Microbial spoilage-by S.D.Mankar types, sources of contamination, factors,Assesment

Types of spoilage, factors affecting the microbial spoilage of pharmaceutical products,
sources and types of microbial contaminants, assessment of microbial contamination and
spoilage.

1. Microbial Spoilage By.S.D.Mankar Assistant Professor Department of Pharmaceutcs Pravara Rural College Of Pharmacy,Pravaranagar

2. Microbial Spoilage Microorganism is integral part of environment. Present in air, food, soil, water etc. Different types of M.O. from environment mainly contaminate the pharmaceutical preparation and spoil it. Such spoilage result in major health & financial problem to users & manufacturer respectively.

3. Definition Pharmaceutical products may be considered to be microbiologically spoiled if low levels of pathogenic microbes or toxic microbial metabolites are present and detectable physical or chemical changes have occurred in the product . Microbial limit std.for non sterile pharmaceutical dosage form are set as per reference std.

4. The amount of water in the formulation is one of the imp.factor . Bcz in presence of water in formulation it support to microbial growth. Nature of formulation- its origin( Plant, Animal, or mineral)

5. Types of microbial Spoilage 1. infection induced by contaminated pharm. Products:- May be contaminated by pathogenic M.O. mainly from raw material or at time of preparation. These contaminated products may cause serious infection to patient when they use this drugs or formulations. 2.Phy & Chemical deterioration of products:- Naturally occurring products sensitive to attack.

6. Types of microbial Spoilage Rate of deterioration depends upon chem. Structure, phy -chem. Properties of env . And the level of microbial contamination present. 3. observable effects of microbial attacks on products:- Microbial spoilage of different dosage forms may be detected by organoleptic test such as unpleasant smell and taste, coloured pigments, and unwanted appearance.

7. Types of microbial Spoilage 4.Ingredients susceptible to microbial attack:- i ) therapeutic agents:- many drugs are capable of gross degradation by wide variety of M.O. ii) surface active agent:- various type of surfactants are susceptible to microbial degradation. Iii) polymers & Humectants:- polymers are subjected to depolymerisation and humectants support to microbial growth unless present in high conc.

8. Types of microbial Spoilage Iv) Fats & Oils:- they are usually attack when dispersed in aq. Formulation V) preservative & disinfectants:- these sub are metabolised readily by many bacteria & fungi. Vi) Sweetening, flavouring & Colouring agent:- All the organoleptic agents are easily support to the growth of M.O.

9. Factors:- The physical & chemical status of pharm. Formulation influences the type & extent of microbial spoilage considerably. 1. Size of innoculum :- Low level of contaminants may be present in a product but it would cause low rate of deterioration. Ing . Contaminated by high level of M.O. causes appreciable microbial degradation.

10. Factors:- 2. Nutritional factors:- microbial spoilage req. appreciable growth of M.O. in dosage form. Many formulations offers considerable nutritional variety for wide range of M.O. E.g. plant or animal source products in formulations. 3.Moisture content:- Dissolved components of an aq. Formulations may form a complex with water molecules via hydrogen and other bonding thus lowering the proportion of water available to contaminants M.O. in the product.

11. Factors:- The high conc. Of solute in formulation indicates that water activity is low. Most of M.O. grow best in high water activity. Hence formulation can be protected from microbial attack by lowering water activity with addition of suitable levels of sugars,PEG , NaCl, or by drying.

12. Factors:- 4) Temperature:- spoilage of pharmaceuticals could occur over the range of about -10 to 60 ˚C. Storage within specific, narrower, temp. ranges will encourage the growth of particular groups of spoilage organism. Syrup & multidose eyedrop preparations are sometimes dispersed woth label “store in cool place” to reduce the

13. Factors:- Risk of contamination. Distilled water for WFI can stored at 80 ˚C prior to sterilization to prevent pyrogen production. 5) pH:- Extremes of pH prevents microbial attack, although growth of mold is commonly observed in solution of dil.HCL . Antacid mixtures, mouth washes and distilled water show growth of pseudomonas species and other gram – ve bacteria.

14. Factors:- Soap emulsion ( pH 8.0 – 9.5) & mixtures of mag.hydroxide or Al. hydroxide gel discourage microbial attack. Acidic conditions favour fungal & yeat growth. 6) Redox Potential:- The ability of microbes to grow in an environment is influenced by its oxidation and reduction balance.( redox potential).

