This presentation gives an overview of : Validation of microbiological methods , Considering some of the limitations and
Key criteria that may be applicable for assessment.
It is process of “Establishing documentary evidence that provide a high degree of assurance that a specific process will consistently produce a product meeting its predetermined specifications and quality attributes”.
In the pharmaceutical industry, it is very important that in addition to final testing and compliance of products, it is also assured that the process will consistently produce the expected results.
Validation is action of proving in accordance with the principles of good manufacturing practices, that any procedure, process, equipment, material, activity or system actually leads to expected results.
Cleaning validation is documented evidence with a high degree assurance that one can consistently clean a system or a piece of equipment to predetermined and acceptable limits.
The primary regulatory concern driving the need for cleaning validation is cross contamination of the desired drug substance either by other API from previous batch runs or by residues from the cleaning agents used.
The prime purpose of validating a cleaning process is to ensure compliance with federal and other standard regulations
1. Cross contamination with active ingredients
Contamination of one batch of product with significant levels of residual active ingredients from previous batch cannot be tolerated.
In addition to the obvious problems posed by subjecting consumers or patients to unintended contaminants, potential clinically significant synergistic interactions between pharmacologically active chemicals are a real concern.
2. Contamination with unintended materials or compounds
While inert ingredients used in drug products are generally recognized as safe for human consumption, the routine use, maintenance and cleaning of equipment's provide the potential contamination with such items as equipment parts, lubricants and chemical cleaning agents3. Microbiological contamination
Maintenance , cleaning and storage conditions may provide adventitious microorganisms with the opportunity to proliferate within the processing equipment.
It is process of “Establishing documentary evidence that provide a high degree of assurance that a specific process will consistently produce a product meeting its predetermined specifications and quality attributes”.
In the pharmaceutical industry, it is very important that in addition to final testing and compliance of products, it is also assured that the process will consistently produce the expected results.
Validation is action of proving in accordance with the principles of good manufacturing practices, that any procedure, process, equipment, material, activity or system actually leads to expected results.
Cleaning validation is documented evidence with a high degree assurance that one can consistently clean a system or a piece of equipment to predetermined and acceptable limits.
The primary regulatory concern driving the need for cleaning validation is cross contamination of the desired drug substance either by other API from previous batch runs or by residues from the cleaning agents used.
The prime purpose of validating a cleaning process is to ensure compliance with federal and other standard regulations
1. Cross contamination with active ingredients
Contamination of one batch of product with significant levels of residual active ingredients from previous batch cannot be tolerated.
In addition to the obvious problems posed by subjecting consumers or patients to unintended contaminants, potential clinically significant synergistic interactions between pharmacologically active chemicals are a real concern.
2. Contamination with unintended materials or compounds
While inert ingredients used in drug products are generally recognized as safe for human consumption, the routine use, maintenance and cleaning of equipment's provide the potential contamination with such items as equipment parts, lubricants and chemical cleaning agents3. Microbiological contamination
Maintenance , cleaning and storage conditions may provide adventitious microorganisms with the opportunity to proliferate within the processing equipment.
Control on Cleanroom Environmental Monitoring (Pharmaceutical)Srinath Sasidharan
A general consideration of Environmental Monitoring in Pharmaceutical manufacturing area. Cleanroom Monitoring Tools and Utilities: Author Sreenath Sasidharan (Geltec Healthcare FZE)
Considering: Environmental monitoring guidance, Background to USP <1116>, Main changes and debates Method limitations, Incident rates, Frequencies of monitoring, Locations of monitoring, Other changes, Regulatory issues and Rapid methods
This presentation includes detail about cleaning levels,equipments for cleaning validation , steps for cleaning method validation and analytical method validation used for cleaning.
Sterility Testing is defined as a testing which confirms that products are free from the presence of viable microorganisms. Sterility testing is very important for medical devices, pharmaceuticals, preparations, tissue materials and other materials that claim to be sterile or free from viable microorganisms.
Contamination Control in Cleanrooms_Dr.A. AmsavelDr. Amsavel A
Basic’s of Contamination
Sources of Contamination
Environment Specification
Elements of Cleanroom Design and Qualification
Definitions
Control of Contaminations
People, Cleaning, Environment & Material
Operation, Monitoring and Control
Documents and Records
The two most commonly used within microbiology are
HACCP (which originated in the food industry) and FMEA
(developed for engineering). This article explores these two
approaches, first with a description of HACCP, followed by a
description and case study of FMEA in sterility testing.
The Changing Landscape of Clinical Research Regulations: Updates and Implicat...ClinosolIndia
The landscape of clinical research regulations is constantly evolving to adapt to the changing needs of the research community, advancements in scientific understanding, and the protection of research participants. Here are some key updates and implications that have shaped the current state of clinical research regulations
Control on Cleanroom Environmental Monitoring (Pharmaceutical)Srinath Sasidharan
A general consideration of Environmental Monitoring in Pharmaceutical manufacturing area. Cleanroom Monitoring Tools and Utilities: Author Sreenath Sasidharan (Geltec Healthcare FZE)
Considering: Environmental monitoring guidance, Background to USP <1116>, Main changes and debates Method limitations, Incident rates, Frequencies of monitoring, Locations of monitoring, Other changes, Regulatory issues and Rapid methods
This presentation includes detail about cleaning levels,equipments for cleaning validation , steps for cleaning method validation and analytical method validation used for cleaning.
Sterility Testing is defined as a testing which confirms that products are free from the presence of viable microorganisms. Sterility testing is very important for medical devices, pharmaceuticals, preparations, tissue materials and other materials that claim to be sterile or free from viable microorganisms.
Contamination Control in Cleanrooms_Dr.A. AmsavelDr. Amsavel A
Basic’s of Contamination
Sources of Contamination
Environment Specification
Elements of Cleanroom Design and Qualification
Definitions
Control of Contaminations
People, Cleaning, Environment & Material
Operation, Monitoring and Control
Documents and Records
The two most commonly used within microbiology are
HACCP (which originated in the food industry) and FMEA
(developed for engineering). This article explores these two
approaches, first with a description of HACCP, followed by a
description and case study of FMEA in sterility testing.
