Diethylstilbestrol detection

Detection and result The chromatogram was dried in the air, immersed in the reagent solution for 5 s and then heated to 120°C for 5 min. cis-Diethylstilbestrol hR( 15 — 20) and trawj-diethylstilbestrol (JtRf 40—45) turned red while ethynylestradiol (hRf 50—55) appeared blue. 

In situ quantitation The absorption photometric analysis was made at A = 540 nm (ethynylestradiol) and X = 605 nm (diethylstilbestrol). The detection limit for ethynylestradiol was 12ng and that for diethylstilbestrol 3 ng per chromatogram zone. 

Certain drugs cannot be used in animals produced for human consumption, including chloramphenicol, clenbuterol, diethylstilbestrol (DES), dimetridazole, ipronidazole, furazolidine, and vancomycin. Severe legal penalties could be incurred if the pharmacist were to provide any of these drugs for use in food-producing animals and subsequent residues were detected in animal tissue. 

Induction of morphological transformation of SHE cells can occur in the absence of detectable induction of gene mutations measured concomitantly in the same cells (Barrett et al, 1983). Diethylstilbestrol and asbestos are two examples of chemicals that incluce cell transformation but not gene mutations (Barrett et al, 1981, 1983). However, a gcxxl correlation is observed between induction of chromosomal mutations (numerical and structural changes) and induction of cell transformation (Barrett etol, 1983 Oshimiueerol, 1984). 

In rodents, some of the urinary metabolites were identified as dienestrol, and as hydroxy- and methoxy-derivatives of dienestrol and diethylstilbestrol. When dietiiylstilbestrol was given orally to ostriches, parent compound and the metabolite dienestrol could be detected in urine (44). Diethylstilbestrol was rapidly excreted, its concentration being above 2 ppb for only 4 days although it could be detected by 18 days. The concentration of dienestrol was just above 2 ppb for 1 day only and could be detected for 3 days only. 

When using photochemical reactions, a gain in fluorescence output or electroactivity not only lowers detection limits but also contributes to confirm an analytical result. Examples are the conversion of diethylstilbestrol to a fluorescent hexahydrophenanthrene (277), and the conversion of fenbendazole to fluorescent species (278). Tlie fluorescence of photoconverted diethylstilbestrol can further be enhanced by a subsequent online postchromatographic derivatization with bisulfite to the highly fluorescent phenanthrenediol (279). Another example of photolytic derivatization is the postchromatographic conversion of penicillins and cephalosporins into electroactive species that can be detected by an amperometric detector (280). 

Use of hormonal-type substances for growth promotion and fattening purposes in food-producing animals is prohibited or restricted in many countries. Administration of diethylstilbestrol, in particular, has been totally banned worldwide. This implies that adequate analytical methods should be available for regulatory control. Due to its sensitivity, radioimmunoassay has have become most important in diethylstilbestrol analysis and represents the final detection step in the EU reference method for stilbene residues analysis (100). 

Methanococcus voltae contains a membrane-bound vanadate-sensitive ATPase [48] that is inhibited by diethylstilbestrol, an inhibitor of eukaryotic P-type ATPases. The purified enzyme is composed of a single subunit (Mr 74 000), forms a covalent acyl-phosphate enzyme intermediate, and is not inhibited by nitrate or bafilomycin [49]. No such ATPase activity has been reported in other archaea. The presence of a second ATPase in M. voltae has been inferred since membranes react with antiserum prepared against the 3 subunit from the V-type ATPase of S. acidocaldarius [50]. Two peptides are detected whose Mr values (51 000 and 65 000) correspond to the masses for the two laigest subunits of the S. acidocaldarius ATPase [51]. There is evidence that ATP synthesis in the M. voltae enzyme is due to the operation of a sodium-translocating ATPase [50]. The relationship of the putative V-like ATPase to the sodium-translocating ATPase has not been established. 

Miner, D.J. Skibic, M.J. Bopp, R.J. Practical aspects of LC/EC determinations of pharmaceuticals in biological media. J.Liq.Chromatogr., 1983, 6, 2209-2230 [plasma column-switching fluorescence detection also diethylstilbestrol, enviradene, enviroxime, enviroxine, hexestrol, pergolide, zinviroxime]... 

