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HPLC Basic Course 2.principle and System Configuration of HPLC (2) - Hitachi High-T

This document discusses the principles of qualitative and quantitative analysis using HPLC. Qualitative analysis involves identifying unknown components by comparing their retention times to a standard. Quantitative analysis requires creating a calibration curve by measuring standard samples at known concentrations and using peak areas to determine unknown concentrations. Attention must be given to uncertainties in qualitative analysis and the need for standard samples for accurate quantitative results.

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142 views2 pages

HPLC Basic Course 2.principle and System Configuration of HPLC (2) - Hitachi High-T

This document discusses the principles of qualitative and quantitative analysis using HPLC. Qualitative analysis involves identifying unknown components by comparing their retention times to a standard. Quantitative analysis requires creating a calibration curve by measuring standard samples at known concentrations and using peak areas to determine unknown concentrations. Attention must be given to uncertainties in qualitative analysis and the need for standard samples for accurate quantitative results.

Uploaded by

matrixworld20
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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HPLC Basic Course 2.

Principle and System Configuration of HPLC(2):Hitachi High-Technologies Corporation

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HPLC Basic Course


2.Principle and System Configuration of HPLC(2)

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HPLC Basic Course

Philosophy of Qualitative analysis and Quantitative analysis by chromatography

UV-Visible/NIR
Spectrophotometer
Fluorescence
Spectrophotometer

Qualitative analysis

A standard sample of aspartame was measured, yielding a peak at 12.5 minutes. This
peak corresponds to aspartame, itself.
Next,
a sample prepared from a beverage was measured under the same conditions,
yielding a number of peaks. The peak appearing at 12.5 minutes can be regarded as
that of aspartame (Figure 1).
The following conditions were the same: the type of fillers, column sizes, column
temperature, composition of the mobile phase, and flow rate.

Atomic Absorption
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Figure 1. Example of aspartame measurement

HPLC Columns
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C18
HPLC Columns
LaChrom Series

Quantitative analysis
The height and area of a peak are proportional to the concentration of the corresponding
component. A calibration curve is created using the standard sample. The
concentration
of aspartame in the beverage can be determined from the peak area of the detected
aspartame.

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HPLC Basic Course 2.Principle and System Configuration of HPLC(2):Hitachi High-Technologies Corporation

Attention should be given to the fact that a qualitative analysis includes many

uncertainties. Other components may have been eluted together with aspartame.
LC and GC systems are good at determining the content of a
certain component in a
sample, rather than the types of the components of a sample. Performance of a
quantitative analysis requires the preparation of a calibration curve. It is very difficult to
perform the qualitative or quantitative analysis of a component for which a standard
sample is not available.

The next chapter is "Configuration of an HPLC system".

Ultra High Performance Liquid Chromatograph ChromasterUltra Rs


High Performance Liquid Chromatograph Chromaster
High Performance Liquid Chromatograph Primaide
HPLC Columns LaChrom Series
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