The Changing Landscape of Clinical Research Regulations: Updates and Implicat...ClinosolIndia
The landscape of clinical research regulations is constantly evolving to adapt to the changing needs of the research community, advancements in scientific understanding, and the protection of research participants. Here are some key updates and implications that have shaped the current state of clinical research regulations
The Impact of Real-World Data in Pharmacovigilance and Regulatory Decision-Ma...ClinosolIndia
Real-world data (RWD) has gained significant importance in pharmacovigilance and regulatory decision-making processes. Real-world data refers to data collected from routine clinical practice, including electronic health records (EHRs), claims databases, registries, and other sources, outside the controlled environment of clinical trials. Here are some key impacts of real-world data in pharmacovigilance and regulatory decision-making
Turning up the Compen-DIAL: Rapid Test Methods for Cell & Gene TherapiesMerck Life Sciences
Watch the presentation of this webinar here: https://bit.ly/3aeCPNB
Find out how we turn up the dial on quality control testing for cell and gene therapies through rapid methods for sterility, mycoplasma, and replication competent virus. We will review the current regulatory expectations as well as the benefits and limitations that come with each method.
Two of the biggest challenges with applying traditional quality control (QC) test methods to cell and gene therapies, is time to results, due to short shelf-life, and availability of sufficient sample, due to small production volumes.
So how can these challenges be overcome while still meeting regulatory expectations?
In this webinar we will discuss and review suitable methods for rapid testing of short-life cell and gene therapies that may also help conserve limited production material. We will look at benefits, limitations, and regulatory expectations for various QC needs including current and future rapid methods for sterility, mycoplasma and replication competent virus.
In this webinar, you will learn:
• Why the shelf life of a cell or gene therapy product may impact your QC testing strategy
• Current regulatory expectations surrounding rapid methods for sterility, mycoplasma and replication competent virus
• Potential impacts of pursuing a non-optimal QC testing strategy
Turning up the Compen-DIAL: Rapid Test Methods for Cell & Gene TherapiesMilliporeSigma
Watch the presentation of this webinar here: https://bit.ly/3aeCPNB
Find out how we turn up the dial on quality control testing for cell and gene therapies through rapid methods for sterility, mycoplasma, and replication competent virus. We will review the current regulatory expectations as well as the benefits and limitations that come with each method.
Two of the biggest challenges with applying traditional quality control (QC) test methods to cell and gene therapies, is time to results, due to short shelf-life, and availability of sufficient sample, due to small production volumes.
So how can these challenges be overcome while still meeting regulatory expectations?
In this webinar we will discuss and review suitable methods for rapid testing of short-life cell and gene therapies that may also help conserve limited production material. We will look at benefits, limitations, and regulatory expectations for various QC needs including current and future rapid methods for sterility, mycoplasma and replication competent virus.
In this webinar, you will learn:
• Why the shelf life of a cell or gene therapy product may impact your QC testing strategy
• Current regulatory expectations surrounding rapid methods for sterility, mycoplasma and replication competent virus
• Potential impacts of pursuing a non-optimal QC testing strategy
Clinical Pharmacology in Orphan Drug DevelopmentE. Dennis Bashaw
This is the fourth talk that I gave in Asia back in May. It was presented at the Konect (Korea National Enterprise for Clinical Trials) 3rd symposia that was held in Seoul at Seoul National University.
This primer is intended for the non-clinician. After reading it, hopefully you will have a slightly better understanding of the complexities of clinical trials.
Modern drug discovery process|| M pharm Pharmacology.pptxRohit chaurpagar
Modern drug discovery is a complex, multidisciplinary process that involves several stages and utilizes advanced technologies. The goal is to develop new therapeutic agents that can safely and effectively treat diseases. Here’s an overview of the key stages in the modern drug discovery process
The regulation of biologicals in AustraliaTGA Australia
View this presentation for information on:
* what biologicals are, including classes and current uses
* the Australian biologicals framework
* new and experimental products
* clinical trials and risk management.
SDTM Training for personnel with Junior and Intermediate level Clinical Trial Experience. Covers summary of most domains. Salient features include order of domain creation, importance of making programming Data/Metadata Driven, Nature of Clinical Raw Data, Summary of the Clinical Trial process with regards to the data flow to arrive at the Study data to be submitted to regulatory authorities like FDA, Importance of deriving ADAM from SDTM and not directly from raw data, Information has been put together from variety of sources including my own programming work.
INTRODUCTION
A PERFECT THERAPEUTIC DRUG
DRUG DISCOVERY- HISTORY
MODERN DRUG DISCOVERY
BIOINFORATICS IN DRUG DISCOVERY
DRUG DISCOVERY BASED ON BIOINFORMATIC TOOLS
BIOINFORMATICS IN COMPUTER-AIDED DRUG DISCOVERY
ECONOMICS OF DRUG DISCOVERY
CONCLUSION
REFERENCES
“Approaches to evaluation of various groups of biological products in Russia”
Provides an overview of the current assessment approaches applicable to biotherapeutics in the Russian Federation
This presentation highlights the reasons which lead to the withdrawal of the 2002 Guidance of the FDA and the current issue with Blend Uniformity and Content Uniformity Determinations.
WHO has recently issued draft document titled "Guidelines on Validation". These guidelines (i.e., the main text included in this working document) cover the general principles of validation and qualification.
These guidelines focus mainly on the overall concept of validation and are not intended to be prescriptive in specific validation requirements. This document serves as general guidance only and the principles may be considered useful in its application in the manufacture and control of starting materials and finished pharmaceutical products (FPPs), as well as other areas. Validation of specific processes and systems, for example, in sterile product manufacture, requires much more consideration and a detailed approach that is beyond the scope of this document. The general text in this document may be applicable to validation and qualification of premises, equipment, utilities, systems, processes, and procedures.
