Speciation determination of arsenic in urine by high-performance liquid chromatography-hydride generation atomic absorption spectrometry with on-line ultraviolet photooxidation

A coupled system for arsenic speciation determination based on high-performance liquid chromatography (HPLC), on-line UV photooxidation and continuous-flow hydride generation atomic absorption spectrometry (HGAAS) was built from commercially available modules with minor modifications to the electronic interface, the software and the gas-liquid separator. The best results were obtained with strong anion-exchange columns, Hamilton PRP X-100 and Supelcosil SAX 1, and gradient elution with phosphate buffers containing KH2PO4-K2HPO4. The on-line UV photooxidation with alkaline peroxodisulfate, 4% m/v K2S2O8-1 mol l-1 NaOH, in a PTFE knotted reactor for 93 s ensures the transformation of inorganic AsIII, monomethylarsonate, dimethylarsinate, arsenobetaine, arsenocholine, trimethylarsine oxide and tetramethylarsonium ion to arsenate. About 32-36 HPLC-UV-HGAAS runs could be performed within 8 h, with limits of detection between 2 and 6 micrograms l-1 As, depending on the species. The method was applied to the analysis of spot urine samples and certified urine reference materials (CRMs). Upon storage at 4 degrees C, reconstituted CRMs are stable for at least 2 weeks with respect to both their total arsenic content and the individual species distribution.