of purified ginsenoside extract of more than 95% which was measured in mass%. A generalized description of the extraction and purification from the rhizome of Panax species using adsorbent resin beads for polymer affinity is diagrammed in Figure 3. The raw material for this extraction process can be the hydroalcoholic solutions containing ginsenosides from the Figure 2, 310, 320, 330. The alcohol is evaporated from this solution 420 and then diluted with distilled water 260 to the original volume to preserve the concentration of triterpene saponin in this aqueous solution without change 420. The appropriate weight of beads of Adsorbent resin (50-75 mg of ginsenoside per gram of adsorbent resin) is washed with water and ethanol before and after being loaded onto a column. The aqueous solution containing the ginsenoside 430 is then loaded onto the column 470 at an expense of 2 to 4 bed volumes per hour. Once the column is fully loaded, the column is washed with water at an expense of 50 ml / hour to remove any impurities from the adsorbed ginsenosides 480. The elution of the adsorbed ginsenosides 490 was achieved with ethanol / water (4 / 1) as an elution solution 290 at an expense of 50 ml / hour and the elution curve for the extract was recorded. The eluate 500 consisting of the purified ginsenoside fraction was analyzed using HPLC. The results of individual experiments can be found in Examples 22 to 25 and Tables 13 to 20. Polysaccharide Extraction Process A generalized description of the extraction of the polysaccharide fraction from the rhizome of Panax species using the percolation processes by aqueous solvent and precipitation by ethanol is diagrammed in Figure 4. The raw material is the residue from the extraction by percolation of the solvent of the ginsenoside (Figure 2). The solvent is distilled water 250, 260, 270. The raw material of the waste can be extracted multiple times in hot aqueous solutions multiple times, for example, water, at approximately 100 ° C, for at least one hour, for at least two hours , for at least three hours, in an extraction vessel. Figure 4 shows 3 extractions three times with water at approximately 100 ° C. The amount of water is generally the same for the first extractions and less for the last extractions. For example, the volume of water for the first extraction 610 and the second extraction 640 was 15 ml / g of raw material residue and for the third extraction 670 was 10 ml / g of raw material residue. After each of the percolations in boiling water the resulting fluid extract is filtered 620, 640, 680. The filtrate is collected as product 710, 720, 730 and was measured for volume and solids content (dry mass weight). The extraction waste material is retained by the filter in the first stage 260 and can then be used as a raw material for the second extraction stage using the same methods, and the process can be repeated in a third step 670. The residue 690 after the third stage it is discarded. Interestingly, the extraction products 710, 720, 730 can be shown to be highly purified polysaccharides (approximately 99% pure the water-soluble Panax polysaccharide species)., insoluble in ethanol) based on a variety of tests which are discussed in Example 30. The various fractions of extracts are dried and stored separately for subsequent recombination in a wide variety of nutraceutical and pharmaceutical formulations derived from species extraction products. of Panax. Many methods are known for the removal of alcohol from the solution. If it is desired to conserve the alcohol for recycling, the alcohol can be removed from the solutions, after extraction, by distillation under reduced or normal atmospheric pressure. Alcohol can be reused. In addition, there are also many methods known in the art for the removal of water from solutions, either aqueous solutions or solutions from which the alcohol was removed. Such methods include, but are not limited to, spray drying the aqueous solutions on a suitable carrier such as, but not limited to, magnesium carbonate or maltodextrin, or alternatively, the liquid may be brought to dryness by freeze drying or drying by refractive window. When carrying out the previously described extraction methods, it was found that more than 80% d weight yield of the essential oil mass in the dried rhizome raw material originally of the Panax species can be extracted in the extract fraction of essential oil ( Stage 1) . Using the methods shown in Figure 2 of more than 98% mass weight yield of the ginsenoside chemical constituents of the original dry rhizome raw material of the Panax species can be extracted into the ginsenoside extract fraction. Furthermore, it appears that more than 99% by weight of the polysaccharide constituents of the dry rhizome raw material originally of the Panax species can be extracted into the polysaccharide fraction. Finally, the methods as set out in the present invention allow the purification (concentration) of the fraction of essential oils, fraction of ginsenosides, and the polysaccharide fraction is as high as 99% of the desired chemical constituents (essentially all essential oils, triterpene saponin, and chemical constituents polysaccharides present in the plant material of original Panax species). The specific extraction environments, extraction rates, solvents, and extraction technology used are often adjusted depending on the profile of the initial chemical constituents of the source material and the level of purification desired in the final extraction products. Specific methods as set forth in the present invention can be readily adjusted by those skilled in the art using no more than routine routine experimentation to adjust a process to consider variations of the samples in the starting materials. For example, in a particular batch of P. ginseng (White Ginseng), the initial concentrations of the essential oil, the ginsenosides and the polysaccharides are determined using methods known to those skilled in the art. A person skilled in the art can determine the amount of change from the initial concentration of the ginsenosides, for example, to the predetermined amounts of ginsenosides for the final extraction product using the extraction methods, as described herein, for achieve the desired concentration in the product of the final P. ginseng composition. Pharmaceutical Compositions In accordance with a further aspect of the invention, the new plant material of extracted Panax species or a new extract composition of Panax species can be further processed to dry, flowable powder. The powder can be used as a dietary supplement that can be added to various edible products. The powder or the final pre-determined single extract compositions of the Panax species are also suitable for use in a fast dissolving tablet. In accordance with a particular aspect of the present invention, compositions of extracted Panax species are produced to have predetermined concentrations of essential oil, gnosenoside, and polysaccharide that are greater than those found in the natural plant material or in products of extracts of conventional Panax species and / or new predetermined profiles of the three main bioactive chemical constituents, where the proportions (profiles) of the amounts (% dry weight) of essential oil / ginsenoside and / or essential oil / polysaccharide and / or ginsenoside / polysaccharide are greater or less than the profiles of the chemical constituents found in the plant material of natural Panax species or extraction products of known Panax species. Said compositions are particularly well adapted for release into the oral cavity of human subjects, for example, via a fast-dissolving tablet. In one embodiment of a method for producing an extract powder of Panax species, a composition extracted from dry Panax species is mixed with a suitable solvent, such as, but not limited to water or ethyl alcohol, together with a grade material. Suitable food using a high shear mixer and then air drying using conventional techniques to produce a powder having very small particles of Panax species particles combined with a food grade carrier. In a particular example, a composition extracted from Panax species is mixed with about twice its weight of a food grade carrier such as maltodextrin having a particle size of between 100 to about 50 microns and a ethyl alcohol solvent using a mixer of high shear stress. Inert carriers, such as silica, preferably having an average particle size in the order of about 1 to about 50 microns, can be added to improve the flow of the final powder that is formed. Preferably, said additions are up to 2% by weight of the mixture. The amount of ethyl alcohol used is preferably the minimum necessary to form a solution with an appropriate viscosity for air drying by spray. Typical amounts are in the range of from about 5 to about 10 liters per kilogram of material extracted from Panax species. The solution of the composition extracted from Panax species, maltodextrin and ethyl alcohol is air-dried by spray to generate a powder with an average particle size comparable to that of the initial carrier material. In a second embodiment, a composition extracted from Panax species and a food grade carrier, such as magnesium carbonate, whey protein, or maltodextrin, are mixed dry, followed by mixing in a high shear mixer. containing a suitable solvent, such as water or ethyl alcohol. The mixture is then dried by freeze drying or refraction window drying. In a particular example, the material of the composition extracted from Panax species is combined with food grade material approximately one and one and one half times by weight of the composition extracted from Panax species, such as magnesium carbonate having a size of average particle of approximately 20 to 200 micrometers. Inert graters such as silica having a particle size of about 1 to about 50 microns may be added, preferably in an amount of up to 2% by weight of the mixture, to improve the flow of the mixture. Magnesium carbonate and silica are then mixed dry in a high shear mixer, similar to a food processor type mixer, operating at hundreds of rpm. The material of the composition extracted from Panax species is then heated until it flows like a heavy oil. Preferably it is heated to approximately 50 ° C. The extracted composition of heated species is then added to the pulverized mixture of magnesium carbonate and silica which are being mixed in the high shear mixer. The mixing preferably continues until the particle sizes are in the range of from about 250 microns to about 1 millimeter. Between about 2 to about 10 liters of cold water (preferably at about 4 ° C) per kilogram of material of the composition extracted from Panax species is introduced into a high shear mixer. The mixture of composition extracted from Panax species, magnesium carbonate, and silica are slowly and increasingly introduced into the high shear mixer while mixing. If necessary, an emulsifying agent such as carboxymethyl cellulose or lecithin may also be added to the mixture. Sweetening agents such as Sucralose or Acesulfame K up to about 5% by weight may also be added at this stage if desired. Alternatively, the Stevia rebaudiana extract, a very sweet-tasting dietary supplement, may be added instead of or in conjunction with a specific sweetening agent (for simplicity, Stevia will be referred to herein as a sweetening agent). After the mixture is completed, it is dried using freeze drying or refraction window drying. The resulting dry flowable powder of the composition material extracted from Panax species, magnesium carbonate, silica and optional emulsifying agent and optional sweetener has an average particle size comparable to that of the initial carrier and a predetermined Panax species extraction composition. According to another embodiment, a material of the composition extracted from Panax species is combined with about an equal weight of food grade carrier such as whey protein, preferably having a particle size of between about 200 to about 1000. micrometers Inert carriers such as silica having a particle size of between about 1 to about 50 microns, or carboxymethyl cellulose having a particle size of between about 10 to about 100 microns, can be added to improve the flow of the mixture. Preferably, the addition of an inert carrier is no more than 2% by weight of the mixture.