The draft on the specific topics, the appendices to this main text, will follow. The following is an overview on the appendices that are intended to complement the text of this working document:
Appendix 1: Validation of heating, ventilation and air-conditioning systems - will be replaced by cross reference to WHO Guidelines on GMP for HVAC systems for considerations in qualification of HVAC systems (update - working document QAS/15.639/Rev. 1)
Appendix 2: Validation of water systems for pharmaceutical use - will be replaced by cross-reference to WHO Guidelines on water for pharmaceutical use for consideration in qualification of water purification systems
Appendix 3: Cleaning validation - consensus to retain
Appendix 4: Analytical method validation - update in process
Appendix 5: Validation of computerized systems - update in process
Appendix 6: Qualification of systems and equipment - update in process
Appendix 7: Non-sterile process validation - update already published as Annex 3, WHO Technical Report Series, No. 992, 2015
Comments on this draft document are due by July 12, 2016.
A presentation on this guidance is given below:
Presentation on New WHO Guidance on Validations
Environmental Monitoring describes the microbiological testing under- taken in order to detect changing trends of microbial counts and micro- flora growth within cleanroom or controlled environments. The results obtained provide information about the physical construction of the room, the performance of the Heating, Ventilation, and Air-Conditioning (HVAC) system, personnel cleanliness, gowning practices, the equipment, and cleaning operations.
Over the past decade, environmental monitoring has become more sophisticated in moving from random sampling, using an imaginary grid over the room and testing in each grid, to the current focus on risk assessment and the use of risk assessment tools to determine the most appropriate methods for environmental monitoring.
This presentation gives current trends in the application of risk assessment to the practice of environmental monitoring.
This presentation is compiled from freely available resources like the websites of FDA, EMA ,WHO and research papers published by experts in this field like Sandle, T Reinmüller, B , Hyde, W,, Costello, E.K., Lauber, C. L., Hamady, M., Fierer, N., Gordon, J.I., Knight, R.
Paper published by T. Sandle on clean room contamination was referred extensively for this presentation. “Drug Regulations” is a non profit organization which provides free online resource to the Pharmaceutical Professional.
Visit http://www.drugregulations.org for latest information from the world of Pharmaceuticals.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
263778731218 Abortion Clinic /Pills In Harare ,sisternakatoto
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micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
There are a number of conditions that present acutely, predominantly with pain and/or swelling
A careful and detailed history and examination, and in some cases, investigations allow differentiation between these diagnoses. A prompt diagnosis is essential as the patient may require urgent surgical intervention
Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
Knee anatomy and clinical tests 2024.pdfvimalpl1234
This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
2. This presentation is compiled from papers
published by experts like Sandle, T , Newby, P ,
Hussong, D.; Mello, R. Friedman, E.M.; Warner,
M.; Shum, S.C.; Adair, F and various
pharmacopeias
“Drug Regulations” is a non profit organization
which provides free online resource to the
Pharmaceutical Professional.
Visit http://www.drugregulations.org for latest
information from the world of Pharmaceuticals.
14-01-2016 2
Drug Regulations : Online
Resource for Latest Information
3. This presentation is compiled from freely available
resource like the website of WHO, specifically the
WHO Guidance on
Quality Risk Management
“Drug Regulations” is a non profit organization
which provides free online resource to the
Pharmaceutical Professional.
Visit http://www.drugregulations.org for latest
information from the world of Pharmaceuticals.
14-01-2016 3
Drug Regulations : Online
Resource for Latest Information
4. 4
This presentation gives an overview
of
◦ Validation of microbiological methods
◦ Considering some of the limitations
◦ Key criteria that may be applicable for
assessment.
Drug Regulations : Online
Resource for Latest Information
5. Analytical methods designed to recover
chemical substances have less variation
On the other hand high variation is seen with
many Micro methods
◦ Especially those based on culture
In spite of this Microbiology has become an
exacting science
5Drug Regulations : Online Resource for Latest Information
6. Micro methods may not meet the exact
demands of analytical ones
Significant progress in rapid microbiological
methods and alternative techniques
Many novel methods use automation and data
capture with statistical analysis
Validation approaches need to consider these
developments
6Drug Regulations : Online Resource for Latest Information
7. Validation, in this context, can be defined as
“Process by which it is established, by
laboratory studies, that the performance
characteristics of a method meet the
requirements for the intended application”
7Drug Regulations : Online Resource for Latest Information
8. Important objectives of validation is
determining
◦ Whether the sample to be examined has any
inherent anti-microbial properties and
◦ Whether the incubation and growth conditions can
recover microorganism that may be present to an
acceptable level.
8Drug Regulations : Online Resource for Latest Information
9. Following should form the basis of validation strategy
and be considered in drawing up assessment criteria
Limit of detection
◦ What is the lowest level of microorganisms that can be detected
Specificity
◦ What range of different microorganisms can be detected
Quantification
◦ The counting accuracy
9Drug Regulations : Online Resource for Latest Information
10. Suitability of criteria will depend upon
◦ The state of the method: automation /
sophistication
◦ Limit quantification or detection required.
10Drug Regulations : Online Resource for Latest Information
11. This presentation provides a set of
criteria and explanation, which can be
considered to build a validation
protocol and experimentation.
11Drug Regulations : Online Resource for Latest Information
12. Broadly fall into three major
categories
12Drug Regulations : Online Resource for Latest Information
13. Qualitative tests
Tests for the presence or absence of microorganisms
◦ e.g. the pharmacopoeia sterility test.
Commonly, qualitative tests are assessed through the
use of turbidity or other growth related changes in a
culture medium
◦ These serve as evidence of the presence of viable
microorganisms in a test sample.
13Drug Regulations : Online Resource for Latest Information
14. Quantitative tests
Tests for the enumeration of microorganisms
Total viable count test
Flow cytometry.
