Study on the Derivatization Process Using N-O-bis-(trimethylsilyl)-trifluoroacetamide,N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide, Trimethylsilydiazomethane for the Determination of Fecal Sterols by Gas Chromatography-Mass Spectrometry

Fecal sterol has been proposed as a chemical indicator of human fecal pollution even when fecal coliform populations have diminished due to water chlorination or toxic effects of industrial effluents. This paper describes an improved derivatization procedure for simultaneous determination of four fecal sterols including coprostanol, epicholestanol, cholesterol and cholestanol using gas chromatography-mass spectrometry (GC-MS), via optimization study on silylation procedures using N-O-bis (trimethylsilyl)-trifluoroacetamide (BSTFA), and N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTBSTFA), which lead to the formation of trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS) derivatives, respectively. Two derivatization processes of injection-port derivatization and water bath derivatization (60 oC, 1h) were inspected and compared. Furthermore, the methylation procedure at 25 oC for 2h with trimethylsilydiazomethane (TMSD) for fecal sterols analysis was also studied. It was found that most of TMS derivatives demonstrated the highest sensitivities, followed by methylated derivatives. For BSTFA or MTBSTFA derivatization processes, the simple injection-port derivatization process could achieve the same efficiency as that in the tedious water bath derivatization procedure.

Use of Caffeine and Human Pharmaceutical Compounds to Identify Sewage Contamination

Fecal coliform bacteria are widely used as indicators of sewage contamination in surface water. However, there are some disadvantages in these microbial techniques including time consuming (18-48h) and inability in discriminating between human and animal fecal material sources. Therefore, it is necessary to seek a more specific indicator of human sanitary waste. In this study, the feasibility was investigated to apply caffeine and human pharmaceutical compounds to identify the human-source contamination. The correlation between caffeine and fecal coliform was also explored. Surface water samples were collected from upstream, middle-stream and downstream points respectively, along Rochor Canal, as well as 8 locations of Marina Bay. Results indicate that caffeine is a suitable chemical tracer in Singapore because of its easy detection (in the range of 0.30-2.0 ng/mL), compared with other chemicals monitored. Relative low concentrations of human pharmaceutical compounds (< 0.07 ng/mL) in Rochor Canal and Marina Bay water samples make them hard to be detected and difficult to be chemical tracer. However, their existence can help to validate sewage contamination. In addition, it was discovered the high correlation exists between caffeine concentration and fecal coliform density in the Rochor Canal water samples, demonstrating that caffeine is highly related to the human-source contamination.

Study on the Derivatization Process Using N-O-bis-(trimethylsilyl)-trifluoroacetamide, N-(tert-butyldimethylsilyl)-N-methyltrifluoroace tamide, Trimethylsilydiazomethane for the Determination of Fecal Sterols by Gas Chromatography-Mass Spectrometry

Fecal sterol has been proposed as a chemical indicator of human fecal pollution even when fecal coliform populations have diminished due to water chlorination or toxic effects of industrial effluents. This paper describes an improved derivatization procedure for simultaneous determination of four fecal sterols including coprostanol, epicholestanol, cholesterol and cholestanol using gas chromatography-mass spectrometry (GC-MS), via optimization study on silylation procedures using N-O-bis (trimethylsilyl)-trifluoroacetamide (BSTFA), and N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTBSTFA), which lead to the formation of trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS) derivatives, respectively. Two derivatization processes of injection-port derivatization and water bath derivatization (60 oC, 1h) were inspected and compared. Furthermore, the methylation procedure at 25 oC for 2h with trimethylsilydiazomethane (TMSD) for fecal sterols analysis was also studied. It was found that most of TMS derivatives demonstrated the highest sensitivities, followed by methylated derivatives. For BSTFA or MTBSTFA derivatization processes, the simple injection-port derivatization process could achieve the same efficiency as that in the tedious water bath derivatization procedure.