This document defines spoilage and describes various types of spoilage that can occur with foods and pharmaceuticals. It notes that spoilage can be caused by microbial, non-microbial, or a combination of factors. Microbial spoilage of pharmaceuticals is defined as deterioration caused by microbial contamination that affects drug safety and quality. The types of microbial spoilage are described as physiological-chemical, chemical, or biological, depending on the nature of changes caused. Several factors that influence the microbial spoilage of pharmaceuticals are also discussed, including nutrition, moisture content, redox potential, temperature, pH, and packaging.
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-V Part-1
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical products, source and type of contaminants. Introduction: Defintion Types of Microbial Spoilage:
1. Infection induced due to contaminated pharmaceutical products: Table no. 1.1 Common pathogens spoiling pharmaceutical products:
2. Physicochemical spoilage –
i) Viable growth ii) Gas production
iii) Colouration / Decolouration
iv) Odour formation
v) Taste change
3. Physical Spoilage:
Cracking of emulsion:
Odor changes
4. Biological spoilage:
Microbial Toxins
Microbial Metabolites
5. Chemical spoilage: Table 1.2 Susceptibility of pharmaceutical ingredients to microbial contamination
Factors affecting microbial spoilage
Size of contaminant inoculum
Nutritional factors
Moisture content
pH
Storage temperature
Redox potential
Packaging design
Sources and Types Of Contamination:
Personnel,
Poor facility design,
Incoming ventilation air,
Machinery and other equipment for production,
Raw material and semi-finished material,
Packaging material,
Utilities,
Different media used in the production process as well as for cleaning and Cleanroom clothing.
Evaluation of Bactericidal and Bacteriostatic (Disinfectant). PHARMACEUTICAL ...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-III Part-5 Evaluation of Bactericidal and Bacteriostatic (Disinfectant). The common methods used for evaluation of a disinfectant are as follows,
Tube Dilution Method.
Agar Plate Method.
Filter Paper & Cup Plate Method.
Ditch-Plate Method.
Phenol Coefficient Method.
The official phenol coefficient tests include,
Rideal-Walker Test (RW Test).
Chick-Martin Test.
United States FDA Test for Phenol Coefficient. (FDA Test)
The US Association of Official Agricultural Chemists Test (FDA Test)
A. Rideal-Walker Test:
Kelsey Sykes Method
Evaluation of Bactericidal and BacteriostaticRajsingh467604
What are disinfectants?
As per the definition given by WHO ( World health organization ) : a disinfectant is a chemical agent, which destroys or inhibits growth of pathogenic microorganisms in the non-sporing or vegetative state.
Why Evaluation?
Evaluation of disinfectants is used to check the ability or efficacy of any disinfectant against specific microorganisms to establish its effectiveness.
Evaluation tests of bactericide.
1. RIDEAL WALKER TEST
This test is also known as the phenol coefficient test,in which any chemical is compared with phenol for its antimicrobial activity.
The result is shown in the form of phenol coefficient.
▪ If a phenol coefficient of a given test disinfectant is less than 1, it means that disinfectant is less effective than phenol.
▪ If a phenol coefficient of a given test disinfectant is more than 1, it means that disinfectant is more effective than phenol.
Procedure
1.1 Different dilutions of the test disinfectant and phenol are prepared and 5 ml of each dilution is inoculated with 0.5ml of the 24 hour growth culture of the organisms.
1.2 All tubes(Disinfectant + organisms & phenol + organisms) are placed in a water bath ( at 17.5° C)
1.3 Subcultures of each reaction mixture are taken and transferred to 5ml sterile broth at an interval of 2.5 minutes from zero to 10 mintues.
1.4 Broth tubes are incubated at 37° C for 2 to 3 days & examined for the presence or absence of the growth.
1.5 Then the Rideal Walker coefficient is calculated :
2. CHICK MARTIN TEST.
CHICK MARTIN test is performed in the much similar way as the RIDEAL Walker test but with a little variation.
Principle : This test is carried out in the presence of organic matter like 3% human feces or dried yeast.
Procedure
2.1 Serial dilutions of test solution and phenol is prepared in distilled water.
2.2 To this 3% yeast suspension is also added.
2.3 To this solution the S. typhi is added
2.4 After contact time of 30 mins the above mixture is transferred to the freshly prepared 10 ml of broth.
2.5 The test tubes are incubated at 37°C for 48 hours.
2.6 Presence or absence of the growth is calculated.
Evaluation tests of Bacteriostatic.
1. Tube dilution & Agar plate Method
1.1 The chemical agent is incorporated into nutrient broth or agar medium and inoculated with test micro-organisms.
1.2 These tubes are incubated at 30° TO 35°C for 2 to 3 days and then the results in the form of turbidity or colonies are observed.
