This document provides an overview of high performance liquid chromatography (HPLC). It defines HPLC as a technique used to separate components of a mixture for identification, quantification or purification. It then describes the main types of HPLC based on the mode of chromatography, principle of separation, elution technique, scale of operation, and type of analysis. The key components of an HPLC instrument are also outlined, including the pump, injector, column, detectors, and various applications of HPLC. Recommendations are provided for operating the system components and ensuring optimal performance.
In this slide contains types of HPLC Columns, Plate theory and Van Deemter Equation.
Presented by : Malarvannan.M (Department of pharmaceutical analysis).
RIPER,anantpur.
HPLC
Chromatography
Mobile Phase & Stationary Phase
CLASSIFICATION OF CHROMATOGRAPHY
Characteristics of HPLC
Purpose
Superiority of HPLC
TYPES OF HPLC TECHNIQYES
Principle
PHASING SYSTEM & (normal vs reversed phase)
INSTRUMENTATION
Flow diagram of HPLC instrument
Advantages of HPLC
Stationary Phase and Mobile Phase Selection for Liquid Chromatography
The presentation focuses on how to choose the appropriate mode of separation, the correct column and highlights the importance of the correct mobile phase. This approach will be applied to a wide selection of compound types ranging from proteins, peptides, glycans to small pharmaceutical molecules and their metabolites. It will also look at specific application areas for monoclonal antibody analysis, namely: titer, aggregation, charge and oxidation variant. Platform methods for biologics characterization are also discussed.
In this slide contains types of HPLC Columns, Plate theory and Van Deemter Equation.
Presented by : Malarvannan.M (Department of pharmaceutical analysis).
RIPER,anantpur.
HPLC
Chromatography
Mobile Phase & Stationary Phase
CLASSIFICATION OF CHROMATOGRAPHY
Characteristics of HPLC
Purpose
Superiority of HPLC
TYPES OF HPLC TECHNIQYES
Principle
PHASING SYSTEM & (normal vs reversed phase)
INSTRUMENTATION
Flow diagram of HPLC instrument
Advantages of HPLC
Stationary Phase and Mobile Phase Selection for Liquid Chromatography
The presentation focuses on how to choose the appropriate mode of separation, the correct column and highlights the importance of the correct mobile phase. This approach will be applied to a wide selection of compound types ranging from proteins, peptides, glycans to small pharmaceutical molecules and their metabolites. It will also look at specific application areas for monoclonal antibody analysis, namely: titer, aggregation, charge and oxidation variant. Platform methods for biologics characterization are also discussed.
The presentation contains basic introduction to mostly used and versatile reversed phase chromatography, its instrumentation, working and applications. It will be useful for you to understand basic concepts about RP-HPLC.
This is a PPT for HPLC which I made for presenting my assigned topic for Practice School during 7 sem of my graduation . Hope it is useful for you guys :)
Chromatography is a laboratory technique for the separation of a mixture. The mixture is dissolved in a fluid called the mobile phase, which carries it through a structure holding another material called the stationary phase.
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)
Muktapishti is a traditional Ayurvedic preparation made from Shoditha Mukta (Purified Pearl), is believed to help regulate thyroid function and reduce symptoms of hyperthyroidism due to its cooling and balancing properties. Clinical evidence on its efficacy remains limited, necessitating further research to validate its therapeutic benefits.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
Local Advanced Lung Cancer: Artificial Intelligence, Synergetics, Complex Sys...Oleg Kshivets
Overall life span (LS) was 1671.7±1721.6 days and cumulative 5YS reached 62.4%, 10 years – 50.4%, 20 years – 44.6%. 94 LCP lived more than 5 years without cancer (LS=2958.6±1723.6 days), 22 – more than 10 years (LS=5571±1841.8 days). 67 LCP died because of LC (LS=471.9±344 days). AT significantly improved 5YS (68% vs. 53.7%) (P=0.028 by log-rank test). Cox modeling displayed that 5YS of LCP significantly depended on: N0-N12, T3-4, blood cell circuit, cell ratio factors (ratio between cancer cells-CC and blood cells subpopulations), LC cell dynamics, recalcification time, heparin tolerance, prothrombin index, protein, AT, procedure type (P=0.000-0.031). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and N0-12 (rank=1), thrombocytes/CC (rank=2), segmented neutrophils/CC (3), eosinophils/CC (4), erythrocytes/CC (5), healthy cells/CC (6), lymphocytes/CC (7), stick neutrophils/CC (8), leucocytes/CC (9), monocytes/CC (10). Correct prediction of 5YS was 100% by neural networks computing (error=0.000; area under ROC curve=1.0).