The whey protein and the inert ingredient are then dry blended in a mixer-type food processor operating at more than 100 rpm. The material of the Panax species extraction composition is heated until it flows as a heavy oil (preferably heated to 50 ° C). The heated Panax species extraction composition is then increasingly added to the whey protein and the inert carrier which are being mixed in the food processor type mixer. The mixture of the Panax species extraction composition and the whey protein and the inert carrier is further mixed until the particle sizes are in the range of about 250 microns to about 1 millimeter. Then, 2 to 10 liters of cold water are placed in a high-shear mixer. (preferably at about 4 ° C) per kilogram of the paste mixture. The mixture of the extracting composition of Panax species, whey protein, and inert carrier is increasingly introduced into the cold water containing the high shear mixer while mixing. Sweetening agents or other flavor additives of up to 5% by weight may be added at this stage, if desired. After completing the mixing, the mixture is dried using freeze drying or refraction window drying. The flowable dry powder resulting from the extracting composition of Panax species, whey protein, inert carrier and optional sweeteners have a particle size of about 150 to 700 microns and a unique predetermined Panax species extraction composition. In a further embodiment a predetermined Panax species extraction composition is dissolved in a C02 SFE fluid which is then absorbed onto a suitable food grade carrier such as maltodextrin, dextrose or starch. Preferably, the SFE C0 is used as the solvent. Specific examples include starting with a composition extracted from new Panax species and adding one by one equivalent to 50% by weight on the material extracted from Panax species of the food grade carrier having a particle size of between about 100. at approximately 150 micrometers. This mixture is placed in a chamber that contains mixing blades and can be compressed and heated. The chamber is compressed with C02 at a pressure in the range of 110 psi to about 8000 psi and adjusted to a temperature in the range of about 20 ° C to about 100 ° C. The exact pressure and temperature are selected to place the C02 in a supercritical fluid state. Once the C02 in the chamber is in the supercritical state, the Panax species extraction composition is dissolved. The mixing blades agitate the carrier powder so that it is in intimate contact with the supercritical C02 which contains the dissolved Panax species extract material. The mixture of supercritical C0, dissolved Panax species extraction material, and the carrier powder is then vented through a hole in the chamber, which is at a pressure and temperature that does not support the supercritical state for C02. The C02 is thus dissipated as a gas. The resulting powder in the collection container is the carrier powder impregnated with the new predetermined Panax species extraction composition. The powder has an average particle size comparable to that of the initial carrier material. The resulting powder is dry and flowable. If necessary, the flow characteristics can be improved by adding inert ingredients to the initial carrier powder such as silica to about 2% by weight as discussed previously. In embodiments wherein the composition of the extract of Panax species with a predetermined composition or profile is to be included in an oral rapid dissolution tablet as described in U.S. Pat. 5, 298,261, the single extract can be used "pure", that is, without some additional components, which are then added in the process of forming the tablet as described in the cited patent. This method then highlights the need to take the unique composition of Panax species extract for a flowable dry powder that is then used to make the tablet. Once a dry powder of the Panax species extraction composition is obtained, such as by means of the methods discussed herein, it can be distributed for use, for example, as a dietary supplement or for other uses. In a particular embodiment the new powder of the Panax species extraction composition is mixed with other ingredients to form a tabletting composition of powder that can be formed into tablets. The tableting powder is first moistened with a solvent comprising alcohol, alcohol and water, or other suitable solvents in an amount sufficient to form a thick, pasty consistency. Suitable alcohols include, but are not limited to, ethyl alcohol, isopropyl alcohol, isopropyl alcohol containing denatured ethyl alcohol, acetone and denatured ethyl alcohol containing acetone. The resulting paste is then compressed in a tablet mold. An automatic tablet molding system, such as that described in U.S. Pat. No. 5,407,339, can be used. The tablets can then be removed from the mold and dried, preferably by air drying for at least several hours at a sufficiently high temperature to release the solvent used to moisten the tabletting powder mixture, typically between about 70 ° C to about 85 ° C. The dry tablet can then be packaged for distribution. The methods and compositions of the present invention comprise compositions comprising unique Panax species extract compositions in the form of a paste, resin, oil, or powder. One aspect of the present invention comprises liquid preparation compositions of unique Panax species extract compositions. Liquid preparations for oral administration may take the form of, for example, syrup solutions or suspensions, or they may be presented as a dry product for reconstitution with water or with another suitable vehicle before administration. Said liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (eg, sorbitol syrup, methyl cellulose, or hydrogenated edible fats); emulsifying agents (for example, lecithin or gum arabic); non-aqueous vehicles (for example almond oil, oily esters or ethyl alcohol), preservatives (for example, methyl or propyl p-hydroxybenzoates or sorbic acid); and artificial or natural colors and / or sweeteners. Compositions of the liquid preparations can be administered to humans or animals in pharmaceutical carriers known to those skilled in the art. Said pharmaceutical carriers include, but are not limited to, capsules, pills, syrups, sprays, rinses, and mouthwashes. One aspect of the present invention comprises compositions of a dry powder Panax species extraction composition. Said dry powder compositions may be prepared in accordance with methods described herein and by means of other methods known to those skilled in the art such as, but not limited to, air mist drying, freeze drying, vacuum drying, and drying by refraction window. The combined dry powder compositions can be incorporated into a pharmaceutical carrier such as, but not limited to, tablets or capsules, or reconstituted in a beverage such as tea.
Although the extraction techniques described herein are discussed in terms of Panax species, it should be recognized that compositions of the present invention may also comprise, in the form of a dry flowable powder or other forms, extracts from other vegetables such as , but it is not limited to turmeric, boswellia, guarana, cherry, lettuce, Echinacia, betel leaves, Areca ca techu, muirá puama, ginger, willow, sum, kava, weed, ginko bilova, mate ', garlic , vine, selenium indicator plant (astragalus) of Arctic root, eucommia, gastropodia, and uncaria, or pharmaceutical or nutraceutical agents. The present invention comprises compositions comprising unique compositions of Panax species extract in tablet formulations and methods of making such tablets. A tableting powder can be formed by adding about 1% to 40% by weight of the sprayed Panax species extract composition, with between 30% to about 80% by weight of a water-dispersible dry absorbent such as, but not limited to, lactose. Other dry additives such as, but not limited to, one or more sweetening, flavoring and / or coloring agents, a binder such as acacia or gum arabic, a lubricant, a disintegrant, and a regulator can also be added to the tableting powder. The dry ingredients are screened at a particle size of between about 50 to about 150 meshes. Preferably, the dry ingredients are screened to a particle size of between about 80 to 100 mesh. The present invention comprises compositions comprising n tablet formulations and methods of making said tablets. Preferably, the tablet has a formulation that results in rapid dissolution or disintegration in the oral cavity. The tablet is preferably a homogeneous composition that rapidly dissolves or disintegrates in the oral cavity to release the contents of the extract for a period of about 2 seconds or less than 60 seconds or more, preferably about 3 to about 45 seconds, and most preferably between about 5 to about 15 seconds. Various formulations for fast dissolving tablets known in the art can be used. Representative formulations are described in U.S. Pat. Nos. 5,464,632; 6, 106, 861; 6, 221, 392; 5,298,261; 6,221,392; and 6,200,604; whose complete contents of each one are expressly incorporated as reference to the present. For example, U.S. Pat. No. 5,298,261 discloses a freeze drying process. This process involves the use of freezing and then drying under vacuum to remove water by sublimation. Preferred ingredients include hydroxyethyl cellulose, such as Natrosol from Hercules Chemical Company, added to between 0.1% and 1.5%. Additional components include maltodextrin (Maltrin, M-500) at between 1% and 5%. These amounts are solubilized in water and used as an initial mixture to which the extracting composition of Panax species is added, along with flavorings, sweeteners, such as Sucralose or Acesulfame K, and emulsifiers such as BeFlora and BeFloraPlus which are extracts of the golden bean. A particularly preferred tabletting composition or powder contains about 10% to 60% by weight of the extract composition of Panax powder species. And about 30% to about 60% of a water soluble diluent. Suitable diluents include lactose, dextrose, sucrose, mannitol, and other similar compositions. Lactose is a preferred diluent but mannitol adds a pleasant cold sensation and additional sweeteners in the mouth. More than one diluent can be used.
A sweetener may also be included, preferably in an amount between 3% to about 40% by weight depending on the desired sweetness. Preferred sweetening substances include sugar, saccharin, sodium cyclamate, aspartame, and Stevia extract used individually or in combination, although alternatively other sweeteners could be used. Flavors such as peppermint, cinnamon, citrus, (for example, lemon or orange), mocha, and others may also be included, preferably in an amount between about 0.001% to about 1% by weight. A colorant may also be added, including natural and / or synthetic colors which are known in the art as safe and acceptable for use in food products or drugs. The colorant, if added may be added in an amount of between about 0.5% to about 2% by weight. Typically this composition for tableting will maintain its shape without the use of a binder. However, if necessary several binders are suitable and can be added in an amount of between about 5% to about 15% as necessary. Preferred binders are acacia or gum arabic. Alternative binders include sodium alginate, Irish moss extract, Panwar gum, Ghatti gum, Isapol shell mucilage, carboxymethyl cellulose, hydroxyethyl cellulose, methyl cellulose, polyvinyl pyrrolidone, VEEGUM® (RT Vandervilt Co., Inc., Norwalk, Conn. .), larch arabogalactam, gelatin, kapa carrageenans, maleic anhydride copolymers with ethylene or methyl ether. A tablet in accordance with this aspect of this invention does not require a lubricant to improve the flow of the powder for the manufacture of the tablet. However, if desired, preferred lubricants include talc, magnesium stearate, calcium stearate, stearic acid, hydrogenated vegetable oils, and carbowax in an amount of between about 2% to about homogeneity, the tablet can alternatively be comprised of 10% by weight. weight of powdered Panax species regions. Similarly, a disintegrant does not appear necessary to produce fast dissolving tablets using the tablet compositions herein. However, a disintegrant may be included to increase the rate at which a resulting tablet dissolves in the mouth. If desired, between about 0.5% to about 1% by weight of a disinfectant can be added. Preferred disintegrants include starches, clays, cellulose, algines, gums, cross-linked polymers (including croscarmellose, crospovidone, and sodium starch glycolate), VEEGUM®HV, agar, bentonite, natural sponge, cation exchange resins, alginic acid, guar gum , citrus pulp, sodium lauryl sulfate in an amount of about 0.5% to about 1% of the total mass of the tablet. Generally it is also unnecessary to regulate the composition of the tablet. However, a regulator can be beneficial in specific formulations. Preferred regulatory agents include mono- and di-sodium phosphates and borates, basic magnesium carbonate and combinations of aluminum and magnesium hydroxide. In a preferred implementation, the tableting powder is made by mixing, in a dry powdered form the various components that were described above, for example, the active ingredient (composition of Panax species extract), diluent, sweetener additive, and flavorings , etc. An excess in the range of about 10% to about 15 of the active extract of the active ingredient can be added to compensate for losses during the subsequent tabletting processing. The mixture is screened through a screen with a mesh size preferably in the range of about 80 mesh to about 100 mesh to ensure a generally uniform particle composition. The tablet can be of any desired size, shape, weight, or consistency. The total weight of the composition composition of the Panax species extract in the form of a flowable dry powder in an individual oral dosage is typically in the range of about 40 mg to about 600 mg. An important consideration is that the tablet is intended to dissolve in the mouth and therefore should not be in a way that encourages the tablet to be swallowed. The largest tablet, the smaller one is probably to be swallowed accidentally, but it will take more time to dissolve or disintegrate. In a preferred form, the tablet is a disk or wafer of about 0.15 inches to about 0.5 inches in diameter and about 0.08 inches to about 0.2 inches in thickness, and has a weight of between about 160 mg to about 1,200 mg. In addition to the disk, the coin or wafer form, the tablet can be in the form of a cylinder, sphere, cube, or other shapes. The tablet is preferably extract composition separated by extract regions of species other than Panax in periodic sequences or not, which may give the tablet a mottled appearance with different colors or shades of colors associated with the extract regions of species of Panax and the extract regions of species other