1.3 The results are recorded and the activity of the given disinfectant is compared.
2. Cup plate method
2.1 Agar is melted and cooled at 45° Celsius.
2.2 Then inoculated with test micro-organisms and poured into a sterile petri plate.
2.3 In the cup plate method, when the inoculated agar has solidified, holes around 8mm in diameter are cut in the medium with a steel cork borer.
2.4 Now the antimicrobial agents are directly placed in the holes.
Assessment of microbial contamination and spoilage. PHARMACEUTICAL MICROBIOLO...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-2
Assessment of microbial contamination and spoilage.
Assessment of microbial contamination and spoilage
1. Physical and chemical changes:
2. Assessment of viable microorganisms in non-sterile products:
3. Sterility test:
4. Estimation of pyrogens:
Microbial Limit Tests:
Total Aerobic Microbial Count:
Membrane Filtration.
Plate Count Methods.
Pour Plate Method.
Surface spread Method.
Most Probable Number(MPN)
Microbiological assay-Principles and methods of different microbiological assay.someshwar mankar
Principles and methods of different microbiological assay. Methods for standardization of
antibiotics, vitamins and amino acids. Assessment of a new antibiotic.
Preservation of pharmaceutical products using antimicrobial agents. PHARMACEU...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-3
Preservation of pharmaceutical products using antimicrobial agents.
Introduction. Ideal Properties of Preservatives:
Antimicrobial Chemical Preservatives
Development of a Preservative System.
Factors affecting efficacy of a preservative: 1. Interaction With components of the formulation
2. Properties of the Preservatives:
3) Effect of Containers.
4) Type of microbes:
5) Influence of pH:
Challenge Test: Efficacy Test of Preservative : Medium used, Choice of test organism:
Preparation of the inoculum:
Procedure:
Interpretation of Results:
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-V Part-1
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical products, source and type of contaminants. Introduction: Defintion Types of Microbial Spoilage:
1. Infection induced due to contaminated pharmaceutical products: Table no. 1.1 Common pathogens spoiling pharmaceutical products:
2. Physicochemical spoilage –
i) Viable growth ii) Gas production
iii) Colouration / Decolouration
iv) Odour formation
v) Taste change
3. Physical Spoilage:
Cracking of emulsion:
Odor changes
4. Biological spoilage:
Microbial Toxins
Microbial Metabolites
5. Chemical spoilage: Table 1.2 Susceptibility of pharmaceutical ingredients to microbial contamination
Factors affecting microbial spoilage
Size of contaminant inoculum
Nutritional factors
Moisture content
pH
Storage temperature
Redox potential
Packaging design
Sources and Types Of Contamination:
Personnel,
Poor facility design,
Incoming ventilation air,
Machinery and other equipment for production,
Raw material and semi-finished material,
Packaging material,
Utilities,
Different media used in the production process as well as for cleaning and Cleanroom clothing.
Evaluation of Bactericidal and Bacteriostatic (Disinfectant). PHARMACEUTICAL ...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-III Part-5 Evaluation of Bactericidal and Bacteriostatic (Disinfectant). The common methods used for evaluation of a disinfectant are as follows,
Tube Dilution Method.
Agar Plate Method.
Filter Paper & Cup Plate Method.
Ditch-Plate Method.
Phenol Coefficient Method.
The official phenol coefficient tests include,
Rideal-Walker Test (RW Test).
Chick-Martin Test.
United States FDA Test for Phenol Coefficient. (FDA Test)
The US Association of Official Agricultural Chemists Test (FDA Test)
A. Rideal-Walker Test:
Kelsey Sykes Method
Evaluation of Bactericidal and BacteriostaticRajsingh467604
What are disinfectants?
As per the definition given by WHO ( World health organization ) : a disinfectant is a chemical agent, which destroys or inhibits growth of pathogenic microorganisms in the non-sporing or vegetative state.
Why Evaluation?
Evaluation of disinfectants is used to check the ability or efficacy of any disinfectant against specific microorganisms to establish its effectiveness.
Evaluation tests of bactericide.
1. RIDEAL WALKER TEST
This test is also known as the phenol coefficient test,in which any chemical is compared with phenol for its antimicrobial activity.
The result is shown in the form of phenol coefficient.
▪ If a phenol coefficient of a given test disinfectant is less than 1, it means that disinfectant is less effective than phenol.
▪ If a phenol coefficient of a given test disinfectant is more than 1, it means that disinfectant is more effective than phenol.
Procedure
1.1 Different dilutions of the test disinfectant and phenol are prepared and 5 ml of each dilution is inoculated with 0.5ml of the 24 hour growth culture of the organisms.
