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1.
Analyst ; 123(5): 1103-7, 1998 May.
Article in English | MEDLINE | ID: mdl-9709494

ABSTRACT

Ractopamine (RCT) is a phenethanolamine member of the family of beta-adrenergic agonists (beta-agonists). This class of compounds have become notable for their properties of enhancing the growth rates of farm animal species but are not licensed for use in Europe. An ELISA procedure employing a polyclonal antibody raised in a goat was developed to detect RCT residues in bovine urine samples. The assay had a high sensitivity (calibration curve mid-point of 22 pg per well), allowing the analysis of urine samples without the need for sample clean-up. In addition, an LC-MS-MS confirmatory procedure was developed which was able to act as a confirmatory procedure for the ELSA results. Four calves were orally treated with RCT (0.1 mg kg-1 body mass for 17 d) and urine samples collected were assayed by both analytical procedures. It was observed that RCT residues were excreted mainly in the form of glucuronides and deconjugation could be achieved using two different sources of the enzyme beta-glucuronidase (Helix pomatia and Escherichia coli). High concentrations of RCT residues were found throughout the medication period (44-473 ng ml-1; LC-MS-MS data) and remained present for several days following removal of the drug from the diet. RCT residues were no longer detectable 2 weeks after withdrawal. Good agreement (r2 = 0.73) was achieved between the ELISA and LC-MS-MS results, especially when sample deconjugation was applied to the urine samples for sets of analyses. The results show that an effective screening and confirmatory system was devised to detect RCT residues in urine samples taken during treatment and close to withdrawal. However, alternative matrices may have to be selected to allow the illegal use of the substance to be detected following prolonged withdrawal times.


Subject(s)
Adrenergic beta-Agonists/urine , Drug Residues , Growth Substances , Phenethylamines/urine , Animals , Cattle , Enzyme-Linked Immunosorbent Assay , Food Contamination
2.
Analyst ; 123(12): 2579-83, 1998 Dec.
Article in English | MEDLINE | ID: mdl-10435303

ABSTRACT

Screening for the presence of anabolic growth promoters in urine samples from cattle grown for meat production can be performed by (semi)quantitative methods such as immuno-, receptor- or cell-based assays or by quantitative methods with mass spectrometric detection which can also include confirmation of compounds. In this study conventional immunoassays used at two different institutes [Veterinary Sciences Division (VSD) in Northern Ireland and TNO Nutrition and Food Research Institute (TNO) in The Netherlands] were compared with the oestrogen radioreceptor assay (ORRA), with GC-MS as the reference method. Urine samples were generated by treating calves (n = 2 per group) intramuscularly with ethynyloestradiol (EE2), diethylstilbestrol (DES) or alpha-zearalanol (zeranol, ZER). Urine samples were collected up to 21 d after administration of the oestrogenic compounds. Samples were screened by enzyme immunoassay or radioimmunoassay and by the ORRA and also by GC-MS. Values found by VSD were lower by a factor of 1-20 than those measured by TNO. These differences could be explained by differences in sample clean-up (immunoaffinity chromatography versus solid-phase extraction) and by differences in cross-reactivities between the antisera used. The ORRA and GC-MS showed similar results for EE2 and DES, but produced lower results (by a factor of ca. 3) for ZER owing to the relatively low affinity of ZER for the oestrogen receptor. The most important finding was that the withdrawal period for calves treated with EE2, DES or ZER was similar for each of the screening methods used. Therefore, it is concluded that the choice of screening method does not affect the probability of finding a positive sample.


Subject(s)
Anabolic Agents/urine , Cattle/metabolism , Drug Residues/analysis , Estrogens/urine , Animals , Immunoassay , Male , Radioligand Assay
3.
Analyst ; 123(12): 2599-604, 1998 Dec.
Article in English | MEDLINE | ID: mdl-10435307

ABSTRACT

The European Union banned the use of anabolic steroids for cattle fattening in 1988. Analytical techniques able to detect trace amounts of the parent drugs and their metabolites are mandatory for the control of abuse. Stanozolol (Stan) is an anabolic steroid that is often found in injection sites and cocktails. However, it has never been detected in tissues (kidney fat, meat) or excreta (urine, faeces) taken during regulatory inspection. The difference between the structure of Stan and the other steroids (a pyrazole ring fused to the androstane ring system) is probably the cause of this phenomenon. In the multi-laboratory study described here, veal calves were treated with intramuscular doses of Stan. In the excreta of these calves the presence, absence and/or concentration of Stan and of its major metabolites 16 beta-hydroxystanozolol and 3'-hydroxystanozolol were determined. For the determination of these analytes the different laboratories used different extraction and clean-up procedures and also evaluated different analytical techniques such as GC-MS (negative chemical ionization) and LC-MS-MS. The aim of this investigation was to explore which analyte should be validated for veterinary inspection purposes.


