Iron-fiber composition, preparation and uses thereof
09566303 ยท 2017-02-14
Assignee
Inventors
Cpc classification
A23V2002/00
HUMAN NECESSITIES
A23V2002/00
HUMAN NECESSITIES
A61K9/70
HUMAN NECESSITIES
A23L33/21
HUMAN NECESSITIES
International classification
Abstract
Compositions comprising ferrous and/or ferric iron compounds and fiber in a complex, methods for preparing such compositions of matter, and the use thereof for treatment of adsorbing certain accessible targets in the gastrointestinal tract and in an extracorporeal system, are provided herein.
Claims
1. A composition suitable for oral administration characterized by: (1) being water-insoluble; (2) having Iron (II) and/or Iron (III) tightly bound to dietary fiber containing a mixture of polysaccharides and/or oligosaccharides and lignin with minimal iron release/availability; (3) with a phosphate binding property.
2. The composition of claim 1, wherein the dietary fiber is a natural composition encompassing lignin and saccharides, or a synthetic composition that contains lignin and saccharides.
3. The composition of claim 1, wherein the iron salt is selected from the group consisting of iron(II) acetate, iron(II) citrate, iron(II) ascorbate, iron(II) oxalate, iron(II) oxide, iron(II) carbonate, iron(II) carbonate saccharate, iron(II) formate, iron(II) sulfate, iron(II) chloride, iron(III) chloride, iron (II) bromide, iron (II) iodide, iron (III) fluoride, iron(II) acetylacetonate, iron (III) phosphate, iron (III) pyrophosphate, and combinations thereof.
4. The composition of claim 1, wherein the iron-fiber complex contains iron at 2 wt % to 50 wt % of the composition.
5. The composition of claim 1, optionally wherein the iron is bonded to the dietary fiber containing saccharides and lignin as a complex via carbon, oxygen, nitrogen or hydrogen bridge bonds.
6. The composition of claim 1, wherein the complex is crystalline, amorphous or comprises microdomains of both amorphous and crystalline regions ranging from 10% to 90% amorphous and 10% to 90% crystalline.
7. The composition of claim 1, wherein the composition has a density of >1 g/ml in its compressed dry form, and has a density of 0.2-0.5 g/ml after being exposed to liquids.
8. The composition of claim 1, wherein the composition is capable of binding to minerals, ions, toxins, metabolites at a wide pH range and optionally wherein the composition is stable at pH 1-12, and remains efficacious at a pH range between 1 to 12.
9. A process for producing an orally administrable composition according to claim 1 comprising the steps of: (a) mixing dietary fiber containing a mixture of polysaccharides and/or oligosaccharides and lignin with an Iron(II) or Iron(III) compound, at a pH <3; (b) maintaining a temperature of reaction mixture of step (a) between ambient and 100 C.; (c) cooling the reaction mixture of step (b) to ambient temperature; (d) adjusting the pH using base until pH>3 with clusters of precipitates formed; and (c) washing the precipitates until pH is neutral; and (f) isolating the composition thereof, wherein the reaction mixture is optionally exposed to pressure during step (b).
10. The process of claim 9, wherein the pH<3 in step (a) of the process can be reached by addition of an acid selected from the group consisting of: hydrogen halides especially hydrochloric acid (HCl), hydrobromic acid (HBr), hydroiodic acid (HI), halogen oxoacids especially hypochlorous acid (HClO), chlorous acid (HClO.sub.2), chloric acid (HClO.sub.3), perchloric acid (HClO.sub.4), and corresponding acids for bromine and iodine, sulfuric acid (H.sub.2SO.sub.4), fluorosulfuric acid (HSO.sub.3F), nitric acid (HNO.sub.3), phosphoric acid (H.sub.3PO4), fluoroantimonic acid (HSbF.sub.6), fluoroboric acid (HBF.sub.4), hexafluorophosphoric acid (HPF.sub.6), chromic acid (H.sub.2CrO.sub.4), and boric acid (H.sub.3BO.sub.3).
11. The process of claim 9, wherein the base used in step (d) of the process is selected from LiOH, KOH, NaOH, NaHCO.sub.3, Na.sub.2CO.sub.3, Ca(OH).sub.2, Mg(OH).sub.2, Li.sub.2CO.sub.3, K.sub.2CO.sub.3, CaCO.sub.3, and MgCO.sub.3.
12. The composition of claim 1, wherein the composition is formulated as a nutritional supplement, a beverage, a snack bar, or a cereal.
13. The composition of claim 1, wherein the composition is formulated as a medicament.
14. The medicament of claim 13, wherein the composition is selected from the group consisting of capsules, sachets, tablets, lozenges, wafers, powders, suspensions in an appropriate liquid.
15. The medicament of claim 13, wherein the medicament is used to adsorb excess phosphate, cholesterol, and toxins from infectious agents.
16. The composition of claim 1, formulated for use according to extracorporeal, ex vivo, or in vitro administration to a subject in need thereof.
17. The composition of claim 1, wherein the composition is formulated as an elemental medical food comprising at least 10 mg of the composition according to claim 1 in a physiological carrier.
18. The elemental medical food of claim 17, formulated as a pill, a tablet, a powder, a bar, a wafer, a suspensions in an appropriate liquid.
19. The elemental medical food of claim 17, further comprising one or more ingredients selected from the group consisting of natural flavors, artificial flavors, major trace and ultra-trace minerals, minerals, vitamins, oats, nuts, spice, milk, egg, salt, flour, lecithin, xanthan gum, and sweetening agents.
20. The composition of claim 1, wherein said complex is formulated as a food supplement suitable for mammals comprising at least 10 mg of the composition according to claim 1.
21. The food supplement of claim 20 formulated as a powder, a bar, a wafer, a suspension in an appropriate liquid.
22. The food supplement of claim 20 comprising one or more additional ingredients selected from the group consisting of natural flavor, artificial flavors, major trace and ultra-trace minerals, minerals, vitamins, oats, nuts, spice, milk, egg, salt, flour, lecithin, xanthan gum, or sweetening agents.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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DETAILED DESCRIPTION
(44) Reference will now be made in detail to representative embodiments of the invention. While the invention will be described in conjunction with the enumerated embodiments, it will be understood that the invention is not intended to be limited to those embodiments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents that may be included within the scope of the present invention as defined by the claims. Thus, there are a variety of suitable formulations of the compositions described herein. These formulations are exemplary and are in no way limiting. Furthermore, one skilled in the art will appreciate that routes of administering the compositions and/or salts thereof include, but are not limited to, oral or alimentary administration. Although more than one route can be used, a particular route can provide a more immediate and more effective response than another route in a given situation.
(45) One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in and are within the scope of the practice of the present invention. The present invention is in no way limited to the methods and materials described.
(46) Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the invention, the preferred methods, devices and materials are now described.
(47) All publications, published patent documents, and patent applications cited in this application are indicative of the level of skill in the art(s) to which the application pertains. All publications, published patent documents, and patent applications cited herein are hereby incorporated by reference to the same extent as though each individual publication, published patent document, or patent application was specifically and individually indicated as being incorporated by reference.
(48) As used in this application, including the appended claims, the singular forms a, an, and the include plural references, unless the content clearly dictates otherwise, and are used interchangeably with at least one and one or more. Thus, reference to a dietary fiber includes mixtures of dietary fibers, reference to an iron complex includes mixtures of iron complexes, and the like.