1.2 All tubes(Disinfectant + organisms & phenol + organisms) are placed in a water bath ( at 17.5° C)
1.3 Subcultures of each reaction mixture are taken and transferred to 5ml sterile broth at an interval of 2.5 minutes from zero to 10 mintues.
1.4 Broth tubes are incubated at 37° C for 2 to 3 days & examined for the presence or absence of the growth.
1.5 Then the Rideal Walker coefficient is calculated :
2. CHICK MARTIN TEST.
CHICK MARTIN test is performed in the much similar way as the RIDEAL Walker test but with a little variation.
Principle : This test is carried out in the presence of organic matter like 3% human feces or dried yeast.
Procedure
2.1 Serial dilutions of test solution and phenol is prepared in distilled water.
2.2 To this 3% yeast suspension is also added.
2.3 To this solution the S. typhi is added
2.4 After contact time of 30 mins the above mixture is transferred to the freshly prepared 10 ml of broth.
2.5 The test tubes are incubated at 37°C for 48 hours.
2.6 Presence or absence of the growth is calculated.
Evaluation tests of Bacteriostatic.
1. Tube dilution & Agar plate Method
1.1 The chemical agent is incorporated into nutrient broth or agar medium and inoculated with test micro-organisms.
1.2 These tubes are incubated at 30° TO 35°C for 2 to 3 days and then the results in the form of turbidity or colonies are observed.
1.3 The results are recorded and the activity of the given disinfectant is compared.
2. Cup plate method
2.1 Agar is melted and cooled at 45° Celsius.
2.2 Then inoculated with test micro-organisms and poured into a sterile petri plate.
2.3 In the cup plate method, when the inoculated agar has solidified, holes around 8mm in diameter are cut in the medium with a steel cork borer.
2.4 Now the antimicrobial agents are directly placed in the holes.
Assessment of microbial contamination and spoilage. PHARMACEUTICAL MICROBIOLO...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-2
Assessment of microbial contamination and spoilage.
Assessment of microbial contamination and spoilage
1. Physical and chemical changes:
2. Assessment of viable microorganisms in non-sterile products:
3. Sterility test:
4. Estimation of pyrogens:
Microbial Limit Tests:
Total Aerobic Microbial Count:
Membrane Filtration.
Plate Count Methods.
Pour Plate Method.
Surface spread Method.
Most Probable Number(MPN)
Microbiological assay-Principles and methods of different microbiological assay.someshwar mankar
Principles and methods of different microbiological assay. Methods for standardization of
antibiotics, vitamins and amino acids. Assessment of a new antibiotic.
Preservation of pharmaceutical products using antimicrobial agents. PHARMACEU...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-VPart-3
Preservation of pharmaceutical products using antimicrobial agents.
Introduction. Ideal Properties of Preservatives:
Antimicrobial Chemical Preservatives
Development of a Preservative System.
Factors affecting efficacy of a preservative: 1. Interaction With components of the formulation
2. Properties of the Preservatives:
3) Effect of Containers.
4) Type of microbes:
5) Influence of pH:
Challenge Test: Efficacy Test of Preservative : Medium used, Choice of test organism:
Preparation of the inoculum:
Procedure:
Interpretation of Results:
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T) Unit-IV Part-3
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic: Introduction:
Principle
Microbiological Assay of Cynocobalamin (Vitamin B12):
Tritrimetric Method.
Turbidimetric Method.
Preparation of Standard Cynocobalmine stock solution:
Preparation of Basal Medium Stock Solution:
Test Solution of the material to be assayed Preparation of inoculum: Procedure of Titrimetric method: Turbidimetric Method: Microbiological assay of Amino acids. Assessment of a New Antibiotic.
Introduction:
MIC of an antibiotic is tested either by one of the following ways,
Liquid Dilution Method.
Solid Dilution Method
Factors affecting action of Disinfectants and Factors Affecting Choice Of Ant...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-III. Factors affecting action of Disinfectants and Factors Affecting Choice Of Antimicrobial Agent: Concentration of the disinfectant.
Chemical Structure of the disinfectant.
Formulation of the disinfectant.
Interfering substances in the environment.
pH of the surrounding.
Potentiation and antagonism of the disinfectants.
Surface Tension.
Temperature.
Time of Contact.
Type and no. of microbes present.
FACTORS AFFECTING CHOICE OF ANTIMICROBIAL AGENT:
Properties of chemical agents
Environment
Types of microorganisms
Intended application
Toxicity agents
Culture state
Sterility testing products (solids, liquids, ophthalmic and other sterile pro...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-6 Sterility testing products (solids, liquids, ophthalmic and other sterile products) according to IP, BP, USP.
Introduction: Test for Sterility. Culture Media. Fluid Thioglycollate Medium (FTM).
Alternative Thioglycollate Medium (ATM).
