Effects of sampling rate and noise filter processing on repeatability assessment in UHPLC with ultraviolet detection based on the ISO 11843-7.

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 1min of baseline when a chromatogram was recorded at sampling rate of 20pointss-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.