This document discusses sample preparation and instrumentation for HPLC analysis. It explains that sample preparation aims to provide a reproducible and homogenous sample suitable for column injection that is free of interference and compatible with the HPLC method. It also describes the main components of HPLC instrumentation, including solvent reservoirs, degassing, the pump, injector, analytical column, detector, and data recorder or processor. The document focuses on providing an overview of important considerations for sample preparation and the basic setup and functions of HPLC instrumentation.
HPLC[ HIGH PERPROMANCE LIQUID CHROMATOGRAPHY OR HIGH PRESSURE LIQUID CHROMAT...Dr. Ravi Sankar
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HPLC[ HIGH PERPROMANCE LIQUID CHROMATOGRAPHY OR HIGH PRESSURE LIQUID CHROMAT...Dr. Ravi Sankar
GASSCHROMATOGRAPHY[GC], ADVANCED STUDY OF THE FOLLOWING AND THEIR APPLICATIONS, INTRODUCTION, THEORY, COLUMN OPERATION,INSTRUMENTATION AND DETECTION,APPLICATIONS AND ADVANTAGES OF GC,PRINCIPLE OF SEPARATION IN GC, HOW GC MECHINE WORKS? COLUMN, DETECTORS.
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Instrumentation of HPLC, principle by kk sahuKAUSHAL SAHU
INTRODUCTION
Instrumentation of HPLC
TYPES OF HPLC
PARAMETERS
APPLICATION
CONCLUSION
REFERENCE
High-performance liquid chromatography ( HPLC) is a specific form of column chromatography generally used in biochemistry and analysis to separate, identify, and quantify the active compounds.
HPLC mainly utilizes a column that holds packing material (stationary phase), a pump that moves the mobile phase(s) through the column, and a detector that shows the retention times of the molecules.
Introduction to chromatography, Definition of Chromatography, Types of column chromatography, Theory of chromatography, Practical considerations in column chromatography , Factors affecting efficiency of a column, Applications.
Instrumentation of HPLC, principle by kk sahuKAUSHAL SAHU
INTRODUCTION
Instrumentation of HPLC
TYPES OF HPLC
PARAMETERS
APPLICATION
CONCLUSION
REFERENCE
High-performance liquid chromatography ( HPLC) is a specific form of column chromatography generally used in biochemistry and analysis to separate, identify, and quantify the active compounds.
HPLC mainly utilizes a column that holds packing material (stationary phase), a pump that moves the mobile phase(s) through the column, and a detector that shows the retention times of the molecules.
Introduction to chromatography, Definition of Chromatography, Types of column chromatography, Theory of chromatography, Practical considerations in column chromatography , Factors affecting efficiency of a column, Applications.
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)
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This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
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The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
Biological screening of herbal drugs: Introduction and Need for
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Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
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http://sandymillin.wordpress.com/iateflwebinar2024
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Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
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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.
1. Presented By Group 11:
Sijawal Ali CHEM19131094
Adnan Sohail Akhtar CHEM19131110
Syed Irshad Bukhari CHEM19131113
Kaleem Ullah CHEM19131116
Muhammad Nazim CHEM19131119
Presented To : Dr.Ahmad Mudassir
2. Introduction:
Sample preparation is an essential part of
HPLC analysis intended to provide a
reproducible and homogenous that is suitable
for injection on the column.
SAMPLE PREPRATION:
3. Aims of Sample Preparation:
Should be
free of
interference
will not
damage the
column
compatible
with HPLC
method
4. Types of Samples:
Sample matrices can be classified as
1) Organic
2) Inorganic.
Further Divided Into:
Solids, semisolids (gels, creams and suspensions)
Liquids
Gases.
5. Gaseous samples are usually by GC but sample which is
labile and thermally unstable or prone to absorb are better
by HPLC.
Compared to gaseous or solids, liquids are much easier to
prepare for HPLC.
Preliminary processing of solids and semisolids, reducing
sample particle size is required.
It is homogeneous allowing more sampling with greater
accuracy and precious.