(49) As used herein, about will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art given the context in which it is used, about will mean up to plus or minus 10% of the particular term.
(50) Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein.
(51) All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., such as) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.
(52) As used herein, the terms comprises, comprising, includes, including, contains, containing, and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, product-by-process, or composition of matter that comprises, includes, or contains an element or list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, product-by-process, or composition of matter.
(53) Disclosed herein are iron compounds complexed to fiber which alter or add further benefit to the nature of dietary fiber. As such, provided herein are iron-fiber compositions having a high content of iron(II) and/or iron (III). Exemplary fibers include natural fibers, man-made fibers, and combinations thereof. The polymer complex can be amorphous, crystalline and contain microdomains of both amorphous and crystalline regions ranging from 10% to 90% amorphous and 10% to 90% crystalline. The location of the iron(II) and iron(III) can be in either the amorphous or crystalline regions or both.
(54) Dietary fiber refers to indigestible portion of plant foods. As used herein dietary fiber includes, but is not limited to non-starch polysaccharides such as arabinoxylans, cellulose, and many other plant components such as resistant dextrins, inulin, lignin, waxes, chitins, pectins, beta-glucans, and oligosaccharides. The dietary fiber may be naturally occurring, synthetic or a mixture thereof.
(55) Exemplary iron compounds include, but are not limited to iron(II) acetate, iron(II) citrate, iron(II) ascorbate, iron(II) oxalate, iron(II) oxide, iron(II) carbonate, iron(II) carbonate saccharate, iron(II) formate, iron(II) sulfate, iron(II) chloride, iron(III) chloride, iron (II) bromide, iron (II) iodide, iron (III) fluoride, iron(II) acetylacetonate, iron (III) phosphate, iron (III) pyrophosphate, and combinations thereof.
(56) The iron-fiber compositions or complexes according one embodiment are oligo- or polynuclear iron compositions in which the iron atoms are bonded to one another via oxygen atoms and/or hydroxyl groups, and wherein the iron is bonded to the fiber as a complex and/or via carbon, oxygen, nitrogen, and/or hydrogen bridge bonds. The hydroxyl bridges also have a high binding affinity for Fe(II) and/or Fe(III). The iron-fiber compositions can also contain water bonded as a complex or via hydrogen bridge bonds.
(57) The iron-fiber compositions according to the invention are characterized by their content of iron(II) and iron(III). This means that some of the iron is present in the oxidation level of 2.sup.+ and some in the oxidation level of 3.sup.+. These are therefore so-called mixed valence compounds, in which the metal is present in several oxidation levels side by side.
(58) In some embodiments, the content of iron(II) and iron(III) in the total iron content of the iron-fiber composition is at least 2 wt. %. For example, the content of iron(II) and iron(III) in the total iron content can be 2 to 50 wt %, or 3 to 50 wt. % or 3 to 25 wt. %, or 20 to 50 wt. %, or 10 to 50 wt %, or 10 to 40 wt. % or 15 to 30 wt %, or about 10 wt %, or about 15 wt %, or about 20 wt %, or about 30 wt %, or about 40 wt %, or any other range or value within those ranges. The content of fiber by weight of the composition is 10 to 98 wt. %, for example, about 10 to 80 wt. %, about 50 to 90 wt %, about 60 to 90 wt %, about 70 to 85 wt %, about 35 to 65 wt. %, about 40 to 60 wt. %, about 45 to 55 wt. %, or about 20%, or about 30%, or about 40%, or about 50% by weight, or any other range or value within those ranges. The iron(II) and iron(III) are on the surface of the fiber and in the bulk fiber; the selected weight ratio of surface vs. bulk iron content can be 10 to 90 wt % or 90 to 10 wt. % and in between. The action of the iron-fiber composition is not affected by the iron at one location compared to another. In some embodiments, the iron-fiber compositions comprise ferrous (Fe.sup.2+) and/or ferric (Fe.sup.3+) compounds and a dietary fiber in a complex or pharmaceutically acceptable salts thereof in a physiologically or pharmaceutically acceptable carrier. The compositions that make up a therapeutic formulation can be mixtures of non-iron containing fibers and iron(II) and iron(III)-containing fibers. As used herein, iron compound, salt, iron-fiber complex or composition thereof, the term iron includes both Iron (II) or Ferrous and Iron (III) or Ferric compounds or combinations thereof.
(59) As used herein, the term liquid includes, but is not limited to water, bodily fluids, aqueous and organic solvents, aqueous and organic solutions.
(60) In one embodiment, a medical food suitable for mammals is provided comprising at least 400 mg of the iron-fiber composition described herein. The medical food can be in the form of a liquid solution; powder, bar, wafer, a suspension in an appropriate liquid or in a suitable emulsion, as detailed below. In some embodiments, the medical food may further comprise one or more additional ingredients selected from the group including, but not limited to natural flavors, artificial flavors, major trace and ultra-trace minerals, minerals, vitamins, oats, nuts, spices, milk, egg, salt, flour, lecithin, xanthan gum and/or sweetening agents.
(61) As used herein the term medical food, as defined in section 5(b) of the Orphan Drug Act (21 U.S.C. 360ee (b) (3)) is a food which is formulated to be consumed or administered enterally under the supervision of a physician and which is intended for the specific dietary management of a disease or condition for which distinctive nutritional requirements, based on recognized scientific principles, are established by medical evaluation.
(62) Formulations suitable for oral administration are described herein for purposes of illustration. Oral formulations can include of (a) liquid solutions, such as an effective amount of the composition thereof dissolved in diluents, such as water, saline, or orange juice; (b) capsules, sachets, tablets, lozenges, and troches, each containing a predetermined amount of the active ingredient, as solids or granules; (c) powders; (d) suspensions in an appropriate liquid; (e) nano or micro particles; and (f) suitable emulsions. Liquid formulations may include diluents, such as water and alcohols, for example, ethanol, benzyl alcohol, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant. Capsule forms can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and corn starch. Tablet forms can include one or more of lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and other excipients, colorants, diluents, buffering agents, disintegrating agents, moistening agents, preservatives, flavoring agents, and pharmacologically compatible excipients. Lozenge forms can comprise the active ingredient in a flavor, usually sucrose and acacia or tragacanth, as well as pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, and the like containing, in addition to the active ingredient, such excipients as are known in the art.
(63) The composition can be consumed at any time during the day, e.g. as a meal, before, during, or after a meal, etc.
(64) The compositions of the invention described herein can be administered to an extracorporeal system to adsorb certain accessible targets in the extracorporeal system in vitro. Furthermore, the compositions of the invention can be administered to a subject in vivo or ex vivo.
(65) The compositions of the invention can be administered to a cell, for example, to a cell of a subject. Subjects include, for example, bacteria, yeast, fungi, plants, and mammals. In some embodiments, the subject is a mammal Mammals include, but are not limited to, the order Rodentia, such as mice, and the order Logomorpha, such as rabbits, the order Carnivora, including Felines (cats) and Canines (dogs), the order Artiodactyla, including Bovines (cows) and Swines (pigs), the order Perssodactyla, including Equines (horses), the order Primates, Ceboids, or Simioids (monkeys), the order Anthropoids (humans and apes). Illustratively the mammal is the human. Furthermore, the subject can be the unborn offspring of any of the forgoing subjects, including mammals (e.g., humans), in which case any screening of the subject or cells of the subject, or administration of compositions to the subject or cells of the subject, can be performed in utero.
