Validation of Analytical and Bioanalytical methodssarikakkadam
Guidelines for Validation of Analytical and Bioanalytical methods as per ICH (Q2R1) and USFDA respectively with an example of Bioanalytical method validation.
The slides are informative of HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY & its thorough components further its advantages and applications. The comparison of HPLC and HPTLC is explained.
Validation of Analytical and Bioanalytical methodssarikakkadam
Guidelines for Validation of Analytical and Bioanalytical methods as per ICH (Q2R1) and USFDA respectively with an example of Bioanalytical method validation.
The slides are informative of HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY & its thorough components further its advantages and applications. The comparison of HPLC and HPTLC is explained.
Ion pair chromatography for pharmacy studentsabhishek rai
Ion-PairChromatography
A GENERALISED OVERVIEW
Chromatography
HPLC
Reverse Phase Chromatography
Ion Pair Chromatography
Ion Pair Reagent
Mechanism of Ion Pair Chromatography
Ion Pair Wash Procedure
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 will help to know about the advance technique to analysis the biological sample in cancer diagnosis and general separation of proteins based upon the molecular weight and helps to analysis the new drug synthesis level
UPLC is an advance analytical technique where it takes advantage of innovation in various technologies such as instrumentation and particle size to achieve dramatic increases in resolution, speed and sensitivity of the liquid chromatography. It operates at higher pressure than that used in HPLC and uses fine particles (less than 2.5µm) & mobile phases at high linear velocities. It can be hyphenated with other techniques such as (MS), (IC), (NMR) and (IR) etc. This is used in all industries and has found application in various fields such as pharmaceutical, food, environmental, forensic, toxicology and pesticide.
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Electron microscopes are used to investigate the ultrastructure of a wide range of biological and inorganic specimens including microorganisms, cells, large molecules, biopsy samples, metals, and crystals. Industrially, electron microscopes are often used for quality control and failure analysis. Modern electron microscopes produce electron micrographs using specialized digital cameras and frame grabbers to capture the image.
Ion pair chromatography for pharmacy studentsabhishek rai
Ion-PairChromatography
A GENERALISED OVERVIEW
Chromatography
HPLC
Reverse Phase Chromatography
Ion Pair Chromatography
Ion Pair Reagent
Mechanism of Ion Pair Chromatography
Ion Pair Wash Procedure
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 will help to know about the advance technique to analysis the biological sample in cancer diagnosis and general separation of proteins based upon the molecular weight and helps to analysis the new drug synthesis level
UPLC is an advance analytical technique where it takes advantage of innovation in various technologies such as instrumentation and particle size to achieve dramatic increases in resolution, speed and sensitivity of the liquid chromatography. It operates at higher pressure than that used in HPLC and uses fine particles (less than 2.5µm) & mobile phases at high linear velocities. It can be hyphenated with other techniques such as (MS), (IC), (NMR) and (IR) etc. This is used in all industries and has found application in various fields such as pharmaceutical, food, environmental, forensic, toxicology and pesticide.
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An electron microscope is a microscope that uses a beam of accelerated electrons as a source of illumination. As the wavelength of an electron can be up to 100,000 times shorter than that of visible light photons, electron microscopes have a higher resolving power than light microscopes and can reveal the structure of smaller objects. A transmission electron microscope can achieve better than 50 pm resolution and magnifications of up to about 10,000,000x whereas most light microscopes are limited by diffraction to about 200 nm resolution and useful magnifications below 2000x.
Electron microscopes are used to investigate the ultrastructure of a wide range of biological and inorganic specimens including microorganisms, cells, large molecules, biopsy samples, metals, and crystals. Industrially, electron microscopes are often used for quality control and failure analysis. Modern electron microscopes produce electron micrographs using specialized digital cameras and frame grabbers to capture the image.
An Over view on Bioassay, structure & principles, types & methods of bioassay. Also mention of other assay's like biotechnology, microbio assay, immunoassay etc.
