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Examining the Capabilities and Benefitsof UHPLC Method Development Scouting

William Hedgepeth, Kenichiro TanakaShimadzu 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-M20APDA

SIL-30AC

LC-30AD

LC-30AD

FCV-34AH

MR180 L

ManifoldColumn

CTO-20AC

System Configuration

Figure 1: Nexera Method Scouting System

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 phases and columns can 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 4Analysis

All conditions can be completed and the results viewed 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 usingNexera 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 scoutingPurpose: 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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