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SOLID PHASE EXTRACTION AND HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH PHOTODIODE ARRAY DETECTION OF CHEMICAL INDICATORS OF HUMAN FECAL CONTAMINATION IN WATER
Citation:
Piocos, E. A. AND A A. de la Cruz. SOLID PHASE EXTRACTION AND HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH PHOTODIODE ARRAY DETECTION OF CHEMICAL INDICATORS OF HUMAN FECAL CONTAMINATION IN WATER. JOURNAL OF LIQUID CHROMATOGRAPHY AND RELATED TECHNOLOGY 23(8):1281-1291, (2000).
Impact/Purpose:
Develop rapid, practical and sensitive biosensor, immunoassay, chemiluminescence and HPLC methods to detect caffeine, urobilin, and coprostanol, using them as non-microbial indicators of human fecal contamination in water.
Description:
Faster and more sensitive analysis of water that is contaminated by human fecal matter is very important for 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 monitoried at 270 nm and 480 nm. The analysis gave linearity and limit of detection down to hundredths of ug/mL.
A solid phase extraction (SPE) protocol was also developed for cafffeine 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.