Note: Descriptions are shown in the official language in which they were submitted.
j~ 1'251~3~28
1 BACKGROUND OF T~E INVENTION
3 This invention relates to absorbent materials adapted for
4 use in body contact applications and, more particularly, to
enzymatic absorbent materials such as bandages and pads that
6 produce a bacteriostatic effect upon contact with body fluids
7 such as serum.
8 Absorbent materials in the form of bandages and pads have
9 long been employed in body contact applications where coverage
and protection of a wound and absorption of its fluids are
11 desirable. Bandaging can serve to physically protect an open
12 wound from the surrounding environment, which is often replete
31 with harmful bacteria, and to absorb fluid materials such as
14 I pus, blood and serum to thereby promote the healing of the
15 ¦ wound. Accordingly, bandages are constructed of strong, yet
lG ! absorbent, materials such that retention of extraneous fluids
17 ¦ can be accomplished without a concommitant loss of strength.
18 ! Absorbent materials such as woven fibers, porous foam
19 i pads, absorbent membranes and solvent-based porous elastomers
have been utilized in the manufacture of bandages and pads~ A
21 wide of variety of raw materials have been employed in an
22 attempt to obtain physically occlusive structures that retain
23 their strength when soaked with fluid from an open or weeping
24 wound or other body fluid source. Materials such as cellulose
and its derivatives (cellulosics), polyester, nylon,
2G polyacrylamide, collagen, polyurethane and polyvinyl alcohol
27 have been fabricated into structures such as woven fibers, foam
28~ ~ pads, poro~ membranes, elastomers and multi-layered combinationsj
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1 of the aforementioned structures. Regardless of the material
2 used, all bandages and pads employed in the protection of ope~
3 wounds must satisfy the requirement of good absorbency in order
4 to be effective.
An absorbent material in contact with an open or weeping
6 wound will retain a substantial amount of blood, blood serum,
7 pus and a variety of wound exudates. Bandages are often
8 constructed in such a way that moisture retained by the absorbent
material in contact with the wound will transpire through the
side of the bandage opposite to that of the source of the
11 fluids, i.e., the open wound. Although structures such as
12 these tend to decrease the amount of moisture retained by the
13 absorbent material, other exudates such as white blood cells,
14 ¦ bacteria, electrolytes, and red blood cells are retained.
15 ¦ These exudates accumulate as a function of the amount of time
lG an absorbent material is in contact with an open or weeping
~7 wound.
18 1l As a result of the retention of such a wide variety of
19 I biological substances, a moist a~sorbent structure can become
20 ~ an effective breeding ground for potentially harmful bacteria.
21 Such an absorbent structure in contact with body fluid of the
22¦¦ type mentioned above can be characterized as a culture support
23 j medium that can give rise to a large number of potentially
24 ¦ harmful bacterial colonies in a relatively small volume. Since
25 I the accumulated bacteria and their waste products remain in
2G ¦ contact with the open or weeping wound, they can leech out of
27 ¦ the absorbent structure and back to the body. In this connection
2~ I symptoms classifiable as toxic shock syndrome have been
51~ 28
1 associated with the use of certain types of feminine hygiene
2 tampons, i.e., internally disposed feminine hygiene absorbent
3 pads.
4 It would, of course~ be advantageous to provide absorbent
materials in the form of bandages, pads, strings and the like
6 that would inhibit bacterial growth in body fluids which are
7 absorbed by or otherwise associated with such absorbent materials
8 or structuresO
9 It is disclosed in the prior art that polymeric wound
10 j dressings for burns may include antibacterial agents,
¦ antibiotics, antifungal agents, proteolytic enzymes as well as
12 I local anesthetics, hormonal compounds and lubricating and
13 ¦ barrier chemicals. See, for example, U.S. Patent 4,122,158
14 ¦ (Schmitt, 1978) and U.S. Patent 4,226,232 (Spence, ~9~0).
15 , It is also disclosed in the prior art that enzymatic
16 I agents can be incorporated into oral products such as toothpaste
17 I and chewing gum for producing hydrogen peroxide during oral
1~ ¦ use.
19 I U.S. Patent 4,150,113 (Hoogendoorn et al., 1979) and U.S.
20¦¦ Patent 4,178,362 (Hoogendoorn et al., 1979) disclose,
21~¦ respectively, an enzymatic toothpaste and an enzymatic chewable
22 I dentifrice containing glucose oxidase which acts on glucose
23 present in saliva and tooth plaque to produce hydrogen peroxide.
