Quantification of Plant Sterols in Pulp and Paper Mill Effluents

Pulp and paper mill effl uents (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 fi sh and other aquatic life inhabiting PPME receiving waters. Plant sterols, or phytosterols, form a constituent of wood extractives that may be released into the effl uents 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 modifi ed 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. Identifi cation and quantifi cation 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 modifi ed technique offered improved method sensitivity and reproducibility, and successfully quantifi ed campesterol, s-sitosterol, s-sitostanol, stigmasterol, stigmastanol, cholesterol, and ergosterol in PPMEs. Primary and secondary treated PPMEs analyses suggested 800 ± 190 μg/L total sterols in primary effl uents, and 211 ± 90 μg/L in biologically treated fi nal effl uents. s-Sitosterol, s-sitostanol, and campesterol alone accounted for about 80% of the total sterols. A general comparison of the sterols in primary and secondary effl uents suggested about 73% removal across the secondary treatment systems sampled.