CA1066624A - Inhibiting production of undesirable products on body surfaces and environs - Google Patents

Abstract

Abstract of the Disclosure Methods, compositions and products are described which inhibit formation of undesirable products on body surfaces and environs resulting from microbial action on lipoidal materials in body secretions. Amino acid compounds are employed.

Description

The present invention is directed to methods and means for controlling undesirable problems arising from microbial action on body fluids secreted or discharged from the body.
'.

One of the long existing problems has been the unpleasant odors of menstrual fluid. Menstrual fluid contains a variety of substances including proteins and lipids. Normally, present in menstrual fluid are a wide number of gram negative and gram positive organisms which may act on these natural products. Bacterial action on proteins has been recognized ~ a8 a source of amine malodor. Bacterial action on lipids could ¦ ~give rise to malodorous materials which are fatty acids. _ ~1 . .
Axillary malodor also has been a long existing problem.
Axillary sweat is composed of secretions of both eccrine and apocrine sweat glands which are present in the axillary area.
While eccrine sweat consists largely of water and salt, apocrine sweat is composed of a variety of substances including protein, carbohydrates and lipids. Numerous microorganisms including Staphylococci and Corynebacteria are present on the dermal surfaces of the axillary region. Microbial decomposition of lipids in apocrine sweat which results in production of lower fatty acids has been established as a primary cause of unpleasant odors in the axillary region. (Borick et al, Antimicrobial Agents Annual - 1960, pp. 647-651, Plenum Press, Inc., N.Y.) ` 1~
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, Another problem, seemingly unrelated, is inflammatory skin disorders such as acne. Sebum is a lipid mixture secreted by the sebaceous glands and having a variety of fatty acids (Baughton et al, J. Invest, Dermat, ~ 49 55, 1959)-Free fatty acids which have their source in the lipids are -~
believed to be primarily responsible for the inflammatory disorder of the skin known as acne vulgaris (Freinkel et al, New Eng. J. Med 273, 850-854, 1965). Microorganisms, especially Corynebacterium acne, an organism present in both normal and diseased follicles, are reported to cause the formation of ` fatty aclds (Scheimann et al, J. Invest, Dermat. 34, 171-174, 1960).

Germicldes and antibiotics have been employed for the control of unpleasant odors of body products such as perspira-' 15 tlon and menstrual fluld and for the alleviation of undesirable ,~ dlsorders such as acne. Thus, hexachlorophene, at one time was a popular constltuent of preparations for controlling perspiration ~.
odors and antibi~otlcs such as tetracycline have been successfully employed in the treatment of acne. However, germicides and ' antibiotics have accompllshed these results by kill of the organisms, thereby disturbing the normal microbial balance.
As is well-known, the kill of non-pathogenic organisms invltes invaslon by opportunlst organlsms such as pathogenic baeterla, yeast or fungi, whose presence may become manifest in febrile, lnflammatory, dermatitic or other undesirable response. Thus, lt ls desirable that the control of unpleasant body odors or of dlsorders such as acne arlsing from microbial action on lipids ' .: :

be accomplished without significant kill of non-pathogenic microbial flora. It is further desirable that the results be accomplished without having adverse effects on human subjects. -' '' Other methods for the control of the undesirable problems have been by the use of products which act on the causative agent after its formation. This approach is generally unsatisfactory tending to require employing relatively large amounts of treating agent and/or over an extended period. Moreover, the results have not been always completely satisfactory both in terms of completeness of ~ control and in terms of avoidance of side reactions. It is ? highly desirable to control the undesirable problems by . .
preventing the formation of the causative agent of these problems.

The present invention is based on the discovery that ;
the foregoing problems as well as a number of other undesirable . , , effects, may be attributed to microorganisms acting in a similar fashion and therefore, that the various problems may be controlled in a similar manner. It has further been discovered that thiæ control may be brought about without significant kill of most bacterial flora.

It has been discovered according to the present invention that many of the undesirable problems such as menstrual malodor, ; , ~ , ~ . - . . , - . : - - ., . ~ , .
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axillary malodor and inflammatory disorders such as acne, caused by formation of undesirable products on body surfaces and environs as a result of microbial action on lipoidal materials in body secretions may be alleviated by inhibiting the formation of the undesirable products by applying to the situs of forma-tion of the undesirable products, an inhibitory amount of an amino acid compound.
More specifically, the invention relates to a method of inhibiting the formation of fatty acids having from 4 to 18 carbon atoms per molecule from the degradation of skin secretions by Corynebacterium comprising applying to the skin, in an amount effective to inhibit the formation of said fatty acids, a composition comprising a carrier suitable for topical application -and, as the sole essential ingredient, an aminopolycarboxylic compound selected from the group consisting of ethylenediamine-, tetraacetic acid, diethylenetriaminepentaacetic acid, N-hydroxy-`~ ethylethylene-diaminetriaacetic acid, triethylenetetraamine-: hexaacetic acid, 1,2-diaminocyclohexane-N,N'-tetraacetic acid, N,N-dihydroxy-ethylethylenediaminediacetic acid, iminodiacetic acid, hydroxy-ethyliminodiacetic acid, nitrilotriacetic acid and their water soluble salts, said aminopolycarboxylic compound -~being present in a concentration of at least about 0.5 weight percent in said composition, based on the weight of the free acid, said composition maintaining the viability of said Corynebacterium.
By " lipoidal materials in body secretion " is meant lipids (as understood in chemical terminology) which are present in body fluids which are secreted, excreted, discharged or exuded.
Thus, "secretions" as herein employed include waste fluids. `~
Typical secretions are sebum, perspiration, menstrual fluid, etc. ~ --. . ~ .

- Lipids of importance in the present context are not only triglycerides but also phospholipids. The triglycerides may be represented by Formula I:
O

~" , I ~j) .
CH-O-C-R -.,, CH2-0-C-R . -wherein in this and succeeding formulas, each R is a hydro-carbon radical derived from fatty acids and may be same or . .
different. The phospholipids of primary concern in the present context are phosphotriglycerides which may be .~ , ' , .
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; PPC 185 . j, . -represented by the formula~
-, e ~:-C~ -O-C-R ;
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CH-O-C-R
o . -C
~ H2-- '-O-B
.'' . ~ i ' ' .',.. ',.','.. ~' ' ' ~ '- :. ' whereln R is as previously defined and B is a residue of an alcohol-amine compound such as an amino alcohol, a hydroxy- - -quaternary ammonium base or a hydroxyamino acid.

. 5 The products which are formed or may be formed by microbial action on lipoidal materials are similar to those ~-whlch may be formed on chemical hydrolysis. Thus, fatty aclds and glycerol are produced from the triglycerides of Formula I as follows~

~ I ~ CHOH + 3 RCOOH -.1 1 ...

.

sinceeach R may be different, various fatty acids may be formed.
Among the acids which may be formed are malodorous lower fatty acids such as butyric, isobutyric, isovaleric, etc. which are -problems in axillary and menstrual malodor. These and other . . .
fatty acids including higher fatty acids (to Clg) may be sources f other undesirable problems such as inflammatory disorders of the skin. ~ _ 10666;Z4 '' :' " ' PPC 185 ~

Fatty acids may also arise from the phospholipids of ~ -Formula II: ~
, . - ' .