24 The patentees note that oral bacteria, through enæymes systems
having SH-GROUPS, effect glycolysis of food products containing
26 ¦ sugars and point out that lactoperoxidase, which is present in
27 I saliva, provides the means for transferring oxygen from hydrogen
28 ¦ peroxide to the oral bacteria resulting in the oxidation of the
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1 ¦ SH-containing enzymes into inactive disulfide enzymes. It is
2 further disclosed that the dentifrice may be formulated with
3 potassium thiocyanate.
4 U.S. Patent 4,269,822 (Pellico et al., 1981) discloses an
antiseptic dentifrice containing an oxidizable amino acid
6 substrate and an oxidoreductase enzyme specific to such substrate
7 ¦ for producing hydrogen peroxide and ammonia upon oral application
8 ¦ of the dentifrice, with pre-application stability being maintainec
9 ~ by limiting the quantity of any water present in the dentifrice.
10 ll
11 I SUMMARY OF THE INVENTION
12 l
13 ¦ In accordance with one aspect of this invention, there is
14 I provided enzymatic absorbent material for body contact applicatior
15 , containing, per gram of material, from about 1.0 to about 1,000
lG I International Units of serum-activated oxidoreductase enzyme
17 ' for producing hydrogen peroxide upon contact of said material
18 I with serum.
19 In accordance with a second aspect of this invention,
20¦ there is provided enzymatic absorbent fiber for conversion into
21' enzymatic absorbent material that is adapted for body contact
22¦ application, wherein the fiber contains, per gram of fiber,
23 I from about 1.0 to about 1,000 International Units of serum-
24 i activated oxidoreductase enzyme for producing hydrogen peroxide
25 ~ upon contact with serum.
27
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1 DETAILED DESCRIPTION
3 The enzymatic absorbent materials of this invention comprise
4 fluid absorbent structures that incorporate serum-activated
oxidoreductase enzyme for producing hydrogen peroxide upon
6 contact with serum. Fluid absorbent structures, in the form of
7 bandages, pads, strips and the like, can be prepared from
8 precursors and by processes well-known in the art to provide
9 appropriate designs and configurations that are adapted for
particular body conLact applications. Structural precursors
11 1 such as woven fibers, porous foam pads, absorbent membranes and
12 I solvent-based porous elastomers can be utilized in the manufactur~
13 I of the ~luid absorbent structures. &auze bandaging alone or
14 secured to adhesive strip and feminine hygiene absorbent pads
15 I and tampons, as well as other externally and internally utilizabl
lG I body contact devices having high absorbency characteristics,
17 I can be advantageously utilized in the practice of this inventionO
18 I The enzymatic absorbent material of this invention, which
19 I incorporates oxidoreductase enzyme, is adapted to be used in
20 I body contact applications that encounter body fluids. These
21 fluids, including blood and tissue serum, contain oxidizable
22 I substrate and other ingredients which undergo an enzymatic
23 I reaction in the presence of oxidoreductase enzyme specific to
24 I the substrate to produce hydrogen peroxide. Oxidoreductase
25 ¦ enzymes which can be utilized in the practice of this invention
2G i and the corresponding oxidizable substrates in serum are set
27 ¦ forth in the following table:
28 I
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1 TABLE A
OXIDOREDUCTASE OXIDIZABLE
ENZYME SUBSTRATE
4 Glucose Oxidase B-D-glucose
Hexose Oxidase Hexose
6 Cholesterol Oxidase Cholesterol
7 Galactose Oxidase D-galactose
8 Pyranose Oxidase Pyranose
Choline Oxidase Choline
Pyruvate Oxidase Pyruvate
11 Oxalate Oxidase Oxalate
12 Glycollate Oxidase Glycollate
13 I D-aminoacid Oxidase D-aminoacid
14 l
15 ¦ In an illustrative enzymatic reaction, glucose oxidase in
lG ¦ the enzymatic absorbent material catalyzes the interaction of
17 I Beta-D-glucose, water and oxygen in the serum to produce hydrogen
18 ¦ peroxide and gluconic acid.