High performance liquid chromatography (HPLC), also known as high pressure liquid chromatography, is essentially a form of column chromatography in which the stationary phase consists of small particle packings (3-50 µm) contained in a column with a small bore (2-5 mm), one end of which is attached to a source of pressurised liquid eluant (mobile phase)
High Performance Liquid Chromatography (HPLC) is a form of column chromatography that pumps a sample mixture or analyte in a solvent (known as the mobile phase) at high pressure through a column with chromatographic packing material (stationary phase).
RP-HPLC Method Development and Validation of Ketoconazole in Bulk and Pharmac...Sunil Vadithya
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June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
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Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
3. INTRODUCTION OF HPLC
HPLC offers a combination of speed, reproducibility and
sensitivity.
HPLC is a form of liquid chromatography used to separate
compounds that are dissolved in solution.
4. TERMINOLOGIES FOR HPLC
Resolving Power: The extent of separation of the
compounds present in the mixture across the column.
Theoretical plates: An imaginary division of the column ,
equal to the length of column.
Stationary phase: The phase which remains fixed in the
column e.g. C18,silica
Mobile phase: carries the sample through the stationary
phase as it moves through the column.
6. COMPOSITION OF A LIQUID
CHROMATOGRAPH SYSTEM
Solvent
Solvent Delivery System (Pump)
Injector
Column
Detectors
Waste Collector
Recorder (Data Collection)
7. SOLVENT
On basis of solvent two types of systems are made:
i. Isocratic system: same mobile phase runs
throughout the elution of sample. e.g. in QC.
ii. Gradient system: strength of mobile phase is
increased with time during sample elution. e.g. for
complex mixtures.
In reverse phase mostly water and acetonitrile or
methanol is used as polar solvents.
The mobile phase must be degassed to eliminate
the formation of air bubbles.
8. PUMP
The role of the pump is to force a liquid (called the
mobile phase) through the liquid chromatograph at a
specific flow rate, expressed in milliliters per min (mL
/min).
9. COLUMN
Considered as “heart of HPLC”.
Its inner is lined with stationary phase, which
separates the sample components of interest using
various physical and chemical parameters.
The stationary phase can be polar or non-polar.
10. INJECTOR
The injector serves to introduce the liquid sample into
the flow stream of the mobile phase.
Manual Injector
Auto sampler
11. DETECTOR:
The detector can see (detect) the individual
molecules that comes out (elute) from the column.
UV-Visible detectors, photo diode array detector,
fluorescence detector are mostly used.
13. PRINCIPLE
The process involves the interaction of the compounds
in the analyte (which travels along with a mobile phase)
across an immobile surface (stationary phase).
The different component in the mixture pass through the
column and differentiates due to differences in their
partition behavior between the mobile phase and the
stationary phase.
The differential wash out or elution of compounds is
basis of HPLC.
14. WORKING
14
Separation in based upon differential
migration between the stationary and
mobile phases.
Stationary Phase - the phase
which remains fixed in the
column, e.g. C18, Silica
Mobile Phase - carries the sample
through the stationary phase as it
moves through the column.
Injector
Detector
Column
Solvents
Mixer
Pumps
High Performance Liquid Chromatograph
Waste
32. ORDER OF CALIBRATION
Detector pump injector column
Detector
i. Wavelength accuracy
ii. Detector linearity
Pump
i. Flow rate accuracy
ii. Pressure test
Injector
i. Injector accuracy
ii. Carryover
iii. Injector linearity
Column
i. Temperature accuracy
34. Wavelength Accuracy:
•Select 3D mode and set the wavelength range
as 200-400nm.
•Inject 20 μl caffine of standard preparation once
into the chromatographic system.
•Extract and record the chromatograms at
wavelengths of 202 to 208nm with an interval
of 1nm and at 269 to 275 nm with an interval of
1nm.
•Note down the height or absorbance.
• Acceptance criteria: The maximum absorbance
should be at 205±2nm and 272±2nm.
35. Detector Linearity:
Standard Preparation: Accurately weigh and transfer
about 60mg of Caffeine into a 100ml volumetric flask.
