This document discusses analytical method validation. It begins with an introduction that defines validation and discusses its importance and regulatory requirements. The document then covers specific validation parameters such as specificity, linearity, accuracy, precision, limit of detection, limit of quantification and more. For each parameter, the document provides definitions, procedures for evaluation, and acceptance criteria. It emphasizes that validation demonstrates a method is suitable for its intended purpose and supports the identity, quality, purity and potency of drug substances and products. The overall summary is that analytical method validation is critical to ensure quality and compliance in the pharmaceutical industry.
Analytical method validation, ICH Q2 guidelineAbhishek Soni
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Analytical Method Validation, ICH Q2 Guideline.
General principles related to the analytical method validation.
Validation of analytical method as per International Council for Harmonisation(ICH) guidelines and the United States Pharmacopeia(USP).
Glossary.
Useful in understanding the terms :
Specificity
Linearity
Range
Accuracy
Precision
Detection limit
Quantitation limit
Robustness
Ruggedness
System suitability testing
Analytical method validation, ICH Q2 guidelineAbhishek Soni
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Analytical Method Validation, ICH Q2 Guideline.
General principles related to the analytical method validation.
Validation of analytical method as per International Council for Harmonisation(ICH) guidelines and the United States Pharmacopeia(USP).
Glossary.
Useful in understanding the terms :
Specificity
Linearity
Range
Accuracy
Precision
Detection limit
Quantitation limit
Robustness
Ruggedness
System suitability testing
Analytical method validation as per ich and usp shreyas B R
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Analytical method validation is a process of documenting/ proving that an analytical method provides analytical data acceptable for the intended use.After the development of an analytical procedure, it is must important to assure that the procedure will consistently produce the intended a precise result with high degree of accuracy. The method should give a specific result that may not be affected by external matters. This creates a requirement to validate the analytical procedures. The validation procedures consists of some characteristics parameters that makes the method acceptable with addition of statistical tools.
In this slide contains Introduction, levels of cleaning, mechanism, sampling method of cleaning validation.
Presented by: P. VENKATESH (Department of pharmaceutical analysis).RIPER, anantapur
What is Validation?
Methods validation is the process of demonstrating that analytical procedures are suitable for their intended use-Guidance for Industry
Validation is a process-risk will determine the effort
High Risk:Total validation
Moderate Risk:Testing,Documentation
Low Risk:Testing the change
Accuracy
ICH defines accuracy of an analytical procedure as the closeness of agreement between the conventional true value or an accepted reference value and the value found.
% Accuracy = Experimental- True Value * 100
True Value
Precision
Precision of analytical procedure is defined as closeness of agreement in values between a series of measurements. As per ICH, precision is considered at three different levels:
Repeatability or intraâassay precision: precision data are obtained by repeatedly analyzing, in one lab on one day, aliquots of a homogeneous sample.
Intermediate precision: precision obtained when the assay is performed by multiple analysts, multiple instruments, and multiple days in one lab.
Reproducibility: precision between laboratories.
Specificity
Specificity is the ability of the method to accurately measure the analyte response in the presence of all potential sample components.
It is very important in the analysis of complex mixtures by GC, HPLC, AA, ICP, etc.
Limit of Detection (LOD)
Limit of Detection (LOD) is the lowest amount of analyte in a sample which can be reliably detected but not necessarily accurately or precisely measured.
Signal/Noise = 2 to 3
Limit of Quantitation (LOQ)
Limit of Quantitation (LOQ) is the lowest amount of an analyte that can be quantitatively determined with suitable precision and accuracy.
Signal/Noise = 10 to 20
Linearity and Range
Linearity of an analytical procedure is its ability (within a given range) to obtain test results which are directly proportional to the concentration (amount) of analyte in the sample.
Range: Interval from the upper to the lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity
Must cover 80-120% of product claims
Usually evaluated from the same data set as linearity, precision, accuracy
Want to learn more about analytical method validation, FDA requirements and best practices to comply with them? ComplianceOnline webinars and seminars are a great training resource. Check out the following links:
ICH, FDA and USP Requirements for Method Validation
How to Validate Analytical Methods and Procedures
Validation of Analytical Methods and Procedures
Eliminate the Confusion - Analytical Method Qualification and Validation
Lifecycle Approach to Analytical Methods with QbD Elements
Analytical Instrument Qualification and System Validation
Lifecycle Approach to Analytical Methods for Drug Products
For details vis
Analytical method validation as per ich and usp shreyas B R
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Analytical method validation is a process of documenting/ proving that an analytical method provides analytical data acceptable for the intended use.After the development of an analytical procedure, it is must important to assure that the procedure will consistently produce the intended a precise result with high degree of accuracy. The method should give a specific result that may not be affected by external matters. This creates a requirement to validate the analytical procedures. The validation procedures consists of some characteristics parameters that makes the method acceptable with addition of statistical tools.
In this slide contains Introduction, levels of cleaning, mechanism, sampling method of cleaning validation.
Presented by: P. VENKATESH (Department of pharmaceutical analysis).RIPER, anantapur
What is Validation?
Methods validation is the process of demonstrating that analytical procedures are suitable for their intended use-Guidance for Industry
Validation is a process-risk will determine the effort
High Risk:Total validation
Moderate Risk:Testing,Documentation
Low Risk:Testing the change
Accuracy
ICH defines accuracy of an analytical procedure as the closeness of agreement between the conventional true value or an accepted reference value and the value found.
% Accuracy = Experimental- True Value * 100
True Value
Precision
Precision of analytical procedure is defined as closeness of agreement in values between a series of measurements. As per ICH, precision is considered at three different levels:
Repeatability or intraâassay precision: precision data are obtained by repeatedly analyzing, in one lab on one day, aliquots of a homogeneous sample.
Intermediate precision: precision obtained when the assay is performed by multiple analysts, multiple instruments, and multiple days in one lab.
Reproducibility: precision between laboratories.
Specificity
Specificity is the ability of the method to accurately measure the analyte response in the presence of all potential sample components.
It is very important in the analysis of complex mixtures by GC, HPLC, AA, ICP, etc.
Limit of Detection (LOD)
Limit of Detection (LOD) is the lowest amount of analyte in a sample which can be reliably detected but not necessarily accurately or precisely measured.
Signal/Noise = 2 to 3
Limit of Quantitation (LOQ)
Limit of Quantitation (LOQ) is the lowest amount of an analyte that can be quantitatively determined with suitable precision and accuracy.
Signal/Noise = 10 to 20
Linearity and Range
Linearity of an analytical procedure is its ability (within a given range) to obtain test results which are directly proportional to the concentration (amount) of analyte in the sample.
