The document discusses high performance liquid chromatography (HPLC). It begins with introducing chromatography and its history. It then discusses why HPLC was developed to overcome limitations of column chromatography, noting that HPLC allows for more efficient separation, uses less solvent, and requires smaller sample sizes. The document goes on to describe key aspects of HPLC, including its instrumentation, components, separation mechanisms, and various modes like reversed phase, normal phase, and ion exchange chromatography. It emphasizes that HPLC is widely used in fields like pharmaceuticals, chemicals, food, and forensics.
Best Practice Maintenance and TroubleshootingYokogawa1
The desire is to achieve an accurate, reliable measurement with a reasonable electrode life expectancy while minimizing the frequency or complexity of required/ needed maintenance. When a quality pH sensor system is undamaged, cleaned, and properly calibrated, it will provide a measurement that is accurate and reliable.
Measurement results obtained from dirty or faulty electrodes can be anything from slow response to an entirely erroneous reading.
This webinar will provide a step-by-step plan for pH troubleshooting. We will focus on checking the complete loop (analyzer, cables, and sensor) and explaining the symptoms (e.g., not responding to changes).
Capturing video in log tells the camera to record more from a scene—meaning you'll be able to recover more from your highlights and shadows and adjust more colors and tones in post-production. In this course, join Rich Harrington as he shows you how to record video in log, process the files, and complete a post-production workflow.
Topics covered include:
Recording options for log
Configuring cameras to recording in log
Monitoring log recordings
Following typical camera workflows
Getting ready for post
Using log files in Premiere and After Effects
Using log files in Final Cut Pro X
Managing and manipulating lookup tables
Working with LOG Footage & Creating Your Own Lookup TablesRichard Harrington
Capturing video in log tells the camera to record more from a scene—meaning you’ll be able to recover more from your highlights and shadows and adjust more colors and tones in post-production. In this course, join Rich Harrington as he shows you how to record video in log, process the files, and complete a post-production workflow.
Sap Tips and Tricks Training for End userArghadip Kar
Sap Tips and Tricks Training for End user
Determine all the Transports related to a particular Object.
Understand preventive measures to avoid releasing Transports with erroneous code.
Understand preventive measures to avoid missing a Transport Migration to Production Environment.
Determine which tables will be used to extract data in SAP.
Download Program code to local file from a respective Package.
Some FAQs and considerations when tuning your PID controller using the https://pidtuner.com tool.
The PID Tuner is a free online application to automate your PID controller tuning by following a simple procedure.
Still there are some important practical aspects to be considered when using the PID Control tuning tool. This presentation shows the reader the best practices for PID tuning.
Best Practice Maintenance and TroubleshootingYokogawa1
The desire is to achieve an accurate, reliable measurement with a reasonable electrode life expectancy while minimizing the frequency or complexity of required/ needed maintenance. When a quality pH sensor system is undamaged, cleaned, and properly calibrated, it will provide a measurement that is accurate and reliable.
Measurement results obtained from dirty or faulty electrodes can be anything from slow response to an entirely erroneous reading.
This webinar will provide a step-by-step plan for pH troubleshooting. We will focus on checking the complete loop (analyzer, cables, and sensor) and explaining the symptoms (e.g., not responding to changes).
Capturing video in log tells the camera to record more from a scene—meaning you'll be able to recover more from your highlights and shadows and adjust more colors and tones in post-production. In this course, join Rich Harrington as he shows you how to record video in log, process the files, and complete a post-production workflow.
Topics covered include:
Recording options for log
Configuring cameras to recording in log
Monitoring log recordings
Following typical camera workflows
Getting ready for post
Using log files in Premiere and After Effects
Using log files in Final Cut Pro X
Managing and manipulating lookup tables
Working with LOG Footage & Creating Your Own Lookup TablesRichard Harrington
Capturing video in log tells the camera to record more from a scene—meaning you’ll be able to recover more from your highlights and shadows and adjust more colors and tones in post-production. In this course, join Rich Harrington as he shows you how to record video in log, process the files, and complete a post-production workflow.
Sap Tips and Tricks Training for End userArghadip Kar
Sap Tips and Tricks Training for End user
Determine all the Transports related to a particular Object.
Understand preventive measures to avoid releasing Transports with erroneous code.
Understand preventive measures to avoid missing a Transport Migration to Production Environment.
Determine which tables will be used to extract data in SAP.
Download Program code to local file from a respective Package.
Some FAQs and considerations when tuning your PID controller using the https://pidtuner.com tool.
The PID Tuner is a free online application to automate your PID controller tuning by following a simple procedure.
Still there are some important practical aspects to be considered when using the PID Control tuning tool. This presentation shows the reader the best practices for PID tuning.
