Determination of endocrine disruptors in environmental waters using poly(acrylamide-vinylpyridine) monolithic capillary for in-tube solid-phase microextraction coupled to high-performance liquid chromatography with fluorescence detection.

A method for the determination of endocrine disruptors, bisphenol A and 17alpha-ethinylestradiol, in environmental water samples was developed using in-tube solid-phase microextraction followed by liquid chromatography and fluorescence detection. A poly(acrylamide-vinylpyridine) monolithic capillary column was applied as the extraction media in view of its greater phase ratio than open-tubular capillaries and thus higher extraction efficiency. After optimizing the extraction conditions, bisphenol A and 17alpha-ethinylestradiol were extracted directly from water samples in a wide dynamic linear range of 0.5-1000 ng mL(-1), with the detection limits obtained as 0.064 and 0.12 ng mL(-1), respectively. The precision of the method was satisfactory with the intraday and interday RSD values smaller than 7.2%. Environmental water samples of different sources were successfully analyzed with the presented method and the monolithic capillary was proved to be robust and reusable in analyzing real water samples.

Behaviour of endocrine disrupting chemicals during the treatment of municipal sewage sludge.

Agricultural application of municipal sewage sludge has been emotionally discussed in the last decades, because the latter contains endocrine disrupting chemicals (EDCs) and other organic micropollutants with unknown fate and risk potential. Bisphenol A (BPA) was chosen as a model substance to investigate the influence of sludge conditioning on the end-concentration of EDCs in sludge. Adsorption studies with radioactive-labelled BPA showed that more than 75% BPA in anaerobically digested sludge is bound to solids (log Kd = 2.09-2.30; log Koc = 2.72-3.11). Sludge conditioning with polymer or iron (III) chloride alone had no influence on the adsorption of BPA. After conditioning with iron (III) chloride and calcium hydroxide desorption of BPA took place. Apparently, it occurred due to the deprotonation of BPA (pKa= 10.3) as the pH-value reached 12.4 during the process. The same behaviour is expected for other phenolic EDCs with similar pKa (nonylphenol, 17beta-estradiol, estron, estriol, 17alpha-ethinylestradiol). This study shows high affinity of BPA to the anaerobically digested sludge and importance of conditioning in the elimination of EDCs during the sludge treatment. Addition of polymer is favourable in the case of sludge incineration. Conditioning with iron (III) chloride and calcium hydroxide shows advantages for the use of sludge as fertiliser.

Suitability of N,O-bis(trimethylsilyl)trifluoroacetamide and N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide as derivatization reagents for the determination of the estrogens estrone and 17alpha-ethinylestradiol by gas chromatography-mass spectrometry.

This paper describes apreviously unreported problem with the use of N,O-bis-(trimethylsilyl)trifluoroacetamide (BSTFA) and N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTBSTFA) to derivatise the natural hormone estrone (E1) and the synthetic estrogen 17alpha-ethinylestradiol (EE2). The resulting trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS) derivatives of EE2 were partially converted to their respective El derivatives. Therefore, these reagents may not be suitable for simultaneous determination of estrogens in environmental samples, which raises questions about the reliability of results from some earlier studies.

Collaborative study of an assay for progestational steroids in individual contraceptive tablets.

A single tablet assay method has been developed for determining norethindrone, norethynodrel, and norgestrel in commercial contraceptive tablets. The steroid is separated from the tablet constituents on a diatomaceous earth column, the chloroform eluate is reacted with isonicotinic acid hydrazide, and the steroid is determined by colorimetry. Norethynodrel is isomerized to a conjugated ketosteroid before the colorimetric determination. Fourteen laboratories collaboratively tested the method. The average recoveries in 3 brands of tablets were as follows: 102.6% for the sample labeled as containing 0.35 mg norethindrone/tablet, 99.4% for the 2.5 mg norethynodrel/tablet, and 98.2% for the 0.5 mg norgestrel/tablet. The assay method has been adopted as official first action.

Collaborative study of the analysis of mestranol in combination with norethindrone or norethynodrel.

PIP: A collaborative study of the analysis of mestranol in combination with norethindrone or norethynodrel by column chromatography and ultraviolet determination is presented. The study consisted of 4 samples which were a commercial norethindrone tablet, a synthetic norethindrone mixture, a commercial norethynodrel tablet, and a synthetic norethynodrel mixture. The samples were analyzed by 13 collaborators. Average mestranol recoveries were 100.4, 99.9, 98.7, and 101.2%, respectively.^ieng

Collaborative study of the colorimetric determination of progestational steroids in contraceptive tablets.

PIP: A collaborative study of a proposed method for the determination of progestational steroids was conducted on commercial brands and simulated tablet mixtures of norethynodrel, norethindrone, and medroxyprogesterone acetate. The proposed method based on a colorimetric determination of the product formed after treatment of the chloroform extract of the steroid with isonicotinic acid hydrazide was adopted as official fist action.^ieng

Metabolism of antifertility steroids. I. Norethynodrel.

PIP: The in vivo and in vitro metabolism of norethynodrel was studied in detail, and the configuration and conformation of the major metabolites were established. In both the in vivo and in vitro studies, the initial concentration of the ethynyl steroidal fractions was obtained by using silver nitrate-silica gel columns, further purified with alumina chromatography. Only traces of unchanged norethynodrel were found. The major ethynyl steroidal metabolites from in vitro rabbit liver homogenate studies were isolated in pure form and were found to be idential to the product obtained by reducing norethynodrel with sodium borohydride. Through use of proton nuclear magnetic resonance spectra (NMR) and mass spectra studies, the configurations of the major metabolites of both the in vivo and in vitro systems were identified as 17alpha-ethynyl-3alpha, 17beta-dihydroxy-delta-estrene and 17alpha-ethynyl-3beta, 17beta-dihydroxy-delta-estrene. In humans, both compounds are found as the beta-glucosiduronates in the ratio of 7:3 in favor of the 3alphaisomer. The conformation of the 3alpha-hydroxyl group was shown to be equatorial, while that of the 3beta-epimer was axial.^ieng