14Drug Regulations : Online Resource for Latest Information
15. Identification tests
Speciation of bacteria using a biochemical test
Morphological and biochemical characterization
Biochemical reactions
◦ Carbon substrate utilization
◦ Characterization of fatty acid composition
◦ Restriction endonuclease banding patterns
◦ Use of 16S or DNA sequence analysis
15Drug Regulations : Online Resource for Latest Information
16. Methods in each category have a different variability
These variations are inherently different from analytical
ones
The wide variability is acknowledged in Ph. Eur. 5.1.6
Chapter relating to alternative microbiological methods
This is particularly with regards to relative broad
ranges.
16Drug Regulations : Online Resource for Latest Information
17. Reason for the variation
Microbiology is a logarithmic science.
Microbiological methods are capable of
distinguishing between 100 and 1000 cells (1
log)
However smaller differences, such as 0.3 or
0.5 of a log can not be distinguished.
17Drug Regulations : Online Resource for Latest Information
18. In other words e.g.
17 CFU is technically no different than 10
CFU; and
1 CFU is different than 10 CFU but not 5 CFU.
Culture based methods provide estimates
rather than exact cell counts.
18Drug Regulations : Online Resource for Latest Information
19. Variability may result in difficulties in comparing
two methods.
In general, only a 50% comparison is achievable
for culture methods.
With rapid and alternative microbiological
methods greater comparability can be achieved
However levels of precision remain in the order
of 15 to 35% relative standard deviation
19Drug Regulations : Online Resource for Latest Information
20. Culture based microbial methods
Validation is limited by the ability of
microorganisms to reproduce under a set of
conditions in relation to
◦ Sample preparation
◦ Cultivation
◦ Incubation.
20Drug Regulations : Online Resource for Latest Information
21. Any method is, therefore, a general indicator
only.
For these methods, as described in the
pharmacopeia, the limit of detection has
never been established quantitatively.
21Drug Regulations : Online Resource for Latest Information
22. Many variables can affect recovery. These
include:
◦ Growth media
◦ The colony forming unit or “CFU” is not a true cell
count
Individual cells are rare in nature, leading to the CFU
being an underestimation of the number of
microorganisms present
22Drug Regulations : Online Resource for Latest Information
23. Many variables can affect recovery. These include:
◦ Incubation conditions (temperature and time),
◦ Nutritional requirement of the organism
◦ Physical condition of the organism
Stressed or sub lethally damaged due to temperature,
humidity, high ionic strength, pH extremes, osmotic shock
(relating to liquid),
◦ Residues of antimicrobial chemicals
23Drug Regulations : Online Resource for Latest Information
24. Many variables can affect recovery. These
include:
◦ Dilution errors
◦ Environmental organisms are unlikely to be recovered when
they are in the exponential growth phase
exponential growth leads to better recovery
◦ Characteristic of the item under test
◦ The types of neutralizers used
24Drug Regulations : Online Resource for Latest Information
25. With non-culture based methods,
these are unlikely to be affected by
the list given earlier
25Drug Regulations : Online Resource for Latest Information
26. Method comparison is required
for
◦ Introduction of a new method
◦ Replacement of one method with
another
26Drug Regulations : Online Resource for Latest Information
27. New methods will most probably
recover grater number of organisms
compared to the older method
27Drug Regulations : Online Resource for Latest Information
28. Positive or higher result with an
alternative method
Negative or lower result with an
established method
Does suggest that the positive result
is a false positive.
28Drug Regulations : Online Resource for Latest Information
29. Similarly because greater numbers of
organisms may be recovered with an
alternative method, does not mean
that the risk is now greater.
This is normally because of process
controls in place.
29Drug Regulations : Online Resource for Latest Information
30. Revision of limits in light of greater
microbial recoveries needs careful
considerations
In evaluating an alternative method a
close match is not necessarily required,
between the alternative method and the
current method.
30Drug Regulations : Online Resource for Latest Information
31. Important consideration
Alternative method should be capable
of allowing an equivalent decision to
be made in relation to sample or
product quality in a consistent
fashion.
31Drug Regulations : Online Resource for Latest Information
32. Define the objective and hypothesis
appropriately.
◦ Hypothesis
This is a statement concerning an expected
outcome of the experiment
The experimental result will prove or
disprove the hypothesis.
32Drug Regulations : Online Resource for Latest Information
33. The objectives for the experiment should
be
◦ Specific
◦ Measurable
◦ Achievable
◦ Realistic
◦ Time based
33Drug Regulations : Online Resource for Latest Information
34. In defining the objectives, the following should be considered:
Validation requirements
Test controls
Microorganisms
Type of data/statistical analysis
◦ this is an area requiring care, especially when comparing compendial methods
with alternative ones
How many times does the experiment need to be run?
How many samples are required?
34Drug Regulations : Online Resource for Latest Information
35. As part of a study, preliminary work method development may be
undertaken when drawing up of a protocol.
This gives some idea about the capability and limitations of the
method
Helps with the development of appropriate assessment criteria.
Doing so also helps to determine if there are aspects within the
protocol that require clarification;
Helps determine the planning of the study
◦ e.g. culture media requirements, number of technicians required and so on
35Drug Regulations : Online Resource for Latest Information
36. Following slides list aspects of
experimental design that need to be
considered and incorporated into the
design stage.
36Drug Regulations : Online Resource for Latest Information
37. Samples
The size of the test sample must be considered.
Importantly, the number of samples must be
representative and of a sufficient number.
A statistical technique may be used to set the number
of samples required.
Care must be taken here since the statistical
technique selected may influence the sample size.
37Drug Regulations : Online Resource for Latest Information
38. Samples
The appropriate volume of sample may be a factor,
particularly with bioburden testing
Ensuring that the sample tested is representative of the
final homogenous bulk is important.
For most validation exercises the number of batches tested
is three (or more).
With areas like environmental monitoring, consideration
should be given to the location of samples, such as the
number of locations within a cleanroom.
38Drug Regulations : Online Resource for Latest Information
39. Samples
Consideration should also be given to:
Testing samples at the end of the shelf-life or expiry time
◦ this may include assessment at interim time points
Degrading samples stored in containers;
Holding samples under “worst case‟ conditions
◦ upper or lower temperatures
Testing samples at the end of any required process hold
times.
