Easy to Pharmacy & learnt. III Sem B.Pharm - 2nd Year.

1. Microbial Spoilage,
Stability of Products &
Animal Cell Culture
Spoilage is a complex reaction such as combination of
microbial & biochemical activities are interact known as
“spoilage”.

2.  INTRODUCTION
• “Microbial spoilage is includes the microbial contamination of pharmaceutical products
with the microbes which lead to spoilage of product affecting the ‘drug safety’ &
‘quality’.”
Any change which refer a product unacceptable for human consumption.
The combination of microbial & biochemical activities may interact.
Spoilage is decreases the ‘stability’ & ‘intensity’ of the products.
• One of the major reason that led to preservation.

3. TYPES OF SPOILAGE
Spoilage are mainly two types:
[A]. Microbial spoilage.
[B]. Non-microbial spoilage.
Microbial spoilage Non-microbial spoilage
Spoilage
Chemical spoilage Enzymatic spoilage Physical spoilage Other spoilage

4.  MICROBIAL
SPOILAGE
• Short term:
Deterioration of pharmaceutical products
by the contaminant microbes such as
bacteria, mould & fungi etc.
It includes the contamination of
pharmaceutical products with the
microbes that leads to spoilage of
the product affecting drug safety &
quality.

5. Non-microbial spoilage
Chemical reaction is caused by foreign material
in the foodstuff that occur in the foodstuff
naturally resulting in rancidity, discoloration or
drying out.
They are further divided as
following;
Chemical spoilage
Physical spoilage
Biological spoilage
Other spoilage

6. Chemical spoilage
• Based on the non-enzymatic chemical reaction occurred with in the foods &
change in flavor.
• These occurs due to various types of chemical reactions, mediated by
contaminating micro-organisms.
Chemical agents foods Off flavor
Trimethylamine Meat & egg. Fishy
Ethanol Fruit juice. Alcoholic
Dimethyl trisulphide Meat , fish. Garlic
Acetic acid, lactic acid Dairy, wine. Souring
Diacetyl Meat. Cheesy

7. ENZYMATIC SPOILAGE
• Based on enzymatic reactions occurs in foods & changes in chemical nature &
resulted rancidity.
• Rancidity also occurred by two types of chemical reactions:
- Hydrolytic rancidity.
- Oxidative rancidity.
Enzymes foods Spoilage action
Lipase Milk, oil. Hydrolytic rancidity.
Thiaminase Meat, fish. Thiamine destruction.
Peroxidases Fruits. Browning.
Proteases Egg, crab. Reduction of shelf life.
lipoxygenases Vegetables. Destruction of vitamin A.

8. Physical spoilage
❑It occurs due to temp., light, humidity & results in mechanical damage of
the food components.
❑These are some changes in physical appearance caused by microbial cell
activities.
Example: - Oxidation of food occurs due to light & changes colour, flavor &
chemical nature.
- In syrup, microbial cell metabolize sugar molecules & due to
which the concentration of syrup may be also affected.

9. Others Spoilage
• It occurs due to insects, rodents, birds & other animals resulting change in
colour, odor & chemical nature.
• Based on rate of spoilage, they are classified in three types:
[1]. High perishable – Meat, fish , eggs & milk etc.
[2]. Semi –perishable : Potatoes, some apples varieties & nutmeats etc.
[3]. Stable non-perishable : Sugar, flour & dry beans etc.

10. ❑ Factors effecting the microbial
spoilage of pharmaceutical products
• The factors that affected the spoilage of products as following;
Types & size of contaminant inoculum.
Nutritional factors.
Water.
Storage temperature.
pH.
Oxidation[-reduction balance (redox potential).
Package design.

11. Types and Size of contaminant
Inoculum
• Low level of contaminants are unable to replicate in products, not cause
appreciable spoilage but higher contaminant are built in protection could be
insufficient & spoilage occur.
• Higher contamination rise if ;
- Raw material usually contaminated.
- Product misuse during administration.
- Problem of the plant-cleaning protocol occurs.