19 , Glucose oxidase is characterized in the literature as a
20 I glycoprotein containing two molecules of flavine-adenine
21 ~ dinucleotide which has a molecular weight of approximately
22 ¦ 150,000, an isoelectric point at pH 4.2 and an optimum pH at 5.5
23 I with a broad pH range from 4 through 7. ,
24 I ' The oxidoreductase enzyme is generally present in the
25 i enzymatic absorbent material in an amount from about 1.0 to
26¦l about 1,000 International Units (hereinafter sometimes abbreviatec
27 I as IU) per gram of material and, preferably, in an amount
2~ from about 10 to about 500 IU per gram of material. The term
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1 International Unit(s) identifies that amount of enzyme that
2 will effect catalysis of l.O micromole of substrate per minute
3 at pH 7.0 and 25C. Oxidoreductase enzymes are supplied in dry
4 or liquid form with the label specifying the concentration in
International Units on a per ~ram or per milliliter basis, as
fi appropriate.
7 In addi~ion to the oxidoreductase enzyme for producing
8 hydrogen peroxide, the enzymatic material can be provided with a
9 second enzyme, namely, a peroxidatic peroxidase for interacting
with hydrogen peroxide and an oxygen-accepting anion in serum
Il for producing an oxidized anionic bacterial inhibitor.
12 Peroxidases which can be used in the practice of this invention
13 include lactoperoxidase, horseradish peroxidase, iodide
14 peroxidase, chloride peroxidase and myeloperoxidase. The
peroxidase is generally present in the enzymatic absorbent
IG material in an amount from about O.l to about lO,OOO
17 International Units per gram of material, and, preferably, in
18 an amount from about lO to about 500 International Units per
19 gram of material.
Oxygen-accepting anions in serum include thiocyanate,
21 chloride and iodide ions which, in the presence of hydrogen
22 peroxide and peroxidase, are oxidized to hypothiocyante,
2.~ hypochlorite and hypoiodite, respectively.
2~ The enzymatic absorbent material described herein may be
'~) augmented by additionally incorporating into the material
'~G supplementary ingredients as, for example, (a) oxidizable
~)7 substrate specific to the oxidoreductase enzyme utilized in the
~8 material, and (b) oxidizable salt such as the thiocyanate,
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1 chloride or iodide salt of sodium, potassium, ammonium, calcium
2 or magnesium or mixtures of such salts.
3 The oxidizable substrate is generally present i~ the
4 enzymatic absorbent material in an amount from about 0.015 to
about 0.6 millimole per gram of material and, preferablyr in an
amount from about 0.025 to about 0.1 millimole per gram of
7 ¦ material. The oxidizable salt is generally present in the
8 ¦ enzymatic absorbent material in an amount from about 0.0001 to
9 1 about 0.01 millimole per gram of material and, perferably, from
10 I about 0.001 to about 0.006 millimole per gram of material. The
11 ¦ term millimole identifies that quantity in grams corresponding
12 I to the molecular weight of the composition divided by one
13 I thousand.
14 I The operable integrity of the enzymatic system can be
15 I affected by catalase which is present in commercial glucose
1~ I oxidase as well as mucous membrane tissue, blood and blood
17 I serum. Catalase, which is extraneous to the enzymatic system
8 I of this invention, competes with lactoperoxidase for hydrogen
19 ' peroxide. In order to reduce loss of hydrogen peroxide through
20 I the presence of catalase, an effective amount of an enzymatic
2,1l inhibitor specific to catalase can be advantageously incorporated
221¦ into the enzymatic absorbent material. An ascorbic salt such
23 ~ as sodium ascorbate, potassium ascorbate, ascorbyl palmitate,
24 I or mixtures thereof can be used as an enzymatic inhibitor which
25 ¦ is specific to catalase. An effective amount of ascorbate salt
2G I for catalase inhibition is from about 0.000001 to about 0.0001
27 I millimole per gram of enzymatic absorbent material. Iron salts
2~ ~ such as ferrous sulfate can be incor~orated into the enzymatic
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1 absorbent material as a potentiator for ascorbate salt in its
2 role as catalase inhibitor.
3 The enzymatic material of this invention may advantageously
4 be formulated with an aminohexose as, for example, an aminoglucose
such as glucosamine, N-acetyl glucosamine or mixtures thereof
6 in order to increase the yield or accumulation of oxidized
7 anionic bacterial inhibitor. The aminoglucose is generally
8 ¦ present in the enzymatic material in an amount from about 0.0001
~ i to about 0.00~ millimole per gram of enzymatic material and,
lO ¦ preferably, in an amount from about 0.0003 to about 0.001
11 ¦ millimole per gram of enzymatic material.