Subject(s)
Anabolic Agents/analysis , Cattle/metabolism , Stanozolol/analysis , Anabolic Agents/administration & dosage , Anabolic Agents/metabolism , Animals , Feces/chemistry , Gas Chromatography-Mass Spectrometry , Injections, Intramuscular , Male , Mass Spectrometry , Stanozolol/administration & dosage , Stanozolol/analogs & derivatives , Stanozolol/metabolism , Stanozolol/urine
4.
Br Poult Sci ; 33(1): 89-100, 1992 Mar.
Article in English | MEDLINE | ID: mdl-1571810

ABSTRACT

1. Two experiments were conducted to examine the effect of feeding D-xylose and L-arabinose on broiler performance, body composition, caecal length and weight, and liver weight. 2. Graded amounts (25, 50 and 75 g/kg) of D-xylose or L-arabinose were added to either a practical type (Exp. A) or a semi-purified (Exp. B) basal diet. As reference, a diet containing 75 g D-glucose/kg was included in both experiments, which were conducted in battery brooders, the birds receiving the isocaloric [on metabolisable energy (ME) basis] diets as dry mash ad libitum from 6 to 27 d of age. 3. A negative dose-dependent effect of both pentose sugars on weight gain and feed utilisation was observed. The same was true for daily food intake of the D-xylose diets. Water intake increased linearly (P less than 0.05) as the dietary concentrations of both pentose sugars was increased. Consequently, dry matter content of the droppings decreased. 4. Fat content of the chick body tended to decrease when either D-xylose or L-arabinose was included in the diets. 5. Caecal weight was increased markedly by feeding L-arabinose. Liver weight was not affected by feeding either D-xylose or L-arabinose to birds. 6. From data for ME intake and gain in body energy it was estimated that utilisation of the ME of both pentose sugars was inferior to that of D-glucose.


Subject(s)
Arabinose/metabolism , Chickens/growth & development , Dietary Carbohydrates/metabolism , Xylose/metabolism , Animal Feed , Animals , Body Composition , Cecum/growth & development , Chickens/physiology , Drinking , Energy Metabolism , Female , Lipids/analysis , Liver/growth & development , Nutritive Value , Organ Size , Proteins/analysis , Weight Gain
5.
J Chromatogr ; 564(2): 429-44, 1991 Apr 05.
Article in English | MEDLINE | ID: mdl-1874847

ABSTRACT

Within the scope of the National Plan for Hormone Control in The Netherlands, a study was performed to develop a system for control of the illegal use of three naturally occurring hormones [oestradiol-17 beta (E2-17 beta), testosterone (T), progesterone (P)] for fattening purposes in animal production. Using a specific high-performance liquid chromatographic-radioimmunoassay method, reference values were established for concentrations of E2-17 beta, T and P and some of their metabolites in blood plasma and urine from untreated male and female veal calves. E2-17 beta levels of both male and female calves were less than 0.01 microgram/l in blood plasma and less than 0.2 microgram/l in urine. For male veal calves levels of T and epitestosterone (epiT) in blood plasma and urine varied widely. The P levels were less than 0.1-0.3 micrograms/l in blood plasma and less than 0.6-10 micrograms/l in urine from both male and female calves. To investigate the effect of anabolic treatment on the hormone levels in plasma and excreta, male veal calves were injected, subcutaneously into the dewlap, with a solution containing 20 mg of E2-17 beta benzoate and 200 mg of T propionate in 5 ml of arachis oil. Only the levels of E2-17 beta and E2-17 alpha in blood plasma and excreta were elevated until about one week after injection, compared with the untreated control calves and the reference values. T and epiT levels were similar in plasma and excreta from both untreated and treated animals.


Subject(s)
Cattle/blood , Chromatography, High Pressure Liquid/methods , Estradiol/blood , Progesterone/blood , Radioimmunoassay/methods , Substance Abuse Detection , Testosterone/blood , Animals , Estradiol/analysis , Estradiol/urine , Feces/chemistry , Female , Male , Progesterone/analysis , Progesterone/urine , Testosterone/analysis , Testosterone/urine
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