This document outlines the validation of an analytical method for the quantification of paracetamol using UV spectrophotometry. It describes the validation parameters that will be tested which include accuracy, precision, linearity, range, limit of detection and limit of quantification, selectivity and specificity, and robustness and ruggedness. The procedure involves preparing calibration standards of paracetamol to generate a linear curve and then testing the method's accuracy by spiking samples. Precision will be evaluated by repeatability, intraday, and interday testing. The document provides the theory and equations needed to calculate the validation parameters.
IPQC?
Its Need
In-Process Quality Control tests for Tablets
Hardness
Friability
Thickness
Disintegration Time
Weight variation
Content uniformity
Dissolution test
Leakage testing for strip and blister packaging
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.
QUALIFICATION OF UV-VISIBLE SPECTROPHOTOMETER, FTIR, DSC, HPLCAnupriyaNR
Analytical method qualification consists of a simplified evaluation of a subset of validation characteristics with a goal to demonstrate that an analytical method is scientifically sound and suitable for its intended use. In contrast to validation, analytical method qualification is performed without predefined acceptability criteria. Qualification may be performed as a prerequisite to method validation, or when an assay for product knowledge has not yet been established as a test for a critical product quality attribute. Qualification of equipment is pre-requisite for validation of the process in which the equipment is being used. Many types of equipment have measuring devices on them. Calibration of measuring devices is a part of qualification. Calibration of measuring devices is important, as the data is often collected through them. If the data collected is not from measuring devices that have been calibrated, the data cannot be relied upon. Thus the whole validation exercise can be questioned.
IPQC?
Its Need
In-Process Quality Control tests for Tablets
Hardness
Friability
Thickness
Disintegration Time
Weight variation
Content uniformity
Dissolution test
Leakage testing for strip and blister packaging
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.
QUALIFICATION OF UV-VISIBLE SPECTROPHOTOMETER, FTIR, DSC, HPLCAnupriyaNR
Analytical method qualification consists of a simplified evaluation of a subset of validation characteristics with a goal to demonstrate that an analytical method is scientifically sound and suitable for its intended use. In contrast to validation, analytical method qualification is performed without predefined acceptability criteria. Qualification may be performed as a prerequisite to method validation, or when an assay for product knowledge has not yet been established as a test for a critical product quality attribute. Qualification of equipment is pre-requisite for validation of the process in which the equipment is being used. Many types of equipment have measuring devices on them. Calibration of measuring devices is a part of qualification. Calibration of measuring devices is important, as the data is often collected through them. If the data collected is not from measuring devices that have been calibrated, the data cannot be relied upon. Thus the whole validation exercise can be questioned.
In Process Quality Control (IPQC) of pharmaceutical dosage form in Pharmaceut...Saad Ahmed Sami
A brief description of in process quality control (IPQC) definition, factors affecting the process and IPQC process in solid, liquid and sterile dosage form . IPQC cover the entire chain of operations from the receipt of raw material in the warehouse to the release of finished products from the warehouse for distribution and or sale. IPQC is a process where quality of a product is ensured that it meets the standard according to regulatory authority guideline.
Definition
Scope of calibration
Scope of validation
Frequency of calibration
Importance/ purpose of calibration
Importance/ advantages of validation
Difference between calibration & validation
QUALIFICATION & VALIDATION.Validation is an essential part of GMP, and an element of QA.Critical steps in the process need to be validated.Need for confidence that the product will consistently meet predetermined specifications and attributes.
IPQC Tests for capsules As per IP, BP & USPPramod Ramane
IPQC- In Process Quality Control Tests for Capsules are
1. Uniformity Of Content
2. Disintigration Test
3. Weight Variation Test
4. Dissolution Test
The tests are with Acceptance limits/Criteria as per Indian Pharmacopoeia (IP), British Pharmacopoeia (BP) & United States Pharmacopoeia (USP)
The drug or drug combination may not be official in any pharmacopoeias.