(66) The amount or dose of a composition should be sufficient to affect a therapeutic or prophylactic response in the subject over a reasonable time frame. The appropriate dose will depend upon the nature and severity of the disease or affliction to be treated or prevented, as well as by other factors. For instance, the dose also will be determined by the existence, nature and extent of any adverse side effects that might accompany the administration of the particular composition. Ultimately, the attending physician will decide the dosage of the composition of the present invention with which to treat each individual patient, taking into consideration a variety of factors, such as age, body weight, general health, diet, sex, composition to be administered, route of administration, and the severity of the condition being treated. An exemplary dose of a composition is the maximum that a patient can tolerate without incurring serious side effects. Typical doses might be, for example, about 0.01 g/kg/day to about 20 g/kg/day.
(67) The compositions can be used for any purpose including, without limitation, the treatment, prevention, or diagnosis of a disease or condition, the screening of compounds that can be used to treat, prevent, or diagnose a disease or condition, or the research of the underlying mechanisms or causes of a disease or condition, which research can be used, for example, in the development of methods to treat, prevent, or diagnose the disease or condition. Without wishing to be bound by any particular theory, it is believed that the compositions of the invention are particularly useful with respect to diseases and conditions involving the adsorption of certain accessible targets in gastrointestinal tract or in the extracorporeal system.
(68) Diagnose, diagnosing, diagnosis, and variations thereof refer to the detection, determination, or recognition of a health status or condition of an individual based on one or more signs, symptoms, data, or other information pertaining to that individual. The health status of an individual can be diagnosed as healthy/normal (i.e., a diagnosis of the absence of a disease or condition) or diagnosed as ill/abnormal (i.e., a diagnosis of the presence, or an assessment of the characteristics, of a disease or condition). The terms diagnose, diagnosing, diagnosis, etc., encompass, with respect to a particular disease or condition, the initial detection of the disease; the characterization or classification of the disease; the detection of the progression, remission, or recurrence or reactivation of the disease; and the detection of disease response after the administration of a treatment or therapy to the individual. The diagnosis of a disease or condition includes distinguishing individuals who have said disease or condition from individuals who do not.
(69) Prognose, prognosing, prognosis, and variations thereof refer to the prediction of a future course of a disease or condition in an individual who has the disease or condition (e.g., predicting patient survival), and such terms encompass the evaluation of disease response to the administration of a treatment or therapy to the individual. Prognosing and variants thereof can also mean predicting evidence of disease (EVD) or no evidence of disease (NED) in the individual at a future preselected time point. The date of prognosing can be referred to as time point 1 (TP1), and the preselected future time point may be referred to as time point 2 (TP2) and can include a specific future date or range of dates, for example post-treatment follow-up.
(70) Evaluate, evaluating, evaluation, and variations thereof encompass diagnosing, treating, prognosing and monitoring of recurrence in a treated individual. Evaluating can include any of the following: 1) diagnosing, i.e., initially detecting the presence or absence of a disease or condition; 2) prognosing at time point 1 (TP1), the future outcome of treatment at time point 2 (TP2), i.e., where TP2 may follow therapy; 3) detecting or monitoring disease progression or recurrence after apparent cure of said disease or condition i.e., wherein monitoring after apparent cure means testing an individual a time point after he or she has received successful treatment, and/or 4) detecting progression from latent infection to active disease.
(71) Treatment, as used herein refers to an intervention performed with the intention of preventing the development or altering the pathology of a disorder. Accordingly, treatment refers to both therapeutic and prophylactic or preventative measures. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented.
(72) Therapy as used herein refers to an intervention performed with the intention of preventing the development or altering the pathology of a disorder. Therapy refers to various methods that target particular diseases with particular disease fighting agents. For example, a targeted therapy might involve providing to a subject in need thereof the iron-fiber composition in a physiologically acceptable carrier for adsorbing undesirable agents including, but not limited to excessive amounts of calcium, cholesterol, phosphate, potassium, sodium, as well as, toxins from infectious agents via in vivo, extracorporeal, ex vivo, or in vitro administration
(73) As used utilized herein, the term pharmaceutically acceptable means approved by a regulatory agency of a federal or a state government or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals and, more particularly, in humans. The term carrier refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered and includes, but is not limited to such sterile liquids as water and oils.
(74) As used herein, the term physiologically acceptable carrier refers to any carrier or excipient commonly used with pharmaceuticals. Such carriers or excipients include, but are not limited to, oils, starch, sucrose and lactose.
(75) A pharmaceutically acceptable salt or salt of an iron-fiber composition is a product of the disclosed composition that contains an ionic bond, and is typically produced by reacting the disclosed compositions with either an acid or a base, suitable for administering to a subject. A pharmaceutically acceptable salt can include, but is not limited to, acid addition salts including hydrochlorides, hydrobromides, phosphates, sulphates, hydrogen sulphates, alkylsulphonates, arylsulphonates, arylalkylsulfonates, acetates, benzoates, citrates, maleates, fumarates, succinates, lactates, and tartrates; alkali metal cations such as Li, Na, K, alkali earth metal salts such as Mg or Ca, or organic amine salts.
(76) A pharmaceutical composition is a formulation comprising the disclosed compositions in a form suitable for administration to a subject. A pharmaceutical composition of the invention is preferably formulated to be compatible with its intended route of administration.
(77) As used herein the term therapeutically effective amount generally means the amount necessary to ameliorate at least one symptom of a disorder to be prevented, reduced, or treated as described herein. The phrase therapeutically effective amount as it relates to the compositions described herein shall mean the dosage that provides the specific pharmacological response for which the composition is administered in a significant number of subjects in need of such treatment. It is emphasized that a therapeutically effective amount that is administered to a particular subject in a particular instance will not always be effective in treating the conditions/diseases described herein, even though such dosage is deemed to be a therapeutically effective amount by those of skill in the art.
(78) Thus, in one aspect a method of treating a disease which benefits from adsorption of certain accessible targets in gastrointestinal tract or in an extracorporeal system is provided. The method comprises administering to a patient in need thereof a therapeutically effective amount of the composition of the instant disclosure. The foregoing method is suitable for use in a subject or patient that is afflicted with a disease or at risk for developing a disease, such as a disease that benefits from adsorption of certain accessible targets in gastrointestinal tract or in an extracorporeal system. Such diseases include, for example, a bone disorder, cardiovascular disease, a cardiovascular complication associated with renal disease, endothelial dysfunction, hyperparathyroidism, hypercalcemia, hyperphosphatemia, an immune disorder, left ventricular hypertrophy, a proliferative disease, proteinuria, renal disease, viral infection, bacterial infection, musculoskeletal disorders, high blood pressure, hypertriglyceridemia, lipid disorders, hyperlipoproteinemia, hyperlipidemia, dyslipidemia, diabetes, hypercholesterolemia, multiple sclerosis, myelodysplastic syndrome, proximal myopathy, premature aging, metabolic syndrome, insulin resistance, obesity. One or more symptoms of the disease is prevented, reduced, or eliminated subsequent to administration of the composition, thereby effectively treating or preventing the disease to at least some degree.
(79) The patient or subject can be any animal, domestic, livestock or wild, including, but not limited to cats, dogs, horses, pigs and cattle, and preferably human patients. As used herein, the terms patient and subject may be used interchangeably.