than Panax. The unique compositions of extract compositions of Panax species may also comprise compositions of Panax species in an amount between about 10 mg and about 750 mg per dose. The essential composition of the new Panax species extract composition may vary wherein the essential oil is in an amount between about 0.1 mg and about 10.0 mg. The total ginsenoside composition of the new compositions of Panax species can vary between about 1.0 mg and about 150 mg per dose wherein the mass% by weight of the ginsenoside constituents in the single extract composition of Panax species is higher in relation to to the weight% of ginsenoside mass found in the plant material of natural Panax species or extracts and beverages of conventional Panax species. The polysaccharide composition of Panax species of the new Panax species extract composition can vary between about 1.0 mg and about 400 mg wherein the mass% weight of the polysaccharide constituents is substantially increased relative to the mass% by weight of the polysaccharide. polysaccharides found in the plant material of Panax species p natural extracts or beverages of conventional Panax species. Finally, the mass weight ratios of the three major beneficial bioactive chemical constituents (essential oil, ginsenosides, and polysaccharides) derived from the Panax species can be altered to produce additional new Panax extract composition profiles for oral delivery in humans using the dose ranges mentioned previously. One exemplary 275 mg tablet contains about 150.0 mg of the single composition of pulverized predetermined Panax species extract, about 12.5 mg of Stevia extract, about 35.5 mg of carboxymethyl cellulose, and about 77.0 mg of lactose (see Example 1). Further exemplary formulations for 300 mg tablets and 350 mg of Panax species extraction composition can be found in Examples 2 and 3. The present invention comprises methods of using compositions comprising unique extracting compositions of Panax species described herein. . Methods of providing dietary supplementation are contemplated. Said compositions may additionally comprise vitamins, minerals and antioxidants. The compositions set forth herein can also be used in the methods of treating various physiological, psychological, and medical conditions. The standardized, reliable and new Panax species extraction compositions of the present invention are used to prevent and treat cardiovascular and cerebrovascular diseases and hypercholesterolemia. The compositions of the present invention can be used to provide cytoprotection and neural protection which are important for the prevention of heart attacks and strokes. The new extracting compositions of Panax species are used to provide potent antioxidant activity to human and animal cells and to cell membranes and protect low density lipoproteins against oxidative damage. The pathologies that are related to damage to the oxygen radical include, but are not limited to, cardiovascular disease, cerebrovascular disease (stroke), arthritis, inflammation, liver disorders, HIV, and cancer. The new extracting compositions of Panax species provide inhibition of platelet agglutination which is important for the prevention of heart attacks and strokes. In addition, the extracting compositions of Panax species of the present invention are used to provide immune enhancement which important protection against infectious diseases, cancer and various liver and lung diseases. Extract compositions of Panax species of the present invention have anti-inflammatory activity and anti-diabetic activity. The new extracting compositions of Panax species are also used to prevent or treat neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. In addition, new extract compositions of Panax species are used to improve memory and knowledge, alleviate chronic fatigue syndromes, and improve male erectile function. These and other related pathologies are prevented or treated by administering an effective amount of the new Panax species extraction compositions of the present invention. The new extracting compositions of Panax species can be administered daily, for one or more times, for the effective treatment of acute or chronic conditions. A method of the present invention comprises administering at least once a day a composition comprising compounds constituting Panax species. The methods also comprise administering said compositions more than once a day, more than twice a day, more than three times a day and in an interval from 1 to 15 times per day. Said administration may be continuously as in each day for a period of days, weeks, months or years, or may occur at specific times to treat or prevent specific conditions. For example, a person can administer extract compositions of Panax species at least once a day for years to improve mental focus, knowledge, and relieve chronic fatigue, or to prevent cardiovascular disease or stroke. All patents, patent applications and references included herein are specifically incorporated by reference in their entirety. It will be understood, of course, that the foregoing concerns only exemplary embodiments of the present invention and that numerous modifications or alterations may be made to it without departing from the spirit and scope of the invention as set forth in this description. Although the exemplary embodiments of the present invention describe in detail methods and compositions for Panax extracts, there are numerous modifications or alterations that may themselves suggest to those skilled in the art for use of the methods and compositions herein for Panax extracts. The present invention is further illustrated by way of the examples contained therein, which are provided for clarity of understanding. Exemplary modalities should not be constructed in any way to impose limitations on the scope of the same. On the contrary, it will be clearly understood that it is possible to resort to several other modalities, modifications, and equivalents thereof which, after reading the description herein, can suggest themselves to those skilled in the art without departing from the spirit. of the present invention and / or the scope of the appended claims. EXAMPLIFICATION EXAMPLE 1. Preparation of the Fraction of Essential Oil from P. notog ± nsßng 30 g of rhizome raw material of P. notoginseng were crushed, passed through a # 20 or # 8 mesh screen, and then the resulting rhizome powders were collected. The ground raw material was loaded in a 250 ml Supercritical Fluid Extraction (SFE) vessel connected to a Supercritical Fluid Extraction Unit for Applied Separations model Spe-ed SFE-2 (Allentown, PA). The non-adsorbed cotton ball was packed at the top and bottom of the extraction vessel to avoid raw botanical flow along with the C02 stream. The oven was preheated to the desired temperature of 89 ° C before the packaged container was loaded. After the vessel was connected to the furnace, the extraction system was tested for leaks by compressing the system with C02 (~ 60 bar), and purged. The system was closed and pressurized to the desired pressure of 400 bar using a pump for liquid C02 activated by air. The system was allowed to equilibrate for approximately 3 minutes. A sampling vial (40 ml) was weighed and connected to the sampling port at room temperature. The extraction was initiated by flowing C02 at a speed of 5.5 - 6.0 g / min, which was controlled by means of a needle valve heated to 90 ° C to prevent plugging of the valve by dry ice during decompression. The ratio of solvent to raw material used was about 17 -18/1 and the extraction time was 90 minutes, the total yield of the essential oil fraction of P. notoginseng was about 0.3% by weight versus the weight of the initial raw material, and the weight in percent of the essential oil in this fraction of essential oil extract was 100%. The raw material residue was removed and used in additional extraction steps to extract the ginsenoside and polysaccharide fractions (see Example 9). The results of this extraction process are shown in Example 18. EXAMPLE 2. Preparation of the Fraction of Essential Oil from P. quinqué folius 30 g of rhizome raw material of P. quinquefoli were crushed, passed through from a # 20 or # 8 mesh screen, and then the resulting rhizome powders were collected. The ground raw material was loaded in a 250 ml supercritical fluid extraction (SFE) vessel connected to a Supercritical Fluid Extraction Unit for Applied Separations model Spe-ed SFE-2 (Allentown, PA). The non-adsorbed cotton ball was packed at the top and bottom of the extraction vessel to avoid raw botanical flow along with the C02 stream. The oven was preheated to the desired temperature of 89 ° C before the packaged container was loaded. After the vessel was connected to the furnace, the extraction system was tested for leaks by compressing the system with C02 (~ 60 bar), and purged. The system was closed and pressurized to the desired pressure of 400 bars using a pump for air-activated C02. The system was allowed to equilibrate for approximately 3 minutes. A sampling vial (40 ml) was weighed and connected to the sampling port at room temperature. The extraction was initiated by flowing C02 at a speed of 5.5 - 6.0 g / min, which was controlled by means of a needle valve heated to 90 ° C to prevent plugging of the valve by dry ice during decompression. The ratio of solvent to raw material used was about 17 -18/1 and the extraction time was 90 minutes, the total yield of the essential oil fraction of P. quinquefolius was about 0.2% by weight versus the weight of the initial raw material, and the weight in percent of the essential oil in this fraction of essential oil extract was 100%. The raw material residue was removed and used in additional extraction steps to extract the ginsenoside and polysaccharide fractions (see Figure 2 and Example 6). The results of this extraction process are shown in Example 19. Example 3. Preparation of the Fraction of Essential Oil from White Ginseng (P. ginseng) 30 g of white ginseng rhizome raw material (P. ginseng) were ground. ), were passed through a # 20 or # 8 mesh screen, and then the resulting rhizome powders were collected. The crushed raw material was loaded in a 250 ml supercritical fluid extraction (SFE) vessel connected to a Supercritical Fluid Extraction Unit for Applied Separations model Spe-ed SFE-2 (Allentown, PA). The non-adsorbed cotton ball was packed at the top and bottom of the extraction vessel to avoid raw botanical flow along with the C02 stream. The oven was preheated to the desired temperature of 89 ° C before the packaged container was loaded. After the vessel was connected to the furnace, the extraction system was tested for leaks by compressing the system with C0 (~ 60 bar), and purged. The system was closed and pressurized to the desired pressure of 400 bar using a pump for liquid C02 activated by air. The system was allowed to equilibrate for approximately 3 minutes. A sampling vial (40 ml) was weighed and connected to the sampling port at room temperature. The extraction was initiated by flowing C02 at a speed of 5.5 - 6.0 g / min, which was controlled by means of a needle valve heated to 90 ° C to prevent plugging of the valve by dry ice during decompression. The ratio of solvent to raw material used was approximately 17 -18/1 and the extraction time was 90 minutes. The total yield of the essential oil fraction of P. ginseng was approximately 0.5% by weight versus the weight of the initial raw material, and the weight in percent of the essential oil in this fraction of essential oil extract was 100% . The raw material residue was removed and used in additional extraction steps to extract the ginsenoside and polysaccharide fractions (see Figure 2 and Example 7). The results of this extraction process are shown in Example 20. EXAMPLE 4. Preparation of the Fraction of Essential Oil from Red Ginseng (P. Ginseng) 30 g of Red Ginseng rhizome raw material were grinded (P. ginseng ), were passed through a # 20 or # 8 mesh screen, and then the resulting rhizome powders were collected. The ground raw material was loaded in a 250 ml supercritical fluid extraction (SFE) vessel connected to a Supercritical Fluid Extraction Unit for Applied Separations model Spe-ed SFE-2 (Allentown, PA). The non-adsorbed cotton ball was packed at the top and bottom of the extraction vessel to avoid raw botanical flow along with the C02 stream. The oven was preheated to the desired temperature of 89 ° C before the packaged container was loaded. After the vessel was connected to the furnace, the extraction system was tested for leaks by compressing the system with C02 (~ 60 bar), and purged. The system was closed and pressurized to the desired pressure of 400 bar using a pump for liquid C02 activated by air. The system was then left to equilibrate for approximately 3 minutes. A sampling vial (40 ml) was weighed and connected to the sampling port at room temperature. The extraction was initiated by flowing C02 at a speed of 5.5 - 6.0 g / min, which was controlled by means of a needle valve heated to 90 ° C to prevent plugging of the valve by dry ice during decompression. The ratio of solvent to raw material used was approximately 17-18/1 and the extraction time was 90 minutes. The total yield of the essential oil fraction of P. ginseng was approximately 0.4% by weight versus the weight of the initial raw material, and the weight in percent of the essential oil in this fraction of essential oil extract was 100% . The raw material residue was removed and used in additional extraction steps to extract the ginsenoside and polysaccharide fractions (see Figure 2 and Example 8). The results of this extraction process are shown in Example 21.
EXAMPLE 5. Preparation of the Fraction Ginsenoside from P. notoginseng The residue of 30 g of crushed rhizome (mesh # 20) after the essential oil was extracted (see Example 1, Step 1) from P. notoginseng , was extracted using a "percolation" process by solvent in three stages. In this method, the residue of 1 extraction of the essential oil (Step 1, Example 1, Figure 1) and 200 ml of solvent for extraction (63% ethanol in water) was charged into four heated flasks in a water bath (50 - 60 ° C) with agitation. The extraction was carried out for two hours. The resulting fluid extract was filtered using a Fisher P8 filter of 20 μm. The filtrate was collected as product and was measured for volume and solids content dry mass that was measured in grams). The extraction residue, material retained on the 20 micron filter, was then used as a raw material for the second extraction stage using the same methods, and the process was repeated in the third stage. The residue was removed and used for Step 4 extraction of the polysaccharide fraction from P. notoginseng (Example 13, Figure 4). The results are tabulated in Tables 5 and 6.
