HPLC instrumentation and chromatography principle  

HPLC instrumentation and chromatography principle

This HPLC chromatography lecture explains the HPLC principle and instrumentation.
 
https://www.youtube.com/embed/pGYXd4k5F9o
 

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INSTALLATION on-site in USA.
Includes:
• Pump sea

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USED AGILENT HP 1200 HPLC Series SYSTEM  

We offer an Agilent Technologies 1200 Series HPLC System in excellent working condition. Includes:
    •    Tested and Verified in Good Working Condition prior to ship
    •    G1322A Agilent 1200 Micro Vacuum Degasser
    •    Your choice Binary or Quaternary Pump
    •    Many AutoSamplers to choose from
    •    G1316B Agilent 1200 Thermostatted Column Compartment
    •    G1315D Agilent 1200/1260 DAD Diode Array Detector
    •    Agilent Solvent tray and bottles
    •    All fittings, cables, an

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Waters Acquity UHPLC  

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principle of hplc:High Performance Liquid Chromatography (HPLC)  

High performance liquid chromatography (HPLC) has become a very versatile and powerful separation and analytical method over the years. It is an advanced form of liquid chromatography (LC).
Instead of introducing the solvent into the column and allowing it to drip down under the influence of gravity, in HPLC the sample is forced through the column under high pressures of nearly 400 atm, resulting in faster and more efficient separation.
This technique is also called high pressure liquid chromatography.

The Principle of HPLC
HPLC follows the same basic principle as chromatography. Different components in the sample have varying affinities to the adsorbent material. This causes a difference in the flow rate for each component which leads to their separation as they come out of the column.

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Agilent 1260 Infinity II LC System  

Infinitely more confident.
The new 1260 Infinity LC sets higher standards in performance and value to give you more confidence in your results. The 600 bar power range combines with 80 Hz UV detector speeds and up to 10 times higher sensitivity. That‘s true UHPLC performance! At the same time you can run your existing HPLC methods completely unchanged. In short, the best solution for any HPLC or UHPLC application.
And within the price range of a typical HPLC!


Raising the HPLC standard 

The 1260 Infinity LC raises the bar in HPLC! With a 600 bar power range up to 5 mL/min for isocratic, quaternary and binary systems  and 200 bar up to 10 mL/min for isocratic and quaternary systems – plus 80 Hz detector speeds and 10 x higher sensitivity, you will reach new levels of versati

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Hplc Agilent Technologies 1260 Infinity II LC System  

Infinitely more confident.
The new 1260 Infinity LC sets higher standards in performance and value to give you more confidence in your results. The 600 bar power range combines with 80 Hz UV detector speeds and up to 10 times higher sensitivity. That‘s true UHPLC performance! At the same time you can run your existing HPLC methods completely unchanged. In short, the best solution for any HPLC or UHPLC application.
And within the price range of a typical HPLC!


Raising the HPLC standard 

The 1260 Infinity LC raises the bar in HPLC! With a 600 bar power range up to 5 mL/min for isocratic, quaternary and binary systems  and 200 bar up to 10 mL/min for isocratic and quaternary systems – plus 80 Hz detector speeds and 10 x higher sensitivity, you will reach new levels of ve

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HPLC Electro Chemical Detector ( ECD )  

There are several different types of ECs. The detection is based on amperometry, polarography, coulometry, and conductrometry. They offer high sensitivity, simplicity, convenience, and wide-spread applicability. It is especially suitable for the use with semi-micro or capillary type system.
Introduction Of Electro Chemical Detector ( ECD )
Electrochemical detection (ECD) for HPLC or UHPLC is an extremely selective and sensitive detection technique that is applied in a number of analyses such as the neurotransmitters dopamine, serotonin and noradrenalin. In combination with the proper electronics, ECD has an enormous linear dynamic range of more then 6 orders of magnitude. This means that concentrations can be measured as low as 50 pmole/L and as high as 100 µmol/L or more.
Fig. 1. HPLC w

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What Is Ultra Performance Liquid Chromatography (UPLC Technology)?  

In 2004, further advances in instrumentation and column technology were made to achieve very significant
increases in resolution, speed, and sensitivity in liquid chromatography. Columns with smaller particles [1.7
micron] and instrumentation with specialized capabilities designed to deliver mobile phase at 15,000 psi [1,000
bar] were needed to achieve a new level of performance. A new system had to be holistically created to perform
ultra-performance liquid chromatography, now known as UPLC technology.
Basic research is being conducted today by scientists working with columns containing even smaller 1-microndiameter
particles and instrumentation capable of performing at 100,000 psi [6,800 bar]. This provides a glimpse
of what we may expect in the future.

