Synthetic steroids

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INTENDED USE

The Evidence Investigator Synthetic Steroids Array is to be used for the simultaneous quantitative detection of multiple related Synthetic Steroid drug residue immunoassays (in parallel) from a single sample. This product is recommended for bovine and equine urine only.

The Evidence Investigator Synthetic Steroids Array is intended for the screening of samples only and not for diagnostic procedures. Positive results should be confirmed by another method.

CLINICAL SIGNIFICANCE

Growth-promoting hormones have been used by the livestock industry for over 30 years to improve an animal's ability to more efficiently utilize nutrients and produce leaner, more affordable meat. Concerns over the use of hormones in food producing animals first arose during the late 1970s and early 1980s, with a number of incidents linking hormone residues in meat to various medical conditions. As a result, the use of growth-promoting hormones in livestock production is prohibited in the European Union(1)and strongly regulated in other countries. Compliance with these regulations is monitored by national monitoring programmes.

PRINCIPLE

The Evidence Investigator Biochip Array technology is used to perform simultaneous quantitative detection of multiple analytes from a single sample.

The core technology is the Randox Biochip, a solid-state device containing an array of discrete test regions containing immobilised antibodies specific to different Synthetic Steroids drug residues. A competitive chemiluminescent immunoassay is employed for the Synthetic Steroids Array. Increased levels of synthetic steroids drug residue in a specimen will lead to decreased binding of synthetic steroids drug residues conjugates labelled with horseradish peroxidase (HRP) and thus a decrease in the chemiluminescent signal emitted.

The light signal generated from each of the test regions on the biochip is detected using digital imaging technology and compared to that from a stored calibration curve. The concentration of analyte present in the sample is calculated from the calibration curve.

Several different immunoassay based multi-analyte arrays have been developed for use on  Evidence Investigator .

The Evidence Investigator Synthetic Steroids Array will quantitatively test for Methyltestosterone, Methandriol, 17β-Clostebol, Gestagens, and Ethinylestradiol simultaneously.

 

Methyltestosterone (MTEST) Assay

INTENDED USE

The Evidence Investigator Methyltestosterone test has been designed for the quantitative measurement of Methyltestosterone and Methylboldenone in bovine and equine urine samples.

This test is for screening use only. Not for use in diagnostic procedures. Positive results should be confirmed using another method.

CLINICAL SIGNIFICANCE

Methyltestosterone and methylboldenone are orally active synthetic derivatives of testosterone(2). In addition to human clinical applications, and illegal use as a growth promoter in cattle(3), methyltestosterone has been used as a masculinising agent in aquaculture (4). Methyltestosterone and methylboldenone are mainly excreted in urine as glucuronides of hydroxylated metabolites(5).

PRINCIPLE

The Evidence Investigator Methyltestosterone assay is a competitive chemiluminescent immunoassay for the detection of Methyltestosterone and Methylboldenone in bovine and equine urine samples.

REFERENCES

1. Directive 2003/74/EC of the European Parliament and of the Council of 22 September 2003. Official Journal of the European Communities; L 262/17.

2. INCHEM, Poisons Information Monographs, Methyltestosterone, (PIM 908).

3. Standardization of Hormone and Veterinary Drug Residue Analysis in Animal Products, Molecule Database. (http://cemu10.fmv.ulg.ac.be/OSTC/default.html).

4. Antiporda J (1986) Preliminary Studies on the Effects of Methyltestosterone on Macrobrachium rosenbergii Juveniles. FAO Corporate Document Repository; NACA/WP/86/46.

5. Van Puymbroeck M, Leyssens L, Vanderzande D, Gelan J, Raus J (1998) Metabolites in feces can be important markers for the abuse of anabolic steroids in cattle. Analyst.; 123(12): 2449-52.

 

Methandriol (MAD) Assay

INTENDED USE

The Evidence Investigator Methandriol test has been designed for the quantitative measurement of Methandriol in bovine and equine urine samples.

This test is for screening use only. Not for use in diagnostic procedures. Positive results should be confirmed using another method.

CLINICAL SIGNIFICANCE

Methandriol is an orally active synthetic derivative of testosterone(2). It is an anabolic androgen, used illegally in animal husbandry (3). Methandriol is often administered as derivatives, such as methandriol dipropionate and methandriol diacetate(3). Methandriol is mainly excreted in urine as sulphate conjugates(4).

PRINCIPLE

The Evidence Investigator Methandriol assay is a competitive chemiluminescent immunoassay for the detection of Methandriol in bovine and equine urine samples.

REFERENCES

1. Directive 2003/74/EC of the European Parliament and of the Council of 22 September 2003. Official Journal of the European Communities; L 262/17.

2. INCHEM, Poisons Information Monographs, Methandriol, (PIM 906).

3. Standardization of Hormone and Veterinary Drug Residue Analysis in Animal Products, Molecule Database. (http://cemu10.fmv.ulg.ac.be/OSTC/default.html).

4. Schänzer W (1996) Metabolism of anabolic androgenic steroids. Clinical Chemistry; 42(7): 1001-1020.

 

17β-Clostebol (CLOST) Assay

INTENDED USE

The Evidence Investigator 17b-Clostebol test has been designed for the quantitative measurement of 17b-Clostebol and 4-chloro-androsten-3, 17-dione (CLAD) in bovine and equine urine samples.