Soybean Casein Digest Medium (SCDM).
Tests for Culture Media:
Sterility of Media.
Growth Promotion Test.
Test for Bacteriostatic and Fungistatic.
Sterility Test Methods. Methods A: Membrane Filtration.
Method B: Direct Inoculation Pyrogen Test Methods. Rabbit Test. LAL Test.
Microbial spoilage-by S.D.Mankar types, sources of contamination, factors,Ass...someshwar mankar
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical products,
sources and types of microbial contaminants, assessment of microbial contamination and
spoilage.
Prof.Mr.Kiran K. Shinde (M.Pharm), Assistant professor (VNIPRC)
Pharmaceutical microbiology (Second year b.pharm) (3rd semester)
Introduction
Types of Spoilage
Factors affecting the Microbial spoilage of pharmaceutical products
Sources and Types of Contamination
Assessment of microbial contamination and spoilage
INCLUDES SPREADING COEFFICIENT AND ITS THEORY AND ALSO FEW OF ITS APPLICATION IN PHARMACEUTICAL FIELD
WILL BE HELPFUL FOR B PHARMACY STUDENTS
INCLUDES HOW IT IS DERIVED AND ALSO HOW IT IS RELATED TO SPREADING OF A CREAM OR OINTMENT ON OUR SKIN
IMPORTANCE OF SPREADING COEFFICIENT
VIRUS PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-2Study of morphology, ...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-2Study of morphology, classification, reproduction/replication and cultivation of Virus. Introduction, Def General characteristics of Viruses: small size characteristic shapes, obligate intracellular parasites no built-in metabolic machinery no ribosomes
only one type of nucleic acid
do not grow in size. Morphology of Virus: Helical, Polyhedral (Icosahedral) Viral Envelop, Complex virus, Classification of virus. Viral Replication LIFE CYCLE OF BACTIRIOPHAGES Lytic cycle: Attachment, Penetration, Biosynthesis, Maturation and Release of progeny Phage Particles. The Lysogenic Cycle, Cultivation of virus : Animal inoculation, Embryonated eggs or chick embryo method and Tissue culture or cell culture: Organ cultures Explant culture and Cell culture. Types of cell culture
1.Primary cell culture: 2. Diploid cell culture (Semi-continuous cell lines):3. Heteroploid cultures (Continuous cell lines):
MULTIPLICATION OF HUMAN VIRUS:1. Attachment of Viral Particles 2. Penetration 3. Uncoating 4. Replication Of Viral Nucleic Acids And Translation Of The Genome 5) Maturation Or Assembly Of Virions. ) 6. Release Of Virions Into The Surrounding Environment
Principles and methods of different microbiological assay, methods for standa...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IV Part-2 Principles and methods of different microbiological assay, methods for standardization of antibiotics.
Introduction: Principles Advantages of Microbial Assay: Disadvantages of Microbial Assay: MICROBIOLOGICAL ASSAY OF ANIBIOTICS PRINCIPLE Media used for antibiotics assay Standard Preparation. Buffer Solutions Preparation of the Sample Solution: Test Organisms Preparation of inoculum: Methods of preparation of test organism suspension: Assay Methods: Method A: Cup-plate or Cylinder Plate Method.
Method B: Turbidimetric or Tube assay Method
Evaluation of the efficiency of sterilization methods.Sterility indicatorsMs. Pooja Bhandare
Evaluation of the efficiency of sterilization methods.Sterility indicators
Sterility criteria: Bioburden ,Sensitivity of microorganisms
Death rate or Survivor curve,D- Value or Decimal reduction time,Z- value or Thermal reduction time, f- value, Q10 Value or Temperature Coefficient, Inactivation Factor:
STERILITY INDICATORS : Physical Indicators, Chemical Indicators
Biological Indicators
1. Physical Indicators: i) Moist heat Indicator ii) Dry heat iii) Radio sterilization iv) Gaseous methods v) Filtration 2.CHEMICAL INDICATORS : I) Browne’s tubes II) WITTNESS TUBES IV) Royce Sachet V) Chemical Dosimeter 3.BIOLOGICAL INDICATORS
MICROBIAL SPOILAGE
TYPES OF SPOILAGE
PHARMACEUTICAL SPOILAGE
MICROBIAL SPOILAGE OF PHARMACEUTICALS
TYPES OF MICROBIAL SPOILAGE
REASON OF CONTAMINATION
FACTORS AFFECTING SPOILAGE OF PHARMACEUTICAL PRODUCTS
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T) Unit-IV Part-3
Microbiological Assay of Vitamin & Amino acid Assessment of a New Antibiotic: Introduction:
Principle
Microbiological Assay of Cynocobalamin (Vitamin B12):
Tritrimetric Method.