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chromatography  

Chromatography is a physical method of separation that distributes components to separate between two phases, one stationary (stationary phase), the other (the mobile phase) moving in a definite direction. The eluate is the mobile phase leaving the column. The eluent is the solvent that carries the analyte.

Chromatography

chromatography.hplchplc.com/ 
 







Chromatography is a physical method of separation that distributes components to separate between two phases, one stationary (stationary phase), the other (the mobile phase) moving in a definite direction. The eluate is the mobile phase leaving the column. The eluent is the solvent that carries the analyte.


Partition Chromatography


chromatography.hplchplc.com/page-461157.html
 
Partition chromatography was one of the first k

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Shimadzu Nexera X2  

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Agilent HPLC 1200  

The Agilent 1200 Series HPLC System was introduced in 2010 with a modular design allowing users to define a configuration ideally suited to meet their HPLC and UHPLC applications and requirements. 

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column chromatography principle  

When a mixture of mobile phase and sample to be separated are introduced from top of the column, the individual components of mixture move with different rates.Those with lower affinity and adsorption to stationary phase move faster and eluted out first while those with greater adsorption affinity move or travel slower and get eluted out last.
The solute molecules adsorb to the column in a reversible manner. The rate of the movement of the components is given as follows
R= Rate of movement of a component / Rate of movement of mobile phase. i.e. it is the ratio of distance moved by solute to the distance moved by solvent.
source:studyread.com/

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Waters ACQUITY Arc HPLC and UHPLC System  

HPLC and UHPLC method compatibility at the flip of a switch

The ACQUITY Arc System is a quaternary-based, modern LC system for scientists working with established methods who are looking for the versatility and robustness required to bridge the gap between HPLC and UPLC while continuing to support validated assays.

True plug-and-play method compatibility and the ability to switch between HPLC and UHPLC separations at the flip of a switch with Arc Multi-flow path Technology
Get mass detection, without changing the way you work, with the ACQUITY QDa Detector
Deploy the benefits of 2.x μm particle columns to boost the chromatographic performance of your methods
On-demand mobile phase creation with Auto●Blend Plus Technology, eliminating the cumbersome and error-prone appr

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Evaporative Light Scattering Detector (ELSD)  

An evaporative light scattering detector (ELSD) is a detector used in conjunction with high-performance liquid chromatography (HPLC). It is commonly used for analysis of compounds where UV detection might be a restriction and therefore compounds does not very efficient absorb UV radiation, such as sugars, antivirals, antibiotics, lipids, phospholipids, terpenoids, and alcohols. ELSDs fall under the category of general-purpose detectors, similar to refractive index detectors (RI).

ELSD | Evaporative Light Scattering Detector

elsd.hplchplc.com/

 

... Scattering Detector. An evaporative light scattering detector (ELSD) is a detector used in conjunction with high-performance liquid chromatography (HPLC).

--------------------------

P

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Column Chromatography is the the first form of techniques developed in chromatography!  

Column chromatography is the prototype or the basic type of chromatography. It was the first form of techniques developed in chromatography. One can easily explain the principle and procedure of chromatography using it.
Other types of chromatography techniques like high performance liquid chromatography (HPLC), gas chromatography (GC), paper chromatography were developed with column chromatography as a module and making slight variations.
In-spite of many advanced methods of chromatography, still this method of chromatography is widely used in research and industry.
This chromatography is basically a type of adsorption chromatography techniques.

Here the separation of components depends upon the extent of adsorption to stationary phase. Here the stationary phase is a polar solid materia

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Principle of HPLC:High Performance Liquid Chromatography (HPLC) : Principle, Types, Instrumentation and Applications  

Chromatography is a technique to separate mixtures of substances into their components on the basis of their molecular structure and molecular composition. This involves a stationary phase (a solid, or a liquid supported on a solid) and a mobile phase (a liquid or a gas). The mobile phase flows through the stationary phase and carries the components of the mixture with it. Sample components that display stronger interactions with the stationary phase will move more slowly through the column than components with weaker interactions. This difference in rates cause the separation of variuos components. Chromatographic separations can be carried out using a variety of stationary phases, including immobilized silica on glass plates (thin-layer chromatography), volatile gases (gas chromatography

