U.S. patent application number 09/947248 was filed with the patent office on 2003-01-23 for dry acid-chitosan complexes.
Invention is credited to Angerer, J. David, Cyron, Donald M., Iyer, Subramanian, Jerrell, Thomas A..
Application Number | 20030018176 09/947248 |
Document ID | / |
Family ID | 22396413 |
Filed Date | 2003-01-23 |
United States Patent
Application |
20030018176 |
Kind Code |
A1 |
Angerer, J. David ; et
al. |
January 23, 2003 |
Dry acid-chitosan complexes
Abstract
The invention is an acid-chitosan complex which is made up of
chitosan, a sufficient amount of one or more acids, and an
effective amount of water. This acid-chitosan is water-soluble in a
dry form. A further aspect of this invention is various methods for
producing such a water-soluble acid-chitosan complex and methods
for using this acid-chitosan complex, particularly for reducing fat
absorption in an animal.
Inventors: |
Angerer, J. David;
(Hockessin, DE) ; Cyron, Donald M.; (Lincoln
University, PA) ; Iyer, Subramanian; (Hockessin,
DE) ; Jerrell, Thomas A.; (Avondale, PA) |
Correspondence
Address: |
Basil S. Krikelis
Arkion Life Sciences
Quillen Building
3521 Silverside Road
Wilmington
DE
19810
US
|
Family ID: |
22396413 |
Appl. No.: |
09/947248 |
Filed: |
September 6, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09947248 |
Sep 6, 2001 |
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09515051 |
Feb 25, 2000 |
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6326475 |
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60121391 |
Feb 25, 1999 |
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Current U.S.
Class: |
536/20 |
Current CPC
Class: |
A61P 3/04 20180101; C08B
37/003 20130101; A61P 3/06 20180101; A61K 47/69 20170801; C08L 5/08
20130101; A61K 47/61 20170801 |
Class at
Publication: |
536/20 |
International
Class: |
C07H 015/22; C08B
037/08 |
Claims
We claim:
1. An acid-chitosan complex comprising chitosan, a sufficient
amount of one or more acids, and an effective amount of water,
wherein said acid-chitosan complex is water-soluble.
2. The acid-chitosan complex of claim 1 wherein the sufficient
amount of acid comprises about 75% or more of stoichiometric, based
on chitosan.
3. The acid-chitosan complex of claim 2 wherein the effective
amount water is about 5% to 130% of the weight of the acid and the
chitosan.
4. The acid-chitosan complex of claim 3 wherein the effective
amount of water is about 5% to 15% of the weight of the acid and
the chitosan.
5. The acid-chitosan complex of claim 1 wherein the acid is
selected from the group consisting of: hydrochloric acid, acetic
acid, lactic acid, glycolic acid, nitric acid, malic acid, pyruvic
acid, citric acid, ascorbic acid, other physiologically acceptable
carboxylic acids, betaine hydrochloride, amine hydrochlorides, and
combinations thereof.
6. The acid-chitosan complex of claim 5 wherein the acid comprises
betaine hydrochloride.
7. An acid-chitosan complex produced by the process comprising the
following steps: (1) forming a homogenous mix of chitosan and a
sufficient amount of one or more acids; and (2) adding an effective
amount of water to the homogenous mix to form a uniform
complex.
8. The acid-chitosan complex of claim 7 wherein the sufficient
amount of acid comprises about 75% or more of stoichiometric, based
on chitosan.
9. The acid-chitosan complex of claim 8 wherein the effective
amount water is about 5% to 130% of the weight of the acid and the
chitosan.
10. The acid-chitosan complex of claim 8 wherein the effective
amount water is about 5% to 15% of the weight of the acid and the
chitosan.
11. The acid-chitosan complex of claim 7 wherein the acid is
selected from the group consisting of: hydrochloric acid, acetic
acid, lactic acid, glycolic acid, nitric acid, malic acid, pyruvic
acid, citric acid, ascorbic acid, other physiologically acceptable
carboxylic acids, betaine hydrochloride, amine hydrochlorides, and
combinations thereof.