~.l II ) fHOH + 2 RCOOH + H3P04 + BOH

. A .
; C~ OH

In preliminary experiments hereinafter described, phospholipids ;
as a source of malodorous fatty acids was established.
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~ 5 It ls further recognized that the undesirable fatty 3 acl~s may arise in other ways as a result of microbial action and control of such fatty aclds are intended to be embraced.
However, the foregoing is believed to be a major source of ~;~ fatty acids.

The expression "situs of formation" refers to the loci where the lipoidal secretions are retained or received and sub~ect to microbial attack.

Normally, the "situs of formation" of primary concern are body or dermal surfaces and environs. These include skin, vagina and materials ln close proxlmlty to body surfaces such as catamenlal devlces, clothing, bed pads, etc. which are lntended to receive or do receive the body fluids.

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; Amino acid compounds useful for the practice of the ~ -present invention generally have in the structure at least one ~ ~
arrangement of the amino group to acid group as follows: -amino C (C)O 1 - acid The acid group is preferably carboxylic, -COOH, but may be sulfonic, -SO20H, or phosphonic, -PO(OH)2. The amino may be substituted and the carbon chain may contain groups such as ~ -hydroxyl -OH, sulfhydryl -SH, and ether -O-. Generally more than one acid containing group is present in the molecule and~the acid containing group may be attached to the same j 10 amino nitrogen. The compounds may be termed "amino polyacid compounds."
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An important group of compounds which inhibits the formation of fatty acids are the aminopolycarboxylic acid -~
compounds. By "aminopolycarboxylic acid compound" is meant an amino acid or water-soluble salt thereof ln which there is two or more substituent groups of the general structure:
I I

~ ( I )o,l COOH
1, ....
in the molecule which are attached to'one or more amino groups.
Where there are more than two such substituent groups, the excess carboxyl groups may be esterified as long as at least two of the ~ foregoing groups are present.
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Preferred salts, especially for certain applications are . .
monovalent salts such as sodium and potassium salts. For ; certain other applications, alkanolamine salts are preferred, especially di- and trialkanolamines such as triethanolamine, diethanolamine, triisopropanolamine, etc. The best known -aminopolycarboxylic acid compounds are the acids and salts Or ethylenediaminetetraacetic (EDTA) acid, diethylenetriamine~
pentaacetic (DTPA) acid and N-hydroxyethylethylenediaminetria-acetic (HEDTA) acid. These compounds are available commercially through trade names such as VERSENE, VERSENEX, VERSENOL, SEQUESTRENE, etc. Other typical compounds in ~ - this class include triethylenetetraaminehexaacetic acid, ,.~ . ~;
~l 1,2-diaminocyclohexane-N,N'-tetraacetic acid, N,N-dihydroxy-ethylethylenediaminediacetic acid, iminodiacetic acid, hydroxyethyliminodiacetic acid, and nitrilotriacetic acid and their salts as well as the propionic acid analogs.
.
Other compounds include N,N'-dihexadecyl ester of ethylenediaminetetraacetic acid, N,N'-dioctadecyl ester of ethylenediaminetetraacetic acid, N,N'-di(15-carboxypentadecyl) ester of ethylenediaminetetraacetic acid, N-(15-carboxy-9-pentadecnyl)iminobis(ethylenenitrilo)tetracetic acid, N-methoxyethyl-iminodiacetic acid, ethylenebis(oxypropylamino-dlacetic acid), ethylenebis(oxyethyliminodiacetic acid), aminoethyl-N-methyliminodiacetic acid, a-mercaptoethylimino-diacetic acid, a-methylmercaptoethyliminodiacetic acid, etc.

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Amino acid compounds of other classes which are of special interest for certain applications are those which also have more than one acid group containing substituent attached to the amino group but in which the acid groups are generally phosphonic or sulfonic. However, these amino acid compounds may have mixed acid groups, i.e., have one or more carboxyl groups in the same molecule as that containing the phosphonic or sulfonic acid groups. Representative amino acid of this type include ethylenediaminotetra(methylene- -~0 phosphonic acid), nitrilotris(methylenephosphonic acid), ~
iminobis(methylenesulfonic acid), aminomethylphosphonic -1 acid-(N,N-diacetic acid), etc.
. ;, . .
'' i ., The amino acid compounds are available as free acids, 1 acid salts and salts. It is preferred that the form of the j5 amino acld compound or combination thereof employed be such ;
that at the situs of operation, the pH be near neutrality, -about 6.2 to about 8.5. However, useful results may be obtained if the pH is at least as high as 4Ø Frequently, it is convenient to prepare the salt or acid salt by mixing the acid and base in the composition.
.. . .

The amino acid compounds are useful in such amounts . . .
as would give a final concentration of at least 0.01% by ~`~ weight of the body secretion. The upper limit is dictated primarily by practical considerations. Generally speaking, ;5 not much advantage is gained by adding an amount which would give a concentration greater than about 0.5% by weight of ,,. ,, ,, ~ :: . ,:, . , :, , - , -` ~ 1066624 ..... -; .

, ', body secretion. The preferred amounts depend on the purpose, place and method of application and on the particular compound. Thus, where microbial population -is expected to be hlgh, such as in menstrual fluid, larger amounts preferably are employed. Thus, for control of menstrual malodor, at least about 0.04% by weight of menstrual fluidis deslrable and 0.10% or more is preferred.
Withln the broad scope, preferred ranges are hereinafter dlsclosed ln the context of the partlcular use and method Or application.
;
', ,.r i The amlno acid compound may be applied to the situs of formation of microblally produced undesirable products i -in varlous ways. In its use for the lnhibition of formation .~,, l .
`j of malodorous materials in menstrual fluid, the amino acid compound ls convenlently applled to receptacles for the fluid such as catamenlal dressings or to such other materials which may receive the fluid such as bed pads, clothing, etc.
By "catamenial dressings" is meant sanitary napkins, tampons and lnterlabial pads which usually consist of a core of one or more layers of highly absorbent, relatively dense materials which have a fluid permeable, soft, knitted, woven or non-woven wrapper. The cores are conventionally made of layers of fibers such as carded cotton webs, air-layered cellulosic ~lber webs, comminuted wood pulp bats, tlssue pulp or like ~25 materlals but may be made of newer synthetlc materials such as synthetlc polymer foams and fibers. Although the amino acld compound may be dlstributed uniformly through the `' 11 .. . . .
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PPC 185 ~
catamenlal dressing, it iæ more useful to place it in that ~ -portion which first contacts the body discharge, Thus, lt is preferably applied to the surface of the absorbent cores of the dressings or to the covers or both in such -a manner that the amino acid compound is present on the surfaces thereof in an amount ranging from about 0.001 g. per square inch to about 0.1 g. per square inch. Such amounts have ; -' - . .
been found to provide the desired concentration in terms of the amount of amlno acld compound to total body secretion.
~0 A preferred range for catamenial dressings is from about 0.004 to o.o6 g. per square inch.
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;' The amlno acid compounds may be applied to the cata-~;l menial dressings during manufacture or use. When applied during manufacture, it may be applied by spraying either an ; aqueous spray or an aerosol spray, padding, soaking, or by . .,~ ,. .
dusting or any known method for applying materials thereto.
An aerosol spray may employ such propellants as dichloro-dlfluoromethanç, trichlorofluoromethane; a solvent spray may employ substantially inert solvents such as isopropanol.
The use of a propellant or inert organic solvent ls preferred over an aqueous solution or suspension to minimize drying of the catamenial dressing subsequent to the appllcation of ;
the amino acid compound. In addition, the amino acid compound may be applied to the dressings prior to use ln ..~
~ dry powder formulations. ~ ~
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The application of the amino acid compound may be made topically to the dermal surfaces preferably in a suitable carrier such that it is distributed on the dermal surfaces in amounts sufficient to provide at least 0.01% by weight of body secretion as hereinbefore specified. The application preferably i8 made prior to the secretion of the body fluid to preclude substantially completely the formation of the undesirable products. However, application may be made sub~equent thereto by minimizing microbial action on the body secretions. Topical - 10 applications to dermal surfaces are suitable for inhibiting malodor development. When the application is to be topical, the :.:
carrier may be in ~olid, liquid, spray or semi-solid form in cosmetic or pharmaceutical carriers intended for topical use.