15. Factors:- Vaccum packing of foodstuffs or the inclusion of oxygen absorbs in the package are to minimize oxygen levels and reduce microbial spoilage. Redox potential may high in viscous emulsions quite high due to appreciable solubility of oxygen in most fats & oils.

16. Factors:- 7) Protective components:- Various components of formulations and products may increase the resistance & longevity of contaminant M.O. M.O. can show more resistant to heat & disinfectant in the presence of polymers such as starch, acacia & gelatin . Adsorption of M.O. on to suspended particles such as kaolin, mg. trisilicate or aluminium hydroxide gel may increase their longevity.Some sub. May increase resistance of M.O.

17. Source & Types of microbial contamination:- The microbial contamination by pharmaceutical product is influence by environment to which they are manufactured & by the materials used in their formulation. 1. Atmosphere:- The number of M.O. in atmosphere depends on activity in environment & the amount of dust which is present.

18. M.O. are carried into atmosphere suspended on particles of dust, skin, clothing's, some are droplets of moisture or sputum following coughing or sneezing. The microbial content of the air may increase during handling of contaminated materials during dispensing, blending, & formulations. Settle plate or diff. air sampling tech. are commonly used for isolation & identification of M.O. present in atmoshphere .

19. 2.Water:- It is main constituent of many and also used for washing and cooling process. Different types of M.O. are present in water & some microbes may contaminate the water from sewage. 3. Raw materials:- Raw materials particularly of natural origin are potentially rich source of M.O.

20. animal- source- gelatin , pancereas Plant source- starch, acacia, tragacanth . 4. Process operator:- M.O. may be transferred to pharm. Formulation from process operators. Natural skin flora M.O. are the main source of contamination. E.g.staphylococcus aureus .

21. Poor personal hygiene. The nasal passages & respiratory tract may contain large numbers of diff. M.O. 5. Equipments :- The equipments should be as simple as possible for the purposes required with minimum of junctions, valves, & pumps to allow cleaning in place by circulation detergents or other chemical antimicrobial agents.

22. 6.Packaging:- The microbial flora of packaging material is dependent upon its composition, nature & storage conditions. Packaging material have dual role and act both to contain the product & to prevent the entry of M.O. or moisture. Packaging materials-glass, cardboards, closures may contain difff . M.O.

23. 7. Building:- diff. moulds & few bacterial species are most common flora of walls & ceilings. They are mainly found in poorly ventilated buildings. they utilize their nutrients from plaster paints. To reduce microbial growth, all walls, floors & ceilings should be smooth & washable. Doors & windows should be fit flush with walls to prevent entry of dust particles & M.O.

24. assessment of microbial contamination and spoilage:- 1. physical & chemical changes:- It is the changes of different pharmaceutical formulations indicate microbial contamination and spoilage. Change in viscosity, pH, emulsion stability and loss of surface activity of formulation indicates microbial spoilage. Measurement of oxygen consumption of the product can indicate the degree of oxidative attack and microbial growth

25. 2. Sterility Test:- Testing which confirms that products are free from the presence of viable microorganisms . Claim to be sterile or free from viable microorganisms . Test is conducted by competent and experienced personnel in an adequately clean room with laminar flow cabinet facilities . All injectables and ophthalmic preparations are sterile hence, these preparations are tested by the sterility test.

26. 3. Assessment of viable microorganisms in non-sterile products :- tested for viable microorganisms for detection of pathogens and total viable counts .

27. 4. Estimation of pyrogens :- Pyrogen a substance, typically produced by a bacterium , which produces fever when introduced or released into the systemic circulation. The lipopolysaccharides and lipoproteins which comprises a major part of the cell wall og Gram – ve bacteria are called endotoxins which are the most commanly called pyrogens . • To test the pyrogens presence, two tests are used 1. RP Test (Rabbit Pyrogen Test) 2. LAL Test (Limulus Amoebocyte Lysate Test

28. 1 . RP Test (Rabbit Pyrogen Test ):- It requires administration of injections to lab animals. Their body temp. is monitored for period of time. 2. LAL Test (Limulus Amoebocyte Lysate Test In which the pyrogen - containing sample causes gel formation in the lysis product of amoebocyte cells of the giant horseshoe crab limulus polyphemus .