ANTIMICROBIAL PRESERVATIVES

Prof. Dr. C. Hanny Wijaya M.Sc

General Information
Microbe/microorganism living cell, very small, and can only be examined under microscope In an appropriate place, it is able to replicate itself every 15-20 minute Microorganisms which have important role in food bacteria, mold and yeast

Natural Preservative-Free Product

Cannot last for long time contains water and a lot of nutrition as microorganisms growing media Preservation important unit in food industry to slow down spoilage Two main classes of preservative: Antioxidant and antimicrobial

Antimicrobial
Substances used to lengthen products shelf life by preventing spoiling microorganism and pathogen Characteristic : - bactericidal - fungi static - fungicidal - bacteriostatic - germicidal

Growth Inhibiting Mechanism of Microbial growth by Antimicrobial Substance

Resulting damage in cell wall lysis Reacting with cell membrane& increasing the permeability of cytoplasm membrane Inhibiting the synthesis of the components of cells building block Inhibit/ inactivate essential intracellular enzymes Inactivate the functions of genetic material

Preservation Requirements with an Ideal Anti-microbe

Has a broad spectrum Not toxic for human and animals Economical Does not cause a difference in aroma and taste Being inactive in preserved food Does not induce the growth of more resistant new strain Performing more killing mechanism

Appraisal Factors in Choosing Appropriate Antimicrobial :

Antimicrobial spectrum of the component used Physical and chemical characteristic of antimicrobial and food product Product storage condition and interaction of product with other processes Initial microbial number on food product before the addition of preservative

Safety and Legality of Antimicrobial

Spectrum of antimicrobial Physical and chemical characteristics of antimicrobial and preservative solubility, pKa, pH and aw Solubility of antimicrobial decide on what kind of food product the antimicrobial can be added to

Safety and Legality of Antimicrobial

pKa decide on the ratio between dissociated and undissociated form of antimicrobial pH - Low pH does not need strong and plenty of antimicrobial - dissociated antimicrobial in its optimum pH high effectiveness minimum aw requirement for microbes to live and replicate

Preservative Types
Legality : GRAS (Generally Recognized as Safe) not toxic like salt, sugar, pepper and vinegar ADI (Acceptable Daily Intake) daily intake limit is set

Varieties of antimicrobials and their status as Food Additives

Organic Preservatives

Benzoic acid and its salt

Formula : C6H5COOH Form : cyrstal or powder Available naturally in food products such as : cranberries, plum, dried plum, cinnamon, ripe clove and most berries

Activity of Benzoic Acid

Antiseptic, antifungal, antipyretic Used in plasticizer forming, resin coating, caprolactam and quantitative analysis standard of alkalimeter method Main function as antimycotic Preventing the aflatoxin production by mold strain of Aspergillus flavus

Sodium Benzoate
A natrium salt Molecular formula : C6H5COONa More soluble in water compared to benzoic acid More stable Odorless Has a sweet, astringent taste Does not have terratogenic dan mutagenic Sodium benzoate is put as GRAS by FDA to the concentration of 0.1%

Benzoate

Effective at pH 2.5 4.0 less effective at pH > 4.5 Mostly applied in high acid food products Products example: carbonated drink, other drinks, syrup, margarines, olives, pickles, relishes, soya sauce, jam, jelly, pie and bread filling, fruit salad, preservatives of fish processing products

Sorbic Acid and its salts

Effective against mold, yeast and various bacterium Do not affect products flavor and aroma Do not affect biovailability Optimum activity at pH < 4.75 Antimicrobial effect of sorbic acid is not significant at pH >6.0 - 6.5

Effectivity of Sorbic Acid

Affected by : - pH - aw - contamination - processes - temperature - other food additives - packaging - initial level and types of microbes which will be prevented - storage length and sanitation

Berries isolated

Crystal or Powder

Sorbic acid

-Prevent fungal growth -Less effective in lowering bacteria effectivity

Sulfur Dioxide VS Sorbic Acid

Use in fruit processing and wine Sulfur dioxide has antimicrobial and antioxidant activity Use of Sulfur Dioxide is less if acid is added Advantage: can prevent the formation of sorbithol and off-odor

Sorbic Acid Derivatives

Sodium sorbate sodium salt of sorbic acid Calcium sorbate salt with antimicrobial and antifungal effect Potassium sorbate potassium salt of sorbic acid which is more soluble compared with the acid. Widely used in cheese products, butter, yoghurt, pickle, dried fruits and cake