Carriers into which the amino acid compound may be incorporated - include lotions, ointments, aerosols, water solutions, creams, .
pulverized mixtures, gel sticks and the like. The various additives and diluents include ointment additives such as polysorbate 80, polyoxyethylene ~orbitan trioleate; surfactants and emulsifiers ~uch as lauryl sulfate, sodium cetyl sulfate, glyceryl monostearate, diethylaminoethyl alkyl amide phosphate, isopropyl myristate, octyl alcohol, glyceryl and glycol esters of stearic acidsS glycols such a~ propylene glycol: other polyhydroxy compounds such a~ glycerol, sorbitol: alcohols uch as ethanol, isopropanol, witch hazel (Hamamelis water);
perfumes; essential oils propellants such as halogenated ' ;

, ;''` ' ' ' " ,~ ~ ' -hydrocarbons, e.g., dichlorodifluoromethane, trichlorofluoro-~:r :' , ethane, etc., carbon dioxide and nitrogen; solid diluents such as calcium carbonate, starch, bentonite, talc; and silicone-type fluids such as polysiloxane fluid. Selection of the particular carrier varies with use. In dermal i -preparations for the control of inflammatory skin conditions, aqueous compositions are desirable. Preferred compositions for such applications are those comprising witch hazel as a . . . . .
carrier component.
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, . , `O In compositions for topical application, the amino ~ acid compound is employed in an amount of at least 0.05% by -- weight. This is desirable in view of the diluting and -possible partial deactivating effect of the carrier. The amino acid compound may be present in larger amounts and ` ;5 may even constitute the ma~or portion of the composition although less deslrable from practical considerations. For ` toplcal appllcatlon, the amino acid compounds are preferably employed in the form of alkanolamine salts. When the carrler ls aqueous, the salt may be formed in the composltlon ` 0 by admixing the free acid and the alkanolamine ln the aqueous ' carrier.
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In preparationsfor the control of inflammatory ; ~ skin conditions, the amino acid compounds employed are ; preferably in the form of acid or water soluble mono-valent salt or mixtures thereof in an aqueous composition in an amount of from about 2 to about 15% by weight when - based on the weight of free acld.

; When incorporating the amino acid compound into a lotion, cream or aerosol, the amino acid compound may be added ln a solvent compatible with the system in which it is incorporated such as water, glycerol, propylene glycol, tripropylene glycol, methyl ether, ethanol, etc. Alternatively, the amino acid compound may be added to the final composition - and intimately admixed therewith. Such would be the preferred method for preparing dust~ng powders as well as aqueous ; 15 solutions such as in witch hazel.
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The compositions above described are applied to the situs of production of undesirable materials by the various ; microorganisms. The microorganisms associated with the ~
` production of materials contributlng to menstrual malodor include Gram Negative organisms such as Proteus mirabilis~
` Klebæiella eumoniae, Aerobacter aerogenes, Escherichia coli, ; Pseudomonas aeruginosa, etc., Gram Positive organisms such as Staphylococcus aureus, Streptococcus faecalis, etc. and yeast such as Candida Albicans, etc. The microorganisms associated :
- 25 with the production of substances contributing to axillary . .

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malodor are those normally present on the skin surfaces of which those of the genera Corynebacterium and Staphylococcus are the most important. Corvnebacteria ~
~; are also associated with the production of acids contri- -buting to the undesirable inflammatory conditions of the skin. By employing the compounds and compositions in ~i~ accordance with the present invention, the action of ~t~ the microorganisms which results in the production of ~ undesirable acids is somehow inhibited or altered. The ,,i 10 am~,ino acid compound is believed to be proceeding by .
removal of the necessary metallic co-factor for the `
~ enzymatic production of fatty acids. HOwever, the inven- :
i tion is not limited to any particular theory and khe ~ . . ~ . . .
~, preventative control of fatty acid production may be achieved without neces,saril.v causing detrimental effect on mlcroblal flora.
.. _ . .
- The effectiveness of the amino acid compound in ` lnhibiting the formation of undesirable fatty acids from :r lipids was determined by gas chromatographic analyses and ~ the effectiveness in odor control was determined by organ-;' 20 oleptic techni~ues.

h~l ~as chromatographic analyses employed comparisons ~-wlth known acids. The method employed was as follows:

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Fatty acids extracted in diethyl ether from acidified test . ~., samples were determined by gas chromatographic procedures using a Hewlett-Packard 7620A instrument and Porapak QS 80/100 mesh in a 6 ft. x 2mm. I.D. glass column. The instrument was pro-S grammed from 135 C to an upper limit of 235 C at the rate of 4 C temperature rise per minute, holding the upper limits for six minutes. Helium was used as a carrier gas at a pressure of 60 PSI and a flow rate of 40 ml./min.

. .
For evaluation of the ~fectiveness Or catamenial dressings in odor control, quantitative organoleptic evaluation method was employed referred to as the Modified Ratio Scale Organoleptic Evaluation method as follows.
. ' .
The Modified Ratio Scale Organoleptic Evaluation Method _ mis method is devised so that data obtained from an organoleptic appraisal panel may produce an evaluation of a sample characterized as an absolute value for odor intensity.
Thus, not only can the difference between two samples of an " ,j odorant placed in different environments (e.g., on a pad . wlth and without deodorizer) be detected, but further, the evaluation will indicate, quantitatively, as to whether the ~' ~0 odor intensities of the samples are strong or weak. For . .
example, one sample may contain a deodorizer which is many times as effective as that contained in a second sample.
. This notwithstanding, both deodorizers may only produce a small decrease in the odor intensity, all of which is indicated '5 by the sub~ect evaluation method. -~

: 1 ~,: - 1066624 The first step in this method is to determine the threshold concentration of the odorant. The method used is described by Fred H. Steiger in Chemical Technology, ~ -Volume 1, pg. 225, April 1971, wherein the determination of the odor threshold concentration for ethylamine is described, applying the Weibull distribution function.
Generally, this procedure requires the gathering of organoleptic data from a panel presented with a series of ;~ samples containing odorant in increasing concentrations in -; 10 order to determine the concentration level at which an arbitrary percentage of the panelists can detect the odor. -For the purposes of the current evaluation that arbitrary percentage is chosen as the cumulative 50% level. As so determined, the threshold concentration of the odorant i8 ~pecific to the odorant and the conditions of the sampling `` procedure.