- Video recording of this lecture in English language: https://youtu.be/kqbnxVAZs-0
- Video recording of this lecture in Arabic language: https://youtu.be/SINlygW1Mpc
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
Basavarajeeyam is a Sreshta Sangraha grantha (Compiled book ), written by Neelkanta kotturu Basavaraja Virachita. It contains 25 Prakaranas, First 24 Chapters related to Rogas& 25th to Rasadravyas.
Basavarajeeyam is an important text for ayurvedic physician belonging to andhra pradehs. It is a popular compendium in various parts of our country as well as in andhra pradesh. The content of the text was presented in sanskrit and telugu language (Bilingual). One of the most famous book in ayurvedic pharmaceutics and therapeutics. This book contains 25 chapters called as prakaranas. Many rasaoushadis were explained, pioneer of dhatu druti, nadi pareeksha, mutra pareeksha etc. Belongs to the period of 15-16 century. New diseases like upadamsha, phiranga rogas are explained.
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
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!
2. INTRODUCTION
DEFINATION OF HPLC :
It is a chromatographic technique used to separate
components of mixture for the purpose to identify, quantify or
purify the individual components of the mixture.
This is widely used in field of biochemistry and analytical
chemistry.
3. Types of HPLC techniques
Based on mode of chromatography.
Based on principle of separation.
Based on elution technique.
Based on scale of operation.
Based on types of analysis.
4. Based on modes of chromatography
Normal Phase mode:
The stationary phase is polar in natural & the mobile phase is
non-polar.
This is not advantageous in pharmaceutical application since most
of the drug molecules are polar in nature and takes longer time to
be eluted and detected.
Reverse phase:
He stationary phase is non-polar & the mobile phase is polar
in nature.
Since most of the drugs and pharmaceuticals are polar in nature,
they are not retained for a longer time and eluted faster.
5. Based on principle of separation
Adsorption chromatography: Separation of components
takes place because of the difference in affinity of compounds
towards stationary phase.
Ion exchange chromatography: An ion is used to separate
a mixture of similar charged ions.
Size exclusion or gel permeation chromatography: A
mixture of components with different molecular sizes are
separated by using gels which acts as sieve.
6. Based on elution technique
Isocratic separation: In this technique, the same mobile
phase combination is used throughout the process of
separation. The same polarity or elution strength is maintained
throughout the process.
Gradient separation: In this technique, a mobile phase
combination of lower polarity or elution strength is used
followed by gradually increasing the polarity or elution
strength.
7. Based on the scale of operation
Analytical HPLC: Where only analysis of the samples are
done. Recovery of the samples is not done
Preparative HPLC: Where the individual fractions of pure
compound can be collected using fraction collector. The
collector samples are reused.
8. Based on the type of analysis
Qualitative analysis: Which is used to identify the
compound, detect the impurities, to find the number of
components, etc.
Quantitative analysis: Which is done to determine the
quantity of the individual or several components in a mixture.
This can be done by comparing peak area of the standard and
sample
11. Pump
The role of the pump is to force a liquid ( called the mobile
phase) through the liquid chromatography at a specific flow
rate, expressed in milliliters per min (mL/min)
Normal flow rates in HPLC are in the 1-2mL/min range.
During the chromatographic experiment, a pump can deliver
a constant mobile phase composition (isocratic) or an
increasing mobile phase composition (gradient).
12. Types of pump
Mainly three types:
Constant flow reciprocating pump
Syringe type pump
Pneumatic pump
13. MIXING UNIT
Mixing unit is used to mix solvents in different proportions and pass
through the column.
There are two types of mixing units.
1.they are low pressure mixing chamber which uses helium for
degassing solvents.
2.high pressure mixing chamber does not require helium for
degassing solvents
Mixing of solvent is done either with a static mixer which is packed
with beads or a dynamic mixer which uses magnetic stirrer and
operates under high pressure.
14. INJECTOR
• The injector serves to introduce the liquid sample into the
flow stear the mobile phase.
• Typical sample volumes are 5-20microliters
• The injector must also be able to withstand the high pressure
of the liquid system.
• An auto sampler is the automatic version for when the user
has many samples to analyze or when manual injection is not
practical.
15. COLUMN
It is the heart of the chromatograph Column length: varies from
5cm to 30cm Column diameter: ranges from 2mm to 50mm
Particle size: from 1µ to 20µ Particle nature: spherical, uniform
sized, porous materials are used.
