Although it is degraded by environmental soil microorganisms, alachlor and its main metabolite, which can difuse into groundwater and disperse in the environment, cause alachlor is also known as an extremely toxic endocrine disrupting chemical and classified as a carcinogen of the B 2 group by the U. S. Environmental 
Protection Agency. The potential toxicity of alachlor has been evaluated in a series of rodent chronic bioassays such as in vitro clastogen in Chinese hamster ovary cells and human lymphocytes. However, when toxic pollutants exist in the environment, an efect on microbial growth ability is usually observed. Szaba et al. studied the in uence of alachlor and zinc on the growth of the filamentous fungus Paecilomyces marquandii and its ability to eliminate alachlor and zinc.
Since then, cytotoxicity studies of herbicides are often carried out in cell culture. In addition to the DNA fragmentation analysis and immunoblot analysis, a simple, fast, Angiogenesis and reliable method to determine alachlor and its metabolite 2,6 DEA in microbial culture medium samples is required in plant pathology studies. For the analysis of chloroacetanilide herbicide metabolites, high performance liquid chromatography is preferred because most of the chloroacetanilide metabolites are ionic compounds, which are not sufciently volatile for analysis by gas chromatography. However, an appropriate pretreatment and the enrichment of target species are required prior to HPLC analysis.
The conventional pretreatment methods for the analysis of alachlor and its metabolites PH-797804 include liquid_liquid extraction, solid phase extraction, pressurized liquid extracLLE is not efcient for polar species and ionic compounds and is under criticism for using large quantities of organic solvents, thereby causing pollution accompanied by health risks, in addi tion to the extensive time consuming cleanup procedures. The use of SPE has eliminated or decreased most of the disadvantages of LLE. However, plugging of the cartridges or disks by high molecular weight species in culture medium limits the application tion, and solid phase microextraction. Although SPME has the advantages of simultaneous solvent free extraction culture medium because the fiber is easily coated by the high molecular weight species present in samples. Therefore, it is vital to investigate a reliable and eco friendly method to extract alachlor and its metabolite 2,6 DEA in culture medium.
In the recent decade, an efcient enrichment c-Met Signaling Pathway method was needed for the analysis of complex matrix liquid samples because and preconcentration, it is not appropriate for sampling in the amount of analyte is at trace or even ultratrace levels. In recent years, as a novel sample preparation technique, hollow fiber liquid phase microextraction has gained con siderable attention for biological and environmental sample analysis because it is simple, efcient, and inexpensive, consumes less organic solvent,and has good sample cleanup ability and high enrichment efciency. HF LPME can be carried out either in two or three phase mode.
Normally, neutral analytes with a high solubility in nonpolar organic solvents can be extracted in a two phase system, and acidic and basic analytes can be CDK extracted Microdialysis is a dynamic molecular sampling technique based on analyte difusion across a semipermeable HF mem brane driven by a concentration gradient. Microdialysis has been applied to isolate components from sample matrix with the advantages of easy operation, speed, and no or less use of organic solvents. Recently, online microdialysis sampling with HF was established as an LPME technique with high enrichment poten tial by controlling the status of the sample solution and the conditions of the perfusion stream. Thus, interference due to the interaction of analyte with sample matrix species can be decreased through the dilution of the sample solution.
Online HPLC with HF microdialysis HSP perfusion sampling provides simplified sample preparation and has been successfully applied in a two or three phase system. cosmetic and polymer wastewater samples, and fermented milk and drinks. However, there is no report related to the application of microdialysis sampling as the cleanup process and enrichment step in the determination of alachlor and its metabolite 2,6 DEA so far. It has the potential to be an alternative to conventional pretreat ment processes in the determination of alachlor in culture medium. In this paper, we report for the first time the applic ability of the microdialysis sampling technique assembled as a hollow fiber membrane liquid phase microextraction online to HPLC is investigated and examined to develop an eco friendly process of enrichment for the determination of alachlor and its metabolite 2,6 DEA in microbial culture medium samples.
In this study, parameters that in uenced the efciency of enrichment, including the material of hollow fiber and its length, the perfusion solvent and its ow rate, the pH, and the addition of salt in sample solution, as well as the chromatographic behaviors, were studied thoroughly to optimize the online HF LPME/HPLC UV technique for the determination of alachlor and 2,6 DEA in culture media. Chemicals and Reagents.