Range: Interval from the upper to the lower concentration (amounts) of analyte in the sample (including these concentrations) for which it has been demonstrated that the analytical procedure has a suitable level of precision, accuracy and linearity
Must cover 80-120% of product claims
Usually evaluated from the same data set as linearity, precision, accuracy
Want to learn more about analytical method validation, FDA requirements and best practices to comply with them? ComplianceOnline webinars and seminars are a great training resource. Check out the following links:
ICH, FDA and USP Requirements for Method Validation
How to Validate Analytical Methods and Procedures
Validation of Analytical Methods and Procedures
Eliminate the Confusion - Analytical Method Qualification and Validation
Lifecycle Approach to Analytical Methods with QbD Elements
Analytical Instrument Qualification and System Validation
Lifecycle Approach to Analytical Methods for Drug Products
For details vis
The Validation Master Plan is a a valuable opportunity to provide an overview of your companyâs validation process, including organization structure, content, and planning.
Validation of Analytical and Bioanalytical methodssarikakkadam
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Guidelines for Validation of Analytical and Bioanalytical methods as per ICH (Q2R1) and USFDA respectively with an example of Bioanalytical method validation.
Method Validation - ICH /USP Validation, Linearity and Repeatability labgo
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Prepared by : Santram Rajput (Technical Manager)
Validation of analytical procedures reinforce the reliability and suitability of a methodology for providing accurate and precise results. This slide show elaborates in detail about the need for method validation with examples, along with that it also covers the factors to be evaluated prior to validation. This slide show further touches upon the characteristics which are of significance in context of the validation procedure.
Analytical method development and validation are one of the very imp aspects in Drug testing and approval process.Here I tried to explain the same with my experience.
Method validation is the process of documenting / proving that an analytical method provides analytical data acceptable for the intended use. A pharmaceutical drug product must meet all its specifications throughout its entire shelf-life. The performance of product characteristics throughout the shelf-life must be tested by analytical method for the productâs chemical, physiochemical, microbiological and biological characteristics. The method of analysis used must be validated. This is required to ensure the productâs safety and efficacy throughout all phases of its shelf-life. Validation is the process of establishing documentary evidence demonstrating that a procedure, process, or activity carried out in testing and then production maintains the desired level of compliance at all stages.
General Principles of Analytical Method of Validation.pdfTamannaKumari8
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Validation is the process of establishing documentary evidence demonstrating that a procedure, process, activity carried out in
testing and then production maintain the desirable level of compliance all stages.
ī´ The process of providing the analytical procedure is acceptable or its intended us.(ICH Q
How the modern concept of a lifecycle model, which is based on process validation and described in ICH guidelines Q8, Q9, and Q10, can be applied to analytical procedures.
Speaker at seminar "The Pharmaceutical quality system: ICH Q8/ICH Q9" - University of Parma, 18 May 2012.
Describing steps, tools, and approaches developed for application of QbD to manufacturing processes that have analogous application to the development and use of analytical methods.
Analytical Method Validation is a process that is used to demonstrate the suitability of an analytical method for an intended purpose.Regulations and quality standards that have an impact on analytical laboratories require analytical methods to be validated.
002. Cephalosporins for students 2023 Prof. P. Ravisankar.pdfDr. Ravi Sankar
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Cephalosporins, Why Cephalosporins? Advantages of cephalosporins over penicillin, Mechanism of action of cephalosporins, Classification of cephalosporins, Structures of some important cephalosporins and cephamycins, Oximinocephalosporins, SAR of cephalosporins,Hydrolytic reactions, degradation and stability of cephalosporins, Uses of cephalosporins, Comparison between 6-APA and 7-ACA and penam and cepham.
1a. Prof. P. Ravisankar, Vignan Pharmacy College, Vadlamudi, Guntur, AP., Ind...Dr. Ravi Sankar
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Antibiotics, characteristics of an antibiotic, Brief historical background of antibiotics, Modern History of antibiotics, Classification of antibiotics, four groups of beta-lactam antibiotics, Classification of Penicillins, Penicillinase resistant Penicillins, Narrow spectrum anti-staphylococcal Penicillins, Amino Penicillins, Nomenclature of Penicillins, Stereochemistry of Penicillins, Hydrolysis of Penicillins, SAR of Penicillins, Mechanism of Penicillins, Beta-lactamase inhibitors, Therapeutic uses of Penicillins, Penicillin advantages, and disadvantages, Hypersensitivity or allergic reactions of Penicillins.
Tetracyclines are Octahydro napthacene derivatives which are bacteriostatic potent broad spectrum antibiotics and are the most widely prescribed form of antibiotic after penicillins.
TETRA means = four
CYCL means = hydrocarbon rings
INE means = derivation.
Tetracyclines are introduced 50 years ago as potent broad spectrum antibiotics.
They are biosynthesized form acetic acid and propionic acid units in microorganisms.
Finally, one of the best ways of reducing cancer is âĻ..
Cancer prevention is an essential component of all cancer control plans because about 40 % of cancers are preventable.
FirstlyâĻPublic education campaigns are important in highlighting the dangers of smoking because possibly as many as 22 % (in the UK 30%) of cancers are caused by smoking.
Secondly, another 30% of cancers are diet-related. Decades of research have clearly shown that by living a healthy life, people can reduce the risk of developing the disease.
Screening tests help in finding Cancer earlier before developing any symptoms.
The benefits of eating high-fiber foods, fruit, and vegetables.
Epigallocatechin gallate, an antioxidant present in green tea, is another potential protective agent.
Finally, The new research funded by the UK and USA which aims to show the number of cancer cases could be prevented by known lifestyle and environmental factors,
like being a non-smoker, keeping a healthy weight, drinking less alcohol, eating a healthy balanced diet, and avoiding being exposed to certain infections or radiation.
Every country, regardless of resource level, can take steps to curb the cancer epidemic, save lives and prevent unnecessary suffering.
Finally, cancer prevention efforts should be preceded by a systematic planning process on cancer control.
Antibiotics definition, Early and modern history, classification of antibiotics, Mechanism of antibacterial action, bacterial cell and drug targets, penicillins nomenclature, degradation reactions of penicillins, medical classification of penicillins, SAR of penicillins, Mechanism of penicillins, Betalactamase inhibitors, Therapeutic uses of penicillins, toxicity of penicillins, Side effects of penicillins.
NOVEL SEPARATION AND QUANTITATIVE DETERMINATION OF LEVOFLOXACIN, PRULIFLOXACI...Dr. Ravi Sankar
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The core AIM of the present study is to develop a novel, rapid, precise and accurate RP-HPLC method for simultaneous separation and quantification of six fluoroquinolones OF LEVOFLOXACIN (LEVO), PRULIFLOXACIN (PRFX), GATIFLOXACIN (GATI), SPARFLOXACIN (SPAR), MOXIFLOXAXIN (MOXI) AND BALOFLOXACIN (BALO) for the the day to day analysis.
The author felt that a novel single method for separation and quantification of all the above said drugs on single chromatographic system without any minor changes in detection wavelength and mobile phase composition.
To develop rapid, sensitive and economical analytical method based on HPLC for separation and estimation of six fluoroquinolones pharmaceutical dosage forms.
To develop method with shorter run time and better sensitivity.
Reducing the solvent consumption to make it more eco-friendly.
Avoid the column damage by minimizing the buffer strength and pH of mobile phase than reported methods.