16. DETECTORS
Detectors are used with high performance
liquid chromatography to detect and identify analytes in the
sample.
UV detectors
Refractive index detector
Flourimetric detector
Conductivity detector
Amperometric detector
17. APPLICATIONS OF HPLC
• Qualitative analysis
• Checking the impurity of a compound
• Presence of impurities
• Quantitative analysis
• Isolation and identification of drugs
• Isolation and identification of mixture of components
• Biopharmaceutical and pharmacokinetic studies
• Stability studies
• purification.
19. How to deal with solvents
• Use clean bottles only.
• Exchange water-based solvents daily.
• Select solvent volume to be used up within 1 – 2 days.
• Use only HPLC-grade solvents and water filtered through 0.2 μm
filters.
• Label bottles correctly with bottle content, and filling date / expiry date.
• Use solvent inlet filters to protect the system from incoming particles.
• Reduce risk of algae growth: use brown bottles for aqueous solvents,
avoid direct sunlight or wrap the bottles in aluminium foil.
20. Daily tasks
• Replace solvents and solvent bottles for mobile phases based on
water/buffer.
• Replace solvents and solvent bottles for organic mobile phase latest
every second day.
• Check presence of seal wash solvent.
• Purge each channel with fresh solvent at 2.5 – 3 mL/min for 5 min.
• Equilibrate your system with composition of your application for 15
min.
• Check the instrument analysis label and calibration label.
21. Weekly tasks
• Change seal wash solvent (10 % / 90 % isopropanol/water) and
bottle.
• Flush all channels with water at 2.5 – 3 mL/min for 5 min to
remove salt deposits if buffer applications were used.
• Inspect solvent filters for dirt or blockages. Clean or exchange if
no flow is coming out of the solvent line when removed from the
degasser inlet.
22. Power-up the system
• Power up the pump
Use new or different mobile phase (as required).
Purge each channel with 2.5 – 3 mL/min for 5 min. Open the purge valve or use the purge
command .
Equilibrate your system with composition of your application for 15 min. Use conditioning for
systems.
• Power up the sampler
Purge the autosampler daily, and before and after sample analysis, especially if you are using
buffers.
Set flow to required value of your application and close the purge valve.
Pump for approximately 10 min.
Use fresh needle wash and/or needle seat backflush solvents like methanol or acetonitrile and
water mixtures without buffer.
Ensure that the vials contain enough sample solution for all injections.
• Power up the detector
Warmup the lamp for at least 1 h.
For RI detectors only: flush the reference and sample side with fresh solvent used for the current
application.
23. Shut-down the system
• Long-term storage of the column
Flush the column with appropriate solvent found in the column manual.
Remove and seal column, and store according to good laboratory practice if
needed.
• Long-term shut-down of the system
Flush system with water to remove buffer.
Remove all samples from the sampler and store according to good laboratory
practice.
Use recommended solvents to store the system.
Power off the system.
24. Recommendations for pumps
• Check pumps performance on regular basis.
• Perform preventive maintenance in the recommended usage
interval.
• Prepare the pump as recommended like described in the power up
section to ensure optimal performance and best life time.
• Use the seal wash function as recommended to ensure optimal
performance and best life time, see below.
25. Recommendations for samplers
• Purge the autosampler after sample analysis.
• Always use fresh wash solvent for the needle or seal wash
function.
• Place the wash solvent reservoir for needle wash (optional: needle
seat flush) into the solvent cabinet.
• Check the drainage routing of the wash port outlet into a waste
container.
• Fill each vial with enough sample solution for all injections.
• Filter, decant, or centrifuge sample to separate from insoluble
solid.
26. Recommendations for detectors
• Warm-up the lamp at least 1 h.
• Keep environment and ambient temperature stable.
• Do not expose the detector to too much air current from the HVAC.
• Use the recommended waste lines for each detector type. Avoid pinching
the waste tube after the cell outlet.
• Ensure that the detector flow cell is bubble free.
• For RI detectors only: flush the reference and sample side with fresh
solvent used for the current application.
• Flush the flow cell after use.
• Use isopropanol to remove organic solvents & milli-q water to remove
salts.
27. Additional Information for pumps
• Purge
Fill the system with fresh or different solvent.
Remove air bubbles in tubes and pump heads.
• Condition
If micro air bubbles persist in the pump head the overall pump
performance may look correct but the pump will perform extra
work and accuracy/precision are negatively affected. To remove
the air efficiently the Condition function can be used.