Turbidimetric Method.
Preparation of Standard Cynocobalmine stock solution:
Preparation of Basal Medium Stock Solution:
Test Solution of the material to be assayed Preparation of inoculum: Procedure of Titrimetric method: Turbidimetric Method: Microbiological assay of Amino acids. Assessment of a New Antibiotic.
Introduction:
MIC of an antibiotic is tested either by one of the following ways,
Liquid Dilution Method.
Solid Dilution Method
Factors affecting action of Disinfectants and Factors Affecting Choice Of Ant...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-III. Factors affecting action of Disinfectants and Factors Affecting Choice Of Antimicrobial Agent: Concentration of the disinfectant.
Chemical Structure of the disinfectant.
Formulation of the disinfectant.
Interfering substances in the environment.
pH of the surrounding.
Potentiation and antagonism of the disinfectants.
Surface Tension.
Temperature.
Time of Contact.
Type and no. of microbes present.
FACTORS AFFECTING CHOICE OF ANTIMICROBIAL AGENT:
Properties of chemical agents
Environment
Types of microorganisms
Intended application
Toxicity agents
Culture state
Sterility testing products (solids, liquids, ophthalmic and other sterile pro...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-6 Sterility testing products (solids, liquids, ophthalmic and other sterile products) according to IP, BP, USP.
Introduction: Test for Sterility. Culture Media. Fluid Thioglycollate Medium (FTM).
Alternative Thioglycollate Medium (ATM).
Soybean Casein Digest Medium (SCDM).
Tests for Culture Media:
Sterility of Media.
Growth Promotion Test.
Test for Bacteriostatic and Fungistatic.
Sterility Test Methods. Methods A: Membrane Filtration.
Method B: Direct Inoculation Pyrogen Test Methods. Rabbit Test. LAL Test.
Microbial spoilage-by S.D.Mankar types, sources of contamination, factors,Ass...someshwar mankar
Types of spoilage, factors affecting the microbial spoilage of pharmaceutical products,
sources and types of microbial contaminants, assessment of microbial contamination and
spoilage.
Prof.Mr.Kiran K. Shinde (M.Pharm), Assistant professor (VNIPRC)
Pharmaceutical microbiology (Second year b.pharm) (3rd semester)
Introduction
Types of Spoilage
Factors affecting the Microbial spoilage of pharmaceutical products
Sources and Types of Contamination
Assessment of microbial contamination and spoilage
INCLUDES SPREADING COEFFICIENT AND ITS THEORY AND ALSO FEW OF ITS APPLICATION IN PHARMACEUTICAL FIELD
WILL BE HELPFUL FOR B PHARMACY STUDENTS
INCLUDES HOW IT IS DERIVED AND ALSO HOW IT IS RELATED TO SPREADING OF A CREAM OR OINTMENT ON OUR SKIN
IMPORTANCE OF SPREADING COEFFICIENT
VIRUS PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-2Study of morphology, ...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IIIPart-2Study of morphology, classification, reproduction/replication and cultivation of Virus. Introduction, Def General characteristics of Viruses: small size characteristic shapes, obligate intracellular parasites no built-in metabolic machinery no ribosomes
only one type of nucleic acid
do not grow in size. Morphology of Virus: Helical, Polyhedral (Icosahedral) Viral Envelop, Complex virus, Classification of virus. Viral Replication LIFE CYCLE OF BACTIRIOPHAGES Lytic cycle: Attachment, Penetration, Biosynthesis, Maturation and Release of progeny Phage Particles. The Lysogenic Cycle, Cultivation of virus : Animal inoculation, Embryonated eggs or chick embryo method and Tissue culture or cell culture: Organ cultures Explant culture and Cell culture. Types of cell culture
1.Primary cell culture: 2. Diploid cell culture (Semi-continuous cell lines):3. Heteroploid cultures (Continuous cell lines):
MULTIPLICATION OF HUMAN VIRUS:1. Attachment of Viral Particles 2. Penetration 3. Uncoating 4. Replication Of Viral Nucleic Acids And Translation Of The Genome 5) Maturation Or Assembly Of Virions. ) 6. Release Of Virions Into The Surrounding Environment
Principles and methods of different microbiological assay, methods for standa...Ms. Pooja Bhandare
PHARMACEUTICAL MICROBIOLOGY (BP303T)Unit-IV Part-2 Principles and methods of different microbiological assay, methods for standardization of antibiotics.
Introduction: Principles Advantages of Microbial Assay: Disadvantages of Microbial Assay: MICROBIOLOGICAL ASSAY OF ANIBIOTICS PRINCIPLE Media used for antibiotics assay Standard Preparation. Buffer Solutions Preparation of the Sample Solution: Test Organisms Preparation of inoculum: Methods of preparation of test organism suspension: Assay Methods: Method A: Cup-plate or Cylinder Plate Method.