To validate the method for different parameters like Accuracy, Precision, Linearity, specificity, Robustness International Conference on Harmonization ICH Q2(R1) guidelines..
To apply the developed RP-HPLC method in the analysis of pharmaceutical formulations.
Analytical method development and validation of novel Levofloxacin, Pruliflox...Dr. Ravi Sankar
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For the first time simple, selective, sensitive RP-HPLC method was developed for the separation and quantitative development of LEVO, PRFX, GATI, SPAR, MOXI and BALO relating to fluoroquinolone anti bacterials in pharmaceutical dosage forms. The most important advantage of developed method was that the 6 separate drugs can be determined on a single chromatographic system without modifications in detection wavelength and mobile phase by RP-HPLC. The chromatographic separation of the selected drugs was carried out on Welchrom C18 column consisting of 250 mm X 4.6 mm, 5 Âĩm particle size utilizing mixture of 10 mM phosphate buffer (pH 3.1): Acetonitrile in the ratio of 70:30,v/v as mobile phase at the flow rate of 1mL/min with detection wave length at 293 nm by using UV spectrophotometric detector with total run time of 10 minutes and 3.613, 4.230, 4.707, 5.497, 5.880 and 6.253 minutes of retention time obtained for LEVO, PRFX, GATI, SPAR, MOXI and BALO respectively. All calibration curves for six drugs showed indicated linearity over a concentration range of 2-10 Âĩg/mL. The results regarding to limit of detection (LOD) and limit of quantitation (LOQ) for LEVO, PRFX, GATI, SPAR, MOXI and BALO were found to be 0.116 Âĩg/mL and 0.348 Âĩg/mL; 0.152 Âĩg/mL and 0.460 Âĩg/mL; 0.084 Âĩg/mL and 0.255 Âĩg/mL; 0.186 Âĩg/mL and 0.558 Âĩg/mL, 0.162 and 0.493, 0.112 and 0.390 respectively. These results clearly show low values of LOD and LOQ. The said developed method was ultimately utilized for quantification of marketed formulation.
NOVEL SIMULTANEOUS SEPARATION AND QUANTITATIVE DETERMINATION OF FOUR SARTANS...Dr. Ravi Sankar
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The core AIM of the present study is to develop a novel, rapid, precise and accurate RP-HPLC method for simultaneous separation and quantification of Hydrochlorothiazide along with four sartans Telmisartan(TELM), Losartan(LOSA), Olmesartan(OLME) and Valsartan(VALS).
To develop Novel methods for separation and quantification of all the above said drugs on single chromatographic system without any minor changes in detection wavelength and mobile phase composition.
Total quality management (TQM), and current Good Manufacturing Practice (cGMP...Dr. Ravi Sankar
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TQM, cGMP, Introduction, Definition, Importance, TQM frame work, Key concepts (Principles) of TQM, specific steps in the cycle, Benefits of TQM, cGMP, principles of GMP, Improtance of GMP, why GMP established?, difference between GMP and cGMP, GMP and cGMP regulations, code of federal regulations.
Tetracyclines BY Dr. P. Ravisankar M. Pharm., Ph.D.Dr. Ravi Sankar
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Tetracyclines by Dr. P. Ravisankar M. Pharm., Ph.D.
Definition
Introduction
Classification
Historical background
Sources
Chemistry
SAR of tetracyclines
Mechanism of action of tetracyclines
Spectrum of activity
Uses of tetracyclines
Side effects of tetracyclines
Dr. P. Ravisankar M. Pharm., Ph.D.
Vitamin A
Vitamin D
Vitamin E
Vitamin K
Definition
Introduction
Classification
Structures,Functions,Deficiency,Diseases,Toxicity and uses.
Introduction to diuretics.
Therapeutic approaches.
Normal physiology of urine formation.
Classification of drugs .
Mechanism of action of Acetazolamide.
Mechanism of action of Thiazides.
Mechanism of action of Loop diuretics.
Mechanism of action of potassium sparing diuretics &aldosterone antagonists.
Smart TV Buyer Insights Survey 2024 by 91mobiles.pdf91mobiles
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91mobiles recently conducted a Smart TV Buyer Insights Survey in which we asked over 3,000 respondents about the TV they own, aspects they look at on a new TV, and their TV buying preferences.
Kubernetes & AI - Beauty and the Beast !?! @KCD Istanbul 2024Tobias Schneck
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As AI technology is pushing into IT I was wondering myself, as an âinfrastructure container kubernetes guyâ, how get this fancy AI technology get managed from an infrastructure operational view? Is it possible to apply our lovely cloud native principals as well? What benefitâs both technologies could bring to each other?
Let me take this questions and provide you a short journey through existing deployment models and use cases for AI software. On practical examples, we discuss what cloud/on-premise strategy we may need for applying it to our own infrastructure to get it to work from an enterprise perspective. I want to give an overview about infrastructure requirements and technologies, what could be beneficial or limiting your AI use cases in an enterprise environment. An interactive Demo will give you some insides, what approaches I got already working for real.
JMeter webinar - integration with InfluxDB and GrafanaRTTS
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Watch this recorded webinar about real-time monitoring of application performance. See how to integrate Apache JMeter, the open-source leader in performance testing, with InfluxDB, the open-source time-series database, and Grafana, the open-source analytics and visualization application.
In this webinar, we will review the benefits of leveraging InfluxDB and Grafana when executing load tests and demonstrate how these tools are used to visualize performance metrics.
Length: 30 minutes
Session Overviewâ
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During this webinar, we will cover the following topics while demonstrating the integrations of JMeter, InfluxDB and Grafana:
- What out-of-the-box solutions are available for real-time monitoring JMeter tests?
- What are the benefits of integrating InfluxDB and Grafana into the load testing stack?
- Which features are provided by Grafana?
- Demonstration of InfluxDB and Grafana using a practice web application
To view the webinar recording, go to:
https://www.rttsweb.com/jmeter-integration-webinar
Builder.ai Founder Sachin Dev Duggal's Strategic Approach to Create an Innova...Ramesh Iyer
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In today's fast-changing business world, Companies that adapt and embrace new ideas often need help to keep up with the competition. However, fostering a culture of innovation takes much work. It takes vision, leadership and willingness to take risks in the right proportion. Sachin Dev Duggal, co-founder of Builder.ai, has perfected the art of this balance, creating a company culture where creativity and growth are nurtured at each stage.
Generating a custom Ruby SDK for your web service or Rails API using Smithyg2nightmarescribd
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Have you ever wanted a Ruby client API to communicate with your web service? Smithy is a protocol-agnostic language for defining services and SDKs. Smithy Ruby is an implementation of Smithy that generates a Ruby SDK using a Smithy model. In this talk, we will explore Smithy and Smithy Ruby to learn how to generate custom feature-rich SDKs that can communicate with any web service, such as a Rails JSON API.
Securing your Kubernetes cluster_ a step-by-step guide to success !KatiaHIMEUR1
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Today, after several years of existence, an extremely active community and an ultra-dynamic ecosystem, Kubernetes has established itself as the de facto standard in container orchestration. Thanks to a wide range of managed services, it has never been so easy to set up a ready-to-use Kubernetes cluster.
