1、 附录 Determination of some endocrine disrupter chemicals in urban wastewatersamples using liquid chromatographymass spectrometry Abstract： An analytical method for the simultaneous determination of trace amounts of fourteen endocrine disrupter chemicals (EDCs) in urban wastewater samples has been dev
2、eloped. The studied compounds were: bisphenol A and its chlorinated derivatives (monochloro, dichloro, trichloro and tretrachlorobisphenol A), three alkylphenols (4-n-nonyl, 4-n-octyl and 4-(tert-octyl)phenol) and six well known phthalate esters (dimethyl, diethyl, di-n-butyl, butylbenzyl, bis(2-eth
3、ylhexyl) and di-n-octyl phthalate). The method involves extraction from the samples and preconcentration of the analytes usingasolid-phase extraction (SPE) procedure and subsequent liquid chromatographic separation with mass spectrometric detection (LC-MS). Bisphenol F was used as a surrogate. Quant
4、ification limits found ranged between 12ngL-1 for diethylphthalate and 69ngL-1for 4-(tert-octyl)phenol. The method was satisfactorily used for determination of these chemicals in urban wastewater samples of Granada City (Spain) and validated using a recovery assay with spiked samples. Keywords: Alky
5、lphenols； Bisphenol A； Bisphenol A chlorinated derivatives； Phthalate esters； Liquid chromatographymass spectrometry (LCMS)； Wastewater analysis 1. Introduction In recent years, a specific category of human madechemicals, called commonly endocrine disrupter chemicals(EDCs), has attracted a great dea
6、l of public and scientificattention because of their suspected carcinogenic and estrogenicproperties. In 1996, the European Union (EU) defined this classof compounds as “exogenous substances that cause adversehealth effects in an intact organism, or its progeny, consequentto changes in endocrinefunc
7、tion”1. Emerging evidence from wildlife and laboratory studiesindicates that some synthetic chemicals may interfere with theendocrine system in a broad variety of ways, including theinitiation of hormones, blocking their action or acceleratingtheir breakdown2. Aware of the problem, both, the EU and
8、the US Environmental Protection Agency (EPA) have authored a“ priority” list of substances for further evaluation of their role in endocrine disruption 3,4 indicating the need to assess the levels and effects of EDCs. A report from the EU 5 proposed a candidate list of 533 substances for further eva
9、luation of their role in endocrine disruption. Moreover, incremental growth in dysfunctions related to male reproductive health has been demonstrated 6, 7. Both, human and wildlife are exposed to complex mixtures of these compounds. Effects are cumulative and it could be that they will only appear i
10、n subsequent generations. Meantime the resulting effects may be irreversible, threatening sustainable human development. Most EDCs are introduced into the environment by way ofanthropogenic inputs and are therefore ubiquitous in aquaticenvironments receiving wastewater effluents. Although it ispossi
11、ble that a large amount of EDCs remain yet unidentified, itis clear that there are some identified families of compoundsshowing potential estrogenic activity. Plastic industry intermediatesas bisphenol A or phthalate esters, detergent degradationcompounds, as 4-n-nonylphenol, 4-n-octylphenol and 4-(
12、tert-octyl)phenol are described in scientific literature. It is nowknown that phthalate esters, alkylphenols and bisphenol A (BPA)are potential endocrine disrupter chemicals 8,9. Let us see someof their uses till now. Bisphenol A is mainly used as a monomer in the preparationof epoxide resins, polyc
13、arbonate plastic and as an antioxidant inpolyvinylchloride 10. Levels of BPA have been found indifferent matrices such as body fluids, canned food andenvironmental samples as wastewaters. The use of chlorinefor disinfecting wastewaters or drinking water has becomewidespread over the years. BPA prese
14、nt in waters may reactwith residual chlorine originally used as a disinfectant,producing chlorinated BPA derivatives 11. PolychlorinatedBPA has been recently identified and biodegradation tests usingactivated sludge revealed that it is not easily biodegraded 12,and chlorinated BPA being more cytotox
15、ic than BPA 13. Alkylphenols such as 4-n-nonylphenol, 4-n-octylphenoland 4-(tert-octyl) phenol are widely used as intermediates toproduce surfactants (anionic and non-ion surfactants) and asstabilizers of ethyl cellulose resin, oil-soluble phenol resin andesters 14,15. Phthalate esters are one of th
16、e most abundant man-madeenvironmental pollutants. They are widely used as additives inthe manufacture of plastics to make them flexible. Because oftheir properties as plasticizers in plastic production and becauseits use has increased significantly in recent years, they are oftenfound in environment
17、al matrices such as waters and soils 16.Six of the most commonly used phthalates are dimethyl, diethyl,di-n-butyl, butylbenzyl, bis(2-ethylhexyl) and di-n-octylphthalate.Bis(2-ethylhexyl) phthalate is the most used and accountsfor up to one quarter of the total production of plasticizers 17.There is
18、 a high industrial activity associated with them andrelatively large amounts of these compounds are released intothe environment, wastewater being the most common way tointroduce these contaminants into the environment 1619. Gas chromatography (GC) and liquid chromatography (LC),after previous conce
19、ntration and clean-up, have been used fordetermining these compounds in water samples. Commonly,their preconcentration and clean-up involves liquidliquidextraction (LLE)with cyclohexane20or dichloromethane21,22, solid-phase extraction (SPE) 1619,2325 or solid-phase microextraction (SPME)26. After an extensivebibliographic revision, SPE seems to be the most