39Drug Regulations : Online Resource for Latest Information
40. Samples
The above points are applicable to many
bioburden and bacterial endotoxin tests.
Any pre-requisite treatment of the
sample, e.g. neutralization, or
microorganisms (promotion of
reproduction) should be considered.
40Drug Regulations : Online Resource for Latest Information
41. Microorganisms
An experiment normally requires a range of
microorganisms.
For all experiments, microorganisms from an
approved culture collection should be used
This ensures uniformity and traceability.
In some cases compendia will indicate the types or
even specific strains of the microorganisms required
◦ e.g the sterility test chapter within the main pharmacopeia
41Drug Regulations : Online Resource for Latest Information
42. Microorganisms
In other cases, the microorganisms will need to
be selected based on professional judgment.
Experiments may be supplemented by
environmental isolates, or 'wildtypes', as
appropriate
Normally appropriate when a culture medium is
used to monitor a production process
42Drug Regulations : Online Resource for Latest Information
43. Microorganisms
In selecting microorganisms it is often a good idea to draw
these across a range of different morphological types.
Suitable categories include:
◦ Gram positive rod and/or a Gram positive spore bearing rod, e.g.
Bacillus sp.
◦ Gram positive cocci e.g. Staphylococcus aureus
◦ Gram negative rod e.g. Pseudomonas aeruginosaiv)
◦ Fungi, (yeast), e.g. Candida albican
◦ Fungi (filamentous), e.g. Aspergillus brasiliensis
43Drug Regulations : Online Resource for Latest Information
44. Microorganisms
In all instances either the specific culture collection
reference must be quoted or the source of the isolate
identified.
For certain Gram-positive bacteria, the protocol
should specify if the organisms should be in the
endospore state.
The type of experiment must be considered when
allocating microorganisms.
44Drug Regulations : Online Resource for Latest Information
45. Microorganisms
A project involving the examination of
water would most likely require
A Gram negative rod (such as
Pseudomonas sp.) and
A coliform (such as Escherichia coli).
45Drug Regulations : Online Resource for Latest Information
46. Microorganisms
In contrast, an experiment conducted at 55°C
◦ as with a test for thermophilic microorganisms
would involve
A thermophile such as Geobacillus
stearothermophilus.
Consideration should also be given to the
storage of cultures.
46Drug Regulations : Online Resource for Latest Information
47. Microorganisms
Sometimes microorganisms are not recovered as expected.
This is a case for carrying method development work in advance.
An example of poor recovery can occur with Gram-negative rods.
With these organisms, desiccation can occur during aerolisation.
This may occur when using an active air sampler or following loss
of moisture content in an exposed settle plate surface.
47Drug Regulations : Online Resource for Latest Information
48. Microorganisms
This effect tends to damage Gram-negative
bacteria more greatly.
This is because soluble cell contents tend to leak
Mechanisms to control the transfer of molecules
and ions in and out of Gram-negative cells in
particular are considerably impaired.
48Drug Regulations : Online Resource for Latest Information
49. Microorganisms
Damage to the mucopeptide
lipopolysaccharide center cell structure also
causes cell damage and loss of viability.
Such damage occurs very shortly after weight
loss to a medium or after aerolisation.
Such cell damage is typically irreversible
49Drug Regulations : Online Resource for Latest Information
50. Microorganisms
Microorganisms used should normally be
prepared from
◦ Cultures which are no more than 24 hours only
and
◦ No more than five passages from the seed lot.
50Drug Regulations : Online Resource for Latest Information
51. Microorganisms
However, some microorganisms require longer cultivation
than 24 hours.
In these instances the culture age must be documented in
advance to ensure that the culture used is as young as it
can be.
With passages (or subcultures), this is to prevent
phenotypic variations from occurring which might influence
the way the microorganism behaves in the presence of the
sample.
51Drug Regulations : Online Resource for Latest Information
52. Microorganisms
It is good practice that the purity of cultures
is confirmed in advance.
This can either be by
◦ Acceptance of a certificate of analysis from the
supplier of the cultures, or
◦ By confirmatory identification conducted by the
recipient laboratory.
52Drug Regulations : Online Resource for Latest Information
53. Microorganisms
In some circumstances, such as when
conducting a challenge test, a post
identification confirmation may also be
deemed necessary.
With some experiments, an attempt may be
made to induce a stressed state to the
microbial population.
53Drug Regulations : Online Resource for Latest Information
54. Microorganisms
This will need to be decided at the time of
writing the test protocol.
The reason for attempting this is based
on the “unstressed” batch culture grown
organisms being are an artificial creation
that rarely exists outside the laboratory.
54Drug Regulations : Online Resource for Latest Information
55. Microorganisms
Stress factors faced by microorganisms in the environment
include:
◦ Desiccation,
◦ Nutrient deprivation,
◦ Nutrient limitation,
◦ Cold shock,
◦ Heat shock,
◦ Exposure to ultra-violet light,
◦ Other types of radiation leading to sublethal damage,
◦ Responses to disinfectant or detergent residues,
◦ Responses to preservative residues,
◦ Osmolality.
55Drug Regulations : Online Resource for Latest Information
56. Microorganisms
Creating a stressed state is difficult in itself
and difficult to verify, given the unknown
effects of causing damage to cells or with
suppressing cellular growth.
In addition, the age of a culture will affect its
recovery and will be important for certain
identification methods.
56Drug Regulations : Online Resource for Latest Information
57. Temperature
Consideration needs to be given over the temperature ranges the
experiment needs to be performed at.
Here there is little value in restricting validation to one
temperature range for testing that takes place over multiple
ranges.
An example here would be testing culture media at 20-250C
Then using it across the range 20-400C and expecting its growth
promoting properties to be consistent.