Table 5. Percolation Method by Solvent in Three Stages for P. notoginseng A = Stage B = Extraction Solvent C = Raw Material (grams) D = Solvent (ml) E = Extraction Time (hours) F = Dry Weight Extracted (grams) G = Performance (%) H = Content of Ginsenosides (%) Table 6. Distribution of the Seven Main Ginsenosides in extracts of P. notoginseng A = Stage; B = Rgl (%); C = Re (%); D = Rf (%) E = Rb? (%); F = Rc (%); G = Rb2 (%); H = Rd (%); I = Total *; J = Performance of the Ginsenoside Stage (%) ** * Total ginsenosides were calculated based on the seven calibrated ginsenosides. There are some other ginsenosides detected (< 5%) but they were not calibrated due to the lack of reference standard. Therefore, the total ginsenoside content is somewhat higher than the number cited in the table. ** The performance of total ginsenoside in the extracts that were accumulated by adding each extraction step. EXAMPLE 6. Preparation of the Ginsenoside Fraction from P. quinqu folius A typical experimental example of solvent extraction in 2 steps of the ginsenoside fraction is as follows: the residue of the 30 g of crushed rhizome (mesh # 20) after that the essential oil (see Example 1, Step 1) was extracted from P. quinqué folius, extracted using a three-step "percolation" process by solvent. In this method, the residue from the extraction of the essential oil (Step 1, Example 1, Figure 1) and 200 ml of solvent for extraction (63% ethanol in water) was charged in four flasks heated in a water bath (50 - 60 ° C) with agitation. The extraction was carried out for two hours. The resulting fluid extract was filtered using a Fisher P8 filter of 20 μm. The filtrate was collected as a product and was measured for volume and solids content (dry mass that was measured in grams). The extraction residue, material retained on the 20 micron filter, was then used as a raw material for the second extraction stage using the same methods, and the process was repeated in the third stage. The residue was removed and used for Step 4 extraction of the polysaccharide fraction of P. quinquefolius (Example 14, Figure 4). The results are tabulated in Tables 7 and 8. Table 7. Percolating Method by Solvent in Three Stages for P. quinquefolius A = Stage B = Extraction Solvent C = Raw Material (grams) D = Solvent (ml) E = Extraction Time (hours) F = Dry Weight Extracted (grams) G = Performance (%) H = Content of Ginsenosides (%) ) Table 8. Distribution of the Seven Main Ginsenosides in extracts of P. quinquefolius A = Stage; B = Rgi (%); C = Re (%); D = Rf (%) E = Rbl (%); F = Rc (%); G = Rb2 (%); H = Rd (%); I = Total *; J = Performance of the Ginsenoside Stage (%) ** *, **, see Table 5 EXAMPLE 7. Preparation of the Ginsenoside Fraction from White Ginseng (P. ginseng) A typical experimental example of solvent extraction in 2 Stages of the ginsenoside fraction are as follows: the residue of the 30 g of crushed rhizome (mesh # 20) after the essential oil was extracted (see Example 1, Step 1) from white ginseng (P. ginseng) , was extracted using a "percolation" process by solvent in three stages. In this method, the residue from the extraction of the essential oil (Step 1, Example 3, Figure 1) and 200 ml of solvent for extraction (63% ethanol in water) was charged in four flasks heated in a water bath (50 - 60 ° C) with agitation. The extraction was carried out for two hours. The resulting fluid extract was filtered using a Fisher P8 filter of 20 μm. The filtrate was collected as a product and was measured for volume and solids content (dry mass that was measured in grams). The extraction residue, material retained on the 20 micron filter, was then used as a raw material for the second extraction stage using the same methods, and the process was repeated in the third stage. The residue was removed and used for Step 4 extraction of the white ginseng polysaccharide fraction (Example 15, Figure 4). The results are tabulated in Tables 9 and 10. Table 9. Percolation Method by Solvent in Three Stages for White Ginseng (P. ginseng) A = Stage B = Extraction Solvent C = Raw Material (grams) D = Solvent (ml) E = Extraction Time (hours) F = Dry Weight Extracted (grams) G = Performance (%) H = Content of Ginsenosides (% Table 10. Distribution of the Seven Main Ginsenosides in White Ginseng Extracts (P. ginseng) A = Stage; B = Rgl (%); C = Re (%); D = Rf (%) E = Rbl (%); F = Rc (%); G = Rb2 (%); H = Rd. { %); I = Total *; J = Performance of the Ginsenoside Stage (%) ** *, **, see Table 5 EXAMPLE 8. Preparation of the Ginsenoside Fraction from Red Ginseng (P. ginseng) A typical experimental example of solvent extraction in 2 Stages of the ginsenoside fraction are as follows: the residue of the 30 g of crushed rhizome (mesh # 20) after the essential oil was extracted (see Example 1, Step 1) from red ginseng (P. ginseng ), was extracted using a solvent "percolation" process in three stages. In this method, the residue from the extraction of the essential oil (Step 1, Example 4, Figure 1) and 200 ml of solvent for extraction (63% ethanol in water) was charged in four flasks heated in a water bath (O - 60 ° C) with agitation. The extraction was carried out for two hours. The resulting fluid extract was filtered using a Fisher P8 20 μm filter. The filtrate was collected as a product and was measured for volume and solids content (dry mass that was measured in grams). The extraction residue, material retained on the 20 micron filter, was then used as a raw material for the second extraction stage using the 'rrms'' methods and the process was repeated in the third stage. The residue was removed and 'used for' Stage 4 extraction of the polysaccharide fraction from > red ginseng '(Example V6, Figure 4). The results are tabulated in Tables JJ 'and 12. • •'. . • ••; Table 11. Percolation Method by Solvent in Three Stages for Red Ginseng (P. ginseng) 1 • 'A = Stage B: = -' Solventé -de Extraction? ', I \. -, = Mátéf-ia 'Prima (grams))' - • "'D = Solvent (ml): E, = Extraction Time (hours) F = Dry Weight Extracted (grams) G = Performance (%) H = Content of Ginsenosides (%) Table 12. Distribution of the Seven Main Ginsenosides in extracts of Red Ginseng (P. ginseng) A = Stage; B = Rgl (%); C = Re (%); D = Rf (%) E = Rbl (%); F = Rc (%); G = Rb2 (%); H = Rd (%); I = Total *; J = Performance of the Ginsenoside Stage (%) ** *, **, see Table 5 EXAMPLE 9. Purification by Polymer Adsorption of the Ginsenoside Fraction from P. notoginseng In a typical experiment (Stage 3, Figure 3) , the fraction of extract, of ginsenoside, percolated by solvent in two stages obtained from the 30 g of original raw material of P. notoginseng, was evaporated first under reduced atmospheric pressure to eliminate the ethanol and then diluted with water to the original volume for keep the concentration of triterpene saponin unchanged. 40 g of ADS-8 adsorbent polymer resin (Nankai University, Tianjin, China) were washed with water and ethanol before and after being loaded onto a column (L = 50 cm x 1 cm D.I., ~ 40 cm3). The aqueous extract fraction of ginsenoside was loaded on the column at an expense of 80 to 100 ml / hr. The optimal expenditure through the resin bed was in the range of 2 to 4 bed volumes / hr. The volume and concentration that passed through the polymeric adsorbent resin was measured and recorded to determine the breaking curve. Once the column was fully charged, the column was washed with 400 ml of water at an expense of 50 ml / hr to remove the impurities of the adsorbed ginsenosides. Elution of ginsenosides was then achieved with 150 ml of ethanol / water (4/1) as an elution solvent at an expense of 50 ml / hour and the elution curve was recorded. For the extract, the loading capacity of the ADS-8 adsorbent resin was approximately 50 to 75 mg of ginsenoside per gram of adsorbent resin. The results of these experiments are tabulated in Tables 13 and 14.
Table 13. Yield of Ginsenoside after column chromatography using ADS-8 resin A = Quantity (g or ml) B = Dry weight (g) C = Yield (% by weight) D = Content of Ginsenoside (%) E = Yield of Ginsenoside (%) Table 14. Comparison of the Distribution of Ginsenoside in Peak Elution with Ginseng Root Raw Material and Percolation Extract by Solvent (% dry weight) EXAMPLE 10. Purification by Polymer Adsorption of the Ginsenoside Fraction from P. quinguerolius In a typical experiment (Stage 3, Figure 3), the ginsenoside fraction of the extract, percolated by solvent in two stages, obtained from the 30 g of raw material of P. quinquefolius, evaporated first under reduced atmospheric pressure to remove ethanol and then diluted with water to the original volume to preserve the concentration of triterpene saponin without change. 40 g of ADS-8 adsorbent polymer resin (Nankai University, Tianjin, China) were washed with water and ethanol before and after being loaded onto a column (L = 50 cm x 1 cm D.I., ~ 40 cm3). The aqueous extract fraction of ginsenoside was loaded on the column at an expense of 80 to 100 ml / hr. The optimal expenditure through the resin bed was in the range of 2 to 4 bed volumes / hr. The volume and concentration that passed through the polymeric adsorbent resin was measured and recorded to determine the breaking curve. Once the column was fully charged, the column was washed with 400 ml of water at an expense of 50 ml / hr to remove the impurities of the adsorbed ginsenosides. Elution of ginsenosides was then achieved with 150 ml of ethanol / water (4/1) as an elution solvent at an expense of 50 ml / hour and the elution curve was recorded. For the extract, the loading capacity of the ADS-8 adsorbent resin was approximately 50 to 75 mg of ginsenoside per gram of adsorbent resin. The results of these experiments are tabulated in Tables 15 and 16. Table 15. Yield of Ginsenoside after column chromatography using ADS-8 resin A = Quantity (g or ml) B = Dry weight (g) C = Yield (% by weight) D = Ginsenoside content (%) E = Yield of Ginsenoside (%) Table 16. Comparison of Ginsenoside Distribution in Peak Elution with Ginseng Root Raw Material and Solvent Percolation Extract (% dry weight) EXAMPLE 11. Purification by Polymer Adsorption of the Ginsenoside Fraction from White Ginseng (P. ginseng) In a typical experiment (Step 3, Figure 3), the ginsenoside fraction of the extract, percolated by solvent in two steps obtained from the 22 g of original raw material of white ginseng (P. ginseng), it was first evaporated under reduced atmospheric pressure to eliminate the ethanol and then diluted with water to the original volume to conserve the triterpene saponin concentration without change. 40 g of ADS-8 adsorbent polymer resin (Nankai University, Tianjin, China) were washed with water and ethanol before and after being loaded onto a column (L = 50 cm x 1 cm D.I., ~ 40 cm3 ). The aqueous extract fraction of ginsenoside was loaded on the column at an expense of 80 to 100 ml / hr. The optimal expenditure through the resin bed was in the range of 2 to 4 bed volumes / hr. The volume and concentration that passed through the polymeric adsorbent resin was measured and recorded to determine the breaking curve. Once the column was fully charged, the column was washed with 400 ml of water at an expense of 50 ml / hr to remove the impurities of the adsorbed ginsenosides. The elution of ginsenosides was then achieved with 150 ml of ethanol / water (4/1) as an elution solvent at an expense of 50 ml / hour and the elution curve was recorded. For the extract, the loading capacity of the ADS-8 adsorbent resin was approximately 50 to 75 mg of ginsenoside per gram of adsorbent resin. The results of these experiments are tabulated in Tables 17 and 18. Table 17. Yield of Ginsenoside after column chromatography using ADS-8 resin A = Quantity (go ml) B = Dry weight (g) C = Yield (% by weight) D = Ginsenoside content (%) E = Yield of Ginsenoside (%) Table 18. Comparison of Ginsenoside Distribution in Elution in Peak with the Raw Material of Ginseng Root and Solvent Percolation Extract (% in dry weight) EXAMPLE 12. Purification by Ginsenoside Fraction Polymer Adsorption from Red Ginseng (P. ginseng) In a typical experiment (Step 3, Figure 3), the ginsenoside fraction of the extract, percolated by solvent in two steps obtained from the 22 g of original raw material of Red Ginseng (P. ginseng), it was evaporated first under reduced atmospheric pressure to eliminate the ethanol and then diluted with water to the original volume to conserve without change the concentration of triterpene saponin. 40 g of ADS-8 adsorbent polymer resin (Nankai University, Tianjin, China) were washed with water and ethanol before and after being loaded onto a column (L = 50 cm x 1 cm D.I., ~ 40 cm3). The aqueous extract fraction of ginsenoside was loaded on the column at an expense of 80 to 100 ml / hr. The optimal expenditure through the resin bed was in the range of 2 to 4 bed volumes / hr. The volume and concentration that passed through the polymeric adsorbent resin was measured and recorded to determine the breaking curve. Once the column was fully charged, the column was washed with 400 ml of water at an expense of 50 ml / hr to remove the impurities of the adsorbed ginsenosides. Elution of ginsenosides was then achieved with 150 ml of ethanol / water (4/1) as an elution solvent at an expense of 50 ml / hour and the elution curve was recorded. For the extract, the loading capacity of the ADS-8 adsorbent resin was approximately 50 to 75 mg of ginsenoside per gram of adsorbent resin. The results of these experiments are tabulated in Tables 19 and 20.
