Abstract

Aromatic amines are widely used as intermediates in the dye, photographic, pharmaceutical and pesticide industries, and as antioxidants in the polymer industry. Because of their toxicity (Shonali and Richard 1990), several of them have been listed on the priority blacklist released by US Environmental Protection Agency in 1970s. Among several types of methods for degrading aromatic amines, biodegradation is one of the most important methods. So far, many species of bacteria have been isolated to biodegrade aromatic amines (Schnell et al. 1989). Accordingly, it is necessary that a simple and sensitive method be founded to determine the residue of aromatic amines in biological samples. HPLC with UV detection is often applied for the analysis of aromatic amines (Lewin et al. 1997). It is essential that an enrichment step be conducted to achieve low detection limit prior to analysis. Liquid–liquid extraction (LLE) and solid-phase extraction (SPE) are conventional pretreatment methods (Smith et al. 2003). However, the procedure of LLE is time-consuming and requires large quantity of toxic solvent that can be harmful to the environment. SPE requires less solvent, but the presence of particulate matter in the sample often results in plugging of the cartridge. Solid-phase microextraction (SPME), which is a rapid and solvent-free extraction, has been widely used for detecting aromatic amines (Wang et al. 2004; Yan and Jen 2003). However, the main drawback of SPME is that the fiber is expensive yet with a limited lifetime. Liquid-phase microextraction (LPME) is an extremely simple, low-cost and virtually solvent-free sample-preparation technique. Now LPME has developed several different forms, for example static and dynamic LPME, hollow-fiber membrane LPME, headspace LPME, and continuous-flow microextraction (CFME). Among these, CFME is a relatively novel LPME method, first reported by Liu and Lee (2000). It possesses such advantages as low cost, simplicity of pretreatment process, little usage of organic solvent and amity to environment. Much research has been carried out so far on microextraction of aromatic amines from water samples (Zhao et al. 2002). However, little information is available on it from Chlamydomonas reinhardtii samples. Therefore, the objective of this research was to use CFME-HPLC technique for analysis of 4-chloroaniline (4-CA) in C. reinhardtii cells. The effects of different factors on CFME efficiencies were also investigated and these factors included the kind of extraction solvent, solvent drop volume, sample flow rate, extraction time and addition amount of salt. To evaluate its real application effect, the optimal method was used to determine 4-CA in C. reinhardtii cells and tap water samples.

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