12 ¦ In an alternative embodiment of this invention, the
13 I oxidoreductase enzyme can be incorporated into absorbent fibers
l~ I of natural or synthetic materials for conversion into enzymatic
~5 ! bandages and pads.
lG ¦ The enzymes of this invention may be advantageously
17 ¦ encapsulated to enhance storage stability in the enzymatic
8 ¦ bandage or pad until utilization of the same. The encapsulating
19 I material can be composed of a water soluble polymer or a
20 ¦ polymer permeable to a substrate specific to the enzyme or
21 ' enzymes contained therein. An illustrative encapsulating
22 material is carboxymethylcellulose.
23 , The enzymatic absorbent materials can be prepared by
24 ¦ various procedures including: (a) solution deposition of the
25 ¦ oxidoreductase enzyme and optional ingredients onto the
26 I absorbent structure, (b) incorporation o~ the oxidoreductase
27 ~ enzyme into a spinning solution that is extruded as fiber
2~ filament ~hich can be converted into bandages and pads, and (c)
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1 incorporation of the oxidoreductase enzyme into the chemical
2 constituency which form poly~eric foams and subsequent
3 injection or solvent deposition of the aforementioned optional
4 ingredients as well as other special purpose additives.
6 RXAMPLE I.
8 This example illustrates the solution deposition of an
oxidoreductase enzyme onto a spun cotton pad and the
0 bacteriostatic effect of the resulting enzymatic pad.
11 An aqueous enzymatic solution was prepared by adding
12 0.2 gm of polyvinylpyrolidone (Mol. Wt. 250,000) and l,000 IU
13 of glucose oxidase (approx. 4.0 mg) to a vessel containing
14 10 ml of distilled water with stirring through the use of
a magnetic stirring bar and electric stirrer. Low turbulence
lG was maintained during stirring to avoid any impairment of the
17 enzyme.
18 A spun cotton pad with dimensions of 3.0 cm by 3. n cm by
19 1.0 cm was impregnated with 1.0 ml of the aqueous enzyme solution
~0 by applying the solution to the pad through a standard medicinal
21 dropper. The residual water was allowed to evaporate from the
22 pad at room temperature (25C). The resulting pad and a
23 non~enzymatic control pad were tested for bacteriostatic
24 properties.
The enzymatic and non-enzymatic pads were placed in separate
26 culture tubes, and approximately 10 ml of human blood serum was
27 added to each of the culture tubes. The tubes were then
28 innoculated with Staphylococcus aureus at a concentration level
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1 of 1.0 x 105 organisms per milliliter of serum and assayed for
2 bacterial coun~s at the end of each 4-hour segment for 24 hours.
3 Cultures to determine counts were prepared from tryptic soy
4 agar and incubated at 35C for 48 hours under aerobic conditions.
The results are set forth in the following table:
7 TABLE I
Glucose Oxidase
9 Enzymatic Pad Non-Enzy~atic Pad
10 Timej hr S. aureus Count/ml S. aureus Count/ml
0 1.2 x 105 1.4 x 105
4 8.6 x 104 2.9 x 105
12 8 6.6 x loA 8.5 x 105
12 5.9 x 104 4.4 x 106
3 16 5.1 x 1~4 9.1 x 106
4.0 x 104 3.6 x 107
1~ 24 3.2 x 104 7.0 x 107
16 EXAMPLE II.
17
18 This example illustrates the preparation of enzymatic
19 fibers by incorporating oxidoreductase enzyme and peroxidatic
peroxidase enzyme into a wet spinning solution that is extruded
21 in fiber form. In addition, this example illustrates the
22 bacteriostatic effect of the resulting enzymatic fiber.
23 To methylene dichloride contained in a 250 ml beaker,
2~ there was added 10 gm of celluose triacetate in pellet form to
~5 bring the total volume of the contents to 100 ml. An aqueous
2G enzymatic solution having a volume of 15 ml was prepared by
27 admixing glucose oxidase (500 IU/ml) and lactoperoxidase
28 (500 IU/ml) with distilled water. The aqueous enzyme solution
~ 1 125~228
1 was added to and admixed with the cellulose triacetate solution
2 for 30 minutes which resulted in an emulsion of the aqueous
3 phase in the organic phase.
4 Fibers were formed from the enzymes/polymer emulsion by
wet spinning technique. A 1~ ml hypodermic syringe was filled
with the emulsion. The syringe was used to extrude the emulsion
7 through a No. 20 hypodermic needle into 100 ml of toluene that
8 was contained in a 150 ml graduated cylinder.