12. An acid-chitosan complex produced by a process comprising the
following steps: (3) dissolving one or more acids in water; and (4)
applying the acid-water mix to chitosan to form a complex wherein
the chitosan is not dissolved; and (5) drying the complex.
13. The acid-chitosan complex of claim 12 wherein the sufficient
amount of acid comprises about 75% or more of stoichiometric, based
on chitosan.
14. The acid-chito san complex of claim 13 wherein the effective
amount water is about 5% to 130% of the weight of the acid and the
chitosan.
15. The acid-chitosan complex of claim 13 wherein the effective
amount water is about 5% to 15% of the weight of the acid and the
chitosan.
16. The acid-chitosan complex of claim 12 wherein the acid is
selected from the group consisting of: hydrochloric acid, acetic
acid, lactic acid, glycolic acid, nitric acid, malic acid, pyruvic
acid, citric acid, ascorbic acid, other physiologically acceptable
carboxylic acids and combinations thereof.
17. A method for reducing the release of trigycerides into the
blood stream of an animal after the animal has digested a
fat-containing substance, the method comprising administering to
the animal an acid-chitosan complex prior to or during digestion of
the fat-containing substance wherein the acid-chitosan complex
comprises chitosan, a sufficient amount of an acid, and an
effective amount of water, wherein said acid-chitosan complex is
water-soluble.
18. The method of claim 17 wherein the acid-chitosan complex is
produced by a process comprising the following steps: (1) forming a
homogenous mix of chitosan and a sufficient amount of one or more
acids; and (2) adding an effective amount of water to the
homogenous mix to form a uniform complex.
19. The method of claim 18 wherein the sufficient amount of acid
comprises about 75% of stoichiometric.
20. The method of claim 19 wherein the effective amount water is
about 5% to 130% of the weight of the acid and the chitosan.
21. The method of claim 20 wherein the effective amount water is
about 5% to 15% of the weight of the acid and the chitosan.
22. The method of claim 17 wherein the acid is selected from the
group consisting of: hydrochloric acid, acetic acid, lactic acid,
glycolic acid, nitric acid, malic acid, pyruvic acid, citric acid,
ascorbic acid, other physiologically acceptable carboxylic acids,
betaine hydrochloride, amine hydrochlorides, and combinations
thereof.
23. The method of claim 17 wherein the acid-chitosan complex is
produced by a process comprising the following steps: (6)
dissolving an acid in water; and (7) applying the acid-water mix to
chitosan to form a complex wherein the chitosan is not dissolved;
and (8) drying the complex.
24. The method of claim 23 wherein the sufficient amount of acid
comprises about 75% of stoichiometric.
25. The method of claim 24 wherein the effective amount water is
about 5% to 130% of the weight of the acid and the chitosan.
26. The method of claim 25 wherein the effective amount water is
about 5% to 15% of the weight of the acid and the chitosan.
27. The method of claim 23 wherein the acid is selected from the
group consisting of: hydrochloric acid, acetic acid, lactic acid,
glycolic acid, nitric acid, malic acid, pyruvic acid, citric acid,
ascorbic acid, other physiologically acceptable carboxylic acids,
betaine hydrochloride, amine hydrochlorides, and combinations
thereof.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 09/515,051 filed Feb. 25, 2000.
FIELD OF THE INVENTION
[0002] The present invention relates to essentially dry complexes
of acid and chitosan which are water-soluble. More specifically,
the invention relates to water-soluble acid-chitosan complexes,
methods for forming such complexes and uses of these acid-chitosan
complexes for, among other things, prevention of fat digestion.
