Sample Preparation Followed by High Performance Liquid Chromatographic (HPLC) Analysis for Monitoring Muconic Acid as a Biomarker of Occupational Exposure to Benzene |
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Authors: | Seyed Jamaleddin Shahtaheri Farhad Ghamari Farideh Golbabaei Abbas Rahimi-Froushani Mohsen Abdollahi |
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Affiliation: | 1. School of Public Health &2. Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, I.R, Iransjtaheri@sphtums.com;4. Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, I.R, Iran |
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Abstract: | Factors affecting solid phase extraction (SPE) of trans,trans-muconic acid (ttMA), as a benzene biomarker, including sample pH, sample concentration, sample volume, sample flow rate, washing solvent, elution solvent, and type of sorbent were evaluated. Extracted samples were determined by HPLC-UV(highperformance liquid chromatography-ultraviolet). The analytical column was C18, UV wave length was 259 nm, and the mobile phase was H2O/methanol/acetic acid run at flow rate of 1 ml/min. A strong anion exchange silica cartridge was found successful in simplifying SPE. There was a significant difference between recoveries of ttMA when different factors were used (p < .001). An optimum recovery was obtained when sample pH was adjusted at 7. There was no significant difference when different sample concentrations were used (p > .05). The optimized method was then validated with 3 different pools of samples showing good reproducibility over 6 consecutive days and 6 within-day experiments. |
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Keywords: | benzene muconic acid sample preparation solid phase extraction biological monitoring chromatography |
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