Table 19. Yield of Ginsenoside after column chromatography using ADS-8 resin A = Quantity (g or ml) B = Dry Weight (g) C = Yield (% by weight) D = Content of Ginsenoside (%) E = Yield of Ginsenoside (%) Table 20. Comparison of the Distribution of Ginsenoside in Peak Elution with Ginseng Root Raw Material and Solvent Percolation Extract (% dry weight) EXAMPLE 13. Preparation of the Fraction Polysaccharide from P. notoginseng In a typical experimental protocol, the residue of 30 g of defatted ginseng root stock powder (fraction of essential oil removed, Step 1, Figure 1, Example 1 ), de-saponinized (removed ginsenoside fraction, Step 2, Figure 2, Example 5) derived from P. notoginseng was loaded into a 500 ml flask. This residue was extracted three times with boiling water for two hours. The volume of water in each case was 500 ml, 500 ml, and then 300 ml. The solutions were freeze-dried to determine the concentration of solids (Table 21). The yield of the extraction was 47.22%, indicating that almost 100% of the polysaccharides were extracted from the raw material P. notoginseng. In addition, the fractions of the polysaccharide extract were highly purified, probably mixtures of polysaccharides of more than 99% of various molecular weights (see Example 30 for analysis).
Table 21. Performance of polysaccharides from P. notoginseng A = Stage B = Extraction Solvent C = Solvent Volume (mi; D = Feeding (g) E = Extraction Time F = Performance (% in Dry Weight) G = Polysaccharide (% yield) EXAMPLE 14. Preparation of the Polysaccharide fraction from P. quinquefolius In a typical experimental protocol, the residue of 30 g of defatted ginseng root stock powder (removed essential oil fraction, Step 1, Figure 1, Example 2), de-saponinized ( ginsenoside fraction removed, Step 2, Figure 2, Example 6) derived from P. quinquefolius was loaded into a 500 ml flask.This residue was extracted three times with boiling water for two hours.The volume of water in each case was 500 ml, 500 ml, and then 300 ml. The solutions were freeze-dried to determine the concentration of solids (Table 22). The yield of the extraction was 18.78%, indicating that almost 100% of the polysaccharides were extracted from the raw material P. quinquefolius. In addition, the fractions of the polysaccharide extract were highly purified, probably mixtures of polysaccharides of more than 99% of various molecular weights (see Example 30 for analysis). Table 22. Performance of polysaccharides from. gui.nguefol.ius A = Stage B = Extraction Solvent C = Solvent Volume (ml) D = Feeding (g) E = Extraction Time F = Yield (% in Dry Weight) G = Polysaccharide (% yield) EXAMPLE 15. Preparation of the Fraction Polysaccharide from White Ginseng (P. ginseng) In a typical experimental protocol, the residue of 30 g of defatted ginseng root stock powder (fraction of essential oil removed, Stage 1, Figure 1, Example 3), De-saponinized (removed ginsenoside fraction, Step 2, Figure 2, Example 7) derived from White Ginseng (P. ginseng) was loaded into a 500 ml flask. This residue was extracted three times with boiling water for two hours. The volume of water used in each case was 500 ml, 500 ml, and then 300 ml. The solutions were freeze-dried to determine the concentration of solids (Table 21). The yield of the extraction was 17.44%, indicating that almost 100% of the polysaccharides were extracted from the raw white ginseng material. In addition, the fractions of the polysaccharide extract were highly purified, probably mixtures of polysaccharides of more than 99% of various molecular weights (see Example 30 for analysis). Table 23. Performance of polysaccharides from White Ginseng (P. ginseng) A = Stage B = Extraction Solvent C = Solvent Volume (ml) D = Feeding (g) E = Extraction Time F = Performance (% in Dry Weight) G = Polysaccharide (% yield) EXAMPLE 16. Preparation of the Fraction Polysaccharide from Red Ginseng (P. ginseng) In a typical experimental protocol, the residue of g of raw material powder of defatted ginseng root (fraction of essential oil withdrawal, Stage 1, Figure 1, Example 4), de-saponinized (ginsenoside fraction removed, Step 2, Figure 2, Example 8) derived from Red ginseng (P. ginseng) was loaded into a 500 ml flask. This residue was extracted three times with boiling water for two hours. The volume of water in each case was 500 ml, 500 ml, and then 300 ml. The solutions were freeze-dried to determine the concentration of solids (Table 24). The yield of the extraction was 47.22%, indicating that almost 100% of the polysaccharides were extracted from the raw material Red Ginseng. In addition, the fractions of the polysaccharide extract were highly purified, probably mixtures of polysaccharides of more than 99% of various molecular weights (see Example 30 for analysis). Table 24. Performance of polysaccharides from Red Ginseng (P. ginseng) A = Stage B = Extraction Solvent C = Solvent Volume (ml) D = Feeding (g) E = Extraction Time F = Performance (% in Dry Weight) G = Polysaccharide (% yield) EXAMPLE 17. Methods by HPLC HPLC analysis of extracts and fractions of ginseng, were carried out using a Shimadzu HPLC system SE0405003 equipped with the following Shimadzu equipment: a SCL-10AVP Controller System, a DGU-14a 4-line vacuum membrane degasser, a unit low-pressure gradient FCV-10ALVP, a LC-10ATVP series piston solvent release unit, a 100-microliter semi-micro blender, a SIL-10AF rapid auto-sampler, a photodiode sorting detector SPD-M10AVP UV-Vis, a column kiln CTO-lOAsvp, a computation program for SP1 Class VP 7.2.1 chromatography. and a fraction collector FRC-10A. The column used in Examples 18-25 was a 5μ C18 300A, 250 x 4.6 mm Jupiter column obtained from Phenomenex, Inc. The solvents used in the HPLC methods, including water, ethanol, methanol, and acetonitrile, were HPLC grade. and were obtained from Sigma-Aldrich, Inc. EXAMPLE 18. Characterization by HPLC of an Essential Oil Fraction from P. notoginseng An essential oil fraction was prepared from P. notoginseng as described in Example 1. HPLC analysis was carried out using the methods and equipment described in Example 17 with the specific conditions described herein. The sample of the essential oil fraction was dissolved in HPLC grade methanol at a concentration of 3 mg / ml. The components of the mobile phase were as follows: the mobile phase component A was phosphate regulator, 0.5% phosphoric acid in water, pH 3.5 ("A"); the mobile phase component B was methanol ("B"); component C of mobile phase was acetonitrile ("C"). The gradient concentration elution program used was as follows: the composition of the initial mobile phase comprised on a volume basis of 50%, 17%, and 33%, respectively, of A, B, and C, at 40 minutes after injection of the sample, the mobile phase composition comprised on a volume basis of 25%, 25%, and 50%, respectively, of A, B, and C. The mobile phase had the linear gradient in 490 minutes which changed from initially 50:17:33 to 25:25:50 from A: B: C and then was retained in these conditions for another 10 minutes. The total analysis time was 50 minutes, the expenditure of the mobile phase was 1 ml and the temperature of the column was controlled at 45 ° C. Peak detection was at 254 nm. The HPLC chromatogram data is given in Tables 25 and 26.
Table 25. Retention times of the HPLC peak for the essential oil fraction of P. notoginseng T. R. = Retention time Table 26. HPLC analysis data from the essential oil fraction of P. notoginseng Table 26. (Continued) TR = Retention Time (min) EXAMPLE 19. Characterization by HPLC of an essential oil fraction from P. quinquefolius An essential oil fraction was prepared from P. quinquefolius as described in Example 2. HPLC analysis was carried out as described in Example 18. The retention times for the peaks designated from the HPLC chromatogram are given in Table 27. Additional data for representative peaks from the HPLC chromatogram are given in the Table 28. Table 27. Peak retention times by HPLC for fractions of essential oil of P. quinquefolius Table 27 (Continued) T. R. = Retention Time Table 28. HPLC data for fractions of essential oil of P. quinquefolius Table 28 (Continued) Table 28 (Continued) T.R. = Retention Time EXAMPLE 20. Characterization by HPLC of an essential oil fraction from White Ginseng (P. ginseng) An essential oil fraction was prepared from White Ginseng (P. ginseng) as described in Example 3 The HPLC analysis was carried out as described in Example 18. The retention times for the peaks designated from the HPLC chromatogram are given in Table 29. The additional data for representative peaks from the HPLC chromatogram are given in Table 30. Table 29. Peak retention times by HPLC for white Ginseng essential oil fractions Table 29 (Continued) T.R. = Retention Time Table 30. HPLC data for fractions of essential oil of White Ginseng (P. ginseng) Table 30 (Continued) EXAMPLE 21. Characterization by HPLC of an essential oil fraction from Red Ginseng (P. ginseng) An essential oil fraction was prepared from Red Ginseng (P. ginseng) as described in Example 4. The HPLC analysis was carried out as described in Example 18. The retention times for the peaks designated to from the HPLC chromatogram are given in Table 31. Additional data for representative peaks from the HPLC chromatogram are given in Table 32.
Table 31. Retention times for peaks by HPLC for fractions of essential oil of Red Ginseng T.R. = Retention Time Table 32. HPLC data for Red Ginseng essential oil fractions Table 32 (Continued) T.R. = Retention Time EXAMPLE 22. HPLC Characterization of the Ginsenoside Fraction from P. notoginseng A ginsenoside fraction was prepared from P. notoginseng as described in Example 9. The HPLC analysis was carried out using the methods and equipment described in Example 17 with the specific conditions described herein. A sample of the ginsenoside fraction was diluted 1/10 in HPLC-grade methanol to produce a final concentration of about 1 mg / ml. The sample injection volume was 10 μl.
