EP1108056A1 - Verfahren zur prüfung von antibakteriellen-verbindungs-zusammensetzungen - Google Patents

Verfahren zur prüfung von antibakteriellen-verbindungs-zusammensetzungen

Info

Publication number
EP1108056A1
EP1108056A1 EP99942498A EP99942498A EP1108056A1 EP 1108056 A1 EP1108056 A1 EP 1108056A1 EP 99942498 A EP99942498 A EP 99942498A EP 99942498 A EP99942498 A EP 99942498A EP 1108056 A1 EP1108056 A1 EP 1108056A1
Authority
EP
European Patent Office
Prior art keywords
bacteria
skin
composition
attachment
visually
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP99942498A
Other languages
English (en)
French (fr)
Inventor
Shamin Alam Ansari
Diana Kalliope Kiozpeoplou-Grina
Thomas Gregory Polefka
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Colgate Palmolive Co
Original Assignee
Colgate Palmolive Co
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US09/141,904 external-priority patent/US5951965A/en
Application filed by Colgate Palmolive Co filed Critical Colgate Palmolive Co
Publication of EP1108056A1 publication Critical patent/EP1108056A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor

Definitions

  • EP 806935 A is directed to the use of a carbohydrate or derivative thereof as an antiadhesive against a host of harmful materials including bacteria, parasites and protozoa on cell surfaces such as skin, mucous membranes, body orifices, interiors or hollow body organs, wounds, eyes and hair.
  • USP 5,416,075 discloses the use of an oil in water emulsion having an amphipathic molecule including a biospecific moiety at the head end of its hydrophilic part. These compositions are applied to the skin. These headgroups inhibit adhesion of bacteria to the skin.
  • a. treating skin with a potential or known inhibition of bacteria attachment composition b. contacting the said skin with a bacteria resulting in the skin having the bacteria attached to it; c. contacting the skin with a bacteria growth supporting medium having optionally therein or optionally later added a compound or mixture thereof which will assist in visually detecting a bacterial colony.
  • bacteria can be visually detectable as a colored grouping without a separate visually detectable medium applied to it.
  • Such bacteria include Serratia marcescens, Straphylococcus aureus, Pseudomonas fluorescens, Pseudomonas aeroginosa, Bacteroides asaccharolyticus and bacteroides melaninogenicus.
  • bacterial growth media have certain component(s) therein which will specifically support the growth of certain bacteria and provide a readily visible color to the bacteria. Furthermore there are certain compound(s) available which when added to the growth media will selectively color certain bacteria, even in the presence of other bacteria.
  • a further aspect of the invention is a method of visually evaluating the comparable effectiveness of potential or known inhibition of bacteria attachment compositions which comprises steps a, b and c above for each bacteria attachment composition being evaluated and comparing the visual quantities of bacteria colonies on the skin for each said composition DETAILED DESCRIPTION OF THE INVENTION
  • bacteria which are inhibited from attaching to the skin include Staphylococcus aureus, Staphylococcus epidermidis, Corynebacterium minutissimum, Escherichia coli, Salmonella choleraesuis and Serratia marcescens as well as other bacteria mentioned in this application.
  • the test is simple to run and provides an easy method of assessing visible quantity of bacteria on the skin.
  • a skin part including but not limited to an ex vivo skin explant or any source from animal and human, synthetic skin and the like, for example the hand, is contacted with a test composition which may inhibit bacteria attachment.
  • a control composition is applied that does not have the component(s) of the test composition which allegedly bring about the inhibition of bacteria attachment.
  • a soap composition having inhibitors of bacteria attachment is applied to the skin (hand).
  • the same soap composition without the component(s) responsible for inhibition of bacteria attachment is applied.
  • Each hand is now contacted with a specific bacterium or various bacteria against which the inhibition of bacteria attachment composition is thought to be effective.
  • a bacteria growth supporting solid medium which will support bacteria growth or a medium to which the bacterial growth nutrient can be readily added.
  • a growth support medium is agar. Incorporated within the medium or added an appropriate time thereafter is an amount of growth nutrient in sufficient quantity to bring about the growth of the various bacteria transferred from the skin. After an appropriate period of time to allow growth to occur, at least partially dependent on the temperature, particular bacteria, and the like, the medium bearing the bacteria is visually assessed.
  • bacteria without any natural color As noted previously, some bacteria do have a natural color. Additionally, color can be imparted to bacterial colonies by the nature of the nutrient growth material employed since many bacteria are capable of producing pigments when grown in medium supplemented with specific nutrients. Such medium can be selective or differential in nature. For example, such nutrient medium will bring about color pigment for the following bacteria: • Escherichia coli will produce colony with a characteristic green metallic sheen on agar containing Eosin Methylene Blue.
  • Such nutrient medium can include such components as peptone, lactose, dipotassium hydrogen phosphate, Eosin Y, Methylene Blue and agar at a pH of 6.8. • Staphylococcus aureus will produce colony with a characteristic yellow color when grown on mannitol salt agar.
  • Such nutrient medium can include the following components per liter of purified water:
  • LB- medium contains Bacto tryptone, yeast extract, NaCl, agar and water.
  • composition Approximate formula per liter purified water Pancreatic digest of Casein 15.0 g
  • enterobacterial species also produce pigments at a particular incubation temperature*:
  • Chromogen a caprylic acid ester, specifically an indolyl caprylate 40-200 mg/ ⁇
  • results are assessed on the basis of the quantity of visually detectable bacteria on the medium - the lesser the number of visually detectable bacteria colonies on the medium, the more advantageous the inhibition of bacteria attachment composition while the greater the number of visually detectable bacteria colonies on the skin, the less advantageous the inhibition of bacteria attachment composition.
  • skin means the top layer of skin (i.e., stratum corneum) and all the components of the stratum, both cellular and acellular (i.e., biomolecules) such as proteins, carbohydrates, lipids and the like.
  • compositions which can potentially or is known to inhibit the attachment of bacteria to skin can be evaluated by this method.
  • compositions disclosed in US provisional applications 60/087,533 and 60/087,532, both incorporated by reference. These compositions are directed to combinations including surfactant(s) with either a silicone such as dimethicone or a hydrocarbonaceous component such as petrolatum, minerol oil, paraffin and the like being present in attachment inhibiting amounts or both a silicone and a hydrocarbonaceous component together in attachment inhibiting amounts.
  • a cationic polymer can also be present with either or both the silicone and hydrocarbonaceous component.
  • fragrance can also be present such as fragrance; antimicrobial materials such as Triclosan or trichlorocarbanilides; and the like.
  • antimicrobial material such as Triclosan or trichlorocarbanilides; and the like.
  • an antimicrobial material such as triclosan or triclocarban can be omitted as well. This method can be used to compare and/or contrast known and unknown antibacteria attachment composition to each other or to a control composition.
  • the transference of bacteria to the skin can occur through any type of contact, for example skin to skin, or skin surface bearing such bacteria, for example doorknob, faucet, telephone, table top and the like. In sufficient density, bacteria can even be transferred to the skin in an airborne manner.
  • Transference of bacteria to the growth supporting medium from skin can occur through simple contact of the surface bearing the bacteria to the surface.
  • An example of such a transfer is pressing a hand on the surface of an agar plate.
  • the bacteria are then allowed to grow into various colonies by incubating at a temperature optimum for bacterial growth and providing nutrients to the growth supporting medium, if not already present. After a period of time necessary to allow such bacteria to grow, at least partially dependent on temperature, pH, type of bacteria and the like, the bacteria are visualized without any assistance with any of the color including media, or compounds previously described. However, these additional method(s) of assisted visualization can be used, if desired.
  • Example 1 Thirty subjects participate in the studies. Subjects undergo a one-week washout period where they refrain from using any products labeled antibacterial such as antibacterial soaps, dishwashing liquids, lotions, creams, talcs, etc. and antidandruff shampoos for one week prior to the beginning of the study and for the duration of the test period.
  • antibacterial soaps such as antibacterial soaps, dishwashing liquids, lotions, creams, talcs, etc. and antidandruff shampoos for one week prior to the beginning of the study and for the duration of the test period.
  • the subject's hands are rinsed with 70% ethanol to remove any contaminating bacteria and allowed to air dry.
  • Each of the subject's hands are washed either four times or once by a technician.
  • the washing procedure consists of a 15 second wash with the bar soap, 45 second lather with the gloved hand and 10 second rinse under running warm tap water.
  • the hands are allowed to air dry before proceeding with the next wash.
  • the subjects gently place each of their hands on the surface of a plastic plate previously contaminated with 200 ⁇ l (approximately 10 6 ) of the marker bacteria, Serratia marcescens (ATCC 14756). Two objects weighing approximately a total of 480 g are placed on top of the hands to help provide even pressure.
  • the hands are left on the plate for 5 seconds.
  • the subjects then, as soon as possible, gently place each of their hands on a pre-poured Microbial Content Agar hand imprint plate to transfer the bacteria.
  • a technician applies gentle pressure to each of the subject's hands and fingers for 10 seconds.
  • the same objects used as weights above are placed on top of the hands to help provide even pressure and the hands are left on the agar plate for 30 seconds.
  • the subjects hands are decontaminated by soaking them in 70% isopropanol for 3 minutes.
  • the agar plates are incubated overnight at 35-37°C.
  • the plates are evaluated by three judges using the following scale:
  • the 3 judges scores for each plate are averaged.
  • a paired t-test is employed using the judge average scores to determine whether significant references existed between products at the 5% significance level.
  • Test Product 2.8 + 1.0 2.0 + 0.8 p-value ⁇ 0.05 ⁇ 0.05
  • the test product has significantly inhibited the attachment of bacteria to the skin.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Toxicology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
EP99942498A 1998-08-28 1999-08-25 Verfahren zur prüfung von antibakteriellen-verbindungs-zusammensetzungen Withdrawn EP1108056A1 (de)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US363898 1982-03-31
US141904 1998-08-28
US09/141,904 US5951965A (en) 1998-08-28 1998-08-28 Method for visually demonstrating the effectiveness of an anti-bacteria attachment composition
US09/363,898 US6165443A (en) 1998-08-28 1999-07-30 Method for visually demonstrating the effectiveness of an antibacteria attachment composition
PCT/US1999/019517 WO2000012751A1 (en) 1998-08-28 1999-08-25 Method for testing anti-bacterial attachment compositions