However, this ease of use means that the subject of security in Kubernetes is often left for later, or even neglected. This exposes companies to significant risks.
In this talk, I'll show you step-by-step how to secure your Kubernetes cluster for greater peace of mind and reliability.
GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using Deplo...James Anderson
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Effective Application Security in Software Delivery lifecycle using Deployment Firewall and DBOM
The modern software delivery process (or the CI/CD process) includes many tools, distributed teams, open-source code, and cloud platforms. Constant focus on speed to release software to market, along with the traditional slow and manual security checks has caused gaps in continuous security as an important piece in the software supply chain. Today organizations feel more susceptible to external and internal cyber threats due to the vast attack surface in their applications supply chain and the lack of end-to-end governance and risk management.
The software team must secure its software delivery process to avoid vulnerability and security breaches. This needs to be achieved with existing tool chains and without extensive rework of the delivery processes. This talk will present strategies and techniques for providing visibility into the true risk of the existing vulnerabilities, preventing the introduction of security issues in the software, resolving vulnerabilities in production environments quickly, and capturing the deployment bill of materials (DBOM).
Speakers:
Bob Boule
Robert Boule is a technology enthusiast with PASSION for technology and making things work along with a knack for helping others understand how things work. He comes with around 20 years of solution engineering experience in application security, software continuous delivery, and SaaS platforms. He is known for his dynamic presentations in CI/CD and application security integrated in software delivery lifecycle.
Gopinath Rebala
Gopinath Rebala is the CTO of OpsMx, where he has overall responsibility for the machine learning and data processing architectures for Secure Software Delivery. Gopi also has a strong connection with our customers, leading design and architecture for strategic implementations. Gopi is a frequent speaker and well-known leader in continuous delivery and integrating security into software delivery.
Dev Dives: Train smarter, not harder â active learning and UiPath LLMs for do...UiPathCommunity
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đĨ Speed, accuracy, and scaling â discover the superpowers of GenAI in action with UiPath Document Understanding and Communications Miningâĸ:
See how to accelerate model training and optimize model performance with active learning
Learn about the latest enhancements to out-of-the-box document processing â with little to no training required
Get an exclusive demo of the new family of UiPath LLMs â GenAI models specialized for processing different types of documents and messages
This is a hands-on session specifically designed for automation developers and AI enthusiasts seeking to enhance their knowledge in leveraging the latest intelligent document processing capabilities offered by UiPath.
Speakers:
đ¨âđĢ Andras Palfi, Senior Product Manager, UiPath
đŠâđĢ Lenka Dulovicova, Product Program Manager, UiPath
The Art of the Pitch: WordPress Relationships and SalesLaura Byrne
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Clients donât know what they donât know. What web solutions are right for them? How does WordPress come into the picture? How do you make sure you understand scope and timeline? What do you do if sometime changes?
All these questions and more will be explored as we talk about matching clientsâ needs with what your agency offers without pulling teeth or pulling your hair out. Practical tips, and strategies for successful relationship building that leads to closing the deal.
Neuro-symbolic is not enough, we need neuro-*semantic*Frank van Harmelen
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Neuro-symbolic (NeSy) AI is on the rise. However, simply machine learning on just any symbolic structure is not sufficient to really harvest the gains of NeSy. These will only be gained when the symbolic structures have an actual semantics. I give an operational definition of semantics as âpredictable inferenceâ.
All of this illustrated with link prediction over knowledge graphs, but the argument is general.
2. ī Validation is a very necessary element of
any firm that falls under the scrutiny of
the governing regulatory agencies The
FDA, under the authority of existing
cGMP regulations guidelines and
directives, considers validation is
necessary and because it makes good
sense of science / engineering.
3. ī Drug substances and drug product
manufacturers must perform
validations, it is very important that
this understanding be shared
throughout the organization
ī The term validation generally to
cover the entire spectrum of cGMP
4. DEFINTION
īThe process to Confirm that the analyticalThe process to Confirm that the analytical
procedure employed for a specific test isprocedure employed for a specific test is
suitable for intended use and that theysuitable for intended use and that they
support the identity, quality, purity andsupport the identity, quality, purity and
potency of the drug substances and drugpotency of the drug substances and drug
products.products.
I. INTRODUCTIONI. INTRODUCTION
5. NEED
ī The process of method development andThe process of method development and
validation has a direct impact on thevalidation has a direct impact on the
quality of these dataquality of these data
ī To trust the methodTo trust the method
ī Regulatory requirementRegulatory requirement
I. INTRODUCTION (Contd.,)
6. Currently available guidelines from variousCurrently available guidelines from various
regulatory bodiesregulatory bodies
ICHICH
USFDAUSFDA
CDERCDER
EPEP
USP XXXIUSP XXXI
I. INTRODUCTION (Contd.,)
7. Analytical procedures to be validatedAnalytical procedures to be validated
I. INTRODUCTION (Contd.,)
ī Identification TestsIdentification Tests
ī Quantitative Tests for ImpuritiesQuantitative Tests for Impurities
Quantitative tests for active moiety in samples ofQuantitative tests for active moiety in samples of
drug substance & drug products and other selecteddrug substance & drug products and other selected
components in drug productcomponents in drug product
8. I. INTRODUCTION (Contd.,)I. INTRODUCTION (Contd.,)
Recommended validation characteristics of the various types ofRecommended validation characteristics of the various types of
proceduresprocedures
9. CONCEPT OF REVALIDATION
When we make any changes inWhen we make any changes in
ī Analytical procedureAnalytical procedure
ī Drug substance (e.g. synthetic route)Drug substance (e.g. synthetic route)
ī Drug product (e.g. composition)Drug product (e.g. composition)
ī The changes may necessitate revalidation ofThe changes may necessitate revalidation of
the analytical proceduresthe analytical procedures
I. INTRODUCTION (Contd.,)I. INTRODUCTION (Contd.,)
10. A few typical cases are presented below whereA few typical cases are presented below where
revalidation is necessaryrevalidation is necessary
ChangeChange Parameters to be considered forParameters to be considered for
revalidationrevalidation
Synthetic routeSynthetic route All parametersAll parameters
Analytical procedureAnalytical procedure All parametersAll parameters
Addition of new impurityAddition of new impurity Specificity, stability, linearity, accuracy,Specificity, stability, linearity, accuracy,
range, LOD & LOD (for new impurityrange, LOD & LOD (for new impurity
only)only)
Composition of drug productComposition of drug product Specificity, stability, Precision andSpecificity, stability, Precision and
accuracyaccuracy
Change in specificationChange in specification Linearity, accuracy and rangeLinearity, accuracy and range
I. INTRODUCTION (Contd.,)
*If solubility differs each other
11. PRE-REQUISITESPRE-REQUISITES
ī AA well-designed experimental matrix (Writtenwell-designed experimental matrix (Written
protocol)protocol)
ī Step-by-step methodology (STP)Step-by-step methodology (STP)
ī Test samples of good quality (should meet the qualityTest samples of good quality (should meet the quality
as per defined specifications)as per defined specifications)
ī Peak purity (preferably >99% purity)Peak purity (preferably >99% purity)
ī Equipments & analytical instruments should be inEquipments & analytical instruments should be in
calibrated state.calibrated state.