In contrast to the above results, DDE has been detected as a mutagen in cultured Chinese hamster ovary cells. The carcinogenic estrogen, diethylstilbestrol (DES), also is missed by all of the mutagenesis-based assays which may reflect the operation of another mechanism of cancer induction in the case of hormone induced cancers. It should be noted, however, that a... 

Natural and synthetic hormones are used worldwide to improve meat production There are, however, possible hazards for the consumer. Therefore, in most European countries the use of hormonal anabolics is restricted or even prohibited by law. Among the banned synthetic anabolics diethylstilbestrol (DES) takes the most important place at the moment. Analytical control is carried out in various ways. Frisch-kom and Smyth have described an HPLC method with electrochemical detection for the determination of growth promoting hormones in meat. 

A method has been developed to detect residual stilbenes such as diethylstilbestrol (DES), dienestrol (DIS), and hexestrol (HS) in animal tissues using solid-phase extraction (SPE) and gas chromatography-mass spectrometry (GC-MS) [153]. The... 

Supported liquid membranes (SLMs) consisting of 5% tri-n-octylphosphine oxide (TOPO) dissolved in di-w-hexylether/n-undecane (1 1) have been used in the simultaneous extraction of a mixture of three stUbene compounds (dienestrol, diethylstilbestrol, and hexestrol) in cow s milk, urine, bovine kidney, and liver tissue matrices [183]. The efficiencies obtained after the enrichment of 1 ng/1 stilbenes in a variety of biological matrices of milk, urine, liver, kidney, and water were 60-70, 71-86, 69-80, 63-74, and 72-93%, respectively. A new method to contribute to the discrimination of polyphenols including resveratrol with synthetic pores was proposed [184]. The work [185] evaluated two types of commonly available chiral detectors for their possible use in chiral method development and screening polarimeters and CD detectors. Linearity, precision, and the limit of detection (LOD) of six compounds (trans-stilbene oxide, ethyl chrysanthemate, propranolol, 1-methyl-2-tetralone, naproxen, and methyl methionine) on four common detectors (three polarimeters and one CD detector) were experimentally determined and the limit of quantitation calculated from the experimental LOD. trans-Stilbene oxide worked well across all the detectors, showing good linearity, precision, and low detection limits. However, the other five compounds proved to be more discriminating and showed that the CD detector performed better as a detector for chiral screens than the polarimeters. 

Th. Reuvers, E. Perogordo and R. Jimenez, Rapid screening method for the determination of diethylstilbestrol in edible animal tissue by column liquid chromatography with electrochemical detection, J. Chromatogr., 1991, 564, 477-484. 

The electrochemical detector (ECD), which is still used for anodic oxidation of phenols and amines offers interesting perspectives. With the aid of a commercial detector we were recently able to detect diethylstil-bestrol in plasma in the low pg-region by an oxidation reaction (see Figure 13). Diethylstilbestrol represents the main metabolite and one of the active principle of fosfestrol, which has been used for treating metastasizing prostate carcinomas for about 30 years. 

The final point - transplacental carcinogenesis - has in a similar manner as the Thalidomide accident not been predicted from animal experiments but was unfortunately first detected in the human population. Diethylstilbestrol, a synthetic female sex hormone, was found to induce adenoma and carcinoma of the vagina in daughters 15 to 20 years after treatment of the mothers for threatened abortion. 

Figure 4 Separation of stilbestrol derivatives on RP precoated plates with different degrees of modification. Plates a. HPTLC precoated plate RP-18W F254s b. HPTLC precoated plate RP-18 F254s. Eluent metha-nol/water 80/20 v/v. Migration distance 5 cm. Chamber normal chamber without saturation. Compounds 1, diethylstilbestrol-dimethyl ether 2, diethylstilbestrol-mono-methyl ether 3, dielhylstilbestrol (all 0.1 %). Application volume 2(X) nl. Detection in-situ evaluation with TLC/HPTLC scanner (Camag) UV 254 nm.

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