Publications (1)

Publication Number Publication Date
EP1108056A1 true EP1108056A1 (de) 2001-06-20

Family

ID=26839555

Family Applications (1)

Application Number Title Priority Date Filing Date
EP99942498A Withdrawn EP1108056A1 (de) 1998-08-28 1999-08-25 Verfahren zur prüfung von antibakteriellen-verbindungs-zusammensetzungen

Country Status (10)

Country Link
EP (1) EP1108056A1 (de)
CN (1) CN1316014A (de)
AU (1) AU772199B2 (de)
BR (1) BR9913193A (de)
CA (1) CA2341411A1 (de)
HU (1) HUP0103440A3 (de)
NO (1) NO20010990L (de)
NZ (1) NZ510081A (de)
TW (1) TWI237060B (de)
WO (1) WO2000012751A1 (de)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6180577B1 (en) * 1998-06-01 2001-01-30 Colgate-Palmolive Company Anti-germ attachment—composition

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4518517A (en) * 1983-03-16 1985-05-21 Colgate-Palmolive Company Non-antimicrobial deodorant cleansing composition
US4812253A (en) * 1985-05-13 1989-03-14 The Procter & Gamble Company Ultra mild skin cleansing composition
IL104399A0 (en) * 1992-01-22 1993-05-13 Mennen Co Deodorant compositions containing materials for inhibiting bacterial adherence,method of use thereof,and method for determining materials that inhibit bacterial adherence
FR2708285B1 (fr) * 1993-07-28 1995-10-20 Rambach Alain Procédé d'identification de microorganismes avec un milieu supplémenté en hydrate de carbone.
DE4328689A1 (de) * 1993-08-26 1995-03-02 Beiersdorf Ag Verfahren zur Detektion und Zählung von Mikroorganismen

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0012751A1 *

Also Published As

Publication number Publication date
TWI237060B (en) 2005-08-01
WO2000012751A1 (en) 2000-03-09
AU5585999A (en) 2000-03-21
HUP0103440A3 (en) 2002-06-28
BR9913193A (pt) 2001-05-15
CA2341411A1 (en) 2000-03-09
NO20010990D0 (no) 2001-02-27
AU772199B2 (en) 2004-04-22
NO20010990L (no) 2001-02-27
HUP0103440A2 (hu) 2002-02-28
NZ510081A (en) 2003-09-26
CN1316014A (zh) 2001-10-03

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