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Antimicrobial Preservation Overview For Formulators and Process Scientists

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Information about Antimicrobial Preservation Overview For Formulators and Process Scientists
Health & Medicine

Published on March 3, 2014

Author: JimMcElroy

Source: slideshare.net

Description

Antimicrobial Preservation Overview For Formulators and Process Scientists
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THE BASICS • THIS PRESENTATION IS FOR THE FORMULATION SCIENTIST THAT HAS LITTLE OR NO MICROBIOLOGY EXPERTISE • IT IS ALSO AN INTRODUCTION TO BASIC PRESERVATIVE CHEMISTRY AND ACTION

COMPOSITION AND STRUCTURE OF MICROORGANISMS BACTERIA BASICALLY COME IN TWO TYPES: • THOSE THAT HAVE A LOT OF PEPTOGLYCAN IN THEIR CELL WALLS AND THOSE THAT HAVE A LITTLE. • WHEN BACTERIA ARE STAINED, IT IS DUE TO A CHEMICAL REACTION BETWEEN THE STAIN AND THE PEPTOGLYCAN. CRYSTAL VIOLET STAIN IS GRAM POSITIVE. • SAFRANIN (WHICH IS RED) INDICATES GRAM NEGATIVE.

CELL MEMBRANES • THE CELL ENVELOPE IS THE RATE LIMITING BARRIER FOR THE PARTITIONING OF : • • • NUTRIENTS WASTE PRODUCTS CHEMICALS • MAIN COMPONENTS ARE PHOSPHOLIPIDS AND PROTEINS • MAJOR PROTEINS ARE HYDROPHOBIC • THE PLASMA STRUCTURE IS STABILIZED BY HYDROGEN BONDING • MG2+ AND CA2+ CATIONS STABILIZE THE MEMBRANE

COMPARISON Property or Constituent Gram Positive Gram Negative 1. Liposacharide Outer Layer Absent Present 2. Polysaccharide High Low 3. Lipids 2-4% 15% or more 4. Solubility of lipids in fat solvents Resistant Less resistant 5. Amino Acids 3-4 Complete range 6. Optimal Growth pH Relatively high Relatively low 7. Nature of toxin Exotoxin Endotoxin

YEAST AND MOLD • YEAST: CELL WALL IS COMPOSED OF TWO OR MORE PROTEIN POLYSACHARIDE COMPLEXES HELD TOGETHER BY COVALENT BONDS • YEAST: MANNAN IS A CELL WALL COMPONENT • MOLD: CELL WALL IS COMPOSED OF TWO OR MORE PROTEIN POLYSACCHARIDE COMPLEXES HELD TOGETHER BY COVALENT BONDS • MOLD: MANNAN IS NOT A CELL WALL COMPONENT

COMPARISON (BACTERIA, YEAST & MOLD) Property Bacteria Yeast, Mold Cell size (μm) 0.5-5 Yeast 20-50 Mold ≥100 Cell wall Teichoic acids, muramic acid peptides Chitin, glucans, manans, and diaminopimelic acid Cytoplasmic membrane No sterols Sterols Cytoplasm No mitochondria and endoplasmic reticulum Mitochondria and endoplasmic reticulum Nucleus Prokaryotic (no membrane) Eukaryotic (membrane)

WHAT IS A PRESERVATIVE? A CHEMICAL SUBSTANCE THAT IS USED TO PRESERVE PHARMACEUTICALS, FOOD OR OTHER ORGANIC MATERIAL FROM DECOMPOSITION OR FERMENTATION BY PREVENTING THE GROWTH OF MICROORGANISMS.

COMMONLY USED PRESERVATIVES INGESTIBLE & TOPICAL • PARABENS • SORBIC ACID • NA, K & CA SORBATE • BENZOIC ACID • NA, K & CA BENZOATE • SODIUM METABISULFITE • PROPYLENE GLYCOL (15-30%) • BHT, BHA • FLAVORS W/ BENZALDEHYDE TOPICAL ONLY • FORMALDEHYDE DONORS • ESSENTIAL OILS • MONOGLYCERIDE • PHENOL • MERCURY COMPOUNDS

PARABENS • PARABENS ARE AMONG THE MOST COMMONLY USED PRESERVATIVES. • THEY ARE RELATIVELY ACTIVE AGAINST A BROAD SPECTRUM OF MICROORGANISMS. • THE METHYL ESTER IS MOST EFFECTIVE AGAINST BACTERIA AND MOLDS • THE ETHYL, PROPYL & BUTYL ESTERS ARE MORE ACTIVE AGAINST YEAST AND MOLDS. • PARABENS ARE MORE EFFECTIVE AGAINST GRAM NEGATIVE THAN GRAM POSITIVE ORGANISMS. • PARABENS DISASSOCIATE AT HIGH PH (8 OR MORE) AND ARE RENDERED INEFFECTIVE.