57Drug Regulations : Online Resource for Latest Information
58. Temperature
Commonly used ranges include:
◦ 2°C - 8°C,
◦ 20°C - 25°C,
◦ 30°C - 35°C,
◦ 36°C - 38°C
◦ 55°C - 60°C
58Drug Regulations : Online Resource for Latest Information
59. Temperature
In addition to the above, dual incubations
across two or more ranges may be
required
This is for the enumeration of bacteria
and fungi with a single-use culture
medium for environmental monitoring
59Drug Regulations : Online Resource for Latest Information
60. Time
As with temperature, consideration needs to be given to the
time period over which the study is to be conducted.
Getting this right is important since the selected time
becomes the maximum run-time for the test.
It is important to ask if incubation time is set long enough
to show the limit of detection?
The time of the validation read must never exceed the time
used by the testing laboratory for the reading of samples.
60Drug Regulations : Online Resource for Latest Information
61. Time
Specification for incubation times the minimum time must
be clearly stated.
For example, “incubate plate at 20-25oC and read at 24
and 48 hours” rather than “incubate plate at 20-25oC and
read after 48 hours.”
Consideration of acceptable tolerance should be stated.
Is “read at 48 hours”, for instance, reading within ±1 hour
of 48 hours or will a wider tolerance be used, and with what
justification?
61Drug Regulations : Online Resource for Latest Information
62. Growth Promotion
A key question here is:
What type of growth promoting conditions is
required for the cultivation of microorganisms?
This would include different types of agars,
broths, dilution reagents and so on.
These may require a separate or first phase
validation.
62Drug Regulations : Online Resource for Latest Information
63. Atmosphere
Whether the microorganism(s) to recover are obligate aerobes,
facultative aerobes/anaerobes, microareophilic, capnophiles, or
obligate anaerobes, should be considered.
A microaerophile, for example, is a microorganism that requires
oxygen to survive, but requires environments containing lower
levels of oxygen than are present in the atmosphere
◦ that is <21% O2; many require 2-10% O2
Once established it will be known whether a specific combination of gases is
required, for example, CO2N.
63Drug Regulations : Online Resource for Latest Information
64. Antimicrobial Activity/Interference
Note should be taken of whether any of the experimental
conditions produce an antimicrobial effect or interfere with
the test in way of enhancement or inhibition.
Attention should be paid to the best method for
neutralizing any anti-microbial properties.
Common methods for neutralization include dilution,
rinsing, filtration or the use of general or specific
neutralizers.
64Drug Regulations : Online Resource for Latest Information
65. pH
Note should be taken of pH conditions.
pH is important to microbial growth and
certain ranges will be unsuitable for some
microorganisms.
65Drug Regulations : Online Resource for Latest Information
66. Growth Phase
The growth phase of a microbial population can
impact upon the accuracy of a method, for example, a
turbidity method.
Ordinarily, in culture, the following growth dynamics
are observed
Cells initially adjust to the new medium (lag phase).
Cells maybe growing by mass, but not in number.
66Drug Regulations : Online Resource for Latest Information
67. Growth Phase
Lag phase is influenced by
◦ Size of the inoculum;
◦ Time to recover from physical damage;
◦ Time required for synthesis of essential coenzymes;
and
◦ Time required for synthesis of new enzymes
necessary to metabolize the substrates present in
the medium.
67Drug Regulations : Online Resource for Latest Information
68. Growth Phase
Cells then start dividing regularly by the
process of binary fission (exponential phase).
Here cells divide at a constant rate,
expressed as generational or doubling time.
Ideal generation times wider according to
different microbial species.
68Drug Regulations : Online Resource for Latest Information
69. Growth Phase
Escherichia coli, for example, will double every
17-20 minutes, whereas Mycobacterium
tuberculosis doubles every 790-930 minutes.
When their growth becomes limited, the cells
stop dividing (stationary phase).
Eventually cells show loss of viability (death
phase).
69Drug Regulations : Online Resource for Latest Information
70. Number of Microorganisms
Some methods will require a minimum number of
microorganisms in order to detect them.
For example, a method linked to microbial
catabolism; adenosine triphosphate (ATP)
growth; bioluminescence etc.
A pre-growth preparatory step may be required
in order to maximize recovery.
70Drug Regulations : Online Resource for Latest Information
71. Single Cultures
Most methods require a single (pure) culture in order
to obtain a valid result.
Mixed cultures can be employed in certain
circumstances
◦ e.g. container closure integrity.
Here, care must be taken that one microorganism
does not significantly out-grow the other.
In general, mixed cultures should be avoided.
71Drug Regulations : Online Resource for Latest Information
72. Enzymes
When detecting presence of specific
microorganisms using a selective
medium, the specificity and selectivity of
the microorganism and media must be
demonstrated using selective and non-
selective microorganisms.
72Drug Regulations : Online Resource for Latest Information
73. In studying a microbiological method, different validation
parameters require assessment.
Not every parameter will be appropriate,
Either due to the nature of the method (qualitative or quantitative)
Or due to its capabilities
A modern, rapid microbiological method, for example, will have
been designed to meet more parameters than an older cultural
method.
Several of the categories have been revisited by microbiologists in
light of the emergence of rapid and alternative microbiological
methods
73Drug Regulations : Online Resource for Latest Information
74. Specificity is the capability of the method to resolve or
measure a range of microorganisms in the presence
of other compounds or microorganisms.
This may be a single microorganism (such as a test
for coliforms) or a range (such as a general bioburden
test).
Freedom from interference from excipients or active
pharmaceutical ingredients, degradation products or
impurities must be noted as part of a recovery
(accuracy) study.
74Drug Regulations : Online Resource for Latest Information
75. Where selecting an appropriate culture medium is
part of the study, the properties of the medium
against selective, non-selective and mixed
cultures must also to be considered.
The microbial challenge should be set above the
limit of detection or quantification, while also
being at level that provides a measure of the
efficacy of the method.
75Drug Regulations : Online Resource for Latest Information
76. For a growth based method, a low number of <100 CFU is
appropriate.
All challenge microorganisms should be recovered.
Where atypical colony morphology is observed, supporting
identification should be considered.