Dissolve and dilute to the volume with mobile phase.
6 linearity solutions are prepared in different
concentrations of the drug.
Procedure: Inject blank, followed by Detector linearity
solutions and record the peak responses of Caffeine
standard plot between the concentration Vs the peak
responses.
Acceptance criteria: The plot should be
linear and regression coefficient (R2) should
not be less than 0.99.
37. Flow Rate Accuracy:
1. Prime all the solvent lines with water.
2. Set the flow rate to 0.500 ml/m.
3.Wait for about 15 m to stabilize the system and ensure that the pressure is
stable.
4.Insert the outlet tubing into a 10 ml volumetric flask and start the stop watch
simultaneously.
5. Stop the stopwatch when the lower meniscus reaches the 10 ml mark on the
flask.
6. Record the elapsed time.
7. Similarly check the flow for 1.0 ml/m and 2.0 ml/m.
Acceptance criteria:
The time taken to collect the water should be with in ± 2.0% of the actual value.
37
38. Pressure Test :
•The first step of the pressure test is to plug the outlet
of the pump using a dead-nut and by setting the
automatic pump shutdown pressure to
6,000 psi.
•The pump-head pressure signal output is connected
to a recorder. Pressurize the pump by pumping
methanol at 1 mL/min. The pressure inside the
pump head increases quickly as the outlet of the
pump is blocked.
•The pressure will gradually rise to the shutdown
pressure if the check valves are able to hold the
mobile phase in the pump chamber as would be
normally expected . If the check valve is not
functioning properly, the pressure will fluctuate at
about 3,000 psi instead of reaching the shutdown
pressure.
38
40. Injector Accuracy:
1. Connect the pump and detector inlet with union.
2. Prepare mobile phase consisting of a mixture of water and Methanol
(70:30)
3. Set a flow rate of 0.5 ml/m and a run time of 1 m.
4. Set the column temperature at 25± 2°C.
5.Fill a standard HPLC vial to 2/3rd with water. Seal the vial properly
with a cap.
6. Weigh the vial and record the weight as W1 grams.
7. Place the vial in the chromatographic system and perform 6 injections
of 50μl volume from this vial.
8. Weigh the vial again and note the weigh after the injections as W2
grams.
9. Calculate the mean volume injected per injection as follows:
•Mean injected volume (μl) = (W1 – W2) × 100/6
•Acceptance criteria: The mean injected volume should be
50.0±1.0 μl. 40
41. Carryover :
• Small amounts of analyte may get carried over from
the previous injection and contaminate the next
sample to be injected.
• The carryover will affect the accurate quantitation of
the subsequent sample.
• The effectiveness of the cleaning can be evaluated
by injecting a blank after a sample that contains a
high concentration of analyte.
• The response of the analyte found in the blank
sample expressed as a percentage of the response
of the concentrated sample can be used to determine
the level of carryover.
41
42. Injector Linearity:
• Standard Preparation: Accurately weigh and transfer
about 60mg of Caffeine into a 100ml volumetric flask.
Dissolve and dilute to the volume with mobile phase.
Transfer 10ml of Standard Preparation into a 100ml
volumetric flask and dilute to the volume with mobile
phase.
• Procedure: Inject 5 μl of the mobile phase as blank
injection. Inject 5 μl, 10 μl, 20 μl, 50 μl and 80 μl of
the Standard Preparation and record the peak areas.
• Plot a curve for the volume injected Vs peak area
• Acceptance criteria: The plot should be
linear and regression coefficient (R2) should
not be less than 0.99.
44. i. It is evaluated with a calibrated digital
thermometer at 30°Cand 60°C .Place the
thermometer probe in the column oven and
set the column oven temperature at
30°C.Wait till the temperature stabilizes.
Record the temperature displayed on the
thermometer. Similarly performs the column
oven temperature accuracy test at 60°C.
Acceptance criteria: The resulting oven
temperature from the thermometer display
should be within ±2°C of the set
temperature.
COLUMN OVEN TEMPERATURE
ACCURACY