(80) In another aspect a method for preparing the disclosed compositions is provided. Generally, in one embodiment an iron salt, or a mixture of iron salts, is mixed together with a fiber carrier under acidic conditions at a pH in the range from about 1.0 to about 6.0 (e.g. from about 1 to about 4, or from about 1 to about 3). To the mixture an alkali salt is added. The resulting solution is purified of excess debris, salts, impurities, etc., by any suitable method to produce an iron-fiber complex with an elemental iron concentration between about 2% to about 50%.
(81) In yet another aspect, the iron-fiber complex is prepared by a process comprising the steps of: (a) mixing one or more fibers and an iron compound, at a pH<3; (b) maintaining a temperature of reaction mixture of step (a) between ambient and 100 C.; (c) cooling the reaction mixture of step (b) to ambient temperature and washing until pH is neutral; and (d) isolating the iron-fiber complex compound formed, wherein the iron content is in an amount of from 2 to 50 wt %.
(82) An acid is used optionally to achieve a pH in a range of about 1 to about 3, said acid is selected from the group of hydrogen halides and their solutions including, but not limited to: hydrochloric acid (HCl), hydrobromic acid (HBr), hydroiodic acid (HI), halogen oxoacids such as hypochlorous acid (HClO), chlorous acid (HClO.sub.2), chloric acid (HClO.sub.3), perchloric acid (HClO.sub.4), and corresponding compounds for bromine and iodine, sulfuric acid (H.sub.2SO.sub.4), fluorosulfuric acid (HSO.sub.3F), nitric acid (HNO.sub.3), phosphoric acid (H.sub.3PO.sub.4), fluoroantimonic acid (HSbF.sub.6), fluoroboric acid (HBF.sub.4), hexafluorophosphoric acid (HPF.sub.6), chromic acid (H.sub.2CrO.sub.4), boric acid (H.sub.3BO.sub.3). Other acids are contemplated herein and are easily identifiable by one skilled in the art.
(83) After fiber and iron compounds are mixed under acidic conditions, a base or an alkali salt can be optionally added to the fiber/iron mixture to alter the pH to be at least 3. In various embodiments, the pH is adjusted to be in the range of greater than about 3 and less than about 12. Alkali carbonates and alkali metal hydroxides are illustrative bases or alkali substances useful herein, though others are contemplated. The base can be selected from the group including, but not limited to LiOH, KOH, NaOH, NaHCO.sub.3, Na.sub.2CO.sub.3, Ca(OH).sub.2, Mg(OH).sub.2, Li.sub.2CO.sub.3, K.sub.2CO.sub.3, CaCO.sub.3, MgCO.sub.3, and Na.sub.2CO.sub.3. The base can comprise any wt. % of the total weight of the iron-fiber mixture, sufficient to alter the pH of the mixture.
(84) The temperature of the reaction mixture is in the range from about 20 C. to about 100 C., for example, about 30 C., about 40 C., about 50 C., about 60 C., about 70 C., about 80 C., about 90 C., or about 100 C. The time interval is in the range from about 60 minutes to about 48 hours, for example, about 2 hours, about 3 hours, about 4 hours, about 6 hours, about 8 hours, about 12 hours, about 18 hours, about 24 hours, about 30 hours, about 36 hours, about 42 hours, or about 48 hours.
(85) The following examples are provided for illustrative purposes only and are not intended to limit the scope of the application as defined by the appended claims. All examples described herein were carried out using standard techniques, which are well known and routine to those of skill in the art.
EXAMPLES
Example 1
(86) Prepared an aqueous solution of FeCl.sub.3 (FeCl.sub.3.6H.sub.2O, Sigma F2877) in water at 0.5 g/ml, at a pH in the range of 1-3).
(87) Mixed 0.5 g dietary fiber (for example, Ultimate Fiber or equivalent) with 12.5 ml of the FeCl.sub.3 solution (pH<3) and allowed the mixture to shake in a shaker for 1 hour (hr) or 24 hrs at room temperature with shaking. Washed with water until the supernatant was clear.
(88) As a control, mixed 0.5 g fiber with 10 ml water. Gently shook the mixture for 24 hrs at room temperature.
(89) Dried the materials using a food dehydrator for 24 hours.
(90) With 0.08 gram of the dried composition, added 1 ml of D-PBS (Invitrogen) containing 10 mM phosphate to each sample and incubated at room temperature for at least 1 hr. Centrifuged and collected the supernatant.
(91) Added 0.5 ml of D-PBS to the precipitate, mixed well, centrifuged and collected the supernatant. Repeated the above process for 5 times.
(92) Determined the phosphate level in the supernatants using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(93)
Example 2
(94) Prepared an aqueous solution of FeCl.sub.3 in water at 0.5 g/ml, preferably at pH in the range of 1-3.
(95) Mixed 0.5 g fiber with 5 g FeCl.sub.3 in 10 ml water. Shook the mixture gently for 24 hrs or 48 hrs at room temperature. Added 0.1 g KOH. Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear (using centrifugation or by filtering through a filter paper).
(96) As a control, mixed 0.5 g fiber with 10 ml water. Incubated the mixture for 48 hrs at room temperature with shaking. Added 0.1 g KOH. Washed with water until the supernatant was at pH=7.
(97) Dried the materials using a food dehydrator for 24 hours.
(98) With 0.08 gram of the dried composition, added 1 ml of D-PBS (Invitrogen) containing 10 mM phosphate to each sample and incubate at room temperature for at least 1 hr. Centrifuged and collected the supernatant.
(99) Added 0.5 ml of D-PBS to the precipitate, mixed well, centrifuged and collected the supernatant. Repeated the above process for 5 times.
(100) Determined the phosphate level in the supernatants using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(101)
Example 3
(102) Mixed 0.5 g or 1 g fiber with 5 g FeCl.sub.3 in 10 ml water. Incubated the mixture for 24 hrs at room temperature with shaking. Added 0.67 g NaOH in 1 ml of water. Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear.
(103) Adjusted the pH with more NaOH until pH=7. Then washed with water for two more times (by centrifugation).
(104) Took 1 g of the wet material. Added 0.5 ml of D-PBS and incubated for 30 mM, mixed well, centrifuged and collected the supernatant.
(105) Added 0.5 ml of D-PBS to the precipitate, mixed well, centrifuged and collected the supernatant. Repeated the above process 3 times.
(106) Added 0.5 ml of D-PBS to the precipitate, incubated for 10 min, centrifuged and collected the supernatant.
(107) Added 0.5 ml of D-PBS to the precipitate, incubated for 30 min, centrifuged and collected the supernatant.
(108) Determined the phosphate level in the supernatants using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(109)
Example 4
(110) Mixed 2 g or 3 g fiber with 5 g FeCl.sub.3 in 30 ml water. Incubated the mixture for 2 hrs at room temperature with shaking. Added 0.3 g KOH (pH 4.5). Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear.
(111) Adjusted the pH with NaOH until pH=7. Then washed with water for two more times (by centrifugation).
(112) Took 1 g of the wet material. Added 0.5 ml of D-PBS and incubated for 30 min, mixed well, centrifuged and collected the supernatant.
(113) Added 0.5 ml of D-PBS to the precipitate, mixed well, centrifuged and collected the supernatant. Repeated the above process for 3 times.
(114) Added 0.5 ml of D-PBS to the precipitate, incubated for 10 min, centrifuged and collected the supernatant.
(115) Added 0.5 ml of D-PBS to the precipitate, incubated for 30 min, centrifuged and collected the supernatant.