This test is for screening use only. Not for use in diagnostic procedures. Positive results should be confirmed using another method.

CLINICAL SIGNIFICANCE

Clostebol is an androgen sex hormone, used both as an anabolic and applied topically in dermatological and ophthalmological treatments(2). Clostebol has been illegally used as an anabolic steroid in animal husbandry and can be administered as clostebol acetate(2). Clostebol acetate is extensively metabolised and as a result, cross-reactivity towards the main urinary metabolites is an important feature in immunological screening(3). The main metabolites identified in urine following intramuscular administration are 17alpha-clostebol, CLAD and 4-chloroandrost-4-en-3alpha-ol-17-one(4).

PRINCIPLE

The Evidence Investigator 17b-Clostebol assay is a competitive chemiluminescent immunoassay for the detection of 17b-Clostebol and 4-chloro-androsten-3, 17-dione in bovine and equine urine samples.

REFERENCES

1. Directive 2003/74/EC of the European Parliament and of the Council of 22 September 2003. Official Journal of the European Communities; L 262/17.

2. Standardization of Hormone and Veterinary Drug Residue Analysis in Animal Products, Molecule Database. (http://cemu10.fmv.ulg.ac.be/OSTC/default.html).

3. Crabbe P, Salden M and Van Peteghem C (2000) Characteristics of antibodies to derivatives of clostebol acetate and their use in sample clean-up and rapid immunological screening of bovine urine. EuroResidue IV; 315-321.

4. Le Bizec B, Montrade MP, Monteau F, Gaudin I, Andre F (1998) 4-Chlorotestosterone acetate metabolites in cattle after intramuscular and oral administrations. Clin Chem.; 44(5): 973-84.

 

Gestagens (GEST) Assay

INTENDED USE

The Evidence Investigator Gestagens test has been designed for the quantitative measurement of Medroxyprogesterone, chlormadinone, megestrol and melegestrol acetates in bovine and equine urine samples.

This test is for screening use only. Not for use in diagnostic procedures. Positive results should be confirmed using another method.

CLINICAL SIGNIFICANCE

Medroxyprogesterone, chlormadinone, megestrol and melengestrol are synthetic analogues of progesterone that are commonly administered orally as acetate derivatives. They are used for synchronisation of oestrous, but have also been used as growth promoters in cattle(2,3,4). Synthetic gestagen residue levels are highest and most persistent in fat. The unchanged parent molecules are used for monitoring purposes(2,3,4).

PRINCIPLE

The Evidence Investigator Gestagens assay is a competitive chemiluminescent immunoassay for the detection of Medroxyprogesterone, chlormadinone, megestrol and melegestrol acetates in bovine and equine urine samples.

REFERENCES

1.Directive 2003/74/EC of the European Parliament and of the Council of 22 September 2003. Official Journal of the European Communities; L 262/17.

2.The European Agency for the Evaluation of Medicinal Products, Committee for Veterinary Medicinal Products, Medroxyprogesterone acetate, Summary Report, EMEA/MRL/0129/96-FINAL July 1996.

3.The European Agency for the Evaluation of Medicinal Products, Committee for Veterinary Medicinal Products, Chlormadinone Acetate, Summary Report, EMEA/MRL/694/99-FINAL May 2000.

4.JECFA (2004) Residues Of Some Veterinary Drugs In Animals And Foods - Melengestrol acetate. FAO Food and Nutrition Paper; FNP 41/16.

 

Ethinylestradiol (EEST) Assay

INTENDED USE

The Evidence Investigator Ethinylestradiol test has been designed for the quantitative measurement of Ethinylestradiol in bovine and equine urine samples.

This test is for screening use only. Not for use in diagnostic procedures. Positive results should be confirmed using another method.

CLINICAL SIGNIFICANCE

Ethinylestradiol is a synthetic estrogen with high oral hormonal potency(2). It is rapidly reabsorbed from the gastrointestinal tract and the presence of an ethinyl at the 17-position reduces hepatic first-passage metabolism(2). It is excreted in urine and some is found in faeces(4). Ethinylestradiol is illegally used in animal husbandry as an anabolic(2). Ethinylestradiol is excreted as parent molecule in faeces, and as glucuronide or sulphate conjugates in urine(3).

PRINCIPLE

The Evidence Investigator Ethinylestradiol assay is a competitive chemiluminescent immunoassay for the detection of Ethinylestradiol acetate in bovine and equine urine samples.

REFERENCES

1. Directive 2003/74/EC of the European Parliament and of the Council of 22 September 2003. Official Journal of the European Communities; L 262/17.

2. Standardization of Hormone and Veterinary Drug Residue Analysis in Animal Products, Molecule Database. (http://cemu10.fmv.ulg.ac.be/OSTC/default.html).

3. Huang CH, Sedlak DL (2001) Analysis of estrogenic hormones in municipal wastewater effluent and surface water using enzyme-linked immunosorbent assay and gas chromatography/tandem mass spectrometry. Environ Toxicol Chem.; 20(1): 133-9.