The components of the mobile phase were as follows: component A of mobile phase was phosphate regulator, 0.5% phosphoric acid in water, pH of 3.5 ("A"); and, component B of mobile phase was acetonitrile ("B"). The concentration gradient elution program used was as follows: the composition of the mobile phase of the initial injection up to 20 minutes comprised in a base volume of 79% and 21%, respectively, of A and B, and, a linear gradient from 20 to 60 minutes with the mobile phase that changed from 79% to 58% of A and 21% to 42% of B. The total analysis time was for approximately 60 -70 minutes, the cost of the mobile phase was 1 ml per minute and the temperature of the column was controlled at 40 ° C. Peak detection was at 203 nm. The retention times for the peaks designated from the HPLC chromatogram are given in Table 33. Additional data for representative peaks from the HPLC chromatogram are given in Table 34.TABLE 33. Retention times for HPLC peaks from the purified ginsenoside fraction of P. no toginseng T.R. = Retention Time Table 34. HPLC data from the purified ginsenoside fraction of P. notoginseng T.R. = Retention Time EXAMPLE 23. Characterization by HPLC of the Ginsenoside Fraction from P. quinquefolius A purified ginsenoside fraction was prepared by affinity adsorption from P. quinquefolius as described in Example 10. The analysis was carried out HPLC as described in Example 22. The retention times for the peaks designated from the HPLC chromatogram are given in Table 35. Additional data for representative peaks from the HPLC chromatogram are given in Table 36. TABLE 35. Retention times for HPLC peaks from the purified ginsenoside fraction of P. quinquefOlius Table 36. HPLC data from the purified ginsenoside fraction of P. quinquefolius EXAMPLE 24. Characterization by HPLC of the Ginsenoside Fraction from White Ginseng (P. ginseng) A purified ginsenoside fraction was prepared by affinity adsorption from White Ginseng (P. ginseng) as described in Example 11. It was carried out the HPLC analysis as described in Example 22. The retention times for the peaks designated from the HPLC chromatogram are given in Table 37. Additional data for representative peaks from the HPLC chromatogram are given in FIG. Table 38 TABLE 37. Retention Times for HPLC Peaks from the Purified Ginsenoside Fraction of White Ginseng Table 38. HPLC data from the purified ginsenoside fraction of White Ginseng EXAMPLE 25. Characterization by HPLC of the Ginsenoside Fraction from Red Ginseng (P. ginseng) A purified ginsenoside fraction was prepared by affinity adsorption from Red Ginseng (P. ginseng) as described in Example 12. It was carried out the HPLC analysis as described in Example 22. The retention times for the peaks designated from the HPLC chromatogram are given in Table 39. Additional data for representative peaks from the HPLC chromatogram are given in FIG. Table 40. TABLE 39. Retention times for HPLC peaks from the purified ginsenoside fraction of Red Ginseng Table 40. HPLC data from the purified ginsenoside fraction of Red Ginseng EXAMPLE 26. Methods of Gas Chromatography ("GC") and Mass Spectroscopy ("MS") Analysis by gas chromatography of ginseng extracts and fractions was carried out using a Hewlett-Packard Model 580 gas chromatograph. The analysis was carried out using a capillary column XTL-5 (30 m extension x 0.25 mm internal diameter, Restek) with a film thickness of 0.25 μm and an expense of 1 ml / min for the helium carrier gas. The gasification temperature was 270 ° C. The column temperature was programmed as follows: 50-140 ° C (retained for 15 minutes) at a rate of 10 ° C / min, then retained at 140 ° C for 15 min, followed by 140-260 ° C at a speed of 15 ° C / min, and then retained at 260 ° C. The total run time was 52 minutes and the fractionation time of the sample was 1:50. The mass spectroscopy analysis was carried out using a Hewlett-Packard Model 5899a mass spectrometer. The temperature of the ion source was 200 ° C, and the ion source was El with an ionization energy of 70 eV. The emission current was 300 mA. The data were collected in full exploration mode from 40 - 600 m / z in the cycles. EXAMPLE 27. Characterization by GC / MS of the Fraction of Essential Oil of P. notoginseng An essential oil fraction of P. notoginseng was prepared, as described in Example 1. The analysis by gas chromatography was carried out as described. described in Example 26. Analysis of the mass spectrum of the GC elution peaks was used to help identify the various chemical constituents. MS data (value and abundance of m / z) is consistent with the presence of the following compounds in the essential oil fraction: +) -spatulenol, CAS No. 6750-60-3; Caffeine, CAS No. 58-08-2; hexadecanoic acid, CAS No. 57-10-3; (-) -Caryophyllene oxide, CAS No. 1139-30-6; ethyl heptanoate, CAS No. 106-30-9; trans methyl ester, trans-octadeca-9, 12-dienoic, CAS No. 2566-97-4; octadec-9-inoic acid methyl ester, CAS No. 1120-32-7; phenylacetylene, CAS No. 536-74-3; ethilentiourea, CAS No. 96-45-7; linoleic acid, CAS No. 60-33-3; 4- methyl-pent-2-enoic acid, CAS No. 10321-71-8; 2- methyl-4-nitroimidazole, CAS No. 696-23-1; 9, 12- octadecadienal, CAS No. 26537-70-2; Mevinfos, CAS No. 7786-34-7; undec-10-inoic acid, CAS No. 2777-65-3; falcarinol ((Z) -1,9-heptadecadiene-4,6-diin-3-ol), CAS No. 21852-80-2; and, [IR- (1, 4β, 4aa, 6β, 8aa)] - octahydro-4, 8a, 9, 9-tetramethyl-1, 6-methane-1 (2H) -naphthol, CAS No. 5986-55- 0 EXAMPLE 28. Characterization by GC / MS of the Fraction of Essential Oil of P. quinquefolius An essential oil fraction of P. quinquefolius was prepared, as described in Example 2. Analysis by gas chromatography was carried out as described. described in Example 26. Mass spectral analysis of peaks eluting from GC was used to help identify the various chemical constituents.
The MS data (value and abundance of m / z) is consistent with the presence of the following compounds in the essential oil fraction: 4,6-diamino- 1, 3, 5-triazin-2 (lH) -one, CAS No. 645-92-1; 2, 2'-methyliminodiethanol, CAS No. 105-59-9; Caffeine, CAS No. 58-08-2; dihydrouracil, 504-07-4; stearic acid (octadecanoic acid), CAS No. 57-11-4; hexadecanoic acid, CAS No. 57-10-3; 4- nitrophenol, CAS No. 100-02-7; linoleic acid, CAS No. 60-33-3; 3- nitrotoluene, CAS No. 99-08-1; 2-, 3- dihydroxypropyl palmitate, CAS No. 542-44-9; Oleic acid, CAS No. 112-80-1; cinnamyl acetate, CAS No. 103-54-8; (9E, 12E) -octadeca- 9, 12-methyl-dienoate (methyl linolelaidate), CAS No. 2566-97-4; and, 7- octadenoic acid, CAS No. 18719-24-9. EXAMPLE 29. Characterization by GC / MS of the Fraction of Essential Oil of White Ginseng (P. ginseng) An essential oil fraction was prepared from Gingeng Blanco (P. ginseng) as described in Example 3. It was The analysis by gas chromatography was carried out as described in Example 26. The mass spectral analysis of the peaks eluting from the GC was used to help identify the various chemical constituents. MS data (value and abundance of m / z) is consistent with the presence of the following compounds in the essential oil fraction: (-) - spatulenol, CAS No. 77171-55-2; (+) - spatulenol, CAS No. 6750-60-3; (-) -carophyllene oxide, CAS No. 1139-30-6; 1-methyl-5-nitro-1H-imidazole, CAS No. 3034-42-2; 2-ethyl-2-methyloxirane, CAS No. 30095-63-7; Caffeine, CAS No. 58-08-2; dihydrouracil, CAS No. 504-07-4; hexadecanoic acid, CAS No. 57-10-3; (9E, 12E) - octadeca- 9, 12-methyl dienoate (methyl linolelaidate), CAS No. 2566-97-4; linoleic acid, CAS No. 60-33-3; undec-10-inoic acid, CAS No. 2777-65-3; phenylacetylene, CAS No. 536-74-3; Sinalbin, CAS No. 27299-07-6; stigmasta-5, 22-dien-3- ß-ol, CAS No. 83-48-7; (3ß, 24S) - stigmst- 5-en-3-ol, CAS No. 83-47-6; stigmas- 5- en- 3- ß- ol, CAS No. 83-46-5; (3ß, 24?) - stigmast- 5- en- 3- ol, CAS No. 19044-06-5; 4-methyl-1, 4-heptadiene, CAS No. 13857-55-1; 9, 12-octadecadienal, CAS No. 26537-70-2; 7, 8-epoxyoctene, CAS No. 19600-63-6; 4- nonino, CAS No. 20184-91-2; 2-cyclopentene-1-undecanoic acid, CAS No. 459-67-6; falcarinol ((Z) -1,9-heptadecadien-4,6-diin-3-ol), CAS No. 21852-80-2; and N-methylcaprolactam, CAS NO. 2556-73-2. EXAMPLE 30. Characterization by GC / MS of the Fraction of Red Ginseng Essential Oil (P. ginseng) An essential oil fraction was prepared from Gingeng Red (P. ginseng) as described in Example 3. It was The analysis by gas chromatography was carried out as described in Example 26. The mass spectral analysis of the peaks eluting from the GC was used to help identify the various chemical constituents. The MS data (value and abundance of m / z) are consistent with the presence of the following compounds in the essential oil fraction: 3-hydroxy-2-methyl-4-pyrone, CAS No. 118-71-8; Pirogalol, CAS No. 87-66-1; [la R- (laa, la, 7aa, 7ba)] - la, 2, 3, 5, 6, 7, 7a, 7b- octahydro-1, 1, 7, 7a-tetramethyl-1H-cyclopropa [a] naphthalene, CAS No. 17334-55-3; [la R- (laa, 4aa, 7a, 7aβ] - decahydro-1, 1, 7-trimethyl-4-methylene-1H-cycloprop [e] azulene, CAS No. 489-39-4, caryophyllene, CAS No. 87 -44-5; [IR- (IR *, 4Z, 9S *)] - 4, 11, 11-trimethylenbicyclo [7.2.1] undec-4-ene, CAS No. 118-65-0; caffeine, CAS No 58-08-2, hexadecanoic acid, CAS No. 57-10-3, 4-methyl-2-phenyl-2-pentenal, CAS No. 26643-91-4; (Z) -9, 17-Octadecanal, CAS No. 56554-35-9, linoleic acid, CAS No. 60-33-3, ethylidenecycloheptane, CAS No. 10494-87-8, Octa-1, 7-diine, CAS No. 871-84-1; - (phenylmethyl) sidnon, CAS No. 16844-42-1, phenylacetylene, 536-74-3, diisopropyl adipate, CAS No. 6938-94-9, 2,3-dihydroxypropyl palmitate, CAS No. 542-44 -9, 9Z, 12Z-octadecadienoic acid (2-linoleoyl glycerol), CAS No. 3443-82-1, and 3-ethenyl-cyclooctene, CAS No. 2213-60-7. EXAMPLE 31. Determination of the concentration of polysaccharides in the Fraction Purified Polysaccharide of Panax species All the dried polysaccharide extracts from P. notoginseng, (Example 13), P. quinquefolius (Example 14), White Ginseng (Example 15) , and Red Ginseng (Example 16) were colorless which indicates that the tannins and other polyphenolic chemical constituents were extracted with ethanol during the extraction of the ginsenosides (Stage 2). When several amounts of salts (up to 100 mg / 2 g (dry mass weight solution) as well as citric acid and acetic acid were added to the solutions of polysaccharide extracts, no precipitation was observed indicating that the protein content In addition, dissolving fractions of dried polysaccharide extracts by freezing in 10 