(116) Determined the phosphate level in the supernatants using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(117)
Example 5
(118) Mixed 4 g or 5 g fiber with 5 g FeCl.sub.3 in 110 ml water. Incubated the mixture for 2 hrs at room temperature with shaking. Added NaOH to pH=9. Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear (pH=7.0).
(119) Took 1 g of the wet material. Added 0.5 ml of D-PBS and incubated for 30 min, mixed well, centrifuged and collected the supernatant.
(120) Added 0.5 ml of D-PBS to the precipitate, mixed well, centrifuged and collected the supernatant. Repeated the above process for 4 times.
(121) Added 0.5 ml of D-PBS to the precipitate, incubated for 30 min, centrifuged and collected the supernatant.
(122) Determined the phosphate level in the supernatants using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(123)
Example 6
(124) Took 1 g of the fiber:FeCl.sub.3=1:1 composition from Example 5. Added 0.5 ml of D-PBS. In one tube, added 2 l of NaOH at 12.5 N. In another tube, added 2 l of concentrated HCl. In another tube, added 2 l of concentrated acetic acid. Incubated for 30 min at room temperature, mixed well, measured pH, centrifuged and collected the supernatant.
(125) Added 0.5 ml of D-PBS to the precipitate. Adjusted the pH by adding NaOH (12.5 N) or concentrated HCl or concentrated acetic acid as described above to the corresponding tubes. Mixed well, determined pH, centrifuged and collected the supernatant.
(126) Added 0.5 ml of D-PBS to the precipitate, mixed well, centrifuged and collected the supernatant immediately. Repeated 3 times.
(127) Added 0.5 ml of D-PBS to the precipitate, incubated for 30 min, centrifuged and collected the supernatant.
(128) Determined the phosphate level in the supernatants using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(129) TABLE-US-00001 Iron:fiber Iron:fiber Iron:fiber Iron:fiber at at 1:1 pH at 1:1 + NaOH pH at 1:1 + HCl pH 1:1 + acetic acid pH Weight of wet composition used in assay 0.88 g 0.89 g 0.93 g 1.02 g Phosphate Bound, Phosphate Bound, Phosphate Bound, Phosphate Bound, Supernatant moles moles moles moles 1 4.89 7 4.66 8 4.71 5 4.90 6 2 4.27 7 3.05 10 4.73 3 4.74 4 3 3.44 7 1.65 10 3.61 6 4.45 7 4 2.92 7 1.65 10 2.80 6.5 2.42 7 5 3.02 7 0.96 9 1.57 7 2.32 7 6 3.30 7 1.49 9 3.47 7 3.89 7 Sum of phosphate 21.84 13.45 20.90 22.72 bound, moles moles phosphate 24.8 15.1 22.5 22.3 bound/g of wet material
Example 7
(130) Mixed 0.5 g fiber with 5 g FeCl.sub.3 in 10 ml water. Incubated the mixture overnight at room temperature. Added 0.1 g KOH. Mixed and incubated at room temperature for at least 1 hr. Washed with water until the supernatant was clear.
(131) As a control, mixed 0.5 g fiber with 10 ml water. Incubated the mixture overnight at room temperature. Added 0.1 g KOH. Mixed and incubated at room temperature for at least 1 hr. Washed with water until the supernatant was clear.
(132) Removed 1 g of the wet fiber mixture from each sample to a column; added D-PBS (Invitrogen) containing 10 mM phosphate. Incubated for at least 1 hr at room temperature. Collect fractions at 1 ml/fraction.
(133) Determined the phosphate level in the fractions using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(134)
Example 8
(135) Mixed 100 g dietary fiber (e.g. Organic Triple Fiber or equivalent) with 500 g of FeCl.sub.3 in 1.5 liter of water. Incubated the mixture overnight at room temperature. Added 10 g KOH. Mixed and incubated at room temperature for at least 1 hr. Washed with water until the supernatant was clear.
(136) As a control, mixed 100 g fiber with 1.5 liter water. Incubated the mixture overnight at room temperature. Added 0.1 g KOH. Mixed and incubated at room temperature for at least 1 hr. Washed with water until the supernatant was clear.
(137) Removed 1 g of the wet material (1 g wet fiber=0.12 g dry fiber) from each sample to a column, added D-PBS (Invitrogen) containing 10 mM phosphate. Incubated for at least 1 hr at room temperature. Collected fractions at 1 ml/fraction.
(138) Determined the phosphate level in the fractions using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(139)
Example 9
(140) Mixed 100 g dietary fiber with 100 g of FeCl.sub.3 in 2.2 liter of water. Incubated the mixture for 24 hrs at room temperature with shaking. Added 46 g NaOH. Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear.
(141) Dried the materials for 24 hours using a food dehydrator.
(142) With 1 gram of the dried composition, added 4 ml of a phosphate solution containing 20.4 mM KH.sub.2PO.sub.4 and 23.9 mM K.sub.2HPO.sub.4 and incubated at room temperature for at least 1 hr. Centrifuged and collected the supernatant.
(143) As a control, mixed 1 g of dietary fiber Metamucil with 4 ml of a phosphate solution containing 20.4 mM KH.sub.2PO.sub.4 and 23.9 mM K.sub.2HPO.sub.4 and incubated at room temperature for at least 1 hr. The liquid portion was completely soaked up by Metamucil. Therefore, added 2 more ml of the phosphate solution containing 20.4 mM KH.sub.2PO.sub.4 and 23.9 mM K.sub.2HPO.sub.4. Centrifuged and collected the supernatant.
(144) Determined the phosphate level in the supernatants using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
Example 10
(145) Took 49 g of the dried fiber:FeCl.sub.3=1:1 composition from Example 9. Mixed the composition with 490 g normal rat chow and 3.23 g KH.sub.2PO.sub.4+1.67 g K.sub.2HPO.sub.4. Ground the mixture until powdery.
(146) As a control, took 49 g of the dried fiber alone composition (no iron during treatment). Mixed the composition with 490 g normal rat chow and 3.23 g KH.sub.2PO.sub.4+1.67 g K.sub.2HPO.sub.4. Ground the mixture until powdery.
(147) Male, Sprague Dawley, rats were placed in metabolic cages with 1 rat per cage. Urine samples were collected for 24 hrs. Blood samples were collected from each rat for serum preparation. The rats were then placed in normal cages. Some rats were provided with the powder rat chow containing fiber alone and KH.sub.2PO.sub.4+K.sub.2HPO.sub.4. The other rats were provided with the powder rat chow containing the iron-fiber composition and KH.sub.2PO.sub.4+K.sub.2HPO.sub.4.
(148) After four days, the rats were placed in metabolic cages with 1 rat per cage. Urine samples were collected for 24 hrs. Blood samples were collected from each rat to prepare serum. The phosphorus/phosphate level was determined in each urine and serum samples. Each group had at least 5 rats.
(149) The serum phosphate was elevated in the rats treated with fiber alone, but not in the rats treated with iron-fiber (
(150) There was no significant difference in the amount of food consumed by the two groups of rats during the treatment.
Example 11
(151) Mixed 5 g Fe.sub.4O.sub.2P.sub.6 (Sigma p6526) in 30 ml water. Adjusted pH by adding HCl (concentrated) until pH at 1. Added 0.5 g fiber. Incubated the mixture for 2 hrs at room temperature with shaking. Added NaOH to neutralize. Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear.