A proper analytical procedure for the drug may not be available in the literature due to patent regulations.
Analytical methods may not be available for the drug in the form of a formulation due to the interference caused by the formulation excipients.
Analytical methods for the quantitation of the drug in biological fluids may not be available.
Analytical methods for a drug in combination with other drugs may not be available.
The existing analytical procedures may require expensive reagents and solvents. It may also involve cumbersome extraction and separation procedures and these may not be reliable.
Understanding of Analytical Method Validation Approach in Pharmaceutical Industry. Analytical method validation Verification is a wide chapter and a huge scope of applicability. In different types of methods, instrument, measurement approach all can effect the validation effort. However the basic fundamental will remains same, the parameters, acceptance criteria, functionality may vary depending upon the type of method, instrument etc.
In Process Quality Control (IPQC) of pharmaceutical dosage form in Pharmaceut...Saad Ahmed Sami
A brief description of in process quality control (IPQC) definition, factors affecting the process and IPQC process in solid, liquid and sterile dosage form . IPQC cover the entire chain of operations from the receipt of raw material in the warehouse to the release of finished products from the warehouse for distribution and or sale. IPQC is a process where quality of a product is ensured that it meets the standard according to regulatory authority guideline.
Definition
Scope of calibration
Scope of validation
Frequency of calibration
Importance/ purpose of calibration
Importance/ advantages of validation
Difference between calibration & validation
QUALIFICATION & VALIDATION.Validation is an essential part of GMP, and an element of QA.Critical steps in the process need to be validated.Need for confidence that the product will consistently meet predetermined specifications and attributes.
IPQC Tests for capsules As per IP, BP & USPPramod Ramane
IPQC- In Process Quality Control Tests for Capsules are
1. Uniformity Of Content
2. Disintigration Test
3. Weight Variation Test
4. Dissolution Test
The tests are with Acceptance limits/Criteria as per Indian Pharmacopoeia (IP), British Pharmacopoeia (BP) & United States Pharmacopoeia (USP)
The drug or drug combination may not be official in any pharmacopoeias.
A proper analytical procedure for the drug may not be available in the literature due to patent regulations.
Analytical methods may not be available for the drug in the form of a formulation due to the interference caused by the formulation excipients.
Analytical methods for the quantitation of the drug in biological fluids may not be available.
Analytical methods for a drug in combination with other drugs may not be available.
The existing analytical procedures may require expensive reagents and solvents. It may also involve cumbersome extraction and separation procedures and these may not be reliable.
Understanding of Analytical Method Validation Approach in Pharmaceutical Industry. Analytical method validation Verification is a wide chapter and a huge scope of applicability. In different types of methods, instrument, measurement approach all can effect the validation effort. However the basic fundamental will remains same, the parameters, acceptance criteria, functionality may vary depending upon the type of method, instrument etc.
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.
To compare filing process of NDA of different countries of India, US and Euro...Aakashdeep Raval
To compare filing process of NDA of different countries of India, US and Europe.
B) Preparation of global list documents of registration of IND and NDA as per USFDA and Europe.
The Gram stain is a fundamental technique in microbiology used to classify bacteria based on their cell wall structure. It provides a quick and simple method to distinguish between Gram-positive and Gram-negative bacteria, which have different susceptibilities to antibiotics
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Ve...kevinkariuki227
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
TEST BANK for Operations Management, 14th Edition by William J. Stevenson, Verified Chapters 1 - 19, Complete Newest Version.pdf
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
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
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.
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!
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
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2 Case Reports of Gastric Ultrasound
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Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
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
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journey
To perform Analytical method validation of Paracetamol Tablets by UV-spectrophotometric method.
1. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
EXPERIMENT NO.: DATE:
AIM: To perform Analytical method validation of Paracetamol Tablets by UV-
spectrophotometric method.
REFERENCES:
1) ICH Q2 (R1) guidelines for validation of analytical procedures.