Method B: Turbidimetric or Tube assay Method
Evaluation of the efficiency of sterilization methods.Sterility indicatorsMs. Pooja Bhandare
Evaluation of the efficiency of sterilization methods.Sterility indicators
Sterility criteria: Bioburden ,Sensitivity of microorganisms
Death rate or Survivor curve,D- Value or Decimal reduction time,Z- value or Thermal reduction time, f- value, Q10 Value or Temperature Coefficient, Inactivation Factor:
STERILITY INDICATORS : Physical Indicators, Chemical Indicators
Biological Indicators
1. Physical Indicators: i) Moist heat Indicator ii) Dry heat iii) Radio sterilization iv) Gaseous methods v) Filtration 2.CHEMICAL INDICATORS : I) Browne’s tubes II) WITTNESS TUBES IV) Royce Sachet V) Chemical Dosimeter 3.BIOLOGICAL INDICATORS
MICROBIAL SPOILAGE
TYPES OF SPOILAGE
PHARMACEUTICAL SPOILAGE
MICROBIAL SPOILAGE OF PHARMACEUTICALS
TYPES OF MICROBIAL SPOILAGE
REASON OF CONTAMINATION
FACTORS AFFECTING SPOILAGE OF PHARMACEUTICAL PRODUCTS
Pharmaceutical Microbiology
What is spoilage?
Types of spoilage
Pharmaceutical spoilage
Microbial spoilage of pharmaceuticals
Types of microbial spoilage
Reason of contamination
Factors affecting spoilage of pharmaceutical products
presented by HAFIZ M WASEEM
university of education LAHORE Pakistan
i am from mailsi vehari and studied in lahore
bsc in science college multan
msc from lahore
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
2. DEFINITION
• The action or process of spoiling, especially the deterioration of
food and other perishable goods.
• Any change which renders a product unacceptable for human
consumption.
• Complex event in which a combination of microbial and
biochemical activities may interact.
• One of the major reason that led to preservation.
• MICROBIAL SPOILAGE: This refers to damage to food,
pharmaceutical products that is caused by micro organisms
(bacteria, moulds and yeasts). Micro organisms can grow in
almost all kinds of food products. As micro-organisms occur
everywhere around us, there is always a risk of microbial
spoilage.
3. TYPES OF SPOILAGE
• MICROBIAL SPOILAGE
• NON-MICROBIAL SPOILAGE
• Based on rate of spoilage: There are three types-
Highly perishable: Meat, Fish, Egg, Milk, Most fruits and Vegetables.
Semi perishable: Potatoes, Some Apple Varities, Nut meats.
Non-perishable or stable: Sugar, Flour, Dry Beans.
• PHARMACEUTICAL SPOILAGE
4. PHARMACEUTICAL SPOILAGE
• Spoilage of pharmaceutical products and drugs are
referred as the changes in the physical and chemical
properties in such a way that the formulation or
therapeutic agent gets deteriorate and become not suitable
for use.
• Pharmaceutical product is a combination of two things:
1.Active drug 2. Formulation additives
If there is any change in their efficacy, action, etc. it is said
to be spoiled product which can not be used.
5. PHARMACEUTICAL SPOILAGE
• There are several factors that cause pharmaceutical
spoilage: Mainly, we study about microbial spoilage
of pharmaceutical products.
PHYSICAL CHEMICAL MICROBIAL
Deterioration of
pharmaceuticals
due to physical
factors like, heat,
temperature,
evaporation, etc.
Deterioration of
pharmaceuticals
due to chemical
reactions like
oxidation,
reduction,
hydrolysis,
photolysis,
ionization, etc.
Deterioration of
pharmaceuticals
due to
contamination of
any microbial cell
like bacteria,
fungi, moulds, etc.
6. MICROBIAL SPOILAGE OF PHARMACEUTICALS
• Microbial spoilage includes the microbial contamination
of pharmaceutical products with the microbes which lead
to spoilage of product affecting the drug safety and
quality and is not intended for use.
• Shortly, microbial spoilage of pharmaceutical products is
the deterioration of pharmaceuticals with contaminant
microbes.
• It decreases the stability and intensity of the product.
7. TYPES OF MICROBIAL SPOILAGE
PHYSIO-CHEMICAL CHEMICAL BIOLOGICAL
VIABLEGROWTH
GAS PRODUCTION
PHYSICAL SPOILAGE
OLFACTORY
COLORATION
HYDROLYSIS
ACETYLATION
DEPOLYMERIZATION
DEGRADATION/
METABOLISATION
RELEASEOFTOXINS
MICROBIAL
METABOLITES
8. PHYSIO-CHEMICAL
In this kind of spoilage, there are some chemical changes
are caused by microbial species and due to these changes
the physical properties are also gets altered or deteriorate,
thus it is called the physico-chemical spoilage.
