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Relation between chromatographic resolution and signal-to-noise ratio in spectrophotometric HPLC detection.

Authors
  • 1
  • 1 Agilent Technologies, Hewlett-Packard Strasse 8, Waldbronn, Germany. [email protected] , (Germany)
Type
Published Article
Journal
Analytical Chemistry
1520-6882
Publisher
American Chemical Society
Publication Date
Volume
85
Issue
10
Pages
4829–4835
Identifiers
DOI: 10.1021/ac4004387
PMID: 23593980
Source
Medline
License
Unknown

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.

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