BACKGROUND OF THE INVENTION
[0003] When chitosan is prepared from the chitin fraction of the
shells of crustcea, such as shrimp, crab and lobster, the final
step is invariably and of necessity a treatment with extremely
strong sodium hydroxide. This is the predominant industrial
treatment that effectively removes acetyl groups from chitin,
converting it into chitosan. Because of this treatment, chitosan is
always found as an aminopolysaccharide with essentially none of the
amino groups being protonated. This can be referred to as the free
base form of chitosan. This natural form of chitosan is not soluble
in water. In order to dissolve chitosan in water-based systems, the
chitosan must be made more hydrophilic. This is done by adding acid
to the water being used in dissolution. The acid reacts with the
amino groups, converting them into (substituted) ammonium ions,
which are much more hydrophilic than the amino groups. When
chitosan is added to this acid-water mix, it becomes protonated. A
fully protonated (i.e., each amino group has reacted with a proton
from the acid) chitosan is the other extreme of chitosan. It should
be noted, however, that essentially no protonation of chitosan will
result if a dry acid and chitosan are merely physically blended. It
is necessary to give the acid molecule mobility, which is done by
the addition of water.
[0004] Chitosan is presently used as a dietary supplement to
prevent some of the ingested fat in a person's diet from being
absorbed and metabolized. It is thus an agent to help control
obesity. When a person takes a dosage of chitosan, the chitosan
exerts a demand on the stomach to produce hydrochloric acid in
order to dissolve it. It is understood in the art that the chitosan
must dissolve to be able to occlude the fat, which can thereafter
be passed through the digestive tract and subsequently expelled
from the body. Since the body's capacity for producing hydrochloric
acid is limited, an agent that supplies part of the necessary
acidity would be beneficial to chitosan's performance.
[0005] Merely treating chitosan with hydrochloric acid in a
manufacturing process to form a water-soluble chitosan, however has
an unintended and devastating side effect. It has been observed
that these hydrochloric acid salts of chitosan undergo
depolymerization upon storage, producing a product with too low a
molecular weight to perform in the desired manner.
[0006] Presently, in the art, water-soluble chitosans are prepared
by making a slurry of the chitosan in water and then adding acid to
the slurry. Alternatively, one may make a solution of the acid and
water and then add the chitosan under effective agitation
conditions. It would be convenient, however, and represent an
advance in the art, were all or part of the acid to be compounded
with the chitosan thus providing a uniform dry complex which can be
a shelf-stable product that will result in a reduced demand on the
stomach for acid. To date, the only way to prepare such a complex
has been achieved, with great difficulty, has been for the
manufacturer to 1) dissolve the chitosan in aqueous acid, 2) filter
the very viscous solution to remove insolubles, if necessary, and
3) spray dry the resulting solution to form a chitosan salt that is
water soluble. Such a process is cumbersome, expensive and
ineffective for an economically viable commercial process.
Therefore, there is a need in the art for a more efficient and
effective method of preparing chitosan salts.
SUMMARY OF THE INVENTION
[0007] It is a primary object of this invention to provide a
water-soluble, acid-chitosan complex.
[0008] It is also an object of the invention to provide a
water-soluble acid-chitosan complex produced by a process
comprising:
[0009] (1) forming a homogenous mix of chitosan and a sufficient
amount of an acid; and
[0010] (2) adding an effective amount of water to the homogenous
mix to form a uniform complex.
[0011] It is a further object of the invention to provide a
water-soluble acid-chitosan complex produced by a process
comprising:
[0012] (1) dissolving an acid in water; and
[0013] (2) applying the acid-water mix to chitosan to form a
complex wherein the chitosan is not dissolved; and
[0014] (3) drying the complex.
[0015] It is an additional object of the invention to provide a
method for reducing the release of triglycerides into the blood
stream of an animal by administering to the animal an effective
amount of a water-soluble acid-chitosan complex.
DETAILED DESCRIPTION OF THE INVENTION
[0016] The applicants have found that the physical blending of dry
acid and chitosan, with the addition of controlled amounts of
water, gives the acid molecules the needed mobility to accomplish
protonation and thus move from a physical mixture to a complex
(further defined below). The result of such a treatment is a state,
or complex, somewhere between full-protonation and no protonation.