12. Nutritional Factors
Microorganisms are utilize the nutritional material as energy source & proliferate
over pharmaceutical products.
The are utilize many formulation components as substrate for biosynthesis, growth
also trace material contained in them.
Demineralized also contain nutritional material which support the growth of
“Pseudomonas bacteria”.
Use in the animal products & crude vegetable materials provided nutritious
environments.

13. Water Activity
• Most of micro-organisms are also grow best in ‘dilute solution’.
• Condensed water film sometime accumulate some pharmaceutical formulation
like ‘tablet’ due to storage in damp atmospheres, ‘supports Fungai’.
• Microorganisms need water to grow.The presence of uncomplexed water in
formulation supports microbial growth.
• Greater the ‘solute concentration’, ‘lessees the activity’.
Microorganisms Water activity
Clostridium Botulinum 0.94
EscherichiaColi 0.93
Salmonella Spp. 0.94
Shigella Spp. 0.96

14. Storage temperature
• The formulation can affected by microbial cell b/w the temperature range
-20 0C to -600C.
• Below -20 0C no microbial contamination is observed & the same as above -600C.
• Water for injection is stores at 800 C before preparation to minimize bacterial growth
& to prevent bacterial activity.
• The temperature determines the spoilage y particular type of microorganisms.

15. pH of Pharmaceutical formulations:
• pH is represent as the potential of hydrogen ions. Some bacterial cells are grown
better in acidic & some are in basic medium, this happen because in favorable pH
the enzymes of bacterial cells are more active to perform their metabolic activities.
• Above pH 8 the spoilage is rare for ‘soap-based emulsions’.
• Some preparations of pH around 5-6 favors growth of moulds but inhibit bacterial
growth.
• Some preparations of pH around 3-4 favors growth of ‘moulds & yeast’.
• Some preparation of neutral pH like mouthwashes, distilled water & antacids.
• Basic pH formulation inhibit bacterial growth i.e. Mg & Al hydroxide gel etc.

16. Redox potential
• The ratio of accepting of electrons to the donating the electron power of
substance, it is a property of any chemical to give or accept electrons.
• Microbial growth influenced by redox reaction.
• Electron transfer is major factor for energy production. If the oxygen is present
in any formulation, then this condition favors microbial growth, if any product
gets infected by microbial cell.
• The redox potential is high in ‘emulations’.

17. Packaging Design
• It have major influence on microbial stability of formulations in controlling
the access of contaminating.
• Multi dose containers are more affected by microbial contamination
because they exposed in environment when the drug withdrawn from
container.
• Wide opening mouth containers that vegetable oils, proteins & vitamins
formulations readily contaminated as they provide large surface area
exposure to the environment.

18. ❑ Sources of microbial
contamination
• Air carrying dust: HEPA filter is used to reduction of duct.
• Skin of the operators: Smooth & covered properly.
• Movement of persons inside the aseptic room: It should be restricted.
• The person walking in the room liberate 5000 bacteria/minutes.
• Operators health: Should be healthy & free from any infections.
• Manufacturing process:The main reason of contamination the lack of dedicated facilities to
manufacture a single product.

19. Types of microbial contaminant
Microbial Contamination
Direct
Contamination
Cross
Contamination
Biological
Chemical
Physical
Biological
Chemical
Physical

20. Types of microbial contamination
• [A]. Direct contamination: Microbial components & poorly maintained heating, ventilation
& air conditioning system.
• [B]. Cross contamination: Process by which microbes are spread.
Direct physical contamination:
Particles, fibers & metal part.
Direct chemical contamination:
Moisture, gases.
Direct biological contamination:
Virus, bacteria fungi.
Cross physical contamination-
Leakage of oil seal.
Cross chemical contamination-
Moisture content increase.
Cross biological contamination-
Improper cleaning of equipment.

21. Assessment of microbial contamination &
spoilage
• Assessment of microbial contamination & spoilage is known as “microbial
quality control”.
• This includes laboratory applications like sterility test, bioburden
determination & product testing etc.