Potassium Sorbates Antimicrobial Activity

Antimicrobial activity of potassium sorbate is 74% more than sorbic acid More widely used than sorbic acid Effective at Ph 6.5 and more effective in lower pH

Parabens
Benzoate derivatives which is alkyl ester of p-hidroxy benzoic acid Colourless & tasteless Relatively not hygroscopic Not volatile Active in basic/ acidic pH Widely used in beer processing, juice, desserm syrup, flavoring and fish processed products

Parabens
More active against yeast and mold Can obstruct membrane transport and electron transport system Low toxicity Easy to be hydrolyzed and conjugated in the body and excreted through urine Considered as GRAS with maximum addition to food 0.1% Has anesthetic effect

Benzene-contained molecules
Has antifungal activity such as diphenyl (E230), sodium ortophenyl phenol (E232), thiabendazole Diphenyl : - relaxing odor - melting point 70C, boiling pont at 255C - causing irritation at skin, eyes and lungs - if inhaled, causing headache, vomitting, gastrointestinal irritation Thiabendazole : - melting point 304C - showing terratogenic activity in mice - used as fungicide

Dimethyl Dicarbonate (DMDC)

CH3-O-O-C-O-C-O-O-CH3 A colorless liquid Slightly soluble in water Very reactive with substances including water, ethanol, alkyl and aromatic amines, sulfhydryl group as an antimicrobial DMDC : highly effective in inactivating mold and yeasts

Dimethyl Dicarbonate (DMDC)

carbonated and non carbonated nonjuice bevereges <250 ppm carbonated and noncarnated juice bevereges < 250 ppm wine : < 200 ppm ready to drink teas : < 250 ppm

LYSOZYME
1,4--N-acetylmuramidase, EC 3.2.1.17 present in avian eggs, mammalian milk, tears and other secretions, insects, fish Hydrolysis of the 1,4 glycosidic bonds between N-acetylmuramic acid and N-acetylglucosamine of peptidoglycan of bacterial cell walls degradation and lysis in hypotonic solutions

Lysozyme
Stable to heat 80oC, 2 min Optimum temperature 55-60oC Lysozyme : most active againts gram positive bacteria, peptioglycan of cell wall is more exposed than gram negative Cheeses : prevent gas formation Seafood, vegetables, salad and pasta

Short-chain Organic Acids and Salts

Acids
Acids as food additives serve a dual purpose
Acidulants Preservatives

Phosphoric acid is used in cola soft drinks to reduce the pH Acetic acid is used to provide tartness in mayonnaise and salad dressings Similar functions are served by organic acids
Citric acid, tartaric, malic, lactic acids

Acids
Straight-chain carboxylic acids, propionic and sorbic acids, are used for their antimicrobial properties Propionic acid is mainly used for its antifungal properties

Inorganic preservatives

Sulfur dioxide
It is a colorless gas It has a strong odor Naturally available in the atmosphere Classified to pollutant gas Can be derived through the burning of sulfur, hydrogen sulfite, or gips It is high in antimicrobial activities and influenced by its pH, concentration, the contact period with food materials, and variety of microbes

Sulfur Dioxide
Antimicrobial mechanisms through distracting the microbe cell component and interact with SH groups of structural protein, enzymes, cofactor, vitamin, nucleic acid, and lipids. Can be used in the wine processing, as paling agent in flour processing, fruits and vegetables fumigation. Can no be utilized in high Thiamin containing foods.

Characteristics of Sulfur Dioxide

Performing a toxicity in high concentration. corrosive Can cause death if directly inhale in air concentration > 33 mg/l

Sodium Sulfite
Air and water vapor sensitive Mutagenic Can cause eyes, skin, and respiration system irritation.

Sodium Hydrogen Sulfite Release sulfur dioxide gas causing asthma and stomach irritation. Sodium Metabisulphite - it has similar odor to sulfur dioxide. - might cause hypersensitivity. Potassium Metabisulphite - In form of white crystal - Mostly used in wine processing. - Causing irritation on eyes, skins, and respiration system.

Nitrite Containing Compounds

Nitrite In the meat curing contributes to formation of colors characteristic, flavor, texture quality, and antimicrobial effects. Nitrite effectiveness depends on its environment (food material)

Potassium Nitrite
Having white to yellows color. It is performed as curing agent and meat preservative Inhibits the botulisms bacteria. Preventing off flavors Maintains the food flavor quality Can cause irritation on eyes, skin, and respiration system.