9 During the extrusion process, the emulsion coagulated as a
]0 fiber in the form of ringlets about 2 cm in diametex which
11 slowly ~ettl^d to the bottom of the gradllated cylinder. The
12 rate of extrusion was controlled to provide a continuous fiber
13 strand of thP ringlets at a rate slow enough to prevent sticking
1~ at the bottom of the cylinder. rJpon completion of the extrusion
16 step, the fibers were removed from the toluene bath and
16 air-dried before testing for bacteriostatic properties.
17 Approximately 9.6 gm of fiber were obtained.
18 The fiber was assayed for activity in respect of both
19 glucose oxidase and lactoperoxidase. An activity of 300 IU per
gram of fiber was obtained for each enzyme.
21 A first control fiber w25 prepared by the foregoing
22 extrusion procedure, except that the extrusion composition did
23 not contain either glucose oxidase or lactoperoxidase. A second
24 control fiber was prepared in accordance with the procedure
25 used in the preparation of the first control fiber, except that
2G the extrusion composition was modified to contain bovine serum
27 albumin, a protein. Neither of the control fibers showed any
28 enzyme activity in tests for either glucose oxidase or
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1 lactoperoxidase. `
2 The fibers described above are of the type that are suitable
3 for the manufacture of woven bandages.
4 The fibers were tested for bacteriostatic properties using
~ Staphylococcus aureus (ATCC 653A) and E. coli (ATCC 25923).
6 Each organism was inoculated into a 20 ml portion of tryptic
7 soy broth and incubated at 35C for 24 hours. Each of the
8 resulting cultures was washed with a phosphate buffer (pH 6.5
9 and 0.01 M) by centrifuging to obtain a suspension of cells,
which was diluted 1:10.
11 Fresh human blood serum samples were prepared in 6 screw
12 cap tubes that held 20 ml of serum in each. Twenty microliters
13 of the diluted bacterial cell suspension were inoculated into
14 each of the tubes containing serum. Thereafter, a 100 mg fiber
sample was placed in each of the inoculated serum tubes which
16 were incubated at 35C for the duration of the testing.
17 Occasional manual agitation of the tubes was undertaken during
18 the test period.
l9 Bacterial counts of the samples were taken at the end of
each 4-hour segment for 24 hours. Tryptic soy agar was used as
21 the plating medium for each sample and the plates were incubated
22 at 35UC for 48 hours under aerobic conditions. The results are
23 set forth in the following table:
27
28
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1 TABLE II
2 2A
3 Enzymatic Fiber
Glucose Oxidase & Lactoperoxidase
: 5 Time, hr.S. aureus Count E. coli Count/ml
6 0 2.0 x 105 1.0 x 105
4 6.2 x 103 3.8 x 104
7 8 2.0 x 103 2.2 x 104
12 1.8 x 103 7.6 x 103
8 16 <1,000 5.1 x 103
<300 1.4 x 103
9 24 <100 ~100
2B
11 Non-Enzymatic Fiber
12 Time, hr. S. aureus CountE. coli Count/ml
3 0 2.0 x 105 1.0 x 105
4 6.6 x 105 9.5 x 105
14 8 2.1 x 106 5.1 x 107
12 8.8 x 106 4.9 x 108
16 2.2 x 107 1.0 x 109
3.7 x 107 1.1 x 109
16 24 5.0 x 107 1.6 x 109
2C
Fiber/Bovine Serum Albumin
19
20 Time, hr. S. aureus CountE. coli Count/ml
.~ o 2.0 x 105 1.0 x 105
: 21 4 4~5 x 105 9.5 x 105
: 8 9.9 x 105 5.3 x 106
22 12 6.1 x 106 8.9 x 106
~; 16 8.4 x 106 1.2 x 107
23 20 1.0 x 107 8.6 x 107 .
24 24 4.2 x 107 2~1 x 109
26 EXAMPLE III.
27
28 This example illustrates a method for incorporating
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1 oxidoreductase enzyme into yieldable, semi-rigid foam and the
2 bacteriostatic properties of the resulting enzymatio foam.
3 An aqueous enzyme solution was prepared by admixing
4 1,000 IU of glucose oxidase (approx. 4.0 mg) with 40 ml of
cold, distilled water, under moderate agitation, until the
6 enzyme dissolved.