The exact composition of the complex is dependent on the amount of
water and acid used and the water solubility of the acid being used
in the treatment. A more soluble acid, a higher amount of acid
and/or the use of higher levels of water will result in a complex
that is nearer to the fully-protonated extreme, whereas the use of
a less soluble acid, lesser amounts of acid and/or lesser amounts
of water will result in a complex that is much less protonated.
[0017] Definitions:
[0018] The following definitions apply throughout:
[0019] The term "acid-chitosan mixture" means the physical blend of
dry acid and chitosan wherein no protonation of any significance of
the chitosan occurs.
[0020] The term "acid-chitosan complex" means the mixture of acid
and chitosan with the addition of controlled amounts of water to
form a uniform, free-flowing mix such that the acid molecules
attain the necessary mobility to accomplish at least partial
protonation of the chitosan, to allow the acid-chitosan combination
to become water-soluble.
[0021] The term "finished moisture level" means the weight percent
of water in an acid-chitosan complex, based on the total weight of
the complex, following the agitation step and before any optional
or necessary drying of the complex is performed.
[0022] The Invention:
[0023] It is the applicants' discovery that a water-soluble
acid-chitosan complex may be easily and effectively produced by
adding varying amounts of one or more acids and a modest amount of
water to a sufficiently-agitated chitosan powder or flake, mixing
to achieve uniformity and, if necessary, drying the resulted
hydrated crumb. By this technique, one can vary the amount of acid
used from a very small amount up to essentially a stoichiometric
quantity, and water-soluble acid-chitosan complexes may be prepared
that range from completely water-soluble to hydratable and soluble
by the addition of lesser quantities of acid than with native
chitosan.
[0024] The technique and amount of water and acid addition are
important to the successful practice of the present technology.
Water is added as a carrier for the acid, allowing it to dissolve,
ionize and penetrate the chitosan particle, and carry out at least
partial protonation, thereby producing a complex of at least
partial salt formation between the acid and the chitosan. If too
little water is added, the acid molecules will not be sufficiently
mobile to penetrate the particle and give a uniform product. If too
much water is added, the hydrated chitosan will begin to approach a
solution; this is not desired because large, hydrated gel masses
may form and the product becomes very difficult to process through
the necessary (in that case) drying and grinding operations
required to produce a finished, marketable product. In this
respect, it is preferred that the amount of water used be in the
range of approximately 5% to 130% of the total weight of the
chitosan and the acid, and more preferable that the amount of water
added be in the range of 5% to 15%.
[0025] The acid to be used must be sufficiently water soluble to at
least partially dissolve in the water used. In the case of less
soluble acids, longer mix times may be necessary to give a more
uniform product. This is generally necessary to form the
water-soluble acid-chitosan complex, otherwise a true complex will
not be formed, and instead, a partial complex and what is
essentially a non-homogeneous mixture of acid and chitosan will
form.
[0026] If the acid is a liquid, it is preferred that it be
dissolved in the process water and thereafter sprayed onto the
chitosan. If the acid is a solid, it may either be dry-blended or,
if it is sufficiently water soluble, it may be dissolved in the
water and the solution sprayed onto the chitosan powder or flake.
Because a major necessity of the process is the diffusion of the
acid, mediated by the water, into the chitosan solid, it may be
easily seen that a period of mixing of the acid with the chitosan
subsequent to the introduction of water to the chitosan is
necessary to assure a homogeneous product.
[0027] The acids that are operative in the process preferably
include, but are not limited to, hydrochloric, acetic, lactic,
glycolic, nitric, malic, pyruvic, citric, ascorbic, and other
physiologically acceptable carboxylic acids. Other acidic
substances, such as betaine hydrochloride or amine hydrochlorides,
such as glycine hydrochloride, which are more acidic than the amino
group on chitosan, are also effective.
