Validation is the process of confirming an analytical method is suitable for its intended purpose. The key steps in validation include assessing accuracy, precision (repeatability and reproducibility), specificity, linearity, range, detection limit, and robustness. Accuracy demonstrates closeness between true and measured values. Precision expresses the degree of variation in measurements. Specificity ensures only the intended analyte is measured. Linearity evaluates the relationship between concentration and response. Range confirms acceptable accuracy and precision within the method's measurable concentrations. [END SUMMARY]
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 intra—assay 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
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 intra—assay 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
A separation technique in which the mobile phase is a gas. Gas chromatography is always carried out in a column.
Separating mixtures of gases or volatile materials based primarily on their physical properties.
This presentation was made to solely for students to make them aware/ understand basics of “Analytical Method Validation”. These slides are part of lectures delivered in M. Pharmacy Curriculum & taken up from various books and websites
A separation technique in which the mobile phase is a gas. Gas chromatography is always carried out in a column.
Separating mixtures of gases or volatile materials based primarily on their physical properties.
This presentation was made to solely for students to make them aware/ understand basics of “Analytical Method Validation”. These slides are part of lectures delivered in M. Pharmacy Curriculum & taken up from various books and websites
Chromatography
Is a technique used to separate and identify the components of a mixture.
Works by allowing the molecules present in the mixture to distribute themselves between a stationary and a mobile medium. Molecules that spend most of their time in the mobile phase are carried along faster.
Gas Liquid Chromatography
Here the mobile phase is an unreactive gas ( eg Nitrogen) flowing through a tube.
And the stationary phase is an involatile liquid held on particles of a solid support.
In the animation below the red molecules are more soluble in the liquid (or less volatile) than are the green molecules.
In practice the Column is contained in a thermostatic oven. (Why ?)
About 1μL of liquid is injected into one end of the column.
As each component reaches the other end it is detected and registered on a chart recorder.
The Retention Time is characteristic of a particular substance. (for the same column, temperature, gas flow etc.)
The area under each peak indicates the relative quantities.
Thin Layer Chromatography
Here the mobile phase is a liquid
Flowing past a thin layer of powder on a solid support.
Substances that are less attracted to the solid or are more soluble in the liquid move faster.
And so move further up the plate by the time that the process has been stopped by taking the plate out of the liqiud. - larger Rf
Rf = distance moved by substance
distance moved by solvent front
For substances that are very soluble in the liquid Rf will be close to ....1
For substances that are rather insoluble in the liquid Rf will be close to 0 ....
khalid hussain 3rd proff Morning
islamia university bhawalpur
Analytical method validation as per ich and usp shreyas B R
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.
This content is suitable for medical technologists/technicians/lab assistants/scientists writing the SMLTSA board exam. The content is also suitable for biomedical technology students and people also interested in learning about test methodologies used in medical technology. This chapter describes test quality assurance (QA) and quality control (QC). Please note that these notes are a collection I used to study for my board exam and train others who got distinctions using these.
Disclaimer: Credit goes to those who wrote the notes and the examiners of each exam question. Please use only as a reference guide and use your prescribed textbook for the latest and most accurate notes and ranges. The material here is not referenced as it is a collection of pieces of study notes from multiple people, and thus will not be held viable for any misinterpretations. Please use at your own discretion.
Systematic error means that your measurements of the same thing will vary in predictable ways: every measurement will differ from the true measurement in the same direction, and even by the same amount in some cases
Random error is a chance difference between the observed and true values of something (e.g., a researcher misreading a weighing scale records an incorrect measurement).
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.
2. Validation?
• Validation is the assessment of a process or
instrument to assure that the process and
instrument is suitable for its intended use
(FDA, 1987).
• Validation enables an efficient and productive
use of the process and instrumental
variables.
• A new assay method, change in operator,
laboratory and equipment than the one in
previous method requires validation.
3. Steps in Validation
REPEATABILITY
PRECISION
ACCURACY Reproducibility
INTERMEDIATE
Sensitivity PRECISION
SPECIFICITY
DETECTION RANGE
LINEARITY QUANTIFICATION
4. Specificit
y
• Specificity is the ability to assess
unequivocally the analyte in the presence of
other components such as impurities,
degradants and matrix etc.
• Lack of specificity of an assay procedure, may
be compensated by other supporting assay(s).
5. • Indicates the linear relationship
between concentration and
response of the detector
6. EVALUATION OF
LINEARITY
• Linearity is evaluated by visual inspection of a plot of
signals as a function of analyte concentration.
• If there is a linear relationship, the data is evaluated by
appropriate statistical methods, for example, linear
regression.
• In some cases, to obtain linearity, the test data may need
mathematical transformation prior to the regression
analysis.
• For the establishment of linearity, a minimum of 5
concentrations are recommended.
7. RANGE
• The specified range is normally derived
from linearity studies and depends on the
intended application of the procedure.
• It confirms that the assay will provide an
acceptable accuracy and precision when
applied to samples containing analyte,
within or at the extremes of the specified
range
8. Applicable Concentration Range
LOL
Instrument response
LOQ => limit of quantitative
measurement
LOQ LOL => limit of linear
response
Useful range
Concentration
9. ACCURACY
• The accuracy of an assay expresses the
closeness or agreement between the true value
and the value found. This is sometimes termed
as trueness
• Within day accuracy (Repeatability)
• Between days accuracy (Reproducibility)
10. PRECISION
• The precision of an assay expresses the degree of
scatter between a series of measurements
obtained from multiple sampling of the same
homogeneous sample under the prescribed
conditions.
• Precision may be considered at three levels:
– Repeatability
– Intermediate precision
– Reproducibility
• The precision is usually expressed as the
variance, standard deviation or coefficient of
variance of a series of measurements.
11. Repeatability
• Repeatability expresses the precision under the
same operating conditions over a short interval
of time.
• Repeatability is also termed as intra-assay
precision.
12. Intermediate precision
• Intermediate precision expresses
within-laboratories variations,
different days, different analysts,
different equipment, etc.
• Typical variations to be studied
include days, analysts, equipment
etc.
18. DETECTION LIMIT
• The detection limit of an analyte in an assay is
the lowest amount of analyte in a sample
which can be detected but not necessarily
quantitated as an exact value
• Several approaches for determining the
detection limit are used, depending on whether
the procedure is a non-instrumental or
instrumental
19. Based on Visual Evaluation
• Visual evaluation may be used for non-
instrumental methods but may also be used
for instrumental methods.
• The detection limit is determined by the
analyzing samples having known
concentrations of analyte and by establishing
the minimum level at which the analyte can
reliably be detected
20. Based on Signal-to-Noise
• This approach is applied to analytical procedures
which exhibit baseline noise.
• Determination of the signal-to-noise ratio is
performed by comparing measured signals of
samples having known low concentration of
analyte with those of blanks , and establishing the
minimum concentration at which the analyte can
reliably be detected.
• A signal-to-noise ratio between 3:1 or 2:1 is
generally considered acceptable for estimating the
detection limit.
21. Based on the Standard Deviation of
the Response and the Slope
The detection limit (DL) may be
expressed as:
DL = 3.3 σ /S
where σ = the standard deviation of
the intercept
S = mean of the slope of the
calibration curve