i The method employed herein for panel evaluation is ' to present each panelist with a series of samples, in a sampling apparatus which consists of an opaque, one pint, S 20 polyethylene Mason jar having a polyethylene screw cap ~.;, fitted onto its neck. The jar is internally lined with a ~; polyethylene bag and a Buchner funnel is fitted about the cap, with the narrow outlet portion of the funnel, below the filter plate of the funnel, extending through the cap and . , into the lined jar. A sample is placed in the ~ar, the jar - ls capped with the Buchner funnel fitted in place and a watch glass is placed across the wide inlet portion of the funnel.
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The sample is then allowed to equilibrate for one hour at ambient conditions. For the purpose of the threshold determination, the samples each comprise a specific concentration of the odorant in a water solution, a total of 3 ml. of solution being placed in the jar. A panel of about 30 women are presented with a series of equilibrated samples of increasing concentration and, starting with a sample at zero concentration (water only), are asked to report the first sample having a detectable odor. The panelists are instructed to sniff each sample in turn, pausing 30 seconds between samplings. The accumulated data is organized to establish the cumulative percentage of the panel which detects an odor at each concentration level corresponding to each sample. The data, so organi~ed, is plotted as described in the aforementioned Steiger article on Weibull Probability Paper with the concentration as the abscissa and the cumulative percentage of the panel as the ordinate. The concentration at 50% is then taken as the s threshold concentration.
';

Having established the threshold value, the Modified Ratio Scale Method is applied by preparing a master curve.
~: Using the same testing apparatus, a series of samples are prepared and presented to the panel wherein the concentra- -tions of odorant can be expressed as multiples of the .... . .
~25 threshold concentration or odor units. One of these samples is 20 times the threshold concentration (20 odor units).
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In accordance with the standard method of Ratio ; Scaling, each panelist is asked to evaluate the set of samples before her and t'o assign a value to the odor inten-sity of each sample in proportion to the intensity of the other samples. The panelists are free to choose whatever scale they wish. For example, a panelist may assign 10 - to the strongest sample. A'sample having half that intensity, '' in accordance with this panelist's evaluation, would then be assigned a value of 5. The accumulated data then consists of a series of evaluations for each panelist, each series ~' being bas~d upon the individual panelists' scale. Arbi-' trarily, a ratio scale value of 100 is assigned to the sample concentration of 20 times threshold concentration. Each of the panelists' evaluations are then proportioned to bring their individual scales to the basis of 100 for 20 times threshold concentration. For example, the evaluations Or a panelist ass1gning a value of 10 to a first sample having " a concentratlon of 20 times threshold concentration will ` be proportioned to show, for that panelist, a value of 100 for the first sample and a value of 50 for tile second sample.
e data is now organized so that, for each sample corres-' ponding to a specific multiple of the threshold concentration, there is a series of Ratio Values, all on the same scale '~ (20 times Threshold Concentration - 100) corresponding to each panelist's evaluation of this sample. The geometric `~ means of the ratio values for each sample is calculated and that value is taken as the Ratio Value for that multiple of threshold concentration.
. . ~

~ 20 ,. . ,: ': '' ' ' f When the log of the Ratio Value is plotted, as the ordinate, against the log of the multiple of threshold concentration, a straight line, fitted to the data points between 3 to 20 times threshold concentration gives an excellent correlation.

It has been discovered that irrespective of which amine - odorantls tested, when a threshold concentration is determined for that specific odorantand the method ~ constructing a ~ Master Curve as described above is followed, the resulting x 10 Master Curves are superimposable between multiples of thres-.~ hold concentration of about 3 to about 20.

m e curve obtained in this way for isobutyric acid, a chemically dissimilar material also was superimposable on the Master Curves for the amines. Thus, the reference odorant ln any test may be chemically dissimilar to the odor constituent or constituents being tested.
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The Master Curve may now be used to evaluate the odor lntensity Or any odorant when placed in any environment such ~ as, for example, on a pad of untreated cellulosic fibers or A~ 20 on a pad of fibers containing deodorizing material and, in addition to obtaining comparisons between the relative inten-slty Or the samples tested, an absolute measure of the intensity each sample may be obtained. To do this, the panel is ;~
presented with a series of samples, one of which is a ~ -standard sample consisting of known concentration of the odorant being tested in an environment identical to that ~, .

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, . - . , - . .. .. -- :. .. , - . . . : -~Q66624 used in producing the Master Curve. Preferably, this standard sample is chosen as having 20 times the threshold concentration and hence, a Ratio Value of 100 on the Master Curve.

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The panelists are again asked to evaluate the ~; series of samples using whatever scale they choose. Based on the value which each panelist gives to the intensity of the standard sample, all other values given by the panelists ~, are proportioned so as to be consistent with the rating of the standard sample, e.g., a panelist assigning a value of 50 to the standard sample and 5 to a second sample of - `unknown intensity will have these values proportioned so the standard will be given a Ratio Value of 100 and the unknown sample given a Ratio Value of 10. By referring to the Master Curve, lt can be determined that the panelists now proportloned Ratio Value Or lO for the second sample is equi~alent to an odor lntensity of a certain number of -multlples of threshold concentration as read from the Master Curve were 1.2, the odor intensity of the sample Or unknown intensity, ln the test environment, is the same as a sample having an odorant concentration of 1.2 times the threshold concentration in the standard environment.
.~ .
Preliminary determinations were made in connection with the menstrual malodor aspect of the present invention to establish the presence of phospholipids in menstrual fluld and to establish malodorous fatty acid production by microbial flora commonly present in menstrual fluid.

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The presence of significant amounts of phospholipids in menstrual fluid was established by a procedure consisting of heating the phospholipids with perchloric acid to oxidize the organic portion of the molecule and convert the phosphorus ~ :- .
to a blue phosphomolybdate which may be read colorimetrically as more fully described on pp. 375-376 of "Bray's Clinicæl Laboratory Methods" by Bauer et al, 17th Edition, C. W. Uosby co., 1969. Ten samples of menstrual fluid were collected in cups and sub~ected to phospholipid analyses. The analyses 0 established the presence of from 2250 to 4140 ppm of phospho-lipids amounting to from 0.225 to 0.4% of the fluid.
To establish the ability of microorganisms commonly present in menstrual fluid to produce malodorous fatty acids, separate samples of sterile human blood were inoculated with different organisms previously isolated from menstrual fluid, ~ollowed by incubation and analysis by gas chromatography as hereinafter described. In the determination, blood was employed ` instead of menstrual fluid because of the availability of sterile blood and the desirability of a sterile substrate to 0 identify fatty acid production as being caused by the organism ~` used in inoculation. The suitability of substituting bloodwas based on the known fact of the presence of triglycerides in both blood and menstrual fluid and on pre~iminary experi-ments on six samples each of human blood and menstrual fluid .:
which showed average total phospholipids to be 2807 ppm and 2771 ppm respectively. The methods employed and the results obtained were as follows:

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: :
- ~0666Z4 ~ ---. --.:

2.5 ml. test samples of sterile whole human blood were separately inoculated with a 0.2 ml. bacterial cell uspension of various test organisms previously isolated by conventional methods from menstrual fluid. The test samples as well as an uninoculated control sample were incubated at 37C. for 24 hours in a constant temperature bath with constant shaking at 200 r.p.m. Thereafter, all samples were sub~ected to gas chromatographic analyses.
The results are seen in Table A.
.... ...