For a non-growth based method, suitable positive and
negative controls should be used to show that any
extraneous matter does not interfere with the detection of
the microorganisms.
76Drug Regulations : Online Resource for Latest Information
77. A new method or test must demonstrate that it is
appropriate for its intended use.
If a method or test is intended to replace an established
method,
◦ Parallel testing of both methods must take place
◦ The collected data compared if possible, ideally by statistical tests
of significance.
In some cases, a direct comparison is not possible
◦ Two different models of particle counter cannot be directly
compared because they will not be sampling the same volume of air
77Drug Regulations : Online Resource for Latest Information
78. Accuracy is the closeness of agreement
between the “measured value” and the “true‟
or “expected” measure or reaction across the
range of the test.
This can be assessed by determining the
recovery of known quantities of a
microorganism that has been added to a
sample.
78Drug Regulations : Online Resource for Latest Information
79. For quantitative tests, this is predicted from:
◦ The dilution of a microbial suspension; or,
◦ By examining for presence/absence; or,
◦ From a taxonomic identification; or,
◦ By comparing the new method to an established
test method
Here the new method must give equivalent or better
results to the established method
79Drug Regulations : Online Resource for Latest Information
80. For enumeration methods, the level of recovery should
reflect the test method.
This is normally by the percentage of microorganisms
recovered by the method.
a. If good recovery is considered to be achievable, then a recovery
level of 50% should be the minimum (this can also be expressed
as a productivity ratio). Where an upper level of recovery is
required, this is normally set at 200% (with a range of 50-200%
quoted).
b. When comparing between two methods, 70% is sometimes set
although other acceptance criteria may be appropriate.
80Drug Regulations : Online Resource for Latest Information
81. Comparison of accuracy can be further
examined by significance testing, such as
Student's t- test or an alternative method.
For example, accuracy can be expressed as:
Accuracy % = (Number of Correct Results in
Agreement/Total Number of Results) x 100
81Drug Regulations : Online Resource for Latest Information
82. For qualitative methods it is recognized that many
hundreds of comparisons may be required if a
negative result is the expected outcome, such as with
the sterility test.
A limitation must be established in relation to the
number of samples;
This is because testing samples until a positive result
is obtained will be impractical.
When comparing two methods, the relative rates of
positive and negative results should be compared.
82Drug Regulations : Online Resource for Latest Information
83. Precision
Precision is the closeness of agreement between
a series of test results or the variation in a series
of test results, when a method is applied
repeatedly to multiple samples.
Precision can be subdivided into:
◦ Repeatability (within test variation)
◦ Intermediate Precision
83Drug Regulations : Online Resource for Latest Information
84. Repeatability (within test variation).
This is the variation in results obtained on
the same sample when assayed repeatedly
with one test or within a short period of
time, by the same technician using the
same reagents and equipment.
84Drug Regulations : Online Resource for Latest Information
85. Repeatability (within test variation).
The key to acceptability is the amount of variation. It may be
expressed as:
a. Standard deviation
b. Coefficient of variation (relative standard deviation)
c. Confidence interval of the mean
d. With specific tests, such as microbial identification systems, other
criteria will be used to determine the similarity of the recovered
organisms.
e. Other statistical techniques like Chi-squared, maybe more appropriate
85Drug Regulations : Online Resource for Latest Information
86. Repeatability (within test variation).
Normally at least 3 replicates are required.
Depending on the types of sample, more than one
determination may be required (for example different dilutions
or a range of microorganisms).
Because the testing technician and consumables are the same,
this approach shows variation in sample as assessed against
the method.
86Drug Regulations : Online Resource for Latest Information
87. Intermediate Precision
◦ This is the variation in results obtained
on the sample when assayed on
several separate occasions by different
technicians using different reagents
and equipment etc.
87Drug Regulations : Online Resource for Latest Information
88. Intermediate Precision
This shows reproducibility. This may be expressed as:
a. Standard deviation,
b. Coefficient of variation (relative standard deviation),
c. Confidence interval of the mean.
d. With specific tests, such as microbial identification systems,
other criteria will be used to determine the similarity of the
recovered organisms.
e. Other statistical techniques like Chi-squared, maybe more
appropriate.
88Drug Regulations : Online Resource for Latest Information
89. Intermediate Precision
A minimum of three determinations should be carried
out.
Appropriate acceptance criteria (such as ≥95%) should
be set for repeatability.
This approach shows variation across people and
reagents, and variability within the method.
89Drug Regulations : Online Resource for Latest Information
90. Range is the interval between the
upper and lower levels of microbial
count, for which the procedure has
been established as suitable with
accuracy, linearity (if appropriate,
where there is requirement to
construct a curve), and precision.
90Drug Regulations : Online Resource for Latest Information
91. For example, the commonly used range for
microorganism recovery is less than 100 CFU for
total count techniques.
Where a range is required, in order to assess the
range of the test, this is covered by diluting a
microbial population; for example, 100 to 106 cells.
With some methods, a regression analysis can be
considered to compare two methods.
91Drug Regulations : Online Resource for Latest Information
92. Robustness is the reliability of a method or test
to withstand small (but deliberate) variations
due to external influence.
For example, different technicians,
instruments, incubation time,ambient
temperature, and reagents.
With rapid methods, robustness can be
undertaken by the method supplier.
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93. However, once in the laboratory, long term
performance must be considered and, if possible,
checked by internal control samples.
For example, the distribution of microorganisms on
a membrane can affect robustness.
Ruggedness is the degree of reproducibility by
testing samples using different testers and
equipment;
this is assessed by coefficient of variation.
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94. This is the lowest number of microorganisms
which can be detected, but not necessarily
quantified (such as a low level challenge) under
the stated experimental conditions.
This test is generally used for rapid and
alternative methods.
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95. Often the amount of sample tested the initial
dilution of the sample and any subsequent dilution
of the sample may determine the limit of detection.
The challenge microorganisms selected should be
of an appropriate range as indicated above).
The challenge can consist of taking each
microorganism and making a serial dilution range.