I. INTRODUCTION (Contd.,)
12. PARAMETERS TO BE EVALUATEDPARAMETERS TO BE EVALUATED
Specificity *Specificity *
Forced degradation study*Forced degradation study*
LOD & LOQ (if applicable)*LOD & LOQ (if applicable)*
Linearity*Linearity*
Precision*Precision*
System Precision (System suitability)*System Precision (System suitability)*
Method Repeatability*Method Repeatability*
Intermediate Precision (or) Ruggedness*Intermediate Precision (or) Ruggedness*
Method Reproducibility**Method Reproducibility**
13. âĸ Accuracy (or) Recovery*
âĸ Solution Stability*
âĸ Robustness*
âĸ System Suitability*
* Included in ICH Guidelines
** Terminology included in ICH guidelines but are not
part of required parameters
14. ī No exact methodology given for each parameterNo exact methodology given for each parameter
ī Only ICH â Q2B and CDER guidelines are provided butOnly ICH â Q2B and CDER guidelines are provided but
not to the extent of 100%not to the extent of 100%
ī Good understanding of each performance characteristicsGood understanding of each performance characteristics
most important. This understanding must be beyond themost important. This understanding must be beyond the
basic definition of each parameterbasic definition of each parameter
ī Understanding must be anchored by sufficient years ofUnderstanding must be anchored by sufficient years of
practical experience and knowledge. It will permit soundpractical experience and knowledge. It will permit sound
and logical decisions, even under the most intenseand logical decisions, even under the most intense
situationssituations
II. METHODOLOGY
15. DEFINITION
ī SpecificitySpecificity of analytical method as its ability to measureof analytical method as its ability to measure
accurately an analyte in the presence of interference, suchaccurately an analyte in the presence of interference, such
as synthetic precursors, excipients, enantiomers andas synthetic precursors, excipients, enantiomers and
known (or likely) degradation product that may beknown (or likely) degradation product that may be
expected to be present in the sample matrix.expected to be present in the sample matrix.
1. SPECIFICITY
16. DISCUSSION
ī SpecificSpecific generally refers to a method that produces agenerally refers to a method that produces a
response for a single analyte onlyresponse for a single analyte only
ī SelectiveSelective refers to a method which provides responses for arefers to a method which provides responses for a
number of chemical entities that may / may not benumber of chemical entities that may / may not be
distinguished from each other. If each response isdistinguished from each other. If each response is
distinguished from all other responses, then the method isdistinguished from all other responses, then the method is
said to be selective.said to be selective.
ī Use of the termUse of the term SelectivitySelectivity is appropriate for the methodsis appropriate for the methods
based on techniques such as HPLC, GC methodsbased on techniques such as HPLC, GC methods..
1. SPECIFICITY
17. 1.SPECIFICITY
EVALUATION
Identification tests: Response for compound of interest only
Assay: Peak purity of analyte peak.
Impurities: Resolution between within the impurity(s) and/or
degradants and form analyte
Peak purity of - analyte peak (for un-spiked)
- analyte and impurity(s) peaks (for spiked samples)
18. 1.SPECIFICITY
ACCEPTANCE CRITERIA
īIdentification Tests: Positive response for compound of interest only
īAssay : No peak should be found at the retention time of analyte peak
and Peak purity of analyte peak should pass.
īImpurities : Should pass Peak purity of -main analyte and Impurity
peaks
No peak should be found at the retention time of analyte/Impurity
īDissolution: No peak should be found at the retention time of analyte.
In case of UV methodology, % difference should be not more than 2.0
19. 1.SPECIFICITY
FORCED DEGRADATION STUDIES
INTRODUCTIONINTRODUCTION
īForced degradation or stress testing is undertaken toForced degradation or stress testing is undertaken to
demonstratedemonstrate specificityspecificity when developing stability-indicatingwhen developing stability-indicating
methodsmethods
īA stability-indicating method is one that accurately quantitatesA stability-indicating method is one that accurately quantitates
the active ingredients without interference from degradationthe active ingredients without interference from degradation
products, process impurities, excipients or other potentialproducts, process impurities, excipients or other potential
impuritiesimpurities
20. ī Address the stability of the compound
ī Establish the degradation pathway
ī Identify the degradation products
ī Validate the stability indicating power of the
analytical procedures used
21. 1.SPECIFICITY / SELECTIVITY (Contd.,)
FORCED DEGRADATION STUDIES
PROCEDUREPROCEDURE
Perform analysis for each stressed (acid / base / peroxide / thermal /Perform analysis for each stressed (acid / base / peroxide / thermal /
photolytic / humidity) sample as per methodologyphotolytic / humidity) sample as per methodology
Normal initial stressed conditions to be appliedNormal initial stressed conditions to be applied
1M HCl1M HCl
1M NaOH1M NaOH
10% H10% H22OO22
105°C/at least 72 Hours105°C/at least 72 Hours
12000 Lux/at least 72 Hours12000 Lux/at least 72 Hours
92% RH/25°C/at least 72 Hours92% RH/25°C/at least 72 Hours
22. ī¨ Initiated at an early stage of development
ī¨ Repeated as methods, processes or formulations
change, so it is an ongoing effort.
