Purification of meta-cresol purple and cresol red by flash chromatography: Procedures for ensuring accurate spectrophotometric seawater pH measurements

Abstract

Impurities in sulphonephthalein indicator salts can result in significant errors in seawater pH determinations. To ensure suitable measurement accuracy and intercomparability on a global basis, impurities must be removed from all indicators used for oceanographic CO2 system analyses. Previous work has described an effective HPLC (high-performance liquid chromatography) procedure for purification of meta-cresol purple, but the technique is labor-intensive, with each HPLC run producing only a small batch of purified indicator. This work describes the use of flash chromatography to more efficiently produce large batches of purified meta-cresol purple (mCP) and cresol red (CR), the preferred indicators for direct water column determinations of seawater pH.Several batches of unrefined mCP and CR of independent origin were prepared by flash chromatography. Indicator purity was then assessed in two ways: by (a) HPLC verification and (b) pH measurements of highly buffered solutions. HPLC chromatograms of the various flash-prepared mCPs indicated that the process did not always result in a completely pure product. In terms of performance, however – i.e., pH measurements of highly buffered solutions – no differences were observed between an HPLC-purified reference mCP and the flash-purified mCPs. HPLC examination of the flash-purified CRs indicated that every product was free of detectable impurities. No differences were seen in comparative pH measurements made with the purified CRs. The flash chromatography procedures outlined in this work are suitable for producing bulk quantities of mCP and CR for use in high-precision spectrophotometric pH measurements in seawater.