TABLE A ~-.- . ~ .. ... .
FATTY ACIDS PRESENT (ppm) Iso-butyric Butyric Iso-valeric . .
Unihoculated Control 0 0 3.5 Gram Ne~ative Bacteria Proteus mirabilis 59.5 9 111.7 Klebsiella pneumoniae 0 0 11.3 / , Aerobacter aerogenes 0 8.7 15.0 ,5 Escherlch~a~coli 0 0 o Pseudomonas aeruginosa 0 0 0 Gram Positive Bacteria ~.
Staphylococcus aureus 13.7 0 50.5 Streptococcus faecalis 0 0 39.1 Yeast . .
Candida albicans 12 o 38.5 ,:
The following examples illustrate the various aspects of the present invention but are not to be construed as limiting:

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:
:
: 1066624 ,, .

EXAMPLE I
. ..
Two menstrual fluid samples were collected and analyses were made initially for fatty acids: isobutyric, butyric and isovaleric by gas chromatography on untreated, unincubated fluid, Thereafter, test samples of about 2 ml.
of each menstrual fluid were prepared. Two sets, each set containing two different samples of menstrual fluid were treated as follows: to one set, disodium ethylenediaminete-, traacetate was added to provide a final concentration thereof i of 0.2% by weight of menstrual fluid ant the other set was left untreated. Both treated and untreated samples were lncubated at 37C. for 24 hours and thereafter analyzed by ga~ chromatography for the fatty aclds previously named.
`~, The results are seen in Table I.
. ,, . , . TABLE I

. FATTY ACIDS (ppm) ---Iso-butyric Butyric Isovaleric i Sample 1 Freshly Obtained Unlncubated, Un-treated Control 0 36 0 ' Incubated l; Untreated 685.7 1014.0 857.1 -.:
`l Ihcubated Na2EDTA Treated 0 16.6 14.3 Sample 2 Freshly Obtained Unincubated ~' Untreated Control O O O
; Incubated Untreated 475.o 307.0 336.0 , .. .
Incubated ~ Na2EDTA Treated4.0 13.0 9.0 .`.', . ' . :. :~
~ .

." . , . .. ~, . . .

EXAMPLE II
Five samples of 2.5 ml. of sterile human blood were inoculated with 0.2 ml. of Proteus mirabilis suspension. --, j .
To four of the samples, disodium ethylenediaminetetraacetate was added to supply amounts ranging from 0.04~ to 0.2%.
.. .
One sample was an inoculated control containing no EDTA -,~.; .j . ..... . . .
compound. In addition, an uninoculated control sample was , , prepared of 2.5 ml. of the blood to which 0.2 ml. of sterile `. dlstilled water was added. The treated and untreated ; lnoculated samples and an uninoculated control sample were 0 then lncubated for 24 hours at 37C. in a constant tempera- -ture water bath with shaking at 200 r.p.m. At the end of this period aliquots were tested for the presence of the ~ aclds by gas chromatography. The results are given in -l Table II.
~ . .. ~ ,, .
r~ ~,I
TABLE II

TREATMENT Fatty Acids (ppm) Na2EDTA Isobutyric Isovaleric Proteus + No Na2 EDTA 102.7 98.2 . .
Proteus + 0.04% Na2 EDTA 28.4 52-3 Proteus + 0.1% Na2 EDTA 6.6 17.5 Proteus + 0.2% Na2EDTA 7.9 17.6 Untreated Sterile Blood O O
, "' '' ' ' .
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.: . -. ....
.' ' ~ .

; ;
. -: -, . . . .
- . ' ' ; '- ' , ;.

: 10666Z4 .

EXAMPLE III

Sterile blood inoculated with Proteus mirabilis was placed on treated and untreated samples of fluff pads and sanitary napkins. To prepare treated samples, a 14 percent aqueous solution of a blend of disodium and tetrasodium ethylenediaminetetraacetate (pH 7) first was applled at various positions and concentrations to samples 4 of fluff pads or samples of sanitary napkins by soaking, spraying or padding on the cover. The samples were there-' after dried and the amount of the ethylenediaminetetraacetate salts dispersed in the samples determined by weighing.
Blood previously inoculated with Proteus mirabilis was added to the various treated samples as well as to an untreated sample and to a dimethylamine reference odorant sample. (Three milliliters were used for fluff pads and ~l~e milliliters were used for the sanitary napkins).
The samples then were incubated at 37C for 24 hours and thereafter sub~ected to organoleptic testing in the ; manner previously described. The results showing significant odor reductlon sre seen ln ~sble III.

. .

, .

.
. .

, ~- 1066624 .. ~. o ~
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~H ~ O P I I N ~ o C
~3 H cq ~3 :
O h O O
h ~ O h p ~1 ~ I I I ~ O ~ H ~ --, .,, ~ ~ ~ ~ O ~ bD
. .~ Cl ~ ,~j h ~ a) 3 ~, ~ ~ c ~ ~1 c H =
O H O ,1 ~ ~ ~ N ~ h ~ h ~ ..
q O a~
h ~ p C P ~d~ C

.. H .H ~ ~ O ~ td tq ~ . H o h o ~a Q) Q' ~¦ H o oC d~ d ~d b ~d ~d H ~ h h ~
H ~ :~ O Cd . .~: ~ a, h ~ ~-. ~q h ~ a 3h ~ ~
~ O h--1 ~ ~ I I 1 5 1 ~ g ~ 6~ ~ h ~ o~ ~h `~ ~ ~ O C ~ ~ :
.. O ~ rl O ~1 ::~ ~ h c)~q ~ ~ ~ ~1 ~ h h ~ h ~ h :~ h ~ I I I I O ~ ~o , ~ ~ ~ ~~O ~ O ~:
.'2 ~ h~ ~
`;' O h rQ H r1 ~ h ~d ~ ~~ = d,~ -1~ ~ P ~ O O O ;~; O O O O O E ,~ ,C O C d E

1 O O O O O O O O O r~ N
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c~ ~D O ~
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"' " ' ' ' ' " ' .~' ~ ' ' ' ' ' ' ~ ' ' .

~. .
In an experiment carried out over a period of 8iX
months, a panel of seven sub~ect~ wore sanitary napkins menstrually for a minimum of four hours or until the dis-charge of menstrual fluid was such that a change of napkin . 5 was necessary. Within thirty minutes after removal, the napkins were inspected and physical data such as weight, ' physical appearance and stain area were recorded. There-; after, the napkins were incubated at 30C. for one hour ant thereafter evsluated for odor.

: 10 Thi8 procedure was repeated by each panelist with a treated and untreated (control) napkins but on alter-nate nths. In some cases, the data was collected over the entire six-month period while in other cases, the ., ~
data was only for two months or for four months. The lS election of product to a given panelist was randomized ~o that factors such as climate or activity changes would have minlmNm influence.

The treated napkins employed had been prepared by removing the non-woven fabric cover of a MODESS* Sanitary Napkin, passing the absorbent tissue inner padding through a ~olution of a blent of disodium and tetrasodium ethylene-d~aminetetraacetate (prepared as described in Example III), drying and replacing in t~e fluid pervious cover. The con-; centration of the salt after drying was 0.004 gram per ~25 square inch of fabric.
: ' ,, * Trademark of JOHNSON & JOHNSON

.' ' . -- ' . ., ' : - 29 .:
- , , . .

` 1066624 . - . .