1.VIABLE GROWTH: In this kind of spoilage, microbial
cells form a visible layer over the surface of pharmaceutical
formulations. This layer or the presence of microbial cell can
be clearly seen by naked eyes.
Example: layer of moulds over syrups or sugar containing
products, creams, ointments.
9. 2. COLORATION: Color change occurs due to the
alteration in components of chemical nature, and these
changes are caused by change in pH of the formulation, redox
of the product, production of some other metabolites by
microorganisms.
Example: Pseudomonas species microorganism metabolize
wide range of metabolites that cause coloration of blue-green,
brown.
Surface decoloration of tablets containing biological products
by some moulds.
10. 3. GAS PRODUCTION: Some microorganisms contaminants
in pharmaceutical formulations produces gases by their
metabolic activities and form gas bubbles and broth/foam over
the formulation.
Formulations containing carbohydrates or starchy material are
more susceptible for gas production.
Example: production of CO2 in syrups caused by osmo-
toleranmt mould and yeasts.
Klebsiella produces gas in creams and ointments containing
vitamins and proteins.
Desulphovibrio oxidise simple organic compounds and
produces hydrogen sulphide in suspensions.
11. 4. PHYSICAL SPOILAGE: These are some changes in
physical appearance caused by microbial cell activities.
Example: In emulsions, some microbial cells cause hydrolysis
of oil phase, cause change in oil-water equillibrium and make
emulsion unstable.Hydrolysis of oils also affect pH of
formulation and alter the stability of emulsions.
In syrups, microbial cells metabolise sugar molecules, and due
to which the concentration of syrup change and product
become unstable.
12. 5. Olfactory spoilage: Spoilage by some organisms and
moulds generates unpleasant smell from the product. This
kind of spoilage mainly caused by microbial cella that
produces sulphur containing gases[SO2, H2S] and fishy
smell due to formation of fatty acids along with odour
generates by amines and alcohol production.
Example: Contamination in syrup of Tolu by penicilin spp.
Produces toluene like smell.
Actinomycetes produces smell of geosmine in water phase
used for formulations.
13. CHEMICAL SPOILAGE
There are various types of chemical spoilage in
pharmaceutical compound, these occurs due to various
types of chemical reactions, mediated by contaminating
microorganisms.
1.HYDROLYSIS: Some bacterial cells contains enzymes
that catalyse hydrolysis of pharmaceuticals.
Example: Atropine hydrolysed by Pseudomonas bacteria.
Aspirin hydrolysed by esterase producing bacteria.
Gelatin hydrolysed by Bacillus and Clostridium species.
2.ACETYLATION: Some microorganisms cellular enzymes
cause acetylation of drugs and cause loss of activity.
14. Example: Chloramphenicol acetylation caused by
staphylococci and streptococci gram +ve bacteria by the
enzyme chloramphenicol acetyl-transferase.
3.DEPOLYMERISATION: It is a process in which the
polymers are degraded to their monomers. A lots of polymers
are used in formulation of many type of pharmaceutical
preparations as diluents, binders, thickening, suspending
agents, etc.
Example: Starch- depolymerize by bacterial amylase
Pectin- depolymerize by bacterial pectinase
Dextran- depolymerize by bacterial dextranase
Cellulase- depolymerize by bacterial cellulase.
15. 4.DEGRADATION: Due to the microbial contamination, the
active therapeutic agents or the formulation ingredients can be
degraded or metabolized.
Examples: Penicilin: degrade by batalactamase containing
bacterial cells
Fatty acids metabolized by some moulds
Prednison degraded by aspergillus species bacteria.
16. BIOLOGICAL SPOILAGE:
Some bacterial cells contaminated the pharmaceuticals and
utilize the various compounds present in that formulation to
perform their metabolic activities. Due to these metabolic
activities, the microbial cells produces certain chemicals
which they release in pharmaceutical preparations. This is
called biological spoilage.
Mainly two types of chemicals released by the
microorganisms:
1.Microbial toxins
2.Microbial metabolites
17. MICROBIAL TOXINS: Several microorganisms produce
toxic molecules that may cause spoilage of pharmaceutical
fomulations. Such as endotoxins produced by some gram –ve
bacterias like E.coli.
Exotoxins by clostridium botulinum.
MICROBIAL METABOLITES: Bacterial metabolites are
the biosynthetic products from microbial cells. Bacterial cells
produces various metabolites which cause product spoilage
bcoz these metabolites are toxic to humans.
Examples are different amines and organic acids from
bacterial cells. Metabolites from fungi and moulds.