(152) Took 0.94 g of the wet material. Added 0.5 ml of D-PBS and incubated briefly, mixed well, centrifuged and collected the supernatant.
(153) Repeated the above process for 5 more times.
(154) Determined the phosphate level in the supernatants using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(155)
Example 12
(156) The Fiber:FeCl.sub.3 composition from Example 9 was sputter coated with Platinum/Palladium and mounted on Aluminum stubs, and examined under Hitachi S3000N Variable Pressure SEM (Scanning Electron Microscope).
(157)
Example 13
(158) Mixed 1 g fiber with 0, 0.2, 0.6, 2 and 5 g FeCl.sub.3 in 20 ml water (pH ranging from 1 to 2.05 with FeCl.sub.3 and pH=7.38 without FeCl.sub.3). Incubated the mixture at room temperature for at least 1 hr. Washed with water until the supernatant was clear. Dried using a food dehydrator.
(159) Removed 0.1 g of the dry composition from each sample, and mixed with 8 ml of water and 2 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, adjusted pH to 7.0 with acetic acid). Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination by the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(160)
(161)
Example 14
(162) Mixed 0.1 g of the Fiber:FeCl.sub.3 composition from Example 13 Tube#4 (fiber:iron at 1:2 ratio) with 2 ml, 4 ml, 6 ml, and 8 ml of the phosphate solution described in Example 13 (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Each tube was added the diluting buffer (the same buffer without phosphoric acid) to a final volume of 8 ml/tube to result in 5 mM, 10 mM, 15 mM and 20 mM of final phosphate concentration. Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination by the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(163)
Example 15
(164) Mixed 1 g fiber with 5 g FeCl.sub.3 in 40 ml water (pH=1.44) per tube. Incubated the mixture at room temperature for at least 1 hr. Added different amounts of NaOH (12.5 N) so that the pH values changed to the following: Tube 1, pH=1.44; Tube 2, pH=1.72; Tube 3, pH=2.14; Tube 4, pH=3.1; Tube 5, pH=7; Tube 6, pH=9.43. Mixed and incubated at room temperature for at least 1 hr. Washed with water until the supernatant was clear. Dried using a food dehydrator.
(165) Removed 0.1 g of the dry composition from each sample, and mixed with 8 ml of water and 2 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(166)
(167)
(168)
(169) Adding 25 mM (final concentration) Tris buffer during the preparation of the iron-fiber composition made no significant differences in the results.
Example 16
(170) Mixed 100 g dietary fiber with 100 g of FeCl.sub.3 in 2.2 liter of water. Incubated the mixture at room temperature for at least 1 hr with shaking. Added 45.4 g NaOH (final pH=10). Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear.
(171) Dried the materials using a food dehydrator.
(172) By inductively coupled plasma optical emission spectrometry (ICP-OES), the iron content in this dry iron-fiber composition was determined to be 15.3%.
(173) Mixed 0.1 gram of the dried iron-fiber composition with the phosphate solution and the diluting buffer as described in Example 14 to a final volume of 20 ml/tube to result in 0 mM, 1 mM, 2.5 mM and 5 mM of final phosphate concentration. Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination by the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(174) As a control, samples containing 0.1 g of sevelamer in powder form in the place of the dried iron-fiber were prepared simultaneously.
(175)
(176)
Example 17
(177) Prepared the phosphate solution and the diluting buffer as described in Example 14 to a final volume of 10 ml/tube at 10 mM of final phosphate concentration. Adjusted the pH to 1.59, 4.39, 7.1, 8.97, and 12.25.
(178) Added 0.1 gram of the dried iron-fiber composition from Example 16. Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination by the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(179)
Example 18
(180) Took dried iron-fiber from Example 16. Mixed the composition with normal rat chow and KH.sub.2PO.sub.4+K.sub.2HPO.sub.4 as in Example 10 except that the amount of the iron-fiber was at 0.2-10% by weight of the total mixture. Ground the mixture until powdery.
(181) As a control, prepared a mixture with sevelamer powder and normal rat chow and KH.sub.2PO.sub.4+K.sub.2HPO.sub.4 with the amount of sevelamer at 0.2-10% by weight of the total mixture. Ground the mixture until powdery.
(182) Male, Sprague Dawley, rats were placed in metabolic cages with 1 rat per cage. Urine samples were collected for 24 hrs. Blood samples were collected from each rat for serum preparation.
(183) Rats were fed with the diet containing high phosphates and different preparations as mentioned above.
(184) After four days, the rats were placed in metabolic cages with 1 rat per cage. Urine samples were collected for 24 hrs. Blood samples were collected from each rat to prepare serum. The phosphorus/phosphate and calcium levels were determined in each urine and serum samples. The serum iron levels were also determined in some serum samples (QuantiChrom Iron Assay Kit by BioAssay System; catalog #DIFE-250).
(185)
(186) No significant difference was observed in the serum calcium concentrations in the rats (
(187)
(188)
(189)
Example 19
(190) Took dried iron-fiber from Example 16. Mixed the composition with normal rat chow (containing 1% calcium and 0.7% phosphate) so that the iron-fiber was at 1 and 3% by weight of the total mixture. Ground the mixture until powdery.
(191) As a control, prepared a mixture with sevelamer powder and normal rat chow with the amount of sevelamer at 1 and 3% by weight of the total mixture. Ground the mixture until powdery.
(192) Male, Sprague Dawley, rats were placed in metabolic cages with 1 rat per cage. Urine samples were collected for 24 hrs. Blood samples were collected from each rat for serum preparation. Rats were fed with normal diet (containing 1% calcium and 0.7% phosphorus in powder form) and the iron-fiber material or sevelamer. After five days, the rats were placed in metabolic cages with 1 rat per cage. Urine samples were collected for 24 hrs. Blood samples were collected from each rat to prepare serum. The phosphorus/phosphate and calcium levels were determined in each urine and serum samples. Each group had at least 6 rats.
(193)
(194)
(195)
(196) Feces samples collected per 24-hr period were ashed at 800 C. for 45 minutes. Weighed 0.1 g of ash from each sample, extracted with 1 ml water by vortexing and shaking at room temperature for at least 60 min Centrifuged and collected supernatant for phosphate determination.
Example 20
(197) Took 0.1 gram of sevelamer or 0.1 gram of the dried iron-fiber composition from Example 16 and mixed with 10 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for at least 24 hrs. Centrifuged and removed 8.6 ml of the supernatant.
(198) Prepared a stock solution of 6 mg cholesterol (water-soluble cholesterol, Sigma C4951) in 1 ml water. Added 0.5 ml (3 mg) to the tubes containing sevelamer or the iron-fiber composition. Incubated at room temperature for at least 30 min with gentle shaking. Removed the supernatant for determining cholesterol using the Stanbio Liquicolor cholesterol assay kit (Catalog #1010-430)
(199)
Example 21
(200) Mixed 1 g fiber with 5 g FeCl.sub.3 in 40 ml water (pH=1.44) per tube. Incubated the mixture at room temperature, or 37 C., or 55 C. for at least 1 hr. Added NaOH (12.5 N) to neutralize. Washed with water until the supernatant was clear and pH was 7. Dried using a food dehydrator.
(201) Removed 0.1 g of the dry composition from each sample, and mixed with 5 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(202)
Example 22
(203) Mixed 1 g fiber with 0, 0.2, 0.6, 2 and 5 g FeCl.sub.3 in 40 ml water. Incubated the mixture at room temperature, or 37 C., or 55 C. for at least 1 hr. Added NaOH (12.5 N) to neutralize. Washed with water until the supernatant was clear and pH was 7. Dried using a food dehydrator.
