Page 24 - LC-SFC_Pharma_Brochure

C190-E202

Effective Analysis Management
Technical Achieved by Method Transfer between

Report HPLC and UHPLC

Akihiro Kunisawa , Daiki Fujimura , Yusuke Osaka 1
1
1

Abstract:

High performance liquid chromatography (HPLC) is widely used for the qualitative and quantitative analysis. In recent years, ultra-high
performance liquid chromatography (UHPLC) has been developed signiÿcantly. Facilities that have both UHPLC and HPLC systems often
apply newly established UHPLC method to ordinary HPLC as well as applying existing HPLC method to UHPLC. During such inter-system
method application, the method must be modiÿed properly to meet the requirements from another system. This modiÿcation is called
“method transfer”. Method transfer requires to modify existing parameters such as °ow rate and/or time pr ogram.
Here, we describe the relationship between column dimensions including particle size of packing material and analytical conditions, then
method transfer from HPLC to UHPLC for high speed analysis and that from UHPLC to HPLC for generalization of analytical conditions as
well as that from Shimadzu’s system to other vendor’s. We also describe the Shimadzu integrated LC system “Nexera-i MT” that supports
method transfer and ACTO (Analytical Conditions Transfer and Optimization) Function equipped in the latest LabSolutions LC software.
Keywords: HPLC, UHPLC, method transfer, ACTO, Nexera-i MT

1. Background
1. Backgr ound
HPLC
In pharmaceutical, food and various industrial fields, HPLC is widely
used for the analysis of target compounds and related impurities. In
recent years, UHPLC system that has more than 100 MPa of pressure
tolerance affords more efficient analysis at ultra-high speed. Due to
these features, UHPLC systems have been introduced in a variety of
facilities where R&D departments often use UHPLC systems to devel-
op an efficient analytical method then modify it to match HPLC analy-
sis. The method transferred are then used by the QC department.
Conversely, an existing HPLC method can be transferred into a
UHPLC method to improve the speed and efficiency of analyses. In
this report, we define method transfer as both directions of method 0.0 10.0 20.0 30.0 40.0 min
modifications from HPLC to UHPLC and from UHPLC to HPLC.
Method transfer
We often face inadequate separation in gradient elution after a (speed improvement)
method transfer from HPLC where the separation is completed, to
UHPLC (Fig.1). Furthermore, even though a method may have been
created successfully using UHPLC analysis, the method transfer from UHPLC
UHPLC to HPLC gives different selectivity of separation, resulting in a
degradation of efficiency for the entire laboratory.
Generally, a method transfer accompanied by employing different
column dimensions including particle size must be required to modify
analytical conditions such as flow rate and time program. However,
to optimize conditions is not easy and method transfer can result in
poor separation compared to those obtained prior to the method
transfer. Keeping a consistent separation pattern before and after
method transfer in terms of parameters, such as resolution and rela-
tive retention times, requires complex calculations based on pre- and
post-transfer data, which include column dimensions (length, internal 0.0 1.0 2.0 3.0 4.0 5.0 6.0 min
diameter, and particle size).
Fig. 1 Problems That Arise During Method Transfer
(Analysis of cephem antibiotics)

1 Analytical & Measuring Instruments Division 1