~ PPC 185 ;:~ The odor data from all sub~ects for this period were pooled and the odor intensity responses for test products compared with a control. The odor response for twenty-five treated and twenty-five untreated napkins were described in odor units of from zero to 21 (maximum). The -untreated napkins had odor units ranging from 3 to 21 with ` five napkins above 15. Ten of the treated napkins had ; odor units of less than 3 and none above 11.
'.', , ,'- .- .
EXAMPIE V '~

~10 Two milliliter portions of dilute human plasma (1 part of plasma to 2 parts of sterile distilled water) were inoculated with Corynebacterium. To three of the - .
~ samples, disodium ethylenediaminetetraacetate was added ; to produce a concentration thereof of 0.1%, 0,2% and 0.5%.
` 15 One sample containing no sodium ethylenediaminetetraacetate served as an untreated control. In addition, an uninocu-lated control sample of unmodified dilute human plasma was prepared. The test and control samples were incubated at 37C. with shaking at approximately 200 r.p.m. for 24 hours.
At the end of this period, the samples were tested for , i free fatty acids by gas chromatography. The resu-lts obtained were a8 follows:

. ~ .

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3 .. . .

:, . .

'4" :10666Z4 .. .

TABLE IV
;
Treatment Fatty Acids (ppm~
Isobutyric Isovaleric Untreated sterile plasma 0 o - Corynebacterium Inoculum -No Na2EDTA 155 564 , ;' Corynebacterium Inoculum 12 22 ~5 + 0.5% Na2EDTA
Corynebacterium Inoculum 25 49 + 0.2~ Na2EDTA
: ~, j Corynebacterium Inoculum 22 41 ! + 0~1% Na2EDTA
, . i ~
.,. ,~
~ ~ EXAMPLE VI
, ~ 10 Disodium ethylenedlaminetetraacetate was added to ; ;~
,1 a two milliliter portion Or a brain-heart-infusion (BHI) 1 broth contalnlng natural axillary flora consisting primarily ~, ~it ` . -of Corynebacterium and Staphylococcus to produce a final concentration of 0.5% by weight. A second two milliliter ` I , ..
Y~115 portlon of inoculated broth contained no disodium i ethylenediamlnetetraacetate. In addition, a two milliliter portlon of sterile BHI broth served as an uninoculated control. The test and control samples were incubated at 37C with shaking at 200 r.p.m. for 24 hours. At the end ¦20 Or this perlod, samples were determined for free fatty ; acids by gas chromatography. The results are seen in ; ~-Table V. ; ~
': ' :
. --' 31 : . . .
~ ~ .
,, : . :

... . ` " - , .;, , ~ . . .. . ~ , . .. . ..

. 10666Z4 PPC 185 :
. ~
TABLE V
:i :
Treatment Fatty Acids (ppm) Isob~yric Isovaleric Sterlle BHI O O
BHI inoculated with : -; underarm culture 32 57 BHI lnoculated with underarm culture + 0.5% Na2EDTA O O
., ., ~:'''.
-' ' :' .
EXAMPLE VII

An aqueous solution containing o.45 percent by weight . . .
of a mixture Na2EDTA and Na4EDTA (prepared as described in . , . ~
Ex. III) was applied to one axilla Or a test sub~ect by means '0 Or a sterile guaze pad whlch had been wetted wlth the solu-1 tion. Sterile distilled water was applied in a similar ,;, ..
~I manner to the other axilla and served as the untreated '~ control. Both axilla were evaluated by olfactory sensing ~ twice dally to determine compartive odor development and ,5 lntenslty. The treated axilla exhibited a substantial deodorant efrect.

,:

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... ~ . : . . .. . , . ., ,:
. ~ . . .

EXAMPLE VIII

A cream suitable for application to the body for inflammatory disorders of the skin is prepared by (1) heating Part A (below) to 70 (2) Part B (below) to 75~C. (3) adding Part B to Part A with agitation and (4) -~: 5 ad~usting the pH to 5.5 with dilute sodium hydroxide.
"' '' .
Part A % Weight Cetyl alcohol 2.5 Stearyl alcohol - 5.o Isopropyl myristate 2.0 ~, .~ Light silicone oil 1 0 . . i -~.~ 10 "Emplex"l 1.5 .~ 2 Methyl paraben 0.15 Propyl paraben3 o.o5 , .
, Part B
b~ . ~
'~ Delonized water 78.8 ~
.3 Disodium salt Or EDTA 4.0 ..
. 15 Propylene glycol 5.0 ;:
. .
,1 lSodium salt Or reaction product Or lactic and stearic ,~ acid (Patco Products, Kansas City, Missouri) - Trademark ;,. .. ..
.~; 'Methyl hydroxybenzoate- Trademark :~
~` 3Propyl hydroxybenzoate - Trademark : ~
,, :- .
- :

33 ~:
,:: -PPC 185 ~
, ; EXAMPLE IX

A lotlon suitable as a skin lotion is preparedby (1) heating Part A (below) to 72C, (2) Part B (below) to 75C (3) adding Part B to Part A with agitation and .

(4) ad~usting the pH to 5.3 with dilute sodium hydroxide.

, _ .
. Part A ~ Wei~ht ;~ 5 Cetyl alcohol 1.9 Stearyl alcohol 3.0 ; ~ Isopropyl myristate 1.3 ~j Llght silicone oil o.8 : ~
"Emplex" 1.1 10 . ~. Nethyl paraben 0.15 ~' Propyl paraben 0.05 . I .
Psrt B
'I , ,~, .il Delonized water ôl-7 ~'l Propylene glycol 3.o _ ~, Dl~odium salt of EDTA 7.0 .j, ..
.~,.. , .;~
t, ' EXAMPl E X

: `: ` ' ' '--,i 15 The compositions described in Examples VIII and IX
:~ may be applied to dermal surfaces to reduce formation of . ,~ .
~`~, f~tty acids by the action of Corynebacterium species on ,`, exuded sebum, sald fatty acids being undesirable in ~-lnrlammatory skin disorders.

~, ",': - ~- .
.~ 34 ~ .

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. . ; , .. . , . ~ . . - , .. ~ -- .. . ~ . , . ~ ,. ...

r- _ ; 1066624 EXAMPLE XI
, A hand and body lotion suitable for malodor control i5 prepared by (1) heating Part A (below) to 82C, (2) heating Part B (below) to 78C, (3) adding Part A to Part B with stirrlng and (4) cooling to 46C and adding Part C. , .. ,~ . ., ~' Part A ~ Weight ;~ Mlneral Oil 3.oO
Glyceryl monostearate5.00 Isopropyl palmitate 3.00 ~ -Amerchol-H-91 1.00 ,, ~ .
Stearic acid 1.50 Propyl paraben 0.05 ' I ~ A
Part B
Methyl paraben 0.15 `~ Onyxide 5002 0.20 ~ '~;
; Propylene glycol - USP~.00 ,; 15 Glycerlne 96% - USP 3.00 Standapol SHC-3013 2.50 ; Dl~odium salt of EDTA5.00 Deionized water 71.35 Part C
; Fragrance 0.25 2 Sterolatum (Amerchol Products, Inc.) - Trademark 2-Bromo-2-nitropropane-1,3-diol (onyx Co.)- Trademark 3 Sulfosuccinate half ester (Henkel Co-)_ Trademark ' -.. . .

~ ~ 35 ~

PPc 185 EXAMPLE XII
~ . .
A body powder suitable for topical application 18 prepared by thoroughly mixing the following:

.
Trisodium ethylenediaminetetraacetate 10 g.
Talc 787 g.
Fragrance 3 g.
; ' ' '' ~''.