[0028] The crux of the invention is that the use of acid and water
with the chitosan and with good agitation allows a reaction between
the acid and the chitosan, resulting in the formation of a true
chemical complex, rather than just an intimate blend of the two
components. The complex can be best described as a salt of
chitosan, acting as a base, with one or more acidic species (see
definition above).
[0029] It is applicants' discovery that mixtures of acids with
chitosan, in amounts that approach or exceed stoichiometric for
salt formation, provide superior performance in the fat-binding
application. In a particular embodiment, acceptable betaine
hydrochloride complexes with chitosan surprisingly show efficacy in
the application and demonstrate good shelf stability, making them
especially suitable for the application.
[0030] Further, the applicants have found a method of preparing
these chitosan salts which results in at least partial salt
formation, taking the formulation from a mixture of chitosan and
acid to the formation of an acid-chitosan complex. This provides
for a uniform product which may be easily formulated into the final
marketable product, but is still stable with regard to molecular
weight of the chitosan over time.
[0031] A preferred embodiment is the complex of chitosan with
betaine hydrochloride. The applicants have found that a dry blend
of 100 parts of chitosan and 75 parts of betaine hydrochloride,
when treated with water according to the invention to form a
complex, forms a homogeneous, water soluble solid. Conversely, when
a dry blend or mixture was prepared of the two compounds in the
same ratio, the two materials separated, due to their large
difference in bulk density, and samples taken from different parts
of the container showed greatly different solubility
properties.
[0032] The preparation of the betaine hydrochloride complex of
chitosan also demonstrates the criticality of the use of controlled
amounts of water. In this case, when 100 parts of chitosan were
treated with 75 parts of betaine hydrochloride and 200 parts of
water (which is equivalent to 115% of the total weight of the
chitosan and the acid), a granular hydrated chitosan/betaine
hydrochloride complex was formed, which was easily dried and ground
to give product. However, when the water was increased to 250 parts
(equivalent to 143% of the chitosan and the acid), the material
became sufficiently hydrated and cohesive to form a stiff, gel-like
mass which actually froze the blades of the Hobart mixer, causing
the motor to burn out. Drying this gummy mass was quite difficult,
as the gel held water for long periods of time. Upon reaching a dry
state, the product formed large, extremely hard pieces of the salt,
making it very difficult to grind.
[0033] In one particular embodiment, wherein the amount of acid is
sufficient to protonate enough of the amino groups of chitosan to
yield a soluble product (i.e. 75% of stoichiometric or higher), the
applicants have found that adding 10% water, based on the sum of
the weights of chitosan and betaine hydrochloride, to a mixture of
chitosan and betaine hydrochloride and mixing after the addition,
gives a dry powder which easily disperses and dissolves in water to
give a viscous chitosan solution.
[0034] In an alternative embodiment, applicants have treated the
chitosan in the manner of the invention with lesser amounts of acid
to give products that disperse in water and, upon the addition of
lesser than usually employed amounts of acid, form a viscous
solution of chitosan.
[0035] Low levels of acid generally yield chitosan complexes that
are not soluble in water (i.e. not enough of the salt has been
formed to render the chitosan water-soluble in its own right). This
reinforces the fact that there must be an adequate number of moles
of acid present to protonate a major fraction of the amine groups
in the chitosan. For example, full solubilization requires at least
about 0.75 moles of acid for each mole of chitosan. The acid
content to be achieved will depend on how much one wants to
decrease the acid demand on the stomach of an ingester of the
complex.
[0036] Another aspect of the invention relates to the use of
water-soluble acid-chitosan complexes in the prevention of fat
digestion and the overall improvement of an animal's health. In
particular it is applicants' discovery that when an effective
amount of a water-soluble acid-chitosan complex, and in particular,
that of chitosan and betaine hydrochloride, is administered to an
animal prior to or during digestion of fat-containing substances,
the triglyceride levels within that animal are reduced to levels
significantly below those observed with the use of chitosan as
known in the prior art. Triglyceride levels are known in the art as
a measure of fat availability within an animal's system. The
following pragraph sets forth a description of trigycerides.