(204) By inductively coupled plasma optical emission spectrometry (ICP-OES), the iron content in the dry iron-fiber composition prepared from incubating 1 g fiber with 5 g FeCl.sub.3 at 55 C. was determined to be 29.3%.
(205) Removed 0.1 g of the dry composition from each sample, and mixed with 5 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(206)
Example 23
(207) Took 0.1 gram the dry iron-fiber composition from Example 22 where the composition was prepared from incubating 1 g fiber with 5 g FeCl.sub.3 at 55 C. Added 5 ml of water. Incubated at 37 C.
(208) As a control, 0.1 g of sevelamer in powder form in the place of the dried iron-fiber was prepared simultaneously.
(209)
(210) Similar results were observed when simulated gastric fluid (0.2% (w/v) NaCl, 0.7% (v/v) HCl, without pepsin) instead of water was used.
(211) The volume (cm.sup.3) of the iron-fiber composition vs. sevelamer at different time points (20 min-180 min): 0.2 vs. 2.0 cm.sup.3. Large swelling volume is associated with GI discomfort. To show the volume at Time 0 more clearly, the volume of iron-fiber or sevelamer at 3 g/sample was also determined (2.7 vs. 4.4 cm.sup.3).
(212) Took 0.1 gram sevelamer or the dry iron-fiber composition from Example 22 where the composition was prepared from incubating 1 g fiber with 5 g FeCl.sub.3 at 55 C. Added 5 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(213) The phosphate-binding property of each sample normalized by per gram of dry iron-fiber was: 0.39 mmol/g of sevelamer vs. 0.35 mmol/g of iron-fiber. The phosphate-binding property of each sample normalized by per ml of final swell volume was: 0.016 mmol/ml of sevelamer vs. 0.172 mmol/ml of iron-fiber.
Example 24
(214) Mixed 5 g fiber with 10 g FeCl.sub.3 in 40 ml water. Incubated the mixture at 55 C. for at least 1 hr. Added NaOH (12.5 N) to neutralize (pH=7). Washed with water until the supernatant was clear and pH was 7. Dried using a food dehydrator.
(215) Removed 0.1 g of the dry composition, and mixed with 5 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for at least 24 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision). The phosphate-binding property of normalized by per gram of dry iron-fiber was 0.59 mmol/g.
(216) By inductively coupled plasma optical emission spectrometry (ICP-OES), the iron content in the dry iron-fiber composition was determined to be 24.5%.
(217) Took 0.1 gram the dry iron-fiber composition. Added 5 ml of simulated gastric fluid (0.2% (w/v) NaCl, 0.7% (v/v) HCl, without pepsin). Incubated at 37 C.
(218) As a control, 0.1 g of unprocessed fiber was treated simultaneously.
(219)
(220) The volume (cm.sup.3) of the iron-fiber composition vs. unprocessed fiber at different time points were: 20 min, 0.2 vs. 1.0 cm.sup.3; 60 min, 0.2 vs. 1.1 cm.sup.3; 120 min, 0.2 vs. 1.2 cm.sup.3; 180 min, 0.2 vs. 1.9 cm.sup.3, 240 min, 0.2 vs. 1.9 cm.sup.3.
Example 25
(221) The iron-fiber sample from Example 24 was analyzed further by the XPS (X-ray Photoelectron Spectroscopy). XPS experiments were performed using the Kratos Axis-165 instrument. Samples were irradiated by a monochromatic AlK.sub. X-ray source (15 kV, 10 mA) at an angle of 30 degrees from the sample surface. Photoelectrons were detected by 8 channeltrons of the concentric hemispherical analyzer over an area of 700300 microns, with a spectrometer take-off angle of zero. The detection was achieved using the constant analyzer energy (CAE) mode.
(222) Survey scans were acquired with a pass-energy of 160 eV, 1.0 eV step-size and 100 msec dwell time; while narrow scans were acquired with a pass-energy of 20 eV, 0.1 eV step-size and 200 msec. All scans were performed with the charge-neutralization system running Charge-referencing were done with the adventitious carbon peak position of 284.8 eV. The XPS analysis chamber base-Pressure was better than 2E-10 Torr, while working-Pressure was better than 3E-9 Torr.
(223)
(224) TABLE-US-00002 Raw Atomic Mass Position FWHM Area Atomic Conc, Conc, Peak BE (eV) (eV) (CPS) RSF Mass % % Fe 2p 710.000 6.100 77697.5 2.957 55.846 6.70 22.67 Cl 2p 197.000 3.481 5920.0 0.891 35.460 1.64 3.52 N 1s 398.000 2.969 2530.0 0.477 14.007 1.31 1.12 C 1s 283.000 4.089 69275.0 0.278 12.011 61.67 44.89 O 1s 530.000 4.179 89832.5 0.780 15.999 28.68 27.80
There is a significantly reduced presence of Cl in proportion to Fe in the material, suggesting that Cl was released and washed away during the process.
(225)
(226) TABLE-US-00003 Peak Position (eV) FWHM (eV) Area 0 284.820 1.218 1751.420 1 286.242 1.757 1512.195 2 288.276 2.162 498.409
Peak 0 is likely associated with Adventitious Carbon or CC bond. Peak 1 likely contains CN, or COH, or COC bonds, which are present in cellulose, arabinoxylan, inulin, beta-glucans and other fiber components. Peak 2 likely contains NCO or CO bonds, which are present in chitin, pectins and other components in natural fiber.
(227)
(228) TABLE-US-00004 Peak Position (eV) FWHM (eV) Area 0 710.585 2.483 992.150 1 712.493 4.138 1346.580 2 717.910 7.824 907.881 Note: The area was calculated from combining 2p1/2 and 2p3/2 for both Fe(3+) and Fe (2+).
The presence of Fe(2+) was calculated to be 42.4% of total Fe (based on the area). The preparation of this material only used FeCl.sub.3.
(229) A search in the patent and literature was conducted on XPS and processed fiber. Examples from the search are shown below.
(230) TABLE-US-00005 References Process XPS results Gustafsson et al., Spruce kraft pulps O 1s, C 1s 2003, Polymer 44: 661 cooked for different times and further OD.sub.0E.sub.1D.sub.1E.sub.2D.sub.2- bleached Bilba and Arsene, Silane coating of fiber O 1s, C 1s, Si 2s, Si 2p 2008, Composites Part A 39: 1488 Wang et al., 2010, Pine chemithermo- C 1s (C1, C2, C3), O1s BioResources 5: 1799 mechanical pulp treated with peracetic acid
Example 26
(231) The iron-fiber sample from Example 24 with the iron-fiber prepared from fiber and FeCl.sub.3 was analyzed further by the Raman Spectroscopy. Samples were dispersed directly onto Silicon substrates for analysis. The Raman spectra were collected using Renishaw in Via Raman instrument equipped with 785 nm laser. The samples were located using a Leica microscope with a 50 objective. The spectrum for the iron-fiber complex under normal condition is shown in
Example 27
(232) Took 0.5 g FeCl.sub.3 (Sigma F2877), FeCl.sub.2 (Sigma 372870), or iron acetate (Sigma 339199), or FeSO.sub.4 (Sigma 215422), or iron(II) ascorbate (Sigma A0207), or iron (III) citrate (Sigma F6129) and mixed with 10 ml water. Adjusted pH if necessary by adding HCl (concentrated) until pH at <3. Added 0.5 g dietary fiber per sample. Incubated the mixture for at least 1 hr at room temperature with shaking. Added NaOH (pH=10). Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear and the pH was at 7. Dried for 24 hours using a food dehydrator.