Fungi and moulds are more specifically grow on the
formulations having Talc, kaolin, and starch.
18. REASON OF CONTAMINATION
• The product gets contaminate due to various reasons like
Accidentally exposure to the environment
Improper storage conditions
Inadequate preparation of formulation
Improper sterilization
19. FACTORS AFFECTING SPOILAGE
OF PHARMACEUTICAL
PRODUCTS• By understanding the influence of environmental parameters on
microorganisms, it may be possible to manipulate formulations to create
conditions which are as unfavorable as possible for growth and spoilage,
within the limitations of patient acceptability and therapeutic efficacy.
• The factors that affects the spoilage of spoilage:
NUTRITIONAL FACTOR
MOISTURE CONTENT
REDOX POTENTIAL
STORAGE TEMPERATURE
pH
PACKAGING DESIGN AND DESIGN
20. •NUTRITIONAL FACTOR:
Presence of nutritional material enable or favours microorganisms
to utilize these nutritional material as energy source and proliferate
over pharmaceutical products.
In any formulation the presence of vegetable/herbal extract or
animal tissue or tissue extract provides nutritional support to
microbial cells.
Demineralized water[prepared by ion exchange method] also
contain some nutritional material which support the growth of
Pseudomonas bacteria.
More complex formulation are more supportive for microbial
growth.
21. •MOISTURE CONTENT[Water Activity]
Microorganisms readily need water or moisture to grow. The
presence of uncomplexed water in any formulation supports
microbial growth. Greater the solute concentration, lesser the
activity.
When solutes dissolved in water they form hydrogen and other
bonds with water and form complexed water, the free or
unbounded water molecules are termed as uncomplexed water,
and this uncomplexed water supports microbial growth.
Condensed water film sometime accumulate over some dry
pharmaceutical formulation like tablets or oils due to the
storage in humid atmosphere, supports fungal i.e. A.glaucus
and yeast growth.
22. CONT…
Moisture films over viscous syrups can also increase water
activity and support growth of fungi and yeast.
Some halophilic bacterial grows in high salt conditions.
Minimal water activity for some microbial cells for growth.
MICROORGANISM WATER ACTIVITY
CLOSTRIDIUM BOTULINIUM 0.94
ESCHERICHIACOLI 0.93
SALMONELLA SPP. 0.94
SHIGELLA SPP. 0.96
23. REDOX POTENTIAL:
• The oxidation-reduction or the Redox potential is defined as
the ratio of the total oxidizing [electron accepting] power to
the total reduction [electron donating power of substance], or
in more easy way it’s a property of any chemical to give or
accept electrons.
• Microbial growth in any environment is influenced by its
oxidation reduction balance i.e. redox reaction. Electron
transfer is major factor for energy production. If the oxygen
is present in any formulation, then this condition favours
microbial growth, if any product gets infected by microbial
cell.
24. PACKAGING MATERIAL AND DESIGN:
• Packaging can have major influence on microbial
contamination and spoilage of pharmaceuticals. Multi dose
containers are more affected by microbial contamination
because they are again and again exposed to environment
when the drug is withdrawn from container.
• Wide opening mouth containers that contain formulations
of vegetable oils, protein, vitamin, animal extracts, are
readily contaminated as they provide large surface area
exposure to the environment.
25. Cont…
• STORAGE TEMPERATURE:
• The pharmaceutical formulation can affected by microbial cell
between the temperature range of -20℃ − 60℃.
• Below -20℃ almost no microbial contamination is observed and
the same as above 60℃ , microbial growth supressed. The reason
for that is inactivation and denaturation of cellular enzymes,
which responsible for cell metabolic activities.
• Water for injection is stored at 80℃ before preparation to
minimize bacterial growth and to prevent bacterial activity to
produce and release pyrogens to the water.
26. pH of Pharmaceutical formulations:
• pH is represent as the potential of hydrogen ions. Some
bacterial cells are grown better in Acidic medium and
some are in basic medium, this happens because in their
favourable pH the enzymes of bacterial cells are more
active to perform their metabolic activities.
• Some preparations of pH around 5-6 favours growth of
moulds but inhibit bacterial growth.
• Some preparations having pH of 3-4 favours growth of
moulds and yeast, i.e. fruit juice flavoured syrups.
• Some preparations of neutral pH like mouthwashes,
distilled water, antacid preparations are contaminated by
bacterial cells i.e. pseudomonas spp.
27. • Basic pH formulations inhibit bacterial growth,
i.e. magnesium and aluminium hydroxide gel,
etc.
• Sometime primary fungal growth occurs in any
product, these fungal cell metabolize the
chemical andproduce acids and raise the pH of
formulations and favours secondary bacterial
growth.