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96. The outcome can be expressed as
presence/absence or enumeration.
Presence/absence is normally qualitative.
An attempt can be made for a semi-
quantitative analysis by varying the microbial
challenge
◦ to develop a limit test, i.e. to detect <100 CFU, <10
CFU etc.
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97. It is recognized that the act of dilution may result in a
greater loss due to lack of homogeneity and the typical
Poisson distribution of microorganisms in liquid;
such as in the evaluation of a raw material by using the
pour plate method and where the limit of detection is
<10cfu/g.
This is further complicated by the impossibility of
obtaining reliable samples containing a single
microorganism.
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98. Therefore, microbiologically limits of detection must
sometimes be considered as theoretical rather than
practically demonstrable.
For this reason, many pharmacopeial` tests require the
use of a low level challenge (<100 CFU) and this is
normally considered sufficient.
For comparing methods Chi-squared is often the
statistic of choice.
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99. This is the lowest level of the sample where the
microbial content can be quantitatively
determined with defined precision and
accuracy.
Again, further complication arises by the
impossibility of obtaining reliable samples
containing a set number of microorganisms.
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100. Therefore, microbiologically limits of detection
must sometimes be considered as theoretical rather
than practically demonstrable.
Many pharmacopeial tests require the use of a low
level challenge (<100 CFU) and this is normally
considered sufficient.
Limits of quantification are normally determined by
3 or more replicates across the range.
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101. This is the ability to elicit results which are
proportional to the concentration of
microorganisms within a given range.
This is measured by correlation coefficient or a
goodness of fit test (such as Chi-Square).
This will only be applicable to enumeration
methods using an analytical system.
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102. For qualitative tests such as the sterility test or
growth of selective media, the use of positive
or negative predictive values may be
appropriate.
This can be expressed as a percentage of the
observed test results against the total or
expected test results.
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103. Rapid and alternative microbiological methods embrace
those methods that are distinct from compendial
methods.
Rapid indicates that the method gives a faster time-to-
result and alternative indicates that the method differs
from one presented in a recognized pharmacopeia
In many cases such methods are more accurate and they
are invariably automated
Although this is not always the case
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104. Given the pace of technological development and commercial
gains, such methods are becoming more common
When considering the introduction of an alternative method, the
following should be considered
Prepare a User Requirement Specification.
Consider instrument qualification; validation requirements of the
alternative technology and aim
◦ e.g. to be equivalent to current methods or to improve upon a current method
Method suitability
◦ e.g. sample volume requirements, critical parameters of compendial test,
suitability for a range of different products to be tested
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105. Sample size and number of samples:
With a new method, it may be appropriate to assess all parameters on one type of
product and then to run method suitability tests only on other products against
the same method.
In terms of results interpretation, non-growth based methods (such as those
looking at metabolic activity or ATP) will provide more accurate cell counts and
these are not comparable with growth-based estimates of colony forming units.
Results from alternative methods, such as cell count, cannot be compared
statistically to CFU results from culture based methods.
Data handling.
Test controls e.g. use of control microorganisms.
Consideration of test limits for samples tested by alternative methods.
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106. With alternative methods the aim is to
verify the detection capability of the
alternative method.
In terms of parameters to review, the
following acts as guidance (based on
USP chapter <1223>)
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108. With the above, a degree of interpretation is required. It follows:
Method suitability: accuracy, precision and specificity (recovery of challenge
organisms) for quantitative methods; for qualitative methods, only
specificity would apply.
Robustness is assessed as above. It is not directly compared with a current
method. Manufacturer‟s data may be considered.
Ruggedness is assessed as above.
It is not directly compared with a current method. Manufacturer‟s data may
be considered.
Limit of detection: this is assessed by using a range of microorganisms.
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109. Approach #1:
Each organism is prepared as a serial dilution,
with the inoculum adjusted to a target of 50%
that shows growth in an existing method.
Both methods should be run over several
replicates.
An appropriate statistical comparison method
is Chi- squared.
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110. Approach #2:
Most Probable Number (MPN) method is used.
For a ten-fold series a range of 10-1 to 10-2
microorganisms is used; or for a two-fold series,
using the range 5 to 10-1 microorganisms.
The alternative and established method should be
run five times using 3 dilutions
◦ the dilutions should provide at least one positive and one
negative dilution
Chi- squared is an appropriate statistical tool for
method comparison.
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111. Where the aim is to show equivalence between
two methods, there are four possible ways to
assess this (again as referenced in USP
<1223>).
The appropriate category should be selected in
advance of conducting the validation and
referenced in the validation protocol.
The options are: ( See next slide )
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113. The options in earlier slide can be interpreted
as:
1. Acceptable procedure: this is not strictly an
equivalence option, it is about the new method
meeting a minimum performance or acceptance
requirement.
◦ The qualification may involve a standard inoculum
with a range of microorganisms to assess recovery,
or a level of ATP.
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114. The options in earlier slide can be interpreted as:
2. Performance equivalence: better or equivalent
results are required from the new method
compared with the existing method.
◦ This is with regard to: accuracy, precision,
specificity, limit of detection, limit of
quantification, robustness and ruggedness
(only those factors that are relevant require
inclusion).
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115. The options in earlier slide can be interpreted as:
3. Results equivalence: this is similar to
performance equivalence expect with the added
requirement that the new method must give
equivalent or better numerical results.
◦ Because the same sample cannot be tested
against two methods, a tolerance value is
required.
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116. The options in earlier slide can be interpreted as:
4. Decision equivalence: this is similar to results
equivalence with the exception that „pass‟ or
„fail‟ results are compared (as with a „growth‟
/ „no growth‟ test result).
◦ It is also possible to consider a Most Probable
Number method for qualitative tests.
◦ When evaluating instruments there should be a one-
sided inferiority hypothesis.
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117. This presentation is compiled from papers
published by experts like Sandle, T , Newby, P ,
Hussong, D.; Mello, R. Friedman, E.M.; Warner,
M.; Shum, S.C.; Adair, F and various
pharmacopeias
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