This PPT covers microbial spoilage.

1. MICROBIAL SPOILAGE
MICROBIAL
SPOILAGE
PREPARED BY:
Bhargavi Mistry
[Assistant Professor,
School of Pharmacy, Rk University]

2. LIST OF
CONTENT
01
02
03
04
05
06
07
Introduction………………………. 3
Types of microbial spoilage………. 4
Factors affecting microbial
spoilage…………………………….
11
Source & types of microbial
contamination……………………..
16
Assessment of microbial
contamination & spoilage………...
23
References………………………… 25
Questions………………………….. 26
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3. ➢ Microorganisms form an integral part of our environment. They are present in the
air, food, soil, water, etc.
➢ Different types of microorganisms mainly contaminate pharmaceutical
preparations and spoil them.
➢ Such spoilage results in major health problems for the users and financial problems
for the manufacturer due to the loss of product or expensive litigation with
aggrieved uses of the medicine.
➢ Microbial spoilage is the contamination of pharmaceutical products by microbial
activity which leads to spoilage of the product affecting drug safety and drug
quality.
INTRODUCTION
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4. Infection induced by
contaminated
pharmaceutical products
Observable effects of
microbial attack
on products
Physical and chemical
deterioration
of products
Ingredients susceptible
to microbial attack
01
01 02
02 03
03 04
04
TYPES OF MICROBIAL
SPOILAGE
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5. ➢ Pharmaceutical products may be contaminated by pathogenic microorganisms mainly
from raw material or at the time of preparations.
➢ These contaminated pharmaceuticals may cause serious infections the patients when
they use these drugs or formulations.
EXAMPLES:
➢ Cholera in a West African country was traced to an oral liquid drug that had been
prepared with contaminated water.
➢ Several children died in the UK from septicemia caused by Pseudomonas
contamination of parental nutritional fluids during their aseptic preparation.
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INFECTION INDUCED BY CONTAMINATED
PHARMACEUTICAL PRODUCT
01

6. PHYSICALAND CHEMICAL
DETERIORIATION OF PRODUCT
➢ Some naturally occurring ingredients are particularly sensitive to attack.
➢ Crude vegetable and animal drug extracts often contain wide assortments of microbial
nutrients besides the therapeutic agents.
➢ The rate of deterioration of ingredients depend upon its chemical structure, physic-
chemical properties of a particular environment and the level of microbial
contamination present.
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02

7. OBSERVABLE EFFECTS OF MICROBIAL
ATTACK ON PRODUCT
➢ Microbial spoilage of different dosage forms may be detected by organoleptic tests.
➢ These spoiled products may release very unpleasant smelling and testing metabolites.
➢ Formulations may become colored green, pink, brown, black or yellow by diffusible
microbial pigments.
➢ Spoiled creams by microbial attack may become lumpy or gritty.
➢ Thickening and suspending agents produces marked reduction in viscosity by
depolymerization.
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03

8. INGREDIENTS SUSCEPTIBLE TO
MICROBIALATTACK
a) THERAPEUTIC AGENTS:
➢ Potent therapeutic, agents such as analgesics (aspirin, paracetamol), alkaloids
(morphine, atropine),barbiturates, steroid esters etc. can be metabolised and serve as
substance for microbial growth.
➢ Localized transformation of steroids has been observed around fungal colonies growing
on the surface of steroids tablets and in steroidal creams.
b) SURFACE ACTIVE AGENTS:
➢ Alkali-metal and soaps of fatty acids (anionic surfactants) are generally stable due to
the slightly ph of the formulations, which is easily degraded in sewage.
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04

9. c) POLYMERS & HUMECTANTS:
➢ Thickening and suspending agents used in pharmacy are subjected to microbial
depolymerization by extracellular enzymes.
➢ Polymers used in plastic packaging are extremely resistant to microbial attack.
➢ Glycerol and sorbitol (humectants) in pharmaceuticals readily support microbial
growth.
d) FATS & OILS:
➢ Fats and oils (hydrophobic substances)are usually attacked extensively when dispersed
in aqueous formulations such as oil-in-water droplets contaminate the bulk phase during
storage.
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10. Prepared by: Bhargavi Mistry
e) PRESERVATIVES & DISINFECTANTS
➢ Most organic preservatives and disinfectants are metabolized readily by many bacteria
and fungi and may serve as growth substrates at concentrations below their effective
‘use’ levels.
f) SWEETENING, FLAVOURING AGENTS & COLOURING AGENTS
➢ Many sugars and other sweetening agents used in pharmacy are ready substrates for
microbial growth.
➢ Aqueous stock solutions of flavouring agents such as peppermint water and
chloroforms water and colouring agents such as amaranth or tratrazine readily support
growth of bacteria and yeasts.
10

11. Size of
inoculum
Nutritional
factors
Moisture
content
Temperature
pH
Redox
potential
Protective
components
01 02 03 04
05 06 07
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FACTORS AFFECTING MICROBIAL SPOILAGE

12. Low levels of contaminants may be present in a product but it would cause low rates of
deterioration. Ingredients contaminated by a high level of microorganisms cause
appreciable microbial degradation.
Most of the organism and inorganic ingredients act as potential carbon or nitrogen
substrates for microbial growth. The complex of many formulations offers considerable
nutritional variety for a wide range of microorganisms.
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01 SIZE OF INOCULUMS
02 NUTRITIONAL FACTORS

13. ➢ The high concentration of the solute in the formulation indicates that water activity is
low.
➢ Condensed moisture films can form on the surface of tablets or bulk oils as a result of
fluctuations in storage temperatures and the moisture content of the air may be
sufficient to initiate fungal growth.
➢ Syrups and multidose eye drop preparations are dispersed with the label ‘store in a cool
place’.
➢ ‘water for injections’ be stored at 80°C prior to sterilization, of prevent pyrogen
production.
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03 MOISTURE CONTENT
04 TEMPERATURE