Open Compute Project (OCP)*Linux*Open Source*IT Infrastrucutre*High Performance Computing (HPC)*Supercomputing*Beowulf Clusters*Datacenters -- Archna Haylock, Community Director for Open Compute Foundation, presented at the Penguin Computing booth theater at SC18 about Open Compute Project membership facts and how to participate and become a member of the OCP community.
Give business owners and non-technical stakeholders an understanding of some key "technical" concepts relating to Magneto in particular and web projects in general. This is to empower them to make decisions and manage operations in such a way that promotes stability and protects their conversion rates.
CHROMacademy Premier Membership offers chemical analysts the opportunity to learn at their own pace or use our tutor supported training courses. Webcasts, Live-on-the-Web and Live @ Your Place offer live training in group settings
This webinar aims to show you how to avoid the common pitfalls that can affect your conversion rate optimization strategy. At the end of this webinar you will be able to:
1) Use A/B Testing best practices to get the most out of it
2) Identify the common mistakes preemptively
3) Appreciate how A/B Testing fits your whole conversion rate optimization strategy (based on real cases, such as Swedoffice and Spotify)
hosted by Simon Dahla from Conversionista Sweden and Martijn Janssen, Partner Manager Optimizely
Why you should stop using the generic term “Technical Debt” and start resolvi...Alex Fedorov
Do you feel like you are accruing all this Technical Debt every week and it slows your whole team down? Do you want to deal with it so badly, but you don’t know where even to start? Did you have “refactoring/rewriting sprints” in the past, and they have failed, and you had to revert?
The first advice is to stop calling it “Technical Debt.” And instead, sit down with your team and figure out the list of specific problems that are causing the most trouble in your software right at this moment. Attack the first problem on the list then only. Repeat on a regular basis.
In this talk, you will learn WHY you should do that, and HOW to do that. Moreover, you are going to be able to apply it tomorrow at your workplace!
Peristaltic pump installation are not so easy. Calculations will help to make the right choice, and will save lots of headage and money. All peristaltic pumps can be applied well, if you don't exceed the capabilities of that selected pump.
How hard can it be to start a software development project? It can be very intimidating to start at the blank page.
This talk offers some ideas how to make getting started less scary and more enjoyable, from the first keyboard stroke to a working usable system.
Open Compute Project (OCP)*Linux*Open Source*IT Infrastrucutre*High Performance Computing (HPC)*Supercomputing*Beowulf Clusters*Datacenters -- Archna Haylock, Community Director for Open Compute Foundation, presented at the Penguin Computing booth theater at SC18 about Open Compute Project membership facts and how to participate and become a member of the OCP community.
Give business owners and non-technical stakeholders an understanding of some key "technical" concepts relating to Magneto in particular and web projects in general. This is to empower them to make decisions and manage operations in such a way that promotes stability and protects their conversion rates.
CHROMacademy Premier Membership offers chemical analysts the opportunity to learn at their own pace or use our tutor supported training courses. Webcasts, Live-on-the-Web and Live @ Your Place offer live training in group settings
This webinar aims to show you how to avoid the common pitfalls that can affect your conversion rate optimization strategy. At the end of this webinar you will be able to:
1) Use A/B Testing best practices to get the most out of it
2) Identify the common mistakes preemptively
3) Appreciate how A/B Testing fits your whole conversion rate optimization strategy (based on real cases, such as Swedoffice and Spotify)
hosted by Simon Dahla from Conversionista Sweden and Martijn Janssen, Partner Manager Optimizely
Why you should stop using the generic term “Technical Debt” and start resolvi...Alex Fedorov
Do you feel like you are accruing all this Technical Debt every week and it slows your whole team down? Do you want to deal with it so badly, but you don’t know where even to start? Did you have “refactoring/rewriting sprints” in the past, and they have failed, and you had to revert?
The first advice is to stop calling it “Technical Debt.” And instead, sit down with your team and figure out the list of specific problems that are causing the most trouble in your software right at this moment. Attack the first problem on the list then only. Repeat on a regular basis.
In this talk, you will learn WHY you should do that, and HOW to do that. Moreover, you are going to be able to apply it tomorrow at your workplace!
Peristaltic pump installation are not so easy. Calculations will help to make the right choice, and will save lots of headage and money. All peristaltic pumps can be applied well, if you don't exceed the capabilities of that selected pump.
How hard can it be to start a software development project? It can be very intimidating to start at the blank page.
This talk offers some ideas how to make getting started less scary and more enjoyable, from the first keyboard stroke to a working usable system.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
High performance liquid chromatography as introduction
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High Performance
Liquid Chromatography
Dr. Deepkumar Joshi
Chemistry Department
Sheth M. N. Science College, Patan
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High Pressure
Liquid Chromatography
Dr. Deepkumar Joshi
Chemistry Department
Sheth M. N. Science College, Patan
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• Introduction & Basics
• Why need of HPLC ?