2) Indian Pharmacopoeia, 2010 vol. III, Govt. of India Ministry of health and Family
Welfare Published by The Indian Pharmacopoeia Commission , Ghaziabad; 1861-
1862.
REQUIREMETNS:
Apparatus: 100ml & 10ml volumetric flasks, pipettes, beakers.
Reagents: Paracetamol API, Paracetamol Tablet, Methanol, Distilled Water.
Equipments:
1) Shimadzu UV-1800
Sr. No.: A114550/08677
2) Shimadzu UV-1800
Sr. No.: A114549/08780
THEORY:
General Description of Paracetamol:
Name: Paracetamol INN or Acetaminophen USAN.
Dose: 500 mg.
Chemical name: N-Acetyl-p-aminophenol.
Structure:
Class: mild analgesic.
Use: over-the-counter analgesic (pain reliever) and antipyretic (fever reducer).It is
commonly used for the relief of headaches and other minor aches and pains and is a major
ingredient in numerous cold and flu remedies. In combination with opioid analgesics,
2. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
Paracetamol can also be used in the management of more severe pain such as post-surgical
pain and providing palliative care in advanced cancer patients. Though Paracetamol is
used to treat inflammatory pain, it is not generally classified as an NSAID because it
exhibits only weak anti-inflammatory activity.
Half-life: 1–4 hours.
Adverse effects: causes gastrointestinal problems or allergic skin reactions. Blood
dyscrasia (e.g. thrombocytopenia), methaemoglobinemia, and hemolytic anemia are very
rare.
Validation Of Analytical Methods
Validation of an analytical method is documented evidence which provide a high
degree of assurance that the given method will consistently produce a product meeting its
predetermined specifications and quality attributes.
Validation parameters as per ICH
1) Accuracy
The accuracy of an analytical method may be defined as the closeness of the test
results obtained by the method to the true value. It is the measure of the exactness of the
analytical method developed. Accuracy may often be expressed as percent recovery by the
assay of a known amount of analyte added. Accuracy may be determined by applying the
method to samples or mixture of excipients to which known amount of analyte have been
added both above and below the normal levels expected in the samples. Accuracy is then
calculated from the test results as the percentage of the analyte recovered by the assay.
Dosage form assays commonly provide accuracy within 3-5% of the true value. The ICH
documents recommend that accuracy should be assessed using a minimum of nine
determinations over a minimum of three concentration levels, covering the specified range
(i.e. three concentrations and three replicates of each concentration).
2) Precision
The precision of an analytical method is the degree of agreement among individual
test results when the method is applied repeatedly to multiple sampling of homogenous
samples. This is usually expressed as the standard deviation or the relative standard
deviation (coefficient of variation). Precision is a measure of the degree of reproducibility
or of the repeatability of the analytical method under normal operating circumstances.
Repeatability:
Repeatability can be defined as the precision of the procedure when repeated by
same analyst under the same operating conditions (same reagents, equipments, settings
3. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
and laboratory) over a short interval of time. Repeatability is also termed as intra-assay
precision.
The ICH documents recommend that repeatability should be assessed using:
a) A minimum of 9 determinations covering the specified range for the procedure (e.g., 3
concentrations/3 replicates each); or
b) A minimum of 6 determinations at 100% of the test concentration.
It is normally expected that at least six replicates be carried out and a table showing each
individual result provided from which the mean, standard deviation and co-efficient of
variation should be calculated for set of n values.
The RSD values are important for showing degree of variation expected when the
analytical procedure is repeated several times in a standard situation. (RSD below 1% for
bulk drugs and RSD below 2% for assays in finished product).
Intermediate precision:
Intermediate precision expresses within-laboratories variations: different days,
different analysts, different equipment, etc.
The extent to which intermediate precision should be established depends on the
circumstances under which the procedure is intended to be used. The applicant should
establish the effects of random events on the precision of the analytical procedure. Typical
variations to be studied include days, analysts, equipment etc. It is not considered
necessary to study these effects individually. The use of an experimental design (matrix) is
encouraged.
