Relation between Chromatographic Resolution and Signal-to-Noise Ratio in Spectrophotometric HPLC Detection

Abstract

Absorption spectrophotometry has been and still is the industry standard for detection in HPLC. Limit of detection (LOD) and linear dynamic range (LDR) are the primary performance requirements and have driven continuous improvement of spectrophotometric HPLC detectors. Recent advances in HPLC column technology have led to low flow-rate HPLC such as capillary HPLC and nanoflow HPLC and put higher demands on optical HPLC signal detection. However, fundamental principles in spectrophotometric HPLC detection have not been reviewed for many years. In particular the relationship between the detector's signal-to-noise ratio (SNR) and band broadening needs to be re-evaluated. In this work, a new quantitative model is presented which allows the calculation of the trade-off made between chromatographic resolution and SNR in spectrophotometric HPLC detection. Modern optics flow cells based on total internal reflection are included and compared to conventional flow cells.

Development and validation of UV- Spectrophotometric and RP-HPLC method for the analysis of raw material and formulations of Aceclofenac

“…The efficiency of HPLC method for both mobile phases (Liang et al, 2007) was determined. It is well established fact that the lower are HETP, the more efficient is chromatographic system, thus enabling the detection of lower concentrations due to the enhanced signal-to-noise ratio of all the peaks in the chromatogram (Kraiczek et al, 2013). Using mobile phase "A", the samples (both standard and commercial) HETPs were calculated to be in the range of 530 to 543, while using mobile phase B, HETPs were evaluated to be in range of 190 to 226.…”

Development and validation of high-performance liquid chromatography method for the simultaneous monitoring of pantoprazole sodium sesquihydrate and domperidone maleate in plasma and its application to pharmacokinetic study

“…The features verified were retention time, tailing factor, peak area, theoretical plates, and resolution. The chromatographic resolution [23] is the difference in retention time between 2 adjacent peaks divided by the average peak width. Equation ( 4) is used for the calculation of resolution.…”

Prediction of Precision for Purity Methods

“…The original UBCI model considers 3 sources of error influencing peak area: error of injection, error of numerical integration, and error introduced by the system noise, estimates of which are rooted in first principles. 10,17,18 For this manuscript, we amended the model with the detector error. While the estimates of the first 3 errors are rooted in the first principles, the estimate of the detector error became empirical.…”