ABSTRACT
After administration of steroids to animals the steroids are partially metabolised in the liver and kidney to phase 2 metabolites, i.e., glucuronic acid or sulphate conjugates. During analysis these conjugated metabolites are normally deconjugated enzymatically with aryl sulphatase and glucuronidase resulting in free steroids in the extract. It is well known that some sulphates are not deconjugated using aryl sulphatase; instead, for example, solvolysis can be used for deconjugation of these aliphatic sulphates. The effectiveness of solvolysis on androgenic steroid sulphates was tested with selected aliphatic steroid sulphates (boldenone sulphate, nortestosteron sulphate and testosterone sulphate), and the method was validated for analysis of androgenic steroids in bovine urine using free steroids, steroid sulphates and steroid glucuronides as standards. Glucuronidase and sulphuric acid in ethyl acetate were used for deconjugation and the extract was purified by solid-phase extraction. The final extract was evaporated to dryness, re-dissolved and analysed by LC-MS/MS.
Subject(s)
Androgens/urine , Glucuronides/urine , Liquid-Liquid Extraction/methods , Steroids/urine , Sulfates/urine , Acetates/chemistry , Animals , Arylsulfatases/chemistry , Cattle , Chromatography, Liquid , Glucuronidase/chemistry , Hydrolysis , Nandrolone/chemistry , Reproducibility of Results , Sulfuric Acids/chemistry , Tandem Mass Spectrometry , Testosterone/analogs & derivatives , Testosterone/chemistryABSTRACT
An intercomparison study of the determinations of glyphosate, chlormequat and mepiquat residues in cereals was performed. Four samples comprising one blank, two incurred and one spiked sample were sent to six participating laboratories. For glyphosate, two laboratories reported considerably lower results than the other four. One of the two laboratories with low results also reported low recoveries. The results of a sample spiked with 0.80 mg kg-1 glyphosate and an incurred sample, ranged from 0.23-0.87 mg kg-1 and 0.11-0.25 mg kg-1 respectively. The strong correlation between the two samples (r2 = 0.95) indicates a systematic between-laboratory variation. Several different principles were used for the analysis of glyphosate using different clean-up techniques and GC/MS, HPLC-fluorescence or LC/MS for detection. The results of the chlormequat residues showed more consistency. All but one laboratory obtained comparable results. However the correlation between the results for the sample spiked with 0.38 mg kg-1 (range: 0.26-0.65 mg kg-1) and the incurred samples (range: 0.19-0.45 and 0.15-0.23 mg kg-1, respectively) again showed a strong correlation (r2 = 0.99 and 0.88) indicating a systematic component. For mepiquat, results above the limit of quantification were only reported for the spiked sample. The results ranged from 0.29-0.92 mg kg-1 (spiked concentration = 0.38 mg kg-1). Three laboratories had results that deviated less than 25% from the fortified concentration. Two laboratories reported results 38% and 141% above the fortified concentration, respectively.