PARABENS HAVE LIMITED SOLUBILITY IN H2O Property Methylparaben Ethylparaben sol. g/100mg sol. g/100mg Propylparaben sol. g/100mg Butylparaben sol.g/100mg Water 18ºC 0.16 0.08 0.023 0.005 Water 25ºC 0.25 0.11 0.04 0.015 Water 80ºC 3.2 0.86 0.45 0.15 Ethanol 22 25 26 110 PG 1.7 0.5 0.4 0.3 Peanut Oil 0.5 1 1.4 5 Mineral Oil 0.01 0.025 0.03 0.1

PARTITION COEFFICIENTS AND EMULSIONS PAY CLOSE ATTENTION TO PRESERVATIVE PARTITIONING IN EMULSION SYSTEMS. PARABENS PREFER THE OIL PHASE THUS LEAVING LITTLE IN THE WATER PHASE WHERE IT IS NEEDED. Water phase Oil droplet Surfactant

PARTITIONING OF PARABENS • PARABENS ARE LIPOPHILIC. • PARABENS WILL PARTITION DISPROPORTIONATELY INTO THE OIL PHASE OF AN EMULSION. • IF LARGE AMOUNTS OF THE PRESERVATIVE PARTITION INTO THE OIL PHASE, IT IS NOT AVAILABLE FOR ANTIMICROBIAL ACTION.

PARTITION COEFFICIENT MEASUREMENT • CW IS THE PRESERVATIVE LEFT IN THE WATER PHASE • C IS OVERALL % OF PRESERVATIVE • KW IS THE PARTITION COEFFICIENT • Ά IS PHASE VOLUME RATIO Cw=C* ά+1 / Kw + 1

PARTITION COEFFICIENTS FOR PARABENS & OTHER PRESERVATIVES Preservative Kw Oil Methylparaben 7.5 Almond Oil 0.1 Mineral Oil 18.0 Isopropyl Myristate 200 Diethyl Adipate Ethylparaben 26.0 Soya Oil Propylparaben 87.0 Soya Oil Butylparaben 280.0 Soya Oil Benzoic Acid 6.10 Soya Oil Phenol 5.60 Arachis Oil 0.07 Mineral Oil 3.3 Almond Oil 0.21 Mineral Oil Sorbic Acid

CHELATING AGENTS AS PRESERVATIVE ENHANCERS ALKALINE EARTH METALS SUCH AS CA+ AND MG+ ARE IMPORTANT FOR THE STABILIZATION OF THE OUTER MEMBRANE OF CELLULAR ORGANISMS. CHELATING AGENTS SEQUESTER THESE IONS. EDTA IS A TYPICAL CHELATING AGENT USED IN FORMULATIONS.

CHELATING AGENTS ARE NOT PRESERVATIVES • PROVIDE A SYNERGISTIC EFFECT FOR THE OVERALL PRESERVATIVE SYSTEM • CONTRIBUTE TO THE PARTIAL SOLUBILIZATION OF THE CELL MEMBRANE WHICH ALLOW PRESERVATIVES A PATHWAY INTO THE CELL.

INGREDIENTS THAT ENHANCE PRESERVATIVE EFFICACY • SOLUTES (SALTS & HIGH CONCENTRATION OF SUGARS) • ESTERS • CATIONIC AND ANIONIC SURFACTANTS • HUMECTANTS (GLYCERIN, PROPYLENE GLYCOL) • PHENOLIC ANTIOXIDANTS (BHT) • CHELATING AGENTS (EDTA) • FRAGRANCES • LOW WATER ACTIVITY

INGREDIENTS THAT HINDER PRESERVATIVE EFFICACY • SUGARS AND ALCOHOL SUGARS • PROTEINS, PEPTIDES, YEAST EXTRACT • NATURAL GUMS & CELLULOSE THICKENERS • PLANT EXTRACTS (ALOE VERA, STARCH,…) • VITAMINS • CLAY COMPOUNDS • HIGH WATER ACTIVITY • SURFACTANTS (TWEEN 80)

MANUFACTURING STEPS THAT ENHANCE PRESERVATIVE EFFICACY • SANITARY CONDITION OF EQUIPMENT • TREATMENT OF RAW MATERIALS • HEAT TREATMENT OF SUB-PHASE OR FINAL BATCH • SUB-MICRON FILTRATION • HOT FILL • ASEPTIC FILL

PACKAGING COMPONENTS AND PRESERVATIVE EFFICACY • DESIGN OF OPENING OR DISPENSER • SUSCEPTIBILITY TO CONTAMINATION FROM ENVIRONMENT • SUSCEPTIBILITY TO CONTAMINATION FROM HUMAN CONTACT

CONCLUSION IT IS IMPORTANT TO TAKE INTO CONSIDERATION ALL ASPECTS OF THE FORMULA BEFORE CONSIDERING A PRESERVATIVE SYSTEM. THIS INCLUDES: • TYPE OF FORMULA (TOPICAL OR INGESTED) • WATER ACTIVITY • ORGANISMS LIKELY TO PROLIFERATE • BULK HANDLING • PACKAGING.

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