Ultra high performance liquid chromatography (UHPLC) has become popular due to the interest in improving productivity and lessening the burden on the environment. Method development systems utilizing UHPLC are also becoming an area of interest. This presentation examines the capabilities & benefits of this technology. For more information, visit www.ssi.shimadzu.com and follow us on Twitter @shimadzussi. Thanks for viewing!

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Examining the Capabilities and Benefits
of UHPLC Method Development Scouting
William Hedgepeth, Kenichiro Tanaka
Shimadzu Scientific Instruments, Columbia, MD., USA

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Introduction
The optimization of analysis conditions in high performance liquid chromatography (HPLC)
requires considerable time and effort.
Particularly with the analysis of impurities and natural substances, methods tend to be
constructed via trial and error.
A search for parameters, centered on columns and mobile phases, is required.
A variety of method development systems have been devised with the objective of
simplifying this process.
Nowadays, ultra high performance liquid chromatography (UHPLC) has become popular,
due to the interest in improving productivity and lessening the burden on the environment.
Method development systems utilizing UHPLC are becoming an area of interest.

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System Requirements
Scouting that can be performed quickly and easily by anyone
Easy configuration of the mobile phases and columns required for scouting
Easy configuration of the equilibration conditions for mobile phase and column switching
Quick creation of schedules for scouting
Desire to see the scouting conditions reflected in the data sampled

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Interested in:
•Investigating conditions using a variety of samples (STD, actual samples, etc.)
•Easily configuring gradient and isocratic conditions
•Reducing preparation time and eliminating configuration errors
•Assessing, in advance, the time required for scouting, as well as the mobile phase
quantities and sample quantities
•Batch management of columns and mobile phases using a database
•Real-time, onscreen confirmation of system status during scouting
•Reducing the data analysis time
•Identifying, at a glance, the conditions (columns, mobile phases, and gradient
conditions) from the name of the data file
•Arranging multiple chromatograms in a single window for comparative investigations
System Requirements

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SPD-M20A
PDA
SIL-30AC
LC-30AD
LC-30AD
FCV-34AH
MR180 µL
Manifold
Column
CTO-20AC
System Configuration
Figure 1: Nexera Method Scouting System

6. 6 / 136 / 13Figure 2: Nexera Method Scouting Interface
System Configuration
Graphical data is provided to support reliable analysis.

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The method scouting system can:
•Search conditions based on a maximum of 6 columns and up to 16 mobile phases.
•Be used with basically all current UHPLC columns (100 MPa valve pressure resistance).
•Easily configure scouting conditions thanks to special software.
•Automatically control everything from system checks to scouting and shutdown.
•Quickly evaluate scouting results via data browser or CLASS-Agent/Agent Report.
System Capabilities

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Benefits of Method Scouting
The clear benefit of a method scouting system is time savings derived from the
automated process.
Columns and mobile phases are placed on the system and set conditions are run without
human intervention - over lunch, overnight, or over the weekend - by automated mobile
phase and column switching.
This allows the method development process to be completed much sooner and frees
the analyst’s time for other tasks.
A significant amount of time is saved by not needing to manually enter methods and
individual injection conditions, made possible by an automated method file and batch
creation software.

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Benefits of Nexera Method Scouting
OvernightColumn 1 Column 2 Column 3 Column 4 Column 5 Column 6
Column 1 Column 2 Column 3 Column 4 Column 5 Column 6
Overnight
Since mobile phasesand columnscan be switched automatically,even overnight,data
can be collected efficiently.
Nexera Method Scouting System
Conventional UHPLC System
Day 1 Day 2 Day 3
Day 1 Day 2
Day 4
Analysis
All conditionscan be completed and the resultsviewed after 4 days.
Overnight
Analysis
Figure 3: Time Savings with Automated System

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Automated Method Scouting of Antibiotics
Investigation of batch analysis conditions for 13 cephem antibiotic compounds using
Nexera Method Scouting and Method Scouting Solution:
Structural Formulas for 13 Cephem Antibiotic Compounds

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Automated Method Scouting of Antibiotics
Step 1: Mobile phase and column scouting
Purpose: To search for the optimal mobile phase and column combination with
universal gradient conditions

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Automated Method Scouting of Antibiotics
Batch analysis conditions for 13 cephem antibiotic compounds are
automatically established with Nexera Method Scouting and Method Scouting
Solution.

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Summary
1. Automated UHPLC method scouting is an underutilized technology that greatly
reduces method development times.
1. Automated scouting can free an analyst’s time to perform other tasks and
increase lab productivity.

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