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Troubleshooting HPLC- Loss in Response for Some, but Not All Analytes  

Patterns in your HPLC chromatography can exhibit telltale signs that point toward probable sources of error.  In this brief series of posts, we will look at possible scenarios that you may encounter in the laboratory and how you might approach resolving those difficulties.
When some, but not all of your peaks are low, and you are analyzing multiple analytes, the number one rule is to look for trends. As mentioned in the previous post for this series, when you have only one analyte, or a handful of very similar analytes, the following may also apply. Again in this post, the assumption is that you are using an established working method and that you see a loss in response for every sample or standard that is injected. If you are only seeing the problem with samples and not your quantitation

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Download Hplc Agilent Technologies 1290 Infinity II LC System Brochure and Guide  

 

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Agilent InfinityLab LC Series Selection Guide

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10-Jun-2016
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Download Agilent 1290 Infinity II LC – More Efficiency, More Free Time
The Agilent 1290 Infinity II LC is the next generation UHPLC, maximizing analytical, instrument and laboratory efficiency.

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ApplicationsArticle ReprintsBrochures

Download Agilent 1290 Infinity II LC – More Efficiency, More Free Time
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Shimadzu Evaporative Light Scattering LC Detector LTII (ELSD-LTII)  

In the history of high-performance liquid chromatographs, which dates to the early 1960s, refractive index detectors (RI detectors) have often been used as general-purpose detectors. RI detectors enable the detection of components that do not possess UV absorbance and give a proportional relationship between the heights of detected peaks and the quantities of detected components. So, in comparison with absorbance detectors (UV detectors), they offer advantages such as the ability to ascertain unknown component quantities and obtain molecular weight distributions for macromolecules. On the other hand, they also have various disadvantages. For example, they cannot be used for gradient analysis, the baselines they produce are susceptible to the influence of fluctuations in the ambie

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Displacement Chromatography  

The basic principle of displacement chromatography is: A molecule with a high affinity for the chromatography matrix (the displacer) will compete effectively for binding sites, and thus displace all molecules with lesser affinities. There are distinct differences between displacement and elution chromatography. In elution mode, substances typically emerge from a column in narrow, Gaussian peaks. Wide separation of peaks, preferably to baseline, is desired in order to achieve maximum purification. The speed at which any component of a mixture travels down the column in elution mode depends on many factors. But for two substances to travel at different speeds, and thereby be resolved, there must be substantial differences in some interaction between the biomolecules and the chromatography m

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Reversed Phase Chromatography (RPC)  

Reversed phase HPLC (RP-HPLC) has a non-polar stationary phase and an aqueous, moderately polar mobile phase. One common stationary phase is a silica which has been surface-modified with RMe2SiCl, where R is a straight chain alkyl group such as C18H37 or C8H17. With such stationary phases, retention time is longer for molecules which are less polar, while polar molecules elute more readily (early in the analysis). An investigator can increase retention times by adding more water to the mobile phase; thereby making the affinity of the hydrophobic analyte for the hydrophobic stationary phase stronger relative to the now more hydrophilic mobile phase. Similarly, an investigator can decrease retention time by adding more organic solvent to the eluent. RP-HPLC is so commonly used that it is oft

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Principle Of Column Chromatography  

Column chromatography in chemistry is a method used to purify individual chemical compounds from mixtures of compounds. It is often used for preparative applications on scales from micrograms up to kilograms. The main advantage of column chromatography is the relatively low cost and disposability of the stationary phase used in the process. The latter prevents cross-contamination and stationary phase degradation due to recycling.
The classical preparative chromatography column is a glass tube with a diameter from 5 mm to 50 mm and a height of 5 cm to 1 m with a tap and some kind of a filter (a glass frit or glass wool plug – to prevent the loss of the stationary phase) at the bottom. Two methods are generally used to prepare a column: the dry method and the wet method.