volumes of ethanol revealed that less than 1% of the fractions of polysaccharide extracts were soluble in ethanol. These polysaccharide extract fractions of Panax species were highly purified, probably more than 99%, mixtures of polysaccharides of various molecular weights.More directly, the amount of carbohydrates was determined in all fractions of purified polysaccharides, using the colorimetric method with Antrone Typically, approximately 0.18 grams of anthrone (CAS No. 90-44-8, obtained a from Sigma-Aldrich), which is also called 9, 10-dihydro-9-oxoanthracene, was mixed with about 50 ml of concentrated sulfuric acid (95.7%, obtained from Fisher). The solution of anthrone and sulfuric acid was stirred and then immersed in a bath of ice water. The calibration of the spectrophotometer (Thermo Spectronic 20D +) was achieved using lactose (CAS No. 63-42-3, obtained from Acros) as a standard. A standard lactose solution (0.05% w / v) was prepared by dissolving approximately 15 mg of lactose in approximately 30 ml of distilled water. Specific volumes of standard lactose solutions (typically about 0, 200, 400, 600, and 800 μl) were pipetted into test tubes, and the volume was adjusted to a final volume of 1 ml using distilled water. The anthrone solution (2 ml), prepared as described above, was added gradually to each of the standard 1 ml lactose samples while the test tube was vigorously shaken. After mixing, the anthrone sample solutions were then stored in an ice bath for 30 minutes, followed by quenching at room temperature. The absorbance of each sample at 625 nm was determined using distilled water as a blank. The absorbance was plotted against the lactose concentration to obtain the standard curve. Samples of the purified polysaccharide fractions (approximately 10 μl) were diluted to 1 ml with distilled water. To the diluted polysaccharide samples, approximately 2 ml of 1 anthrone reagent was added with vigorous stirring. After mixing the solutions of polysaccharide-anthrone was stored for 30 minutes in an ice bath, and then warmed to room temperature. The absorbance at 625 nm was determined for each of the anthrone polysaccharide sample solutions. The amount of carbohydrates in each sample was determined using the standard lactose curve prepared as described above. Typically, using this method, it was determined that all the polysaccharide fractions derived from all the Panax species studied contained more than 90% by weight of carbohydrate components. EXAMPLE 32. Composition in Dosage Form of Extract of P. notoginseng An extract of P. notoginseng was prepared in accordance with the present invention and was used to prepare a composition in dosage form suitable for tablets, capsules, or powder for addition to water or another solution as a drinkable solution. The composition in dosage form was prepared according to the formulation given in Table 41, wherein the quantities given are the quantities per individual dosage form. TABLE 41. Formulation composition of P. notoginseng. Extract of P. notoginseng 150.0 mg Essential Oil (2 mg, 1.3% dry weight); Total Ginsenosides (38 mg, 25.3% dry weight); Polysaccharides (110 mg, 73.3% dry weight) Stevioside (Stevia Extract) 12.5 mg Carboxymethyl cellulose 35.5 mg Lactose 77.0 mg Total 275.0 mg The new extract of P. notoginseng comprises an essential oil, ginsenosides, and polysaccharides in mass percentage greater than that found in the natural rhizome material or conventional extraction products. Formulations can be made in any oral dosage form and administered daily or up to 15 times a day as needed for the desired physiological, psychological and medical effects (improved memory and knowledge, relief of chronic fatigue syndrome, improvement of erectile function male) and medical effects (anti-platelet anti-agglutination oxidation, prevention and treatment of cardiovascular and cerebrovascular diseases, anti-hypercholesterolemia, cytoprotection, nervous system protection, neurological degenerative diseases such as the prevention and treatment of Parkinson's disease and Alzheimer's disease, anti-inflammatory, immune improvement, anti-viral, lung disease, diseases and liver protection, hypoglycemic and anti-diabetes, and prophylaxis and cancer treatment). The dosage composition as provided in Table 41 can be compressed into a tablet, used in a gelatin capsule, or used in a fast dissolving tablet. EXAMPLE 33. Composition for Dosage Forms with Extract of P. quinquefolius An extract of P. quinquefolius was prepared in accordance with the present invention and was used to prepare a composition in dosage form suitable for tablets, capsules, or powder for addition to water or another solution as a drinkable solution. The composition in dosage form was prepared according to the formulation given in Table 42, wherein the amounts given are the amounts for the individual dosage form. TABLE 42. Formulation composition of P. quinquefolius. Extract of P. quinquefolius 150.0 mg Essential Oil (2 mg, 1.3% dry weight); Total Ginsenosides (20 mg, 13.3% dry weight); Polysaccharides (128 mg, 85.4% dry weight) Vitamin C 15.0 mg Sucralose 35.0 mg Powdered gold beans (10: 1) 50.0 mg Mocha flavor 40.0 mg Flavor (chocolate, cherry, mocha, etc.) 10.0 mg Total 300.0 mg The Golden Bean powder 10: 1 refers to the water content (10 parts of golden bean to 1 part of water). It is used as a binder. The new extract composition of P. quinquefoli us comprises an essential oil, ginsenosides, and chemical constituents polysaccharides in mass percentage greater than that found in natural plant material or conventional extraction products. The formulation can be made in any oral dosage form and safely administered up to 15 times a day as needed for the desired physiological, psychological and medical effects (see Example 1 above). The dosage composition as provided in Table 42 can be compressed into a tablet, used in a gelatin capsule, or used in a rapid dissolving tablet. EXAMPLE 34. Composition for Dosage Forms with White Ginseng Extract (P. ginseng) A White Ginseng extract (P. ginseng) was prepared in accordance with the present invention and was used to prepare a composition in dosage form suitable for tablets. , capsules, or powder for addition to water or another solution as a drinkable solution. The composition in dosage form was prepared in accordance with the formulation given in Table 43, wherein the amounts given are the amounts for the individual dosage form. TABLE 43. Formulation Composition of White Ginseng (P. ginseng). White Ginseng Extract (P. ginseng) 150.0 mg Essential Oil (5.0 mg, 3.3% dry weight); Total Ginsenosides (45.0 mg, 30.0% by dry weight); Polysaccharides (100.0 mg, 66.7% dry weight) Vitamin C 15.0 mg Sucralose 35.0 mg Gold bean powder (10: 1) 30.0 mg Flavor (Strawberry) 60.0 mg Base-X M-500 54.0 mg Base-X Xanthan Gum 1.0 mg Total 350.0 mg The new composition of White Ginseng extract (P. ginseng) comprises an essential oil, ginsenosides, and chemical constituents polysaccharides in percentage by mass weight greater than that found in natural plant material or conventional extraction products.
Note also the change of profile in the composition of the extract of White Ginseng (the ratio of the oil / ginsenosides in raw material was 1 / 6.4 and in the composition of the extract is 1/5, the ratio of essential oil / polysaccharides in matter premium was 1/35 and the composition of the extract is 1/20, and the proportions of ginsenosides / polysaccharides was 1 / 5.4 and the composition of the extract is 1/4). The formulation can be made in any oral dosage form and safely administered up to 15 times a day as needed for the desired physiological, psychological and medical effects (see Example 1 above). The dosage composition as provided in Table 43 may be compressed into a tablet, used in a gelatin capsule, or used in a rapid dissolving tablet. Although the invention has been described in detail with particular reference to specific embodiments, it will be obvious to the person skilled in the art that various modifications and variations may be made to the present invention in light of the foregoing without departing from the scope or spirit of the invention. invention. Other embodiments of the invention will be obvious to those skilled in the art considering the specification and practice of the invention described herein. It is envisaged that the specification and examples are considered exemplary only, with a scope and true spirit of the invention which is indicated by the following claims.
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Claims (30)

NOVELTY OF THE INVENTION Having described the present invention, it is considered as novelty, and therefore the content of the following is claimed as property: CLAIMS
1. - A composition characterized in that it comprises a ginsenoside in an amount greater than 10% by weight. 2. The composition according to claim 1, characterized in that the ginsenoside comprises R0, Ri, R2, Ra, Rb ?, Rb2, Rc, R, Re Rf Rg ?, Rg2,
Rg3 Rg5, Rhl, R 2 r Rh4 r Rsl / Rs2, Rs3 r ° Rs.
3. The composition according to claim 1, characterized in that the amount of ginsenosides is greater than 15, 20, 25, 30, 35, 40, 45 or 50% by weight.
4. The composition according to claim 1, characterized in that it comprises a polyacetylene.
5. The composition according to claim 4, characterized in that the polyacetylene comprises pananaxinol, panaxidol, panaxitriol, acetylpanaxidol, chloropapaxidol, panaxidolchlorohydrin, panaxin, ginsenoin A, ginsenoin B, ginsenoin C, ginsenoin D, ginsenoin E, ginsenoin F, ginsenoin G, Ginsenoin H, Ginsenoin I, Ginsenoin J, or Ginsenoin K.
6. - The composition according to claim 4, characterized in that the polyacetylene comprises pananaxinol, panaxidol, panaxitriol, acetylpanaoxidol, chloropapaxidol, panaxidolchlorohydrin, panaxin, ginsenoin A, ginsenoin B, ginsenoin C, ginsenoin D, ginsenoin E, ginsenoin F, ginsenoin G, Ginsenoin H, Ginsenoin I, Ginsenoin J, and Ginsenoin K.
7. - The composition according to claim 4, characterized in that the ginsenoside comprises R0, Ri, R2, Ra, Rbi, Rb2, Rc, Rd, Re, f Rgi, Rg2,
Rg3 Rg5, Rhl, Rh2, h, Rsl, Rs2, Rs3, ° Rs4 • 8.- The composition according to claim 4, characterized in that the amount of ginsenosides is greater than 15, 20, 25, 30, 35, 40 , 45 or 50% by weight.
9. The composition according to claim 4, characterized in that the amount of polyacetylene is 1, 2, 4, 6, 8, or 10% by weight.
