RESUMEN
The use of gestagens in animal fattening is prohibited within the European Union. Recently, the use of spectrometric methods for the detection and confirmation of banned substances was made obligatory. Therefore, conventional high-performance liquid chromatographic (HPLC) methods have been superseded. It has been possible to couple a previously described HPLC method for the determination of acetyl-gestagens in kidney fat to tandem mass spectrometry (LC-MS/MS). The decision limits CCalpha and the detection capability CCbeta are found to be below the minimum required performance limit (MRPL) established for medroxyprogesterone acetate (MPA) at 1 microg kg(-1). The calculated values for CCalpha are as follows: megestrol acetate (MGA)--0.15 microg kg(-1), melengesterol acetate (MLA)--0.15 microg kg(-1), chlormadinone acetate (CMA)--0.37 microg kg(-1) and for medroxyprogesterone acetate (MPA)--0.24 microg kg(-1). The CCbeta values for these compounds have been determined as 0.19, 0.19, 0.47 and 0.32 microg kg(-1), respectively.
Asunto(s)
Cromatografía Liquida/métodos , Riñón/metabolismo , Espectrometría de Masas/métodos , Progestinas/análisis , Calibración , Química Orgánica/métodos , Acetato de Clormadinona/análisis , Cromatografía Líquida de Alta Presión/métodos , Humanos , Acetato de Medroxiprogesterona/análisis , Acetato de Megestrol/análisis , Acetato de Melengestrol/análisis , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Solventes/químicaRESUMEN
A liquid chromatographic method was developed for determination of 3 progestogens-melengestrol acetate, megestrol acetate, and chlormadinone acetate-found in edible tissue at concentrations between 10 and 1000 ppb. These progestogens are commonly used as feed additives to control herd estrus and to improve feed efficiency. Rendered fat was extracted with acetonitrile, washed with hexane, and dried. The remaining lipids were saponified with sodium hydroxide and precipitated with magnesium chloride. The progestogens were extracted from the basic solution with hexane, dried, and cleaned up on a cyanopropyl solid-phase extraction column in the normal-phase mode. The eluate was dried and reconstituted with acetonitrile-water (7 + 3, v/v). Chromatography was performed on a 5 microns high-carbon load C18 column with acetonitrile-water (7 + 3, v/v) at 1 mL/min and UV detection at 291 nm. Recoveries from fortified samples ranged from 84 to 116%. The limit of quantitation was 10 ppb for both beef and pork. The detection limit was 3 ppb.
Asunto(s)
Tejido Adiposo/química , Acetato de Clormadinona/análisis , Cromatografía Liquida , Residuos de Medicamentos/análisis , Acetato de Megestrol/análisis , Acetato de Melengestrol/análisis , Acetonitrilos/química , Tejido Adiposo/metabolismo , Animales , Bovinos , Hexanos/química , Cloruro de Magnesio/química , Estándares de Referencia , Reproducibilidad de los Resultados , Hidróxido de Sodio/química , Espectrofotometría Ultravioleta , Porcinos , Agua/químicaRESUMEN
A new extraction method for the acetylgestagens medroxyprogesterone acetate (MPA), chloromadinone acetate and megestrol acetate, from kidney fat, has been developed. The method is a combination of matrix solid phase dispersion and solid phase extraction and is simpler and safer than previous methods, especially as it can be automated. The recovery was estimated as 59 +/- 5% (mean +/- standard deviation) for MPA. For screening purposes detection can be achieved using a commercially available enzyme immunoassay kit giving detection limits in the range of 1.0-2.0 ng g-1.
Asunto(s)
Tejido Adiposo/química , Acetato de Clormadinona/aislamiento & purificación , Acetato de Medroxiprogesterona/aislamiento & purificación , Megestrol/análogos & derivados , Mataderos , Automatización/métodos , Acetato de Clormadinona/análisis , Cromatografía Líquida de Alta Presión/métodos , Técnicas para Inmunoenzimas , Riñón , Acetato de Medroxiprogesterona/análisis , Megestrol/análisis , Megestrol/aislamiento & purificación , Acetato de Megestrol , Técnica de Dilución de Radioisótopos , Juego de Reactivos para Diagnóstico , TritioAsunto(s)
Griseofulvina/análisis , Esteroides/análisis , Betametasona/análisis , Fenómenos Químicos , Química , Acetato de Clormadinona/análisis , Cromatografía , Estabilidad de Medicamentos , Efedrina/análisis , Estradiol/análisis , Fermentación , Fluocinolona Acetonida/análisis , Métodos , Microquímica , Pomadas , Excipientes Farmacéuticos , Progesterona/análisis , Solventes , Espectrofotometría Ultravioleta , ComprimidosAsunto(s)
Anticonceptivos Orales/normas , Estabilidad de Medicamentos , Solubilidad , Acetato de Clormadinona/análisis , Etinilestradiol/análisis , Diacetato de Etinodiol/análisis , Linestrenol/análisis , Medroxiprogesterona/análisis , Megestrol/análisis , Mestranol/análisis , Noretindrona/análisis , Norgestrel/análisis , Agua/análisisRESUMEN
PIP: Infrared spectrometry was applied to the determination of mestranol, chlormadinone, estradiol benzoate, progesterone, testosterone propionate, and methyltestosterone. Except for mestranol, all of the steroids were determined in the range of 1656-1735 cm (-1) on the ground of the absorption of the carbonyl groups. Mestranol can be determined at 3308 cm (-1) (ethyl group variations) or at 1502 cm (-1) (variations of the aromatic ring system). The asorption band 1495 cm (-1) has been shown to be suitable for assaying estradiol benzoate. A relative error of .58% to 3.26% has been charged for the methods used. Methyltestosterone and progesterone can also be assayed by using the absorption bands reflecting the vibrations of the methyl groups, at 1361 cm (-1) and 1380 cm (-1). However, the assays at the latter wave lengths are found to be less accurate, with relative error at 3.49% and 4.41% and precision error at 1.19% and 1.39%, respectively. All measurements were performed in a medium of highly purified chloroform.^ieng