14. ➢ Extremes of pH prevent microbial attack, although growth of mold is commonly
observed in solutions of dilute hydrochloric acid.
➢ Antacid mixtures, mouth washes and distilled water show growth of Pseudomonas
species and other Gram- negative bacteria at neutral pH.
➢ The ability of microbes to grow in an environment is influenced by its oxidation -
reduction balance (redox potential), Vacuum packing of foodstuffs or the inclusion of
oxygen absorbers in the package are to minimize oxygen levels and reduce microbial
spoilage.
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05 pH
06 REDOX POTENTIAL

15. ➢ Various components of formulations and products may increase the resistance and
longevity of contaminant microorganisms.
➢ Microbes can show more resistant to heat or dessication in the presence of some
polymers such as starch, acacia or gelatin.
➢ Adsorption of microorganisms onto suspended particles such as kaolin, magnesium
trisilicate of aluminum hydroxide gel may increase their longevity.
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07 PROTECTIVE COMPONENTS

16. Atmosphere Water Raw materials
Equipments Packaging Buildings
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SOURCES & TYPES OF MICROBIAL CONTAMINATION

17. ➢ Microorganisms are carried into the atmosphere suspended on particles of dust, skin,
clothing, droplets of moisture or sputum following coughing or sneezing.
➢ The microbial content of the air may be increased during the handling of contaminated
materials during dispensing, blending and formulations.
➢ Micro- organisms commonly isolated from the atmosphere are bacteria and fungi e.g.
Staphylococcus spp., Streptococcus spp., Clostridium Spp., Bacillus spp., Penicillium
spp., Aspergillus spp., etc.
➢ The microbial count of air or atmosphere may be reduced by chemical disinfection,
ultraviolet light, filtration and gaseous agents.
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ATMOSPHERE

18. ➢ Water is one of the main constituents of many products and it is also used for washing
and cooling process.
➢ Different types of microorganisms are present in fresh water and some microbes may
contaminate the water from sewage e.g. Pseudomonas spp., Proteus spp.
➢ Water may be treated by ultraviolet light, filtration or stored at elevated temperature to
prevent microbial growth.
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WATER

19. ➢ Raw materials, particularly of natural origin are a potentially rich sourceof
microorganisms.
➢ Raw materials mainly contain bacteria and fungi e.g. Bacillus spp., Pseudomonas spp.,
Lactobacillus spp. etc
➢ heat treatment, filtration and other sterilization processes are mainly applied for
reduction of the microbial load in raw materials.
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RAW MATERIALS

20. ➢ The equipment should be as simple as possible for the purposes required with minimum
of junctions, valves and pumps to allow cleaning in place by circulation of detergents or
other chemical antimicrobial agents.
➢ The types and extent of growth will depend on the source of the contamination,
environmental conditions and nutrients.
➢ Equipments may be sterilized or disinfected by heat, gaseous agents or chemicals.
Many tanks, small Equipments and reaction vessels are sterilized by steam under
pressure (autoclave).
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EQUIPMENTS

21. ➢ The microbial flora of packaging materials is dependent upon its composition and
storage conditions.
➢ Packaging material have a dual role and act both to contain the product and to prevent
the entry of microorganisms or moisture.
➢ Glass containers, cardboard boxes and closure liners carry different types of micro-
organisms if not treated by any methods
e.g. Bacillus spp., Penicillium spp., etc.
➢ Dry heat at 170°C is often used for sterilization of vials and ampoules.
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PACKAGING

22. ➢ Different molds and a few bacteria species are the most common flora of walls and
ceilings e.g. Aspergillus niger, Penicillium spp. etc.
➢ They are mainly found in poorly ventilated buildings.
➢ Microorganisms utilize most of their nutrients from the plaster paint.
➢ To reduce microbial growth, all walls, floors and ceilings should be smooth and
washable.
➢ Doors and windows should be fit flush with walls to prevent entry of dust particlesand
microorganisms.
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BUILDINGS

23. ➢ Physical and chemical change of different pharmaceutical formulation and spoilage.
➢ Change in viscosity, pH, emulsion stability and loss of surface activity of formulations
indicate microbial spoilage.
➢ It is important that material which are to be tested for sterility are not subject to
contamination from the operator or the environment during the course of the test.
➢ It is essential that a sterility test a is conducted by competent andean room with laminar
flow cabinet facilities.
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ASSESMENT OF MICROBIAL CONTAMINATION & SPOILAGE
01 PHYSICAL & CHEMICAL CHANGES
02 STERILITY TESTING

24. ➢ Non-sterile products are tested for viable microorganisms for detection of pathogens
and total viable counts.
➢ A pyrogen is a material which causes rise in body temperature when injected into a
patient.
➢ Two main procedures are used for the detection of pyrogens:
1) BP pyrogen test
2) Test LAL test (Limulus Amoedbocyte Lysate test)
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03 ASSESMENT OF VIABLE MICROORGANISMS
IN NON-STERILE PRODUCTS
04 ESTIMATION OF PYROGENS

25. Prepared by: Bhargavi Mistry 25
REFERENCES
1) Dr. Imtiyaz Wani, Pharmaceutical Microbiology, PV books, , First colour edition,
Page no. 203 to 215
2) Dr. Chandrakant Kokare, Pharmaceutical Microbiology – Principle and Applications,
Eighth edition, Page no. 14.1 to 14.18

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QUESTIONS
1) Write a short note on types of microbial spoilage.
2) Explain in brief about different factors affecting microbial spoilage of pharmaceutical
products.
3) Explain different sources and types of microbial contamination of pharmaceutical
products.

27. THANK YOU
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