ī¨ Evaluate the each unique formulation before formal
stability begins
23. EVALUATIONEVALUATION
Assay:Assay: % Difference of assay for Control (Un-stressed) and each Stressed% Difference of assay for Control (Un-stressed) and each Stressed
samplessamples
Peak purity of analyte peak for Control and stressed samplePeak purity of analyte peak for Control and stressed sample
Impurities:Impurities: Peak purity of analyte peak for Control and each Stressed samplePeak purity of analyte peak for Control and each Stressed sample
ACCEPTANCE CRITERIAACCEPTANCE CRITERIA
Assay:Assay: Peak purity of analyte peak in Control and each Stressed samples shouldPeak purity of analyte peak in Control and each Stressed samples should
passpass
Impurities:Impurities: Peak purity of analyte peak in Control and each Stressed samplesPeak purity of analyte peak in Control and each Stressed samples
should passshould pass
1.SPECIFICITY / SELECTIVITY (Contd.,)
FORCED DEGRADATION STUDIES
24. Points to rememberPoints to remember
If the degradation media degrades the drug substance/drug product to too great extentIf the degradation media degrades the drug substance/drug product to too great extent
or do not degrade the drug substance/drug product at all, then alternative action shouldor do not degrade the drug substance/drug product at all, then alternative action should
be taken (e.g., change the strength of the degradation medium or exposure time or applybe taken (e.g., change the strength of the degradation medium or exposure time or apply
heat over a period of time to achieve minimum level of degradation)Â heat over a period of time to achieve minimum level of degradation)Â
Different numerical values were proposed for the extent of degradation in recentDifferent numerical values were proposed for the extent of degradation in recent
literatureliterature
* Minimum 5%* Minimum 5%
Some compound may not necessarily degrade under a given stress condition. NoSome compound may not necessarily degrade under a given stress condition. No
further stressing is advised in these casesfurther stressing is advised in these cases
Over stressing may lead to the formation of secondary degradants, causes disturbance toOver stressing may lead to the formation of secondary degradants, causes disturbance to
thethe selectivityselectivity of the method, requires further developmentof the method, requires further development
1.SPECIFICITY / SELECTIVITY (Contd.,)
FORCED DEGRADATION STUDIES
25. Hence, the forced degradation studies should mimic the conditions to which theHence, the forced degradation studies should mimic the conditions to which the
drug substance / drug product is actually exposed during its shelf lifedrug substance / drug product is actually exposed during its shelf life
A simple logic behind these studies is, ability to separate the analyte ofA simple logic behind these studies is, ability to separate the analyte of
interest from its degradation impurities formed, if any, caused byinterest from its degradation impurities formed, if any, caused by
quality of chemicals used for its manufacturing, processing,quality of chemicals used for its manufacturing, processing,
packaging, storage, shipment, and/or any unexpected exposurepackaging, storage, shipment, and/or any unexpected exposure
during shelf life of the drug substance/drug productduring shelf life of the drug substance/drug product
1.SPECIFICITY / SELECTIVITY (Contd.,)
FORCED DEGRADATION STUDIES
26. 2. LIMIT OF DETECTION AND LIMIT OF QUANTIFICATION
(LOD & LOQ)
DEFINITIONDEFINITION
LOD:LOD: Lowest amount of analyte in a sample which can be detected but notLowest amount of analyte in a sample which can be detected but not
necessarily quantitated, under the stated experimental conditions (LOD)necessarily quantitated, under the stated experimental conditions (LOD)
LOQ:LOQ: Lowest amount of analyte in a sample which can be quantitativelyLowest amount of analyte in a sample which can be quantitatively
determined with suitable precision and accuracy (LOQ)determined with suitable precision and accuracy (LOQ)
27. Different approaches suggested by ICH, USP & EP
Several approaches are given in the ICH guidelines for estimating LOD and LOQSeveral approaches are given in the ICH guidelines for estimating LOD and LOQ
Depending on the procedure : instrumental / non-instrumentalDepending on the procedure : instrumental / non-instrumental
ApproachApproach LODLOD LOQLOQ
Visual inspectionVisual inspection Minimum level detectableMinimum level detectable Minimum level quantifiableMinimum level quantifiable
Signal-to-Noise ratioSignal-to-Noise ratio 2:1 or 3:12:1 or 3:1 10:110:1
SD of response (SD of response (ĪĪ) &) &
Slope (S)Slope (S)
[3:3 x[3:3 x ĪĪ] / S] / S [10.0 x[10.0 x ĪĪ] / S] / S
RSD CriteriaRSD Criteria Concentration at whichConcentration at which
RSD10 to 33.0%RSD10 to 33.0%
Concentration at which RSDConcentration at which RSD
â¤â¤10.0%10.0%
As per ICH and USP, other approaches suggested above are also
acceptable
2. LOD & LOQ (Contd.,)
28. PROCEDUREPROCEDURE
SD of response (SD of response (ī˛ī˛) & Slope (S)) & Slope (S): Prepare linearity curve with a series of related: Prepare linearity curve with a series of related
substance(s) solutions at different concentrations (3 concentrations below 50 % ofsubstance(s) solutions at different concentrations (3 concentrations below 50 % of
specification level and 3 more concentrations above 50 % specification level)specification level and 3 more concentrations above 50 % specification level)
RSD criteria:RSD criteria: Prepare a series of related substance(s) solutions of differentPrepare a series of related substance(s) solutions of different
concentrations below to specification level (generally about 10%, 20%, 30%, 40% andconcentrations below to specification level (generally about 10%, 20%, 30%, 40% and
50% of specification concentration) and inject six replicate injections into HPLC.50% of specification concentration) and inject six replicate injections into HPLC.
Precision should be established (if predicted from other than RSD criteria) at LOQ andPrecision should be established (if predicted from other than RSD criteria) at LOQ and
LOD level as per ICH, USP & EP guidelinesLOD level as per ICH, USP & EP guidelines
** Prepare the solution at predicted concentration (for LOQ/LOD) and inject in to sixPrepare the solution at predicted concentration (for LOQ/LOD) and inject in to six
replicates as per methodologyreplicates as per methodology
2. LOD & LOQ (Contd.,)
29. 2. LOD & LOQ (Contd.,)
EVALUATIONEVALUATION
SD of response (SD of response (ī˛ī˛) & Slope (S)) & Slope (S):: Calculate residual standard deviation on response âCalculate residual standard deviation on response âī˛ī˛ââ
(also called residual standard deviation on Y- intercept or residual standard error or mean(also called residual standard deviation on Y- intercept or residual standard error or mean
square error or residual sum of squares) and slope (S) obtained from regression datasquare error or residual sum of squares) and slope (S) obtained from regression data
LOD = 3.3 xLOD = 3.3 x ī˛ī˛ / S LOQ = 10 x/ S LOQ = 10 x ī˛ī˛ / S/ S
  RSD criteria:RSD criteria: Determine RSD for impurity response for each concentration and declareDetermine RSD for impurity response for each concentration and declare
the LOQ concentration in âÂĩg/mLâ at which RSD of six replicate injections isthe LOQ concentration in âÂĩg/mLâ at which RSD of six replicate injections is â¤â¤ 10.0%10.0%
and similarly, LOD concentration in âÂĩg/mLâ at which RSD of six replicate injections isand similarly, LOD concentration in âÂĩg/mLâ at which RSD of six replicate injections is
betweenbetween >> 10.0% and10.0% and â¤â¤ 33.0%33.0%
ACCEPTANCE CRITERIAACCEPTANCE CRITERIA
(if predicted from other than RSD criteria)(if predicted from other than RSD criteria)
RSD of six replicate injections isRSD of six replicate injections is â¤â¤ 10.0% for LOQ and between10.0% for LOQ and between >> 10.0% and10.0% and â¤â¤ 33.0%33.0%
30. 3. LINEARITY
DEFINITIONDEFINITION
TheThe LinearityLinearity of an analytical procedure is its ability (within a given range) toof an analytical procedure is its ability (within a given range) to
elicit (obtain) test results that are directly proportional to the concentrationelicit (obtain) test results that are directly proportional to the concentration
(amount) of analyte in the sample(amount) of analyte in the sample