Sodium Nitrite
It is Na NO2 granules White to darker yellows in color. Main functions preventing the growth and toxic production from Clostridium botulinum toxic, carcinogenic and irritating

Sodium Nitrate & Potassium Nitrate

It is a natural mineral in form of granules or crystal (white) Over consumption of nitrite and nitrate can cause the high concentration of both in our blood circulation.

Hexamine
Crystalline granules (CH2)6N4 white in color, and having antimicrobial activities Produced by reacting formaldehyde carcinogenic caiMenyebabkan iritasi pada kulit, mata dan saluran pernafasan

Phosphate and its derivatives

The functions of phosphate in food processing : - pH stabilizer (buffer) - acidity regulator - sequestering of metal - complex formation with polyelectrolyte organic (protein, pectin, starch)

Phosphate and its derivatives

The functions of phosphate in food processing : - de-flocculation - peptisation - emulsification - nutrition supplement - anti caking - antimicrobial preservative - leavening agent

Phosphate and its derivatives

Antimicrobial activity in positive gram bacteria is having a direct influence Over consumption of phosphate can reduce the bioavailability of calcium, iron, and other minerals.

Hydrogen Peroxide
Groups of bleaching and oxidation agent. Having antimicrobial characteristic. Effectiveness depends on the species of microbes. Mostly used in: - dairy and cheese preservation - preparation of pasteurized food product - enhancing flavor, aroma, and color of fish products Utilization legality from FDA is around concentration 0.05% for dairy processing to make cheese.

Sodium Chloride (salts)

Main utilization as adjunct In its pure condition is colorless. Main source sea water though drying process. Decrease the solubility of O2 and absorb water from the media. The antimicrobial activities of NaCl depend on : pH, temperature, thermal application, concentration, species and number of microorganism, food product, availability of preservative.

Biologically obtained preservatives

Grapefruit Seed Extract (GSE)

Very safe to commonly use as disinfectant. Performing an antimicrobial activity, antiseptic, antibacterial, and antioxidant. Produced from the extract of seed and grapefruit. The mixture of 60% GSE in 40% glycerin vegetable having storage period up to 79 years. Seldom to use as preservative in commercial food products.

Lactic Acid Bacteria (LAB)

It is classified to natural agent and able to inhibit the growth of some pathogens. The conditions that support the LABs growth : - availability of carbohydrate - decrease in oxygen concentration - decrease in ability of providing culture starter - sufficient production of lactic acids

The Mechanism of preserving by Lactic Acid Bacteria (LAB) in meat products)

Decreasing pH up to below 4.5 that prevents the growth of unwanted bacteria. The formation of organic acids material that can not be disassociate in a substantial amounts. Substrate buffering capacity Nutrient competition towards others disadvantageous bacteria. Antibiotic production and bacteriocin. Lowering the red ox potential.

Nicins
It is produced during the growth of Lactococcus lactis & Streptococcus lactis It has antibiotic activity Very effective against positive gram bacteria and spore forming bacteria. Popular to use in dairy and cheese products.

Natamycin/pimaricin
First isolated in 1955 from Streptomyces natalensis C33H47NO13; MW 665,7 Da a polyene macrolide antibiotic. Amphoteric low solubility in water (30-100 mg/L) active againts nearly all molds and yeasts No effect on bacteria and viruses.

Natamycin/pimaricin
Use in cheese making : 200-300 ppm baking industry : 5o ppm fruits and meats : 50-100 ppm The ADI 0,3 mg/kg body weight/day (FAO/WHO) Intravenous route : toxic Nausea, vomiting, diarrhea in human given orally >1000 mg/day

Other preservatives

Sucrose (sugars)
Effective to be an inhibiting agent for microbial growth. The minimum dose is 3% or 30 gram/kg material. Esther sucrose can impede the growth of lactic acid bacteria and S. cerevicae within concentration of 0.2 %

Carbon Dioxide
The mechanism of antimicrobial activities is assumed because of its exclusion ability that can reject the existence of oxygen. This antimicrobial activity influences molds and psychotropic bacteria. Resistant to Lactic acid bacteria (LAB) and anaerobes bacteria. Popular in its utilization on Modified Atmosphere Packaging (MAP).

Prohibited Preservatives for Food Additives.

Formaldehyde
CH2O liquids Colorless Can be utilized as disinfectant and fungicide. Performing a toxic activity when inhaled, swallowed, and absorbed. Mutagenic

Borax and Borate Acids

Considered as high risk preservatives Borate acids can cause failure in fetus development and reproduction system damage Causing irritation on skin, eyes and respiratory system in fair concentration

Warning !!! What is listed in this presentation is only a part of the whole information. You should learn more and develop your curiousity like this cat.