Reproducibility:
Reproducibility means the precision of the procedure when it is carried out under
different conditions-usually in different laboratories-on separate, identical samples taken
from the same homogenous batch of material. Comparisons of results obtained by
different analysts, by the use of different equipments, or by carrying out the analysis at
different times can also provide valuable information.
3) Linearity
The 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.
A linear relationship should be evaluated across the range of the analytical procedure. It
may be demonstrated directly on the drug substance (by dilution of a standard stock
4. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
solution) and/or separate weighing of synthetic mixtures of the drug product components
using the proposed procedure.
Linearity should be evaluated by visual inspection of a plot of signals as a function of
analyte concentration or content. If there is a linear relationship, test results should be
evaluated by appropriate statistical methods, for example, by calculation of a regression
line by the method of least squares. The correlation coefficient, y-intercept, slope of the
regression line and residual sum of squares should be submitted. A plot of the data should
be included. In addition, an analysis of the deviation of the actual data points from the
regression line may also be helpful for evaluating linearity.
The linear range of detectability that obeys Beer’s law is dependent on the compound
analyzed and the detector used. The working sample concentration and samples tested for
accuracy should be in the linear range. The claim that the method is linear is to be justified
with additional mention of zero intercept by processing data by linear least square
regression. Data is processed by linear least square regression declaring the regression co-
efficient and b of the linear equation y= ax + b together with the correlation coefficient of
determination r. For the method to be linear the r value should be close to1.
For the establishment of linearity, a minimum of 5 concentrations is recommended. Other
approaches should be justified.
4) Range
The range of an analytical procedure is the interval between the upper and 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.
The specified range is normally derived from linearity studies and depends on the intended
application of the procedure. The following minimum specified ranges should be
considered:
For the assay of a drug substance or a finished (drug) product: normally from 80 to 120%
of the test concentration;
For content uniformity, covering a minimum of 70 to 130% of the test concentration,
unless a wider more appropriate range, based on the nature of the dosage form (e.g.,
metered dose inhalers), is justified;
For dissolution testing: +/-20 % over the specified range;
e.g., if the specifications for a controlled released product cover a region from20%, after 1
hour, up to 90%, after 24 hours, the validated range would be 0-110% of the label claim.
5. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
5) Limit of Detection and Limit of Quantitation
Limit of Detection (LOD): - The limit of detection is the parameter of limit tests. It is the
lowest level of analyte that can be detected, but not necessarily determined in a
quantitative fashion, using a specific method under the required experimental conditions.
The limit test thus merely substantiates that the analyte concentration is above or below a
certain level.
Limit of Quantitation (LOQ): - Limit of quantitation is a parameter of quantitative
assays for low levels of compounds in sample matrices such as impurities in bulk drugs
and degradation products in finished pharmaceuticals.
The limit of quantitation is the lowest concentration of analyte in a sample that may be
determined with acceptable accuracy and precision when the required procedure is
applied. In many cases, the limit of quantitation is approximately twice the limit of
detection.
Several approaches for determining the detection limit are possible, depending on whether
the procedure is a non-instrumental or instrumental.
I. Based on Visual Evaluation
Visual evaluation may be used for non-instrumental methods but may also be used
with instrumental methods. The detection limit is determined by the analysis of
samples with known concentrations of analyte and by establishing the minimum
level at which the analyte can be reliably detected.
II. Based on Signal-to-Noise
The determination of the limit of detection of instrumental procedures is carried
out by determining the signal-to-noise ratio by comparing test results from the
samples with known concentration of analyte with those of blank samples and
establishing the minimum level at which the analyte can be reliably detected. A
signal-to-noise ratio of 2:1 or 3:1 (For LOD) and 10:1(For LOQ) is generally
accepted. The signal-to-noise ratio is determined by dividing the base peak by the
standard deviation of all data points below a set threshold. Limit of detection is
calculated by taking the concentration of the peak of interest divided by three times
the signal-to-noise ratio.