EXAMPLE XIII
.

The composition described in Example XII may be applied to dermal surfaces to inhibit formation of malodorous fatty aclds by the action of Corynebacterium :
specles on exuded sebum.
, -' ' EXAMPLE XIV ;

A sanitary napkin i8 prepared having an absorbent core of comminuted wood pulp and a nonwoven porous cover, ana having on the upper surface o~ the wood pulp core sodlum nitrilotriacetate in an amount of 1.0 milligram per square inch of surface area. In its preparation, the nltrllotriacetate 18 applied to the absorbent core in ;
dry 8tate from an aerosol spray employing dichlorodifluoro- ;
methane as a propellant after which the nonwoven cover i8 ~ - .
placed around the core.

~,. . .

.. . ..
. ~

. . .

EXAMPLE XV

A sanltary napkin simllar to that described in Example XIV except that both the upper surface of the wood pulp core and the fluid pervious cover has applied thereto sodium N-hydroxyethyiethylenediaminetriacetate (HEDTA salt) at a rate of 1.6 mg. per square inch of surface area.

EXAMPIE XVI
~: ,': . ' ~ A catamenial tampon is prepared having an il absorbent compressed, cylindrical core of tissue pulp and short rayon fibers. The leading one-half Or the ~,; 10 surface of the core has applied thereto 1.5 mg. per square inch of a neutral blend of di- and tetrasodium ethylenedlaminetetraacetate. The salt is applied to the core surface in a dry state from an aerosol spray employing dlchlorodifluoromethane propellant. A nonwoven cover is wrapped around the core and a withdrawal string 18 knotted around the core at its trailing end.

EXA~PLE XVII

catamenial tampon similar to that described in ~' Example XVI but wherein the leading two-thirds surface Or both the absorbent core and fluid pervious cover have applied thereto l.0 mg. per square inch of trisodium ethylenediamine tetraacetate.

. ' ; :

. :`

. ~ .

~ EXAMPLE XVIII

; A cream stick was prepared by admixing at about 60C the following materials in the indicated proportions:

Weight in Component Grams , Ozokerite wax ~ 48.o ; Carnauba wax 32,0 Candellila wax 64.o f~ '~ Emcol 249-3K 42.0 ` Tenox 42 0.8 ., ' . -1 Benton M-203 80.0 ~' Talc 120.0 -I 10 Propyl paraben o.8 Neutral blend of Na2EDTA 40.0 plus Na4EDTA~

! 1 Alkoxyl~ted alcohol (Witco Chemlcal Company) - Trademark ~ , ~ ..
20~ Butylated hydroxy anisole, 20~ butylated hydroxy toluene, 60% corn oll (Eastman Chemical Products Inc.
- Trademar~
Modi~led bentonite (National Lead Company) _ Trademark t` 4 Prepared as described in the Footnote 5 to Table III.

' Thereafter the mixture was poured into chilled molds to provide a final product in chilled stlck form.
. . .
. .
Test panellsts were provlded with two coded cream ;
stlcks for axillary deodorant use, one stick being of the above composition and the other stlck of similar composition but not contalning the blend of the two sodium ethylene-..

A~ 38 .. .

~, . . .

dlaminetetraacetate salts. The test panellsts were not lnformed as to which stick contalned the EDTA salts.

, The subJects applied one stick to one axilla and ; the other stick to the other axilla and a~ter each ~5 application the subJects made qualitative evaluations on 7 odor intensity. The resuits are given in Table VI.
' ' ~ ', TABLE VI --Odor Intensity by Sel~-Evaluation Panelist Control Axilla Test Axilla A Strong No odor -., . - .
~ B Very Strong No Odor C Very Strong No Odor r~
-~1 D Strong No Odor E Strong No Odor ~15 F ~ Very Strong Sllght G Very Strong Sllght ~~
H Strong Slight I Very Strong Slight J Very Strong Slight K Strong Slight ;: L Strong Slight M Very Strong Slight . ~ :
EXAMPLE XIX
.
An aerosol composition is prepared by first ;
admixing the ~ollowlng materials ln the indicated prQportions:

-'- . ..
39 :~:

. ~ - : . . ., . ~ : . -` 1066624 .
.; . Parts by Weight Mlcropulverlzed Talc 2.50 Mlcropulverized Na3EDTA a.50 ., . Fragrance 0.16 . , Anhydrous ethanol 0.20 .~ 5 Isopropyl myrlstate 0.60 ';c ::
.. . .
Thereafter, the mlxture 18 placed ln vessels .:. sultable ror pressurlzatlon (cans) and the followlng 1.~ ~, . . .
~ propellants added ln the lndlcated proportions:

;,i. ` Freon ll 47.02 ~ 10 Freon 122 47.20 ; , .~ :L . .
Trlchloromonorluoromethane (E.I. duPont de Nemours & Co.) Dlchlorodl~luoromethane (E.I. duPont de Nemours & Co.
- Tra ema~k .'' . ' . '' ' ~ .

. '~ .
.

.. i ... .
. ~ -.

-~ .

PPC 185 ~. . -. EXAMPLE XX

In an experlment carried out ln a manner slmilar to that descrlbed in Example V, test sample~ were prepared by adding .-to samples Or sterlle plasma inoculated with ~rynebacterium, ~.
one of the follo~ing aminopolycarboxylic acid compounds: - -dlsodlum ethylenedlamlnetetraacetate (Na2EDTA), N-hydroxy~
.: ethylethylenediamlnetriacetic acld (HEDTA), diethylenetrlamine-~ pentaacetlc acid (DTPA), ethylene glycol bis(amlnoethyl ether~
.. tetracetlc acld (EGTA), and a trlethanolamine salt of .
ethylenediamlnetetracetlc ac~d (TEDTA) prepared by adding . :
trlethanolamlne to aqueous ethylenedlaminetetraacetlc acld to :
a pH of 6.3. The amount of amlnopolycarboxylic acid compound.
added to the samples was that surficlent to provlde ln the test medla 0.2% by welght Or the amlnopolycarboxylic acld ... . .. ..
compound (calculated as the free acld). In addltlon two control samples were prepared: One control s~mple was Or unmodlrled sterlle piasma (unlnoculated and contalnlng no ~ :
aminopolycarboxyllc acld compound) and the second control aample was an.lnoculated sample containlng no aminopolycarboxylic ; acid compound and served as untres,ted control. The samples wore incubated and tested for free ratty aclds as pre~lously .. descrlbed. m e results obtalned were as rollows: -.
" ~ " ' :: ' .
., . ~, .

` 4l ~ ` :

. .
~, ~ 10666Z4 TABLE VII
. , .