10. The composition according to claim 4, further characterized in that it comprises an essential oil selected from the group consisting of (+) spatulenol; (-) spatulenol; caffeine; hexadecanoic acid; (-) - caryophyllenium oxide; ethyl heptanoate; trans methyl ester, trans-octadeca-9, 12-dienoic; octadec-9-inoic acid methyl ester; phenylacetylene; ethylentiourea; linoleic acid; 4- methyl-pent-2-enoic acid; 2- methyl-4-nitroimidazole; 9, 12- octadecadienal; mevinfos; undec-10-inoic acid; falcarinol ((Z) -1,9-heptadecadien-4,6-diyl-3-ol); [IR- (la, 4β, 4aa, 6β, 8aa)] - octahydro-4, 8a, 9, 9-tetramethyl-1,6-methano-1 (2H) -naphthol; 4,6-diamino- 1, 3, 5-triazin-2 (1H) -one; 2, 2'-methyliminodiethanol; dihydrouracil; stearic acid; 4- nitrophenol; 3- nitrotoluene; 2,3-dihydroxypropyl palmitate; oleic acid; cinnamyl acetate; 7- octenoic acid; 1-methyl-5-nitro-1H-imidazole; 2-ethyl-2-methyloxirane; (9E, 12E) - octadeca- 9, 12-methyl-dienoate; sinalbin, stigmasta- 5, 22- dien- 3-4- ol; (3ß, 24S) - stigmast- 5- in- 3- ol; stigmast- 5-in-3 ß- ol; (3ß, 24?) - stigmast- 5- in- 3- ol; 4-methyl-1, 4-heptadiene; 9, 12- octadecadienal; 7, 8-epoxyoctene; 4- nonino; 2-cyclopentene-1-undecanoic acid; 3-hydroxy-2-methyl-4-pyrone; pyrogallol; [laR- (laa, 7a, 7aa, 7ba)] - la, 2, 3, 5, 6, 7, 7a, 7b-octahydro-1, 1, 7, 7a-tetramethyl-1H-cyclopropa [a] naphthalene: [la R- (laa, 4aa, 7a, 7aβ, 7ba)] - decahydro-1, 1, 7-trimethyl-4-methylene-1H-cycloprop [e] azulene; caryophyllene; [IR- (1R *, 4Z, 9S *)] - 4, 11, 11-trimethyl-8-methylenebicyclo [7.2. OJundec- 4- eno; 4- methyl-2-phenyl-2-pentenal; (Z) - 9, 17, octadecadienal; ethylenedicycloheptane; octa-1, 7- diine; 3- (phenylmethyl) sidnon; diisopropyl adipate; 2-, 3- dihydroxypropyl palmitate; acid (2-linoleoyl glycerol) 9Z, 12Z, octadecadienoic; and 3-ethenyl-cyclooctene.
11. The composition according to claim 1, characterized by comprising a polysaccharide.
12. The composition according to claim 11, characterized in that the polysaccharide comprises glucose, arabinose, galactose, rhamnose, xylose, or uronic acid.
13. The composition according to claim 11, characterized in that the polysaccharide comprises glucose, arabinose, galactose, rhamnose, xylose, and uronic acid.
14. The composition according to claim 11, characterized in that the ginsenoside comprises R0, R1 f R2, Ra, Rb ?, Rb2, Re, Rd, Re, Rf Rg ?, Rg2, Rg3 Rg5, Rl, Rh2, h, Rsl, s2, Rs3, O Rs.
15. The composition according to claim 11, characterized in that the amount of ginsenosides is greater than 15, 20, 25, 30, 35, 40, 45 or 50% by weight.
16. The composition according to claim 11, characterized in that the amount of polysaccharide is 25, 30, 35, 40, 45, 50, 55 or 60% by weight.
17. The composition according to claim 11, further characterized in that it comprises an essential oil selected from the group consisting of (+) spatulenol; (-) spatulenol; caffeine; hexadecanoic acid; (-) - caryophyllenium oxide; ethyl heptanoate; trans methyl ester, trans-octadeca-9, 12-dienoic; octadec-9-inoic acid methyl ester; phenylacetylene; ethylentiourea; linoleic acid; 4- methyl-pent-2-enoic acid; 2- methyl-4-nitroimidazole; 9, 12- octadecadienal; mevinfos; undec-10-inoic acid; falcarinol ((Z) -1,9-heptadecadien-4,6-diyl-3-ol); [IR- (1, 4β, 4aa, 6β, 8aa)] - octahydro-4, 8a, 9, 9-tetramethyl-1, 6-methano-1 (2H) -naphthol; 4,6-diamino- 1, 3, 5-triazin-2 (1H) -one; 2, 2'-methyliminodiethanol; dihydrouracil; stearic acid; 4- nitrophenol; 3- nitrotoluene; 2,3-dihydroxypropyl palmitate; oleic acid; cinnamyl acetate; 7- octenoic acid; 1-methyl-5-nitro-1H-imidazole; 2-ethyl-2-methyloxirane; (9E, 12E) - octadeca- 9, 12-methyl-dienoate; sinalbin, stigmasta- 5, 22- dien- 3-4- ol; (3ß, 24S) - stigmast- 5- in- 3- ol; stigmast- 5-in-3 ß- ol; (3ß, 24?) - stigmast- 5- in- 3- ol; 4-methyl-1, 4-heptadiene; 9, 12- octadecadienal; 7, 8-epoxyoctene; 4- nonino; 2-cyclopentene-1-undecanoic acid; 3-hydroxy-2-methyl-4-pyrone; pyrogallol; [laR- (laa, 7a, 7aa, 7ba)] - la, 2, 3, 5, 6, 7, 7a, 7b-octahydro-1, 1, 7, 7a-tetramethyl-1H-cyclopropa [a] naphthalene: [la R- (laa, 4aa, 7a, 7aβ, 7ba)] - decahydro-1, 1, 7-trimethyl-4-methylene-1H-cycloprop [e] azulene; caryophyllene; [IR- (1R *, 4Z, 9S *)] - 4, 11, 11-trimethyl-8-methylenebicyclo [7.2.0] undec-4-ene; 4- methyl-2-phenyl-2-pentenal; (Z) - 9, 17, octadecadienal; ethylidenecycloheptane; octa-1, 7- diine; 3- (phenylmethyl) sidnon; diisopropyl adipate; 2-, 3- dihydroxypropyl palmitate; acid (2-linoleoyl glycerol) 9Z, 12Z, octadecadienoic; and 3-ethenyl-cyclooctene.
18. The composition according to claim 1, characterized in that it comprises a polyacetylene and a polysaccharide.
19. The composition according to claim 18, characterized in that the ginsenoside comprises R0, Ri, R2, Ra, Rbi, b2, Rc, d, Re, Rf Rgi, Rg2, Rg3 Rg5, Rhl, Rh2, Rh, Rsl , Rs2, Rs3, or Rs4.
20. - The composition according to claim 18, characterized in that the amount of ginsenosides is greater than 15, 20, 25, 30, 35, 40, 45 or 50% by weight.
21. The composition according to claim 18, characterized in that the polyacetylene comprises pananaxinol, panaxidol, panaxitriol, acetylpanaoxidol, chloropapaxidol, panaxidolchlorohydrin, panaxin, ginsenoin A, ginsenoin B, ginsenoin C, ginsenoin D, ginsenoin E, ginsenoin F, ginsenoin G, Ginsenoin H, Ginsenoin I, Ginsenoin J, or Ginsenoin K.
22. The composition according to claim 18, characterized in that the polyacetylene comprises pananaxinol, panaxidol, panaxitriol, acetylpanaoxidol, chloropapaxidol, panaxidolchlorohydrin, panaxin, ginsenoin A, ginsenoin. B, Ginsenoin C, Ginsenoin D, Ginsenoin E, Ginsenoin F, Ginsenoin G, Ginsenoin H, Ginsenoin I, Ginsenoin J, and Ginsenoin K.
23. - The composition according to claim 18, characterized in that the amount of polyacetylene is 1, 2, 4, 6, 8, or 10% by weight.
24. The composition according to claim 18, characterized in that the polysaccharide comprises glucose, arabinose, galactose, rhamnose, xylose or uronic acid.
25. - The composition according to claim 18, characterized in that the polysaccharide comprises glucose, arabinose, galactose, rhamnose, xylose and uronic acid.
26. The composition according to claim 18, characterized in that the amount of polysaccharide is 25, 30, 35, 40, 45, 50, 55, or 60% by weight.
27. The composition according to claim 18, further characterized in that it comprises an essential oil selected from the group consisting of (+) spatulenol; (-) spatulenol; caffeine; hexadecanoic acid; (-) - caryophyllenium oxide; ethyl heptanoate; trans methyl ester, trans-octadeca-9, 12-dienoic acid; octadec-9-inoic acid methyl ester; phenylacetylene; ethylentiourea; linoleic acid; 4- methyl-pent-2-enoic acid; 2- methyl-4-nitroimidazole; 9, 12- octadecadienal; mevinfos; undec-10-inoic acid; falcarinol ((Z) -1,9-heptadecadien-4,6-diyl-3-ol); [IR- (la, 4β, 4aa, 6β, 8aa)] - octahydro-4, 8a, 9, 9-tetramethyl-1,6-methano-1 (2H) -naphthol; 4,6-diamino- 1, 3, 5-triazin-2 (1H) -one; 2, 2'-methyliminodiethanol; dihydrouracil; stearic acid; 4- nitrophenol; 3- nitrotoluene; 2,3-dihydroxypropyl palmitate; oleic acid; cinnamyl acetate; 7- octenoic acid; 1-methyl-5-nitro-1H-imidazole; 2-ethyl-2-methyloxirane; (9E, 12E) - octadeca- 9, 12-methyl-dienoate; sinalbin, stigmasta- 5, 22- dien- 3-4- ol; (3ß, 24S) - stigmast- 5- in- 3- ol; stigmast- 5-in-3 ß- ol; (3ß, 24?) - stigmast- 5- in- 3- ol; 4-methyl-1, 4-heptadiene; 9, 12- octadecadienal; 7, 8-epoxyoctene; 4- nonino; 2-cyclopentene-1-undecanoic acid; 3-hydroxy-2-methyl-4-pyrone; pyrogallol; [la R- (laa, 7a, 7aa, 7ba)] - la, 2, 3, 5, 6, 7, 7a, 7b-octahydro-1, 1, 7, 7a-tetramethyl-1H-cyclopropa [a] naphthalene; [la R- (laa, 4aa, 7a, 7aβ, 7ba)] - decahydro-1, 1, 7-trimethyl-4-methylene-1H-cycloprop [e] azulene; caryophyllene; [IR- (1R *, 4Z, 9S *)] - 4, 11, 11-trimethyl-8-methylenebicyclo [7.2.0] undec-4-ene; 4- methyl-2-phenyl-2-pentenal; (Z) - 9, 17, octadecadienal; ethylenedicycloheptane; octa-1, 7- diine; 3- (phenylmethyl) sidnon; diisopropyl adipate; 2-, 3- dihydroxypropyl palmitate; acid (2-linoleoyl glycerol) 9Z, 12Z, octadecadienoic; and 3-ethenyl-cyclooctene.
28. The composition according to claims 1, 4, 11, or 18, further characterized in that it comprises a pharmaceutical carrier.
29. - A method of extracting a Panax species characterized in that it comprises, sequentially extracting a plant material of the Panax species to produce an essential oil fraction, a ginsenoside fraction and a polysaccharide fraction, wherein the fraction of essential oil is derived from extracting plant material raw material by means of the extraction of supercritical carbon dioxide, the ginsenoside fraction is extracted from the remainder of the extraction of essential oil by means of hydroalcoholic extraction, and the polysaccharide fraction is derived from the extraction by hot water of the remnant of ginsenoside extraction.
30. The method according to claim 29, characterized in that the extraction of the ginsenoside comprises: a) contacting a remnant of a raw material material from an extraction of essential oil by means of supercritical carbon dioxide with a hydroalcoholic mixture per a sufficient time to extract ginsenosides to form an aqueous solution of extracted ginsenosides; b) passing the extracted aqueous ginsenoside solution through a column of adsorbent resin wherein the ginsenosides are adsorbed; and c) eluting ginsenosides from the adsorbent resin.
MXMX/A/2008/003885A 2005-09-19 2008-03-19 Compositions and methods comprising panax species MX2008003885A (en)

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