III. Based on the Standard Deviation of the Response and the Slope
The detection limit may be expressed as:
LOD = 3 * σ / S
LOQ = 10 * σ / S
6. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
Where, σ = the standard deviation of the intercept
S = the slope of the calibration curve
6) Selectivity and Specificity
The selectivity of an analytical method is its ability to measure accurately and
specifically the analyte of interest in the presence of components that may be expected to
be present in the sample matrix.
If an analytical procedure is able to separate and resolve the various components of a
mixture and detect the analyte qualitatively the method is called selective. On the other
hand, if the method determines or measures quantitatively the component of interest in the
sample matrix without separation, it is said to be specific.
Hence one basic difference in the selectivity and specificity is that, while the former is
restricted to qualitative detection of the components of a sample, the latter means
quantitative measurement of one or more analyte.
Selectivity may be expressed in terms of the bias of the assay results obtained when the
procedure is applied to the analyte in the presence of expected levels of other components,
compared to the results obtained on the same analyte without added substances. When the
other components are all known and available, selectivity may be determined by
comparing the test results obtained on the analyte with and without the addition of the
potentially interfering materials. When such components are either unidentified or
unavailable, a measure of selectivity can often be obtained by determining the recovery of
standard addition of pure analyte to a material containing a constant level of the other
components.
7) Robustness and Ruggedness
Robustness: - The robustness of an analytical method is a measure of its capacity to
remain unaffected by small but deliberate variation in method parameters and provides an
indication of its reliability during normal usage.
The evaluation of robustness should be considered during the development phase and
depends on the type of procedure under study. It should show there liability of an analysis
with respect to deliberate variations in method parameters.
Ruggedness: - The ruggedness of an analytical method is the degree of reproducibility of
test results obtained by the analysis of the same samples under a variety of normal test
conditions such as different laboratories, different analysts, using operational and
environmental conditions that may differ but are still within the specified parameters of
the assay.
7. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
For the determination of ruggedness, the degree of reproducibility of test result is
determined as function of the assay variable. This reproducibility may be compared to the
precision of the assay under normal condition to obtain a measure of the ruggedness of the
analytical method.
PROCEDURE:
Linearity
Stock solution:
Stock solution (100g/ml) of PCM was prepared by dissolving 0.01 g PCM in 100 ml
volumetric flasks and completing the volume distilled water.
Selection of analytical wavelength for PCM:
The solution of PCM was prepared in Water at a concentration of 15 µg /ml from the stock
solution. It was scanned in the wavelength range of 200-400 nm. Maximum absorbance
was obtained at 245 nm. This analytical wavelength was selected for determination of
PCM.
For calibration curve:
An aliquots of stock solution of PCM (0.5, 1, 1.5,2, and 2.5 ml) were pipettes out in 10ml
volumetric flasks and further diluted to attain concentration of about 5, 10, 15, 20, 25
µg/ml.
Now plot the graph of Absorbance Vs Concentration. Find out the r2
value from the
Graph.
Accuracy
API: From the stock solution (100µg/ml) pipette out 1 ml of solution in 3 volumetric
flasks, further add 80,100 & 120% of solution from the stock solution to each test tube
respectively. Similarly 2 other sets are prepared.
Measure the absorbance of each set i.e. total 3 concentrations of 3 sets. Take the Mean of
each concentration of each set. Find out the % recovery, Standard Deviation & % Relative
Standard Deviation.
TABLETS: From the stock solution of PCM tablets (100µg/ml) pipette out 1 ml of
solution in 3 volumetric flasks, further add 80,100 & 120% of solution from the Std. stock
solution to each volumetric flasks respectively. Similarly 2 other sets are prepared.
Measure the absorbance of each set i.e. total 3 concentrations of 3 sets. Take the Mean of
each concentration of each set. Find out the % recovery, Standard Deviation & % Relative
Standard Deviation.
8. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
Precision
Repeatability: Select the middle concentration i.e. 15µg/ml and carry out the repeatability
by taking the absorbance of the solution 6 times.
Calculate the mean of the absorbance and find the Standard Deviation and % Relative
Standard Deviation.
Intraday precision: Prepare 3 concentrations from the stock solution (i.e. 5, 15 &
25µg/ml), make another 2 sets similarly. Measure the absorbance of each concentration of
each set at the time interval of 2 hours.
Calculate the Mean, Standard Deviation & % Relative Standard Deviation.
Interday precision: Prepare 3 concentrations from the stock solution (i.e. 5, 15 &
25µg/ml), make another 2 sets similarly. Measure the absorbance of each concentration of
each set for 3 days.
Calculate the Mean, Standard Deviation & % Relative Standard Deviation.
LOD & LOQ
LOD & LOQ are found by using the following equation.
LOD = 3.3 * σ / S
LOQ = 10 * σ / S
Where, σ = the standard deviation of the intercept
S = the slope of the calibration curve
Robustness and Ruggedness
Robustness: It is carryout by doing deliberate variation in method parameters is done (i.e.
change in wavelength). Absorbance of any one concentration (i.e. 15µg/ml) is measured at
2 different wavelengths i.e. 244.8&245.2 nm. And calculate the % Assay or % Drug
Recovery. Repeat the procedure three times.
Ruggedness: It is carried out by the analysis of the sample (i.e. 15µg/ml) under a variety
of normal test conditions such as different laboratories, different analysts, using
operational and environmental conditions that may differ but are still within the specified
parameters of the assay.
Instruments used:
1. Shimadzu UV-1800
Sr. No.: A114550/08677
2. Shimadzu UV-1800
Sr. No.: A114549/08780
9. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
Measure the Absorbance and calculate Mean, Standard Deviation, % Relative standard
deviation & % Assay.
CALCULATIONS:
Equations
Std. Deviation =
Where, ∑= Sum across the value
X= Number of values
X= mean or Average
n = Number of values
% RSD = Std. Deviation * 100 / Mean
Linearity:
Concentrations
(µg/ml)
Set
1
Set
2
Set
3 Mean
Std.
Deviation
%
RSD
5 0.385 0.391 0.379 0.385 0.006 1.558
10 0.798 0.819 0.811 0.809 0.010 1.309
15 1.159 1.152 1.157 1.156 0.003 0.311
20 1.591 1.587 1.579 1.585 0.006 0.385
25 1.932 1.946 1.924 1.934 0.011 0.575
Mean % RSD 0.827
y = 0.0777x + 0.0069
R² = 0.9989
0
0.5
1
1.5
2
2.5
0 5 10 15 20 25 30
Absorbance
Concentration(µg/ml)
Calibration curve of PCM (set 1)
10. Drug Regulation & Regulatory Authorities
Dept. Of Quality Assurance & Regulatory Affairs
L. J. Institute of Pharmacy, Ahmedabad.
Accuracy(Std. addition method):
Sr.
No. Concentration 1 2 3 Mean
Std.
Deviation % RSD
% Drug
recovery
1 80% 1.215 1.202 1.195 1.204 0.01015 0.84293 85.92 %
2 100% 1.295 1.327 1.326 1.316 0.01819 1.38248 84.62 %
3 120% 1.425 1.472 1.465 1.454 0.02536 1.74398 84.62 %
Mean of % RSD 1.32313
85.03 %Average of % Drug Recovery
y = 0.0776x + 0.0156
R² = 0.9986
0
0.5
1
1.5
2
2.5
0 5 10 15 20 25 30
Absorbance
Concentration(µg/ml)
Calibration curve of PCM (set 2)
y = 0.0772x + 0.0126
R² = 0.9986
0
0.5
1
1.5
2
2.5
0 5 10 15 20 25 30
Absorbance
Concentration(µg/ml)
Calibration curve of PCM (set 3)