[0037] A triglyceride is a primary form of fat transported within
an animal's body. Triglycerides are found as a normal component in
an animal's bloodstream. They are compounds (esters) of fatty acids
and glycerol that bind to proteins and form low-density lipoprotein
(LDL) and very-low density lipoprotein (VLDL). Normally,
triglyceride levels rise immediately after eating. In particular,
after an animal eats, its body digests the fats from the food and
then releases triglycerides into the bloodstream. The triglycerides
are transported throughout the body of the animal to give the
animal energy or they are easily stored as fat. Thus, as is known
in the prior art, a reduction in the level of triglycerides is
directly correlated to a reduction in the fat available to the body
through the digestive process.
[0038] The liver also produces triglycerides and converts some into
cholesterol. Further, there is a link between triglyceride levels
and the development of coronary heart disease. High triglyceride
levels are an important predictor of myocardial infarction. LDL and
VLDL contain large amounts of cholesterol and triglycerides that
can adhere to the arteries in the form of fatty plaques. Therefore,
a reduction in triglycerides has several beneficial aspects.
[0039] The administration of the acid-chitosan complex to an animal
also has a beneficial effect on the levels of aspartate
aminotransferase (AST) and alanine aminotransferase (ALT) in that
animal. The levels of AST and/or ALT in an animal relate to
characteristics of liver function. For example, ALT levels are
present in kidney and muscle as well as liver, and are used to
confirm that AST elevations are of liver origin.
[0040] It is Applicants' discovery that with the administration of
a water-soluble acid-chitosan complex, and particularly chitosan
and betaine hydrochloride, the ALT and AST levels were generally
found to be significantly lower than that in commercially available
chitosan and even the positive control of the drug Xenical.TM..
Water-soluble acid-chitosan complexes are found to be less
hepatotoxic than commercially available Chitosan or Xenical.TM. in
animals being fed such products.
[0041] The advantageous properties of this invention can be further
observed by reference to the following examples which illustrate
the invention.
EXAMPLES
Example 1
[0042] Sixty five grams of ground chitosan (-60 mesh), available
from DCV, Inc., Wilmington, Del., was added to a Hobart mixer.
Hydrochloric acid (6.5 g on a 100% acid basis) was added to enough
water to make 150 ml total solution. The mixer was started at
medium speed and the acid solution was sprayed onto the mix over a
5-minute period. Mixing was continued for 15 minutes following the
addition of acid to yield a homogeneous, moist crumb. At that
point, the moist acid-chitosan complex was put into an aluminum
tray and dried overnight in a convection oven, set at 65.degree. C.
When dry, the material was re-ground to -60 mesh. The grinder used
was a Tekmar grinder with cooling jacket, Model No. A-10.
[0043] To test water solubility, a 1.5 g portion was put into a
beaker with 150 g of water. A magnetic stirring bar was placed in
the beaker and the solution was mixed for one hour. The solid
dissolved to give a viscous solution.
Example 2
[0044] Sixty-five (65) grams of chitosan flake was added to a
Waring blender. Solid betaine hydrochloride (48 g) was added to the
chitosan. The mixer was started at low speed and 100 ml of water
was sprayed on over a 5-minute period. Following the addition of
water, the blend was mixed for an additional 10 minutes to form the
homogeneous acid-chitosan complex. The moist complex was dried
overnight at 55.degree. C. and was then ground to -20 mesh using
the Tekmar grinder. 2 g of this was added to water and stirred to
give a viscous solution.
Example 3
[0045] Seventy five (75) pounds of chitosan was added to a Henschel
mixer. Solid betaine hydrochloride (56 lbs.) was added to the
chitosan. The mixer was started at low speed and 10 lbs. Of water
was sprayed on over a 2 minute period. Following the addition of
water, the blend was mixed for 15 minutes to form the homogenous
acid-chitosan complex, which was then screened through a 20 mesh
screen and packaged. Two grams of this was added to 100 ml of
water. A gel was formed within 1 minute with a pH of approximately
3 with no additional acid added.
