Abstract
Pulp and paper mill effluents (PPMEs) may contain high levels of otherwise naturally occurring organic pollutants such as plant sterols, which are suspected endocrine disrupting chemicals. Exposure to such chemicals may cause various physiological and morphological abnormalities that have been reported in the fish and other aquatic life inhabiting PPME receiving waters. Plant sterols, or phytosterols, form a constituent of wood extractives that may be released into the effluents during the pulping and paper making processes. Isolation and analysis of sterols from the complex mixture of PPMEs is challenging and standard analytical protocols do not exist. The need for having a reliable method for analyzing a particular environmental contaminant such as plant sterols cannot be overemphasized. In the present study a technique was modified for reliable analysis of PPME sterols. The technique involves liquid-liquid extractions using methyl-t-butyl ether and trimethyl-silylation derivatizations of the extracted sterols. Identification and quantification of the PPME sterols were accomplished by gas chromatography and mass spectrometry. Analytical problems were resolved by conducting multiple extractions, drying the sterol extracts, and redissolving and silylating the extracts at an increased derivatization temperature of 70°C. This shortened the suggested incubation period from 12 to 4 h. The modified technique offered improved method sensitivity and reproducibility, and successfully quantified campesterol, β-sitosterol, β-sitostanol, stigmasterol, stigmastanol, cholesterol, and ergosterol in PPMEs. Primary and secondary treated PPMEs analyses suggested 800 ± 190 µg/L total sterols in primary effluents, and 211 ± 90 µg/L in biologically treated final effluents. β-Sitosterol, β-sitostanol, and campesterol alone accounted for about 80% of the total sterols. A general comparison of the sterols in primary and secondary effluents suggested about 73% removal across the secondary treatment systems sampled.
