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C190-E204
Technical Improving Method Transfer
Report by Adjustment of Gradient Delay
Akihiro Kunisawa , Daiki Fujimura , Yusuke Osaka 1
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1
Abstract:
In facilities that have number of high performance liquid chromatography (HPLC) systems, an existing method that gives proper result by
one HPLC system is often applied to other HPLC systems (method transfer). However, due to the difference in system volume, pump
characteristics, and liquid delivery mechanisms among systems, method transfer can yield different results even though the same method
is used.
In this report, we ÿrst explain the gradient delay due to the system volume difference and the effect of the delay on separation. We then
show an example of method transfer with adjustment of gradient delay using alkylphenones and UV absorbents as samples. We also
describe the integrated liquid chromatograph (LC) system "i-Series" that supports to optimize method transfer and the newly designed
function of "Analytical Conditions Transfer and Optimization (ACTO) " equipped in the latest version of LabSolutions software.
Keywords: HPLC, method transfer, ACTO, i-Series
ound
1. Background
1. Backgr
HPLC is used for the analysis of target compounds and their related System A
impurities in a variety of applications including pharmaceutical and
food products. Facilities that use HPLC systems create methods using
their own original analytical conditions and/or specified testing regu-
lations. The validated methods are then used with a number of other
HPLC systems in many cases. In such situations, reproducibility
(compatibility) among systems is an important factor as well as re-
peatability of measurements. Even when using the same method, dif-
ferent HPLC systems can give different chromatograms (Fig. 1). Par-
ticularly in gradient elution, retention times, resolution and other fac-
tors will be largely affected as a result of method transfer. For exam- 0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 min
ple, while an existing method may succeed in separating a target
compound from co-existing impurities in one system, the same
method may not succeed in separating these compounds in other Method Transfer
systems. So it is often required to optimize analytical conditions for
each individual system, which is an extremely time-consuming pro-
cess. Such variations in retention time and separation are caused by
difference in system volume and pump performance among systems
(see section 2 for details). Especially in ultrahigh-speed analysis, even Unable to obtain the same System B
small difference in system volume can cause great difference in analy- chromatogram
sis results due to small volume of dedicated column.
Further, in pharmaceuticals, food and other fields where the test
methods are specified by regulations, changes in analytical conditions
are not permitted, which may be an issue.
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Fig. 1 Problems in Method Transfer
1 Analytical & Measuring Instruments Division 1