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Besides microbiological methods, fecal pollution of surface waters is estimated by gas chromatographic (GC) determination of sterols present in human and animal sewage effluents. The most frequently used biomarkers for the evaluation of contamination levels include coprostanol, cholesterol, dihydrocholesterol, stigmasterol, β-sitosterol, and stigmastanol. Although several GC techniques are used to measure these compounds in aquatic systems, the analytical performance of GC-mass spectrometric (MS) determination of these sterols has not been systematically characterized. Therefore, the aim of this work is to validate a simple and rapid GC-MS method for the simultaneous analysis of six sterols, considering all parameters and requirements defined by Good Laboratory Practice. Following liquid-liquid extraction of spiked surface water samples, the extracts are silylated and analyzed by GC-MS. The method is evaluated for linearity and limits of detection and quantitation, as well as for precision, extraction efficiency, and stability. The assay is linear up to 160 ng; the limits of detection and quantitation are 5–10 ng and 20 ng, respectively. The within- and between-day precision ranged from 1% to 9% and 1% to 16%, respectively. The extraction efficiency was 65–80%. The stability studies indicate that the sterols in surface water samples begin to degrade after 24 h of refrigerated storage. However, three freeze/thaw cycles could be performed without their decomposition. The method is applied to the analysis of surface water and wastewater samples. The technical advantages make this GC-MS analysis suitable for routine environmental monitoring of fecal pollution in aquatic systems.

Method Validation for the Simultaneous Determination of Fecal Sterols in Surface Waters by Gas Chromatography-Mass Spectrometry