HPLC-DAD Purification and Characterization of Meta-Cresol-Purple for Spectrophotometric Seawater pH Measurements

High-quality pH measurements are essential for observing ocean acidification and interpreting its chemical and ecological effects. Spectrophotometric measure is the preferred method for pH analysis of seawater because of its high reproducibility and excellent precision. Meta-cresol purple (mCP) is one of the most used indicator dyes for spectrophotometric pH determination. Impurities in indicator salts can significantly bias spectrophotometric pH determinations, and therefore the use of purified mCP is recommended. This work describes the use of a fast, simple, and green-friendly High-Performance-Liquid Chromatography (HPLC) procedure for purification of mCP dye solution. A C18 reversed phase column was used, and different compositions of the mobile phase were considered. The selected mobile phase was 85% Milli-Q water and 15% CH3CN pH 8.0 that allowed reasonable run time and a suitable purified mCP retention time (4.65 & PLUSMN; 0.005 min). Aliquots of purified mCP were mixed and evaporated by rotary evaporator that resulted in the fastest preconcentration procedure among those considered in this study. Purified mCP was used for measuring the pH in seawater samples collected in the Ross Sea (Antarctica) and the values ranged between 8.202 and 7.983, with the highest values at the surface.

Automated summary

High-quality pH measurements are crucial for studying ocean acidification. The spectrophotometric analysis using indicator dyes like meta-cresol purple (mCP) is preferred, but impurities in indicator salts can bias results. This study introduces a fast and green-friendly HPLC method for purifying mCP, which was successfully applied for pH measurement in seawater samples from the Ross Sea.