• Why Pressure ? Why Performance?
• Concept & Scope of HPLC
• Instrumentation & Working
• Separation Mechanism
Today’s Discussion
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• Chromatography is an analytical
method used to separate
compounds physically prior to
measurement
• Satisfies the purpose of separation
and quantification of target
Introduction & Basics
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History of Chromatography
M. Tswett – A Russian Botanist
Introduction & Basics
Petroleum ether
CaCO3
Chlorophylls
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Some Important Term(s)
• Stationary phase
• Mobile phase
• Chromatogram
• Retention Time
Introduction & Basics
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Column Chromatography
Column : Diameter – 5cm
: Length – 1 metre
Packing Material : Silica
Particle Size : 100 micrometer
Introduction & Basics
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Drawbacks of Column
Chromatography
• Separation is not efficient
• More time consumption
• Wastage of Solvent
Introduction & Basics
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Overcoming the drawbacks
• Lesser time consumption
• Less solvent
• Efficient separation
• High sensitivity & resolution
• Small sample size (ppm / ppb)
Why need HPLC ?
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• Pharmaceuticals
• Bio-chemicals
• Food products
• Industrial chemicals
• Forensic chemistry
• Environmental science
Scope of HPLC
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• Separation of compounds take place
as they move at different rates in the
column
Concept - Mechanism
2
1
column
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• Due to different interactions
between sample and the stationary
phase, molecules move at different
rate
Concept - Mechanism
Stationary Phase
Stronger
interaction
Weaker
interaction
Mobile Phase
2
1
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Instrumentation
Pump
Injector
Column
Oven
Detector
Mobile Phase
Data
processor
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Components of HPLC instrument
• Solvent delivering reservoir
• High pressure pumps
• Sample injector
• Column oven
• Detectors
Instrumentation
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Possible configuration
• Isocratic system
• Low-pressure gradient system
• High-pressure gradient system
Instrumentation
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Isocratic System
Instrumentation
Simple system with one pump and one solvent reservoir.
If more than one solvent is used, solvents should be premixed.
Data
processor
Pump
Injector
Column
Oven
Detector
Mobile Phase
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Low-pressure Gradient System
Instrumentation
•One pump used to control 4 reservoirs
•Mixing is done before pump.
.
low
pressure
gradientvalve
Data
processor
A B DC
Pump
Injector
Column
Oven
Detector
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• Excellent gradient accuracy.
• 2-3 pumps required - one pump per solvent used.
Data
processor
pump
pump
pump
A
B
C
Injector
Column
Oven
Detector
Mixer
High-pressure Gradient System
Instrumentation
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Detectors
• UV-Vis detector
• PDA detector
• Fluorescence detector
• Mass Spectrometer detector
Instrumentation
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Chromatogram
tR : Retention time
A : Area
h : Height
tR
Signal
Time
Peak
h
A
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• Reverse phase chromatography
• Normal phase chromatography
• Ion exchange chromatography
• Size exclusion chromatography
• Affinity chromatography
Working
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Working
Stationary phase: Non-polar property
Mobile phase : Polar property
This combination is defined as
Reversed Phase Mode
Reversed phase
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Working
Stationary phase: polar property
Mobile phase : Non-Polar property
This combination is defined as
Normal Phase Mode
Normal phase
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Working in Rev. Phase
• C18 (ODS) type
• C8 (octyl) type
• C4 (butyl) type
• Phenyl type
• TMS type
• Cyano type
C18H37Si O Si
CH3
CH3
Non-polar
Reversed phase HPLC
• Stationary phase: Non-polar property
• Mobile phase: Polar property
Stationary Phase
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Mobile phase
• Water + Organic solvent
–Methanol
–Acetonitrile
–Tetrahydrofuran
Working in Rev. Phase
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Hydrophobic Interaction
Separation Mechanism
A
B
B
B
B
B
A
A
A
A
A
B
Support
particle
Nonpolar
bonded phase
Interstitial area
(mobile phase)
Less polar analyte
More polar analyteB
A
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• Sample having carbon (alkyl) chain(s)
or aromatic ring(s) are more
hydrophobic
• Sample having –COOH, -NH2, -OH are
less hydrophobic
Separation Mechanism
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Retention Time & Hydrophobicity
Separation Mechanism
OH
OH
C18 (ODS)
Strong
Weak
1
1
2
2
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Effect of Stationary phase
Separation Mechanism
C18 (ODS)
Strong
C8
sample
sample
sample
C4
Medium
Weak
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THANK YOU
deepjoshi359@yahoo.co.in
http://deepjeejoshi.wix.com/deepjoshi
HPLC