Example 4
[0046] A sample of untreated chitosan (2 g at 6.7% moisture =1.87 g
chitosan on a 100% basis) was placed in 200 ml of water. The
initial pH of the slurry was 7.3. One normal (1N) hydrochloric acid
was added dropwise, with stirring, until a solution was formed and
the final pH was 3.0. It was found that 7.75 ml of acid were
required, or 4.144 meq/gm of 100% chitosan.
[0047] In a like manner, chitosan from the same original lot that
had been treated in the manner of the invention such as to form the
acid-chitosan complex with a 10% ratio of 100% hydrochloric acid to
chitosan (2 g at 1.4% moisture, representing 1.79 g of 100%
chitosan) was dissolved and adjusted to pH 3.0. While the pH of the
water before introduction of the chitosan was observed to be 7.30,
the pH drifted down to 5.7 before any acid was added and the
chitosan was observed to visibly begin to hydrate, indicating
partial solubility ascribed to partial formation of the
acid-chitosan complex. It was found that the treated sample
required only 5.4 ml of acid (corresponding to 3.016 meq/gm of 100%
chitosan) to lower the pH to 3.0. This is direct confirmation that
partial complex formation had occurred and that there was a reduced
acid demand to effect solution.
Example 5
[0048] The purpose of this study was to (1) to compare the fat
binding ability of Betasanne.TM. (chitosan and betaine
hydrochloride complex prepared by any of the methods described in
the above examples) with that of various commercially available
chito sans; (2) to optimize the conditions for maximum absorption
of fat from the diet using Betasanne.TM. or commercially available
chitosan; and (3) to compare the efficacy of fat binding using
Betasanne.TM. with that of commercially available chitosan.
[0049] Study Design
1TABLE 1 Group Number of Animals Test Article 1 12 Mixture of
Chitosan and 5% Vitamin C 2 12 Chitosan 3 12 Chitin 4 12 Betasanne
.TM. 5 12 Mixture of Chitosan and 1% Vitamin C 6 12 High Fat Diet 7
12 Xenical .TM. 8 12 Low Fat Diet
[0050] All rats were fed a basal diet for 5 days prior to
initiation of dosing on Day 0. At Day 0, animals in Groups 1 to 5
and 7 received the appropriate formulated diet. The diet was
available ad libitum. Animals in Group 6 received only the basal
diet throughout the study and animals in Group 8 received Purina
Diet #5001 without the addition of additional fat throughout the
study. The total amount of the diet consumed by each animal was
determined every 3 to 4 days.
[0051] Results
[0052] Table 2 below outlines differences in weight gain and
triglyceride, AST and ALT levels in the blood of the rats used in
the trial as sert forth above.
2 TABLE 2 Chitosan + Chitosan + High Low 5% Vit C Betasanne
Chitosan 1% Vit C Xenical Fat Fat Weight Gain (gms) 419 444 464 458
425 469 452 Triglycerides (mg/dL) 133 119 157 141 107 172 111 ALT
(IU/L) 52 46 54 48 54 41 38 AST (IU/L) 80 63 73 64 104 67 83
Bilirubin (mg/dL) 0.4 0.3 0.5 0.5 0.4 0.6 0.2 (Note: For all number
values in Table 2, the lower the number, the more effective the
product)
[0053] Conclusion and Summary
[0054] On the basis of equal weighting for all the properties that
were measured in this study, the two groups which used an
acid-chitosan complex as set forth in the invention showed the
greatest effects (i.e. Betasanne and Chitosan+5% ascorbic acid).
The animals administered Betasanne or Chitsan+5% ascorbic gained
less weight than did those animals administered either commercially
available chitosan or a high fat diet. Similar results were
observed for triglyceride levels.
[0055] As for ALT and bilirubin levels, Betasanne is the closest to
the high fat and low fat diet, thus showing that it is the least
hepatotoxic.
* * * * *