Methods to reduce particle size are blending, chopping,
cutting, grinding and milling etc.
6. Drying a Sample:
Drying the sampling is usually necessary for
reliable for reliable assay.
Filtration:
Particle should be removed prior to injection
because of their adverse effect on column.
HPLC filters in the range of 0.25 to 2μm.
8. i. Binary System:
Two reservoirs or bottles will be there.
ii. Quaternary System:
Two reservoirs or bottles will be there.
Four bottles or reservoirs will be there.
HPLC grade solvents are used.
These solvents are highly pure.
(2) Degasser:
For removal of gases with are dissolve in our mobile phase
or solvent is done by vacuum pump.
9. (3) Solvent Mixing Valve:
For mixing of the solvents.
All solvents are run by the pipe diameter of this pipe is ⅛ inch
made up of PTFE (poly tetra floro ethylene) is also called Teflon.
(4) HPLC Pump:
There are two types.
1. Constant pressure pump
2. Constant flow rate pump
1) Constant Pressure Pump:
Constant pressure pump means pressure of the pump will be
constant so flow rate may vary and this will not give proper
separation.
10. 2) Constant Flow Rate Pump
Flow rate will be constant and pressure may be change
means according to our column resistance pressure will be
change and due to that pressure change our flow rate will
not effect.
Means column resistance will be higher and pressure will
be higher to maintain flow rate, if column resistance will
be lower pressure will also be lower to maintain constant
flow rate.
Generally, we are using 1 ml/min flow rate in HPLC. One
problem which face in HPLC.
11. Back Pressure:
Due to resistance of the column.
Low back pressure in new column.
High back pressure in used column.
It is due to blockage of the pores of stationary phase.
(5) Precolumn/Guard Column
Precolumn used to protect analytical column. So, any type
of concentration present in our solvent will be removed by
precolumn.
Internal diameter is 4-5 mm and length are 2-10 cm.
12. (6) Sample Injector:
There are three types.
1) Septum injector
2) Stop flow septum less injection
3) Microvolume sampling
1) Septum Injector:
Auto selling septum will be there, we will injection our sample
by using micro string.
2) Stop Flow Septum Less Injection:
In this type of injection first we will stop the flow of mobile
phase then we will open the upper part of our column after that
we will put our sample here.
13. (7) Analytical column:
3) Microvolume Sampling:
This is modern sample injector. By using this sample
injector highest accuracy will be achieved and auto
sampling technique is use by using this sample injection
method.
Length 10-30 cm
Internal diameter 4-5 mm
External diameter 6.35 mm
14000-15000 psi pressure is applied during packing.
14. The separation is performed inside the column.
The recent columns are often prepared in a stainless steel
housing, instead of glass columns.
This column is made up of stainless steel is a very solid
alloy which can resist the high pressure.
The packing material generally used is silica or polymer
gels compared to calcium carbonate.
The eluent used for LC varies from acidic to basic
solvents.
Most column housing is made of stainless steel since
stainless is tolerant towards a large variety of solvents.
15. (8) Column Heater:
The LC separation is often largely influenced by the column
temperature.
In order to obtain repeatable results, it is important to keep
consistent temperature conditions.
Also for some analysis, such as sugar and organic acid,
better resolutions can be obtained at elevated temperatures
(50 to 80°C).
Thus columns are generally kept inside the column oven
(column heater).
16. (9) Detectors:
Various detectors use in HPLC.
1. Refractive Index Detector
2. Conductivity Detector
3. UV Visible Detector
4. Fluorescence Detector
5. Mass Detector
6. Amperometric Detector
17. (10) Recorder:
The change in eluent detected by a detector is in the form of
an electronic signal, and thus it is still not visible to our eyes.
In older days, the pen (paper)-chart recorder was popularly
used. Nowadays, a computer-based data processor (integrator)
is more common.
There are various types of data processors; from a simple
system consisting of the in-built printer and word processor
while those with software that are specifically designed for an
LC system which not only data acquisition but features like
peak-fitting, baseline correction, automatic concentration
calculation, molecular weight determination, etc.