7 An isocyanate terminated, prepolymer solution was prepared
8 by reacting stoichiometric amounts of 80/20 2,4-/2,8
9 tolylenediisocyanate and Carbowax*l,000 polyoxyethylene polyol
(Mol. Wt. 1,000) at 135C for 5 hours in the presence of a
11 catalyst comprising 2-ethylhexanoic acid and tin octoate. The
12 resulting urethane prepolymer was diluted to 90~ solids with
13 acetone.
14 To 24.5 gm of the acetone solution of prepolymer, there
was added 0.22 gm of Tween*80 !a polyoxye~hylene derivative of
16 sorbitan fatty acid ester having a molecular weight of about
17 1,309) and the viscous mixture was stirred by hand for
approximately 30 seconds. Twenty milliliters of the aqueous
19 enzyme solution was added to the prepolymer composition and the
20 ¦ resulting paste-like mixture was stirred to obtain a dispersion
21 of the ingredients. The remaining aqueous enz~me solution was
22 then added to the reaction mixture which was again stirred to
23 obtain dispersion of the ingredients. Upon completion of the
24 enzyme addition and dispersion steps, the reacting mass was
poured into a Petri dish (100 mm diameter) and allowed to
26 remain at ambient temperature for 5 minutes at which time the
27 reaction was substantially complete.
2~ There was obtained from the reaction a white, spongy, wet,
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semi-rigid, enzymatic foam that was approximately 1.2 to 1.4
~ centimeters thick. The wet foam was re~oved from the dish and
3 stored in a refrigerator at approximately 5C, with the foam
4 remaining damp during storage. One gram of this foam material
(dry weight) was assayed for glucose oxidase activity using
6 glucose as a substrate, horseradish peroxidase and o-dianisidine
as a chromogen. The activity of the one gram piece of foam was
8 approximately 12 IU per gram, or about 60% of the original
activity.
Five grams of enzymatic foam (dry weight) were placed in a
11 large culture tube containing 10 ml of whole pig's blood. The
12 blood was inoculated with approximately 200,000 or~anisms of
13 S. aureus. A control tube was prepared by placing 5 grams of
14 ¦non-enzymatic foam in a culture tube containing 10 ml of whole
15 Ipig's blood which was inoculated with approximately 200,000
16 ¦organisms of S. aureus. Bacterial counts, per one ml of blood,
17 were taken at the end of each 4-hour segment for 24 hours. The
18 results are set forth in the following table:
19
Table III
21 Glucose Oxidase
22 Enzymatic Foam Non-Enzymatic Foam
23 Time, hr. S. aureus Count E. coli Count/ml
0 2.0 x 105 2.0 x 105
241 4 9.6 x 104 3.5 x 105
8 8.2 x 104 5.8 x ln5
25 12 5.0 x 1043 3.9 x 106
16 6.6 x 10 1.7 x 107
26 20 3.4 x 1033 6.8 x 107
27 24 2.2 x 10 9.5 x 1~7
28 By incorporating a spermicidal composition in the
1;~5~
1 enzymatic foam, a contraceptive flexi~le foam pad is obtained.
3 EXAMPLE IV.
This example illustrates an enzymatic pad which is
6 self-contained with respect to oxidoreductase enzyme and
7 corresponding oxidizable substrate for producing hydrogen
8 peroxide in the presence of serum.
9 An enzymatic solution was prepared by dissolving, with
stirring, glucose oxidase (5,000 IU) and glucose (5.0 gm) in
11 methylene dichloride (50 ml). One millimeter of the enzymatic
12 solution was added to a spun cotton pad similar to that described
13 in Example I to effect solution deposition of the ingredients.
1~ .
EXAMPLE V.
lG
17 This example illustrates an enzymatic pad which is
18 self-contained with respect to oxidoreductase enzyme, oxidizable
substrate, peroxidatic enzyme and oxidizable salt for producing
an oxidized anionic bacterial inhibitor in the presence of
21 serum.
22 An enzymatic solution was prepared hy dissolving, with
23 stirring, glucose oxidase (l,000 IU), glucose (l0 gm),
2~ lactoperoxidase (500 I~), and potassium thiocyanate in
methylene dichloride (5U ml). One milliliter of the enzymatic
26 solution was added to a spun cotton pad similar to that described
27 in Example I to effect solution deposition of the ingredients.
28 In view of the fore~oing description and examples, it will
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1 become apparent to those of ordinary skill in the art that .
2 equivalent modifications thereof ~ay be made without departing
5 l ¦from the Eirit and scope of this invention.
8~
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