For the dry

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Shimadzu Nexera XR  

The Most Accessible UHPLC Available 
Developed with expandability and compatibility in mind, the Nexera XR ultra high performance liquid chromatograph enables more customers to make use of high-speed, high-resolution systems. Configure the optimal system to meet the specific analysis objective by selecting from among a wide range of highly accurate and reliable modules. The next milestone in the evolution of liquid chromatography, the Nexera XR promises to become an indispensable tool in laboratories in a variety of fields, including pharmaceuticals, biochemistry, chemistry, environmental, and foods.
Expanding the Possibilities of UHPLC :


Superb Functionality

Accuracy and reproducibility of solvent delivery and injection volume are particularly important performance aspects. Shim

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Shimadzu Prominence nano  

The Most Accessible UHPLC Available 
Developed with expandability and compatibility in mind, the Nexera XR ultra high performance liquid chromatograph enables more customers to make use of high-speed, high-resolution systems. Configure the optimal system to meet the specific analysis objective by selecting from among a wide range of highly accurate and reliable modules. The next milestone in the evolution of liquid chromatography, the Nexera XR promises to become an indispensable tool in laboratories in a variety of fields, including pharmaceuticals, biochemistry, chemistry, environmental, and foods.
Expanding the Possibilities of UHPLC :


Superb Functionality

Accuracy and reproducibility of solvent delivery and injection volume are particularly important performance aspects. Shim

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1290 HPLC  

The Agilent 1290 Infinity LC system provides the highest levels of speed, resolution, flexibility and sensitivity for any LC and LC/MS application

 

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Partition Chromatography  

Partition chromatography was one of the first kinds of chromatography that chemists developed. The partition coefficient principle has been applied in paper chromatography,thin layer chromatography, gas phase and liquid–liquid separation applications. The 1952Nobel Prize in chemistry was earned by Archer John Porter Martin and Richard Laurence Millington Synge for their development of the technique, which was used for their separation of amino acids. Partition chromatography uses a retained solvent, on the surface or within the grains or fibers of an "inert" solid supporting matrix as with paper chromatography; or takes advantage of some coulombic and/or hydrogen donor interaction with the stationary phase. Analyte molecules partition between a liquid stationary phase and the eluent.

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waters hplc  

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Troubleshooting HPLC- Loss in Response for Some, but Not All Analytes  

Just as in GC applications, patterns in your HPLC chromatography can exhibit telltale signs that point toward probable sources of error.  In this brief series of posts, we will look at possible scenarios that you may encounter in the laboratory and how you might approach resolving those difficulties.
We occasionally get calls or emails from folks that are seeing lower than expected response for peaks of interest. This may be a matter of method development if you have not done this analysis previously. Suggestions in the post are provided assuming you are working with an established method and have had successful results over an extended period of time (until recently). These suggestions also assume that the response is low for quantitation standards as well as in prepared QC samples. If

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Troubleshooting HPLC- Loss in Response for all Analytes  

Just as in GC applications, patterns in your HPLC chromatography can exhibit telltale signs that point toward probable sources of error.  In this brief series of posts, we will look at possible scenarios that you may encounter in the laboratory and how you might approach resolving those difficulties.
We occasionally get calls or emails from folks that are seeing lower than expected response for peaks of interest. This may be a matter of method development if you have not done this analysis previously. Suggestions in the post are provided assuming you are working with an established method and have had successful results over an extended period of time (until recently). These suggestions also assume that the response is low for quantitation standards as well as in prepared QC samples. If t

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Troubleshooting HPLC- Tailing Peaks  

As in the previous posts for this series, these suggestions assume that you are working with an established method and had successful results in the past. If a method is rugged, it is developed to minimize tailing by using a stationary column phase that is base-deactivated as well as appropriate modifiers and pH adjustment in the mobile phase as needed.  Keep in mind that if the method is not as rugged as it should be, change(s) in the method might be advised if tailing continues to be an ongoing problem.

With troubleshooting, as always, it is critical to note when the change occurred and how it might correlate to changes in the system.  Equally important is to change one thing at a time to identify the source(s) of difficulty. The following are examples of things that could contribute

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principle of hplc:How Does High Performance Liquid Chromatography Work?/what is Detector,Chromatogram?  

How Does High Performance Liquid Chromatography Work?

The components of a basic high-performance liquid chromatography [HPLC] system are shown in the simple diagram in Figure E.
A reservoir holds the solvent [called the mobile phase, because it moves]. A high-pressure pump [solvent delivery system or solvent manager] is used to generate and meter a specified flow rate of mobile phase, typically milliliters per minute. An injector [sample manager or autosampler] is able to introduce [inject] the sample into the continuously flowing mobile phase stream that carries the sample into the HPLC column. The column contains the chromatographic packing material needed to effect the separation. This packing material is called the stationary phase because it is held in place by the column hardware.

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