Range: The interval between the upper and lower level( Including these level)Range: The interval between the upper and lower level( Including these level)
that have been demonstrated to be determined with precision, accuracy andthat have been demonstrated to be determined with precision, accuracy and
linearity using this method as writtenlinearity using this method as written
DISCUSSIONDISCUSSION
In order to determine the quantity of any analyte present in unknown sample,In order to determine the quantity of any analyte present in unknown sample,
some kind of relation ship (mathematical/empirical) between concentration andsome kind of relation ship (mathematical/empirical) between concentration and
response is essentialresponse is essential
ANALYTE ------ PROCEDURE ------ANALYTE ------ PROCEDURE ------ī ī RESPONSERESPONSE
ICH and USP encouraging Linear and non-Linear relation shipsICH and USP encouraging Linear and non-Linear relation ships
31. PROCEDUREPROCEDURE
Prepare a series of solutions (not less than five is recommended) with standard /Prepare a series of solutions (not less than five is recommended) with standard /
reference samples in the specified concentration range and analyze them as perreference samples in the specified concentration range and analyze them as per
methodmethod
Assay:-Assay:- 80% to 120% of test concentration80% to 120% of test concentration
CU :-CU :- 70% to 130% of70% to 130% of test concentrationtest concentration
Dissolution:-Dissolution:- Âą 20% of expected release (Q) for immediate releaseÂą 20% of expected release (Q) for immediate release
0 to 120% (for extended release)0 to 120% (for extended release)
Impurities:-Impurities:- LOQ to 200% of specificationLOQ to 200% of specification
3. LINEARITY (Contd.,)
32. 3. LINEARITY (Contd.,)
EVALUATIONEVALUATION
SlopeSlope
- indicates sensitivity of the method- indicates sensitivity of the method
InterceptIntercept
- indicates response for no analyte (interference)- indicates response for no analyte (interference)
Residual sum of squaresResidual sum of squares
- indicates uncertainty of intercept(in blank response)- indicates uncertainty of intercept(in blank response)
Correlation CoefficientCorrelation Coefficient
- indicates the relation ship chosen is correct- indicates the relation ship chosen is correct
ACCEPTANCE CRITERIAACCEPTANCE CRITERIA
Correlation Coefficient should be not less than 0.999 for assay, CU, dissolution testCorrelation Coefficient should be not less than 0.999 for assay, CU, dissolution test
methods and 0.99 for impurities test methodmethods and 0.99 for impurities test method
33. 4. PRECISION (Contd.,)
DEFINITIONDEFINITION
Closeness of agreement (degree of scatter) between a series of measurementsCloseness of agreement (degree of scatter) between a series of measurements
obtained from multiple sampling of the same homogenous sample under theobtained from multiple sampling of the same homogenous sample under the
prescribed conditions.prescribed conditions.
Precision may be considered at three levels.Precision may be considered at three levels.
1.1. System precision (System suitability)System precision (System suitability)
2.2. Method RepeatabilityMethod Repeatability
3.3. Intermediate precisionIntermediate precision
4.4. ReproducibilityReproducibility
34. 4. PRECISION (Contd.,)
DISCUSSIONDISCUSSION
īļ Repeatability:Repeatability: precision under same operating conditions (with-in a laboratoryprecision under same operating conditions (with-in a laboratory
over a short period of time using the sameover a short period of time using the same analyst with the same equipment)analyst with the same equipment)
īļMeasurement / Injection repeatability (System Precision)Measurement / Injection repeatability (System Precision)
īļMethod repeatability (Method Precision)Method repeatability (Method Precision)
īļIntermediate precisionIntermediate precision: precision under different laboratory conditions (within-: precision under different laboratory conditions (within-
laboratory variation, as on different days, or with different analysts, or equipmentslaboratory variation, as on different days, or with different analysts, or equipments
within the same laboratory)within the same laboratory)
īļReproducibilityReproducibility: precision between laboratories / intermediate precision can be: precision between laboratories / intermediate precision can be
considered during the standardization of a procedure before it is submitted to theconsidered during the standardization of a procedure before it is submitted to the
pharmacopoeiapharmacopoeia
As per CDER guidelines, it is not normally expected if intermediate precision isAs per CDER guidelines, it is not normally expected if intermediate precision is
accomplishedaccomplished
35. 4.PRECISION (Contd.,)
PROCEDUREPROCEDURE
īŋ Six replicate measurements/injections of standard preparation (System Precision)Six replicate measurements/injections of standard preparation (System Precision)
īŋ Six replicate analysis of the samples through the complete analytical procedure fromSix replicate analysis of the samples through the complete analytical procedure from
sample preparation to final result (Method Precision)sample preparation to final result (Method Precision)
īŋ Three different matrices at 80%, 100% and 120%(Triplicate preparation atThree different matrices at 80%, 100% and 120%(Triplicate preparation at
triplicate concentration)triplicate concentration)
īŋ Six replicate analysis of the samples through the complete analytical procedure fromSix replicate analysis of the samples through the complete analytical procedure from
sample preparation to final result by two different analysts, columns, instruments,sample preparation to final result by two different analysts, columns, instruments,
different daysdifferent days (Ruggedness)(Ruggedness)
36. 4.PRECISION (Contd.,)
ACCEPTANCE CRITERIA
ProcedureProcedure
%RSD%RSD
System PrecisionSystem Precision Method PrecisionMethod Precision RuggednessRuggedness
APIAPI DPDP APIAPI DPDP APIAPI DPDP
AssayAssay 1.0/2.01.0/2.0 1.0/2.0*1.0/2.0* 1.01.0 2.02.0 1.01.0 3.03.0
DissolutionDissolution NANA 1.0 / 2.0*1.0 / 2.0* NANA 6.06.0 NANA 5.05.0
ImpuritiesImpurities 5.0*5.0* 5.0*5.0* 10.010.0 10.010.0 10.010.0 15.015.0
NA â Not applicable
API â Active Pharmaceutical Ingredient
DP â Drug Product
* : As given in STP
37. 5. ACCURACY
DEFINITIONDEFINITION
īThe accuracy of an analytical procedure expresses theThe accuracy of an analytical procedure expresses the
closeness of agreement between the value, which is acceptedcloseness of agreement between the value, which is accepted
either as a conventional true value or an accepted reference valueeither as a conventional true value or an accepted reference value
and the value foundand the value found
38. 5.ACCURACY (Contd.,)
PROCEDURE
Assay/Dissolution:- Known amount of drug substance spiked with synthetic
mixtures of drug product components (excipients) â minimum of three levels
īŧ80%, 100% & 120% of test concentration-assay;
īŧÂą 20% of expected release-dissolution), each level is triplicate
Impurities:- drug substance/drug product spiked with known amounts of impurities
- minimum of three levels
īŧLOQ level to 200% of specification and
īŧAt LOQ level, each level is triplicate
EVALUATION
- Recovery from amount added and amount found
- Precision (%RSD) at each level (for three replicate preparations)
39. 5.ACCURACY (Contd.,)
ACCEPTANCE CRITERIA
Assay:- Recovery should be between 98% to 102%( Depends upon the your
Strength)
Dissolution:- 95% to 105%
Impurities:- if, Specification is ⤠0.2% : 85% to 115%
if, Specification is > 0.2% : 90% to 110%
At LOQ level : 80% to 120%
A simple logic behind this performance characteristic is whether
the procedure is capable of estimating a true value or not
40. 7. STABILITY
DISCUSSIONDISCUSSION
īļ It is often essential that solutions (standards, test samples) be stable enoughIt is often essential that solutions (standards, test samples) be stable enough
to allow for delays covering instrument break downs / overnight analysesto allow for delays covering instrument break downs / overnight analyses
īļ A minimum of 12 Hrs, 18 Hrs, or 24 Hrs is routinely recommended forA minimum of 12 Hrs, 18 Hrs, or 24 Hrs is routinely recommended for
chromatographic methods for which vialed solutions may remain on an auto-chromatographic methods for which vialed solutions may remain on an auto-
samplers at ambient temperatures due to various delays.samplers at ambient temperatures due to various delays.