Fatty Aclds Produced (ppm) Isobutyric -Isovaleric Untreated Sterile Plasma 0 0 ! ,, . .
Corynebacterium Inoculu~
No AminopolycarboxyIic ~!'.' ~ Acld Compound 105 ~ 187 :r ' . _ ~
Corynebacterium Inoculum 0 0 + 0.1% Na2EDTA
.
Corynebacterium Inoculum 0 0 + 0.1% HEDTA
Corynebacterlum Inoculum 0 0 + 0.1% DTPA
~'' . ' Corynebacterium Inoculum 0 0 " + 0.1% EGTA
Corynebacterium Inoculum 0 0 + 0.1% TEDTA

. . .
EXAMPLE XXI ~
:.
i5 Sanltary napklns are prepared havlng an amlnopolycarboxy-lic acld compound impregnated on the upper surface of the absQrbent , coro and fluld pervious cover ln a manner slmllar to that ; described in Examples xrv and XV except that-they are modlfled ' wlth respect to the amlnopolycarboxyllc acld compound and . ..
~ amount employed per square inch o~ sur~ace area as follows:
Disodlum ethylene glycol bis(aminoethyl ether)-tetracetate, 3 mg. per square inch.
~ . . .
`~ Sodium iminodiacetate, 2 mg. per s~uare inch.
Ethylenediaminetetra(methylenephosphonic acid)-i5 neutralized with dlethanolamine to pH 6.3, 1.5 mg. per Bquare inch.
Pentasodium nitrllotrl6(methylenephosphonate), 2 mg. per square inch.
Ethylenedlamlnetetra(methylenesulronic acid) neutra-~ lized with triethanolamin~ to pN 6.3, 2 mg. per square inch.
' ~0666Z~
-- -- .

EXAMPLE XXI_ A cream stick is prepared ln the manner described ln Example XVIII by admixing the following materials ln ; the lndicated proportions~
Welght ln Com~onent Grams Ozokerite wax 48.o Carnauba wax 32.0 Candellila wax 64.o . . .
Emcol 249-3K * 42.0 . .
Tenox 4 * o.8 Benton M-20 * .80.0 , 10 Talc 120.0 Propyl paraben * o .8 Triethanolamlne salt Or ethylanedlamlnetetraacetic acid S-. :
The stlck. may be applied to the dermal surraces Or the 8xllla to inhlbit formation o~ odorous fatty aclds by actlon Or -~
CorYnebacterium on llpoldsl materials. :
. ~. ~ . . ..
~ ' .
T~`YA ~T lP YYSTT
~l .
Composltions suit8ble for toplcal appllcation to inhiblt ratty acid production from lipoldal materlals by Corynebacterlum :.
may be prepared by lntimately admixlng the components speclfled below ln the amounts indlcated to obtain treating compositions:
* Trademark : . .
.
. ' 43 . .
.~ ~ , .

~0666Z4 ~
,"'' .
; PPC 185 .
Composition A

; N-Hydroxyethylethylene-dlaminetetraacetlc acld10 part by welght ~: Trlethanolamlne10 parts by weight Wltch hazel80 parts by welght Composltlon ~

. 5 Ethylene glycol bls(aminoethyl- ~ -. ~ ether)tetraacetlc acld13 parts by welght~
; Trlethànolamlne 13.8 part~ by welght ; Witch hazel 73.2 parts by weight . .; , Compositlon C .:
. .
Ethylenedlamlnetetraacetlc acld 10 parts by welght ~:
Trilsopropanolamlne 17.7 parts by welght Witch hazel 72.3 parts by welght .. . .
,.~ . '' -EXAMPLE xxrY

A composltlon suitable for appllcatlon to dermal tlssue 18 prepared by lntimately admixlng the rollowing:

~i5 Ethylenedlamlnetetracetlc acld 10 parts by welght Triethanolamlne 13.8 parts by welght Nltch hazel 76.2 parts by welght ffl ls composltlon may be applled to dermal tlssues to inhiblt development of lnflammatory skin condltlons resultlng from _ -` ~066624 .- . :.

fatty acids arising from secreted llpoidal materialæ by the action of Corynebacterium thereon. In such use daily .. ..
application is made by swabbing the affected area of the dermal surface and repeating such applicatlon until~the ~ -condltion ls alleviated. Further application may be made on the appearance of development Or the undesirable ~-condltionæ to prevent further development of æald condltlons.
.; , ~ ~.

EXAMPLE XXV

The compositlon of Example XXIV was employed to - --~0 determine efflcacy ln inhlbltlng undeslrable lnflammatory condltlons Or the skin typical Or acne. The undeslrable ., , ; ...... ...
`` condltlons may be those de8crlbed as the exlstence of erythema and the presence Or papules and pustules.

In the determlnatlon, six sub~ects wlth similar degree ~ 15 f nflammatory condltlons on both sides of the face applied (~ the abo~e composltlon to one slde of the face after r~rst making determlnatlons on the number of ~rythem~us papules and pustules on each slde of the face, herein termed "leslons."
The other slde of the face served as control. The appllcatlons were made twlce dally to one slde of the face arter rlrst --separately cleaning each ~lde of the face, and the appllcation was contlnued for a period of three weeks. At the end thls perlod the percent change ln the number Or leslons on t~e treated and untreated æides of the face were determlned.
~25 The results were as follows:

L~ ~ ~ , '`. ' :10666Z4 TABLE VII
Percent Change Panelist Treated Untreated 1 70% Reduction ~.6% Reduction 2 70% n - 55% Increase 3 5~ 1 7~
4 62% 11 20% n 62% ~ 3% Reductlon 6 59% " 5~ 1! s ."'' ' ;
~,.
Although ln the past, undeslrable products resulting .. . from microbial action on body secretions have been lnhlblted by kill o~ microorganlsm~, lt has now been discovered that by a use Or the composltlons Or the present lnvention, the desired : . inhlbltory actlon may be accompllshed wlthout the neces~ity .. . .
:~ o~ kill Or the organisms. Thus, ror example, when growth o~
.~ Protous mirabilis in heparinized blood at 37 with and . 15 ~ithout added disodtum ethylenedlaminetetraacetaee was ~ollowed over a 24-hour period, there was found to be no detrimontal erfect on the growth o~ organisms as can be een in the ~ollowing tabl-:
. Bacterial Count Blood Inoculated 20With Proteus Mirabllls 0 Hours ~ 24 Hours No Na2EDTA 100 x 1~ ~~. 1& x 107 ~:: 2~ Na2EDTA 100 x 1~ 231 x 107 --., ~ . ~ .

Claims (8)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A composition suitable for application to dermal tissue to inhibit the formation of undesirable microbially produced fatty acids consisting of a carrier suitable for topical applica-tion and, as the sole active ingredient, an aminopolycarboxylic compound selected from the group consisting of ethylenediamine-tetraacetic acid, diethylenetriaminepentaacetic acid, N-hydroxy-ethylethylenediaminetriaacetic acid, triethylenetetraaminehexa-acetic acid, 1,2-diaminocyclohexane-N,N'-tetraacetic acid, N,N-dihydroxyethylethylenediaminediacetic acid, iminodiacetic acid, hydroxyethyliminodiacetic acid, nitrilotriacetic acid and their water soluble salts; said aminopolycarboxylic compound being present in a concentration of at least about 0.5 weight percent in said composition, based on the weight of the free acid, said composition maintaining the viability of Corynebacterium.

2. A composition according to Claim 1, wherein the car-rier is an aqueous composition.

3. A composition according to Claim 2, wherein the aque-ous composition includes witch hazel.

4. A composition according to Claim 1, wherein the amino carboxylic compound is a water-soluble salt of ethylenediamine-tetraacetic acid.

5. The composition of Claim 4, wherein said salt is an alkali metal salt.

6. The composition of Claim 5, wherein said alkali metal salt is a sodium salt.

7. The composition of Claim 6, wherein said salt is a di-sodium ethylenediaminetetraacetic salt.

8. The composition of Claim 7, wherein said salt is a trisodium-ethylenediaminetetraacetic salt.