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Updated: Sep 30, 2025

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry UPLC-HRMS
Published on: May 20, 2013
Effects of sampling rate and noise filter processing on repeatability assessment in UHPLC with ultraviolet detection
Koichi Machida1, Ryo Watanabe1, Akira Kotani2
1School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan.
Abstract:
The present study examines whether short measurement time and noise filter processing in an ultra-high-performance liquid chromatography with ultraviolet detection (UHPLC-UV) contribute to limitations for repeatability assessment based on the ISO 11843 part 7 (ISO 11843-7), which can stochastically provide a measurement standard deviation (SD) caused by baseline noise (SB). In this study, ergosterol was used as an example in UHPLC-UV analysis. From the results of power spectrum analysis of baseline noise, 1024 consecutive digital data points provided a suitable SB. Thus, it was found that an SB can be obtained from about 1 min of baseline when a chromatogram was recorded at sampling rate of 20 points s-1 in the present UHPLC-UV system. The relative SDs (RSDs) of the peak area obtained by the ISO 11843-7 were within 95% of the confidential intervals of the RSDs obtained by repetitive measurements, indicating the ISO 11843-7 is applicable to estimate repeatability in a UHPLC-UV system. In a similar way, we found that the RSD of the peak area obtained from a chromatogram with noise filter processing in UHPLC-UV could also be estimated by the ISO 11843-7. In conclusion, we experimentally demonstrate that short measurement time and noise filter processing are not limitations for repeatability assessment based on the ISO 11843-7.
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