41. 7. STABILITY (Contd.,)
PROCEDUREPROCEDURE
Prepare test sample as per procedure and analyze at initial and at different timePrepare test sample as per procedure and analyze at initial and at different time
intervals by keeping the sample at room temperature ( 25°C) / refrigeratorintervals by keeping the sample at room temperature ( 25°C) / refrigerator
condition (2-8°C)condition (2-8°C)
EVALUATIONEVALUATION
% Difference from initial response to specified interval for analyte / each impurity% Difference from initial response to specified interval for analyte / each impurity
ACCEPTANCE CRITERIAACCEPTANCE CRITERIA
% Difference is not more than% Difference is not more than
Assay, CU :Assay, CU : 2.02.0
Dissolution :Dissolution : 3.03.0
Impurities :Impurities : 10.010.0
42. 7. STABILITY (Contd.,)
A simple logic behind this study is to determine the periodA simple logic behind this study is to determine the period
of time, a solution can be held before analysis withoutof time, a solution can be held before analysis without
compromising accuracycompromising accuracy..
43. 8. ROBUSTNESS
DEFINTIONDEFINTION
Measure of its capacity to remain unaffected by small, but deliberate variationsMeasure of its capacity to remain unaffected by small, but deliberate variations
in method parameters and provides indication of its reliability during its normalin method parameters and provides indication of its reliability during its normal
usageusage
DISCUSSIONDISCUSSION
varying method parameters within a realistic range and the quantitativevarying method parameters within a realistic range and the quantitative
influence of the variables is determined, and, if the influence of the parameter isinfluence of the variables is determined, and, if the influence of the parameter is
within a previously specified tolerance, then, the parameter is said to be withinwithin a previously specified tolerance, then, the parameter is said to be within
the methodâs robustness rangethe methodâs robustness range
According to ICH guidelines, robustness should be considered early in theAccording to ICH guidelines, robustness should be considered early in the
development stage of a method, but it is not required to be included as part of adevelopment stage of a method, but it is not required to be included as part of a
registration application.registration application.
44. 8. ROBUSTNESS (Contd.,)
Typical variations include under validation programmeTypical variations include under validation programme
ī Flow rate (Flow rate (īĄīĄ 10%)10%)
ī Wavelength (Wavelength (īĄīĄ 2 nm)2 nm)
ī Mobile phase composition, generally, Organic composition (Mobile phase composition, generally, Organic composition (īĄīĄ 2 (or) 5%)2 (or) 5%)
ī Temperature (Temperature (īĄīĄ 5° C)5° C)
ī pH of the mobile phase (pH of the mobile phase (īĄīĄ 0.2 units)0.2 units)
45. 8. ROBUSTNESS (Contd.,)
PROCEDUREPROCEDURE
Assay:Assay:Analysis of Resolution (if applicable), Standard & Test samples (2Analysis of Resolution (if applicable), Standard & Test samples (2
replicates) by proposed analytical methodology and the method operated atreplicates) by proposed analytical methodology and the method operated at
variable conditions.variable conditions.
Impurities:Impurities:Analysis of Resolution & Test sample by proposed analyticalAnalysis of Resolution & Test sample by proposed analytical
methodology and the method operated at variable conditions.methodology and the method operated at variable conditions.
EVALUATIONEVALUATION
Assay:Assay:System Suitability parameters at all variable conditionsSystem Suitability parameters at all variable conditions
% Assay of samples at all variable conditions% Assay of samples at all variable conditions
Impurities:Impurities: System Suitability parameters at all variable conditionsSystem Suitability parameters at all variable conditions
RRTs at all variable conditionsRRTs at all variable conditions
( monitor the separation at each variable condition)( monitor the separation at each variable condition)
46. 8. ROBUSTNESS (Contd.,)
ACCEPTANCE CRITERIAACCEPTANCE CRITERIA
Assay:Assay:System Suitability criteria should meet at each variable conditionSystem Suitability criteria should meet at each variable condition
Overall RSD for % assay results obtained at STP condition and variable conditionOverall RSD for % assay results obtained at STP condition and variable condition
for each variabilityfor each variability
Impurities:Impurities:System Suitability criteria should meet at each variable conditionSystem Suitability criteria should meet at each variable condition
47. III Time management in validation
There are no official guidelines on the sequence of validation
experiments and the optimal sequence can be depending on the
method itself. Based on experience, for HPLC method the
following sequence has been proven to be useful for time
management
If the method is proved as stable andIf the method is proved as stable and
robust under method developmentrobust under method development
(Pre validation) programme(Pre validation) programme
In case of stability and robustnessIn case of stability and robustness
data is not available with methoddata is not available with method
development datadevelopment data
SpecificitySpecificity StabilityStability
LinearityLinearity RobustnessRobustness
LOD & LOQ (if applicable)LOD & LOQ (if applicable) SpecificitySpecificity
PrecisionPrecision LinearityLinearity
AccuracyAccuracy LOD & LOQ (if applicable)LOD & LOQ (if applicable)
RangeRange PrecisionPrecision
StabilityStability AccuracyAccuracy
RobustnessRobustness RangeRange
48. IV VALIDATION REPORT
Every DMF / ANDA / COS data package submitting for US FDA and
European community etc., should consist method validation data. For
easy to review, method validation report is usually attached to package.
īĄ Generally method validation report should have
ī Objective and scope of the method
ī Molecule details (IUPAC name, CAS No. Molecular Formulae,
Molecular weight and its Molecular Structure etc.,)
ī Detailed list of chemicals, reagents, reference standards
ī Listing of equipment and its functional and performance requirements
ī Methodology followed
ī Validation data (parameter wise â procedure, results, conclusion etc.,)
ī Summary of data (results in brief â parameter wise)
ī Summary of report (overall view of validation exercise, any critical
issues, recommendations etc., for the application of method)
49. IV. VALIDATION REPORT (Contd.,)
Instrument out puts, which should represent critical method
parameters
ī Specificity and LOD â for Identification Test (Generally,
photographs)
ī Selectivity / Specificity data (discriminating chromatogram, peak
purity data, blank and placebo chromatograms and stressed samples
chromatograms)
ī Linearity graphs
ī Resolution and related system suitability chromatograms
ī any out puts which are significant