SOLID PHASE EXTRACTION AND HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH PHOTODIODE ARRAY DETECTION OF CHEMICAL INDICATORS OF HUMAN FECAL CONTAMINATION IN WATER

Abstract

Faster and more sensitive analysis of water that is contaminated by human fecal matter is very important for public health. The current microbiological methods to assess water quality do not meet this need. Alternate non-microbial human fecal indicators have been proposed by various researchers. The high performance liquid chromatography (HPLC) analysis with photodiode array detection (PDA) of three human fecal indicators, caffeine, urobilin, and coprostanol was developed. Both caffeine and urobilin were analyzed simultaneously using a linear gradient protocol and monitored at 270 nm and 480 nm. The analysis gave linearity and limit of detection down to hundredths of μg/mL.

A solid phase extraction (SPE) protocol was also developed for caffeine and urobilin. Coprostanol was analyzed using HPLC with PDA detection with post-column derivatization. The derivatization was achieved by acylation of coprostanol with p-nitrobenzoyl chloride.

ACKNOWLEDGMENTS

Drs. Carl Selikar and Tom Ridgeway of the University of Cincinnati and Roger Macomber, now of Pepperdine University, are gratefully acknowledged for their helpful discussions. This research was supported, in part, by an appointment to the Post Graduate Research Participation Program administered by the Oak Ridge Institute for Science and Education through an inetragency agreement between the U. S. DOE and the U. S. EPA.

Dr. Piocos is currently with the Procter and Gamble Company, Cincinnati, Ohio 45201, U.S.A.