(233) Removed 0.1 g of the dry composition from each sample, and mixed with 5 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for 3 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(234) The phosphate-binding property of each sample normalized by per gram of dry iron-fiber was: fiber containing FeCl.sub.3 (0.38 mmol/g), fiber containing FeCl.sub.2 (0.57 mmol/g), fiber containing iron acetate (0.48 mmol/g), fiber containing FeSO.sub.4 (0.20 mmol/g), fiber containing iron(II) ascorbate (0.42 mmol/g), fiber containing iron (III) citrate (0.43 mmol/g).
Example 28
(235) Took 0.5 g FeCl.sub.3, or 0.5 g iron acetate, or the mixture of 0.25 g FeCl.sub.3 plus 0.25 g iron acetate and mixed with 10 ml water. Checked pH and adjusted pH if necessary by adding HCl (concentrated) until pH at <3. Added 0.5 g dietary fiber per sample. Incubated the mixture for at least 1 hr at room temperature with shaking. Added NaOH to neutralize. Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear and the pH was at 7. Dried for 24 hours using a food dehydrator.
(236) Removed 0.1 g of the dry composition from each sample, and mixed with 5 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for 24 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(237) The phosphate-binding property of each sample normalized by per gram of dry iron-fiber was: fiber containing FeCl.sub.3 alone (0.50 mmol/g), fiber containing iron acetate alone 0 (0.54 mmol/g), fiber containing the mixture of FeCl.sub.3 and iron acetate (0.52 mmol/g).
Example 29
(238) Took 0.5 g FeCl.sub.2, or 0.5 g FeSO.sub.4, or the mixture of 0.25 g FeCl.sub.2 and 0.25 g FeSO.sub.4, or the mixture of 0.25 g FeCl.sub.2 and 0.25 g iron acetate and mixed with 10 ml water. Checked pH and adjusted pH if necessary by adding HCl (concentrated) until pH at <3. Added 0.5 g dietary fiber per sample. Incubated the mixture for at least 1 hr at room temperature with shaking. Added NaOH to neutralize. Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear and the pH was at 7. Dried for 24 hours using a food dehydrator.
(239) Removed 0.1 g of the dry composition from each sample, and mixed with 5 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for 3 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(240) The phosphate-binding property of each sample normalized by per gram of dry iron-fiber was: fiber containing FeCl.sub.2 alone (0.54 mmol/g), fiber containing FeSO.sub.4 alone (0.20 mmol/g), fiber containing the mixture of FeCl.sub.2 and FeSO.sub.4 (0.54 mmol/g), fiber containing the mixture of FeCl.sub.2 and iron acetate (0.44 mmol/g).
Example 30
(241) Took 0.5 g FeCl.sub.3, or the mixture of 0.45 g FeCl.sub.3 plus 0.05 g FeCl.sub.2, or the mixture of 0.40 g FeCl.sub.3 plus 0.10 g FeCl.sub.2, or the mixture of 0.25 g FeCl.sub.3 plus 0.25 g FeCl.sub.2, or the mixture of 0.10 g FeCl.sub.3 plus 0.40 g FeCl.sub.2, or 0.5 g FeCl.sub.2 and mixed with 10 ml water. Checked pH (<3). Added 0.5 g dietary fiber per sample. Incubated the mixture for at least 1 hr at room temperature with shaking. Added NaOH to neutralize. Mixed and incubated at room temperature for at least 1 hr with shaking. Washed with water until the supernatant was clear and the pH was at 7. Dried for 24 hours using a food dehydrator.
(242) Removed 0.1 g of the dry composition from each sample, and mixed with 5 ml of a 20 mM phosphate solution (1.37 ml of 85% phosphoric acid, 3.18 g of sodium carbonate and 4.68 g of NaCl in 1 liter of water, pH=7.0). Incubated at room temperature for 3 hrs. Centrifuged and collected the supernatant for phosphate determination using the phosphate colorimetric assay (Catalog #K410-500 from Biovision).
(243) The phosphate-binding property of each sample normalized by per gram of dry iron-fiber was: fiber containing FeCl.sub.3 alone (0.33 mmol/g), fiber containing FeCl.sub.3:FeCl.sub.2 at 9:1 (0.39 mmol/g), fiber containing FeCl.sub.3:FeCl.sub.2 at 4:1 (0.49 mmol/g), fiber containing the mixture of FeCl.sub.3:FeCl.sub.2 at 1:1 (0.51 mmol/g), fiber containing the mixture of FeCl.sub.3:FeCl.sub.2 at 1:4 (0.45 mmol/g), fiber containing FeCl.sub.2 alone (0.51 mmol/g).
Example 31
(244) The iron-fiber sample from Example 29 with the iron-fiber prepared from 0.5 g FeCl.sub.2+0.5 g fiber was analyzed further by the XPS (X-ray Photoelectron Spectroscopy). XPS experiments were performed using the Kratos Axis-165 instrument. Samples were irradiated by a monochromatic AlK.sub. X-ray source (15 kV, 10 mA) at an angle of 30 degrees from the sample surface. Photoelectrons were detected by 8 channeltrons of the concentric hemispherical analyzer over an area of 700300 microns, with a spectrometer take-off angle of zero. The detection was achieved using the constant analyzer energy (CAE) mode.
(245) Survey scans were acquired with a pass-energy of 160 eV, 1.0 eV step-size and 100 msec dwell time; while narrow scans were acquired with a pass-energy of 20 eV, 0.1 eV step-size and 200 msec. All scans were performed with the charge-neutralization system running Charge-referencing were done with the adventitious carbon peak position of 284.8 eV. The XPS analysis chamber base-Pressure was better than 2E-10 Torr, while working-Pressure was better than 3E-9 Torr.
(246)
(247) TABLE-US-00006 Raw Atomic Mass Position FWHM Area Atomic Conc, Conc, Peak BE (eV) (eV) (CPS) RSF Mass % % Fe 2p 709.000 5.043 60320.0 2.957 55.846 5.04 17.89 Cl 2p 196.000 4.205 2660.0 0.891 35.460 0.71 1.61 C 1s 283.000 4.129 70540.0 0.278 12.011 60.82 46.47 O 1s 530.000 3.724 108132.5 0.780 15.999 33.43 34.03
(248)
(249) TABLE-US-00007 Peak Position (eV) FWHM (eV) Area 0 284.777 1.215 1210.937 1 286.284 1.274 1304.316 2 287.734 2.306 606/741
(250)
(251) TABLE-US-00008 Peak Position (eV) FWHM (eV) Area 0 710.532 2.686 1021.598 1 712.819 3.633 707.039 2 718.741 5.311 219.676 3 732.217 5.095 135.074 Note: The area was calculated from combining 2p1/2 and 2p3/2 for both Fe(3+) and Fe (2+).
The presence of Fe(2+) and Fe(3+) were calculated to be 59% and 41% of total Fe (based on the area), respectively. The preparation of this material only used FeCl.sub.2. The presence of Fe (3+) suggests oxidation during the process.
(252) Various embodiments of this invention are described herein. Variations may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, the inventors contemplate all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.