C. Van Poucke

Analytical possibilities for the detection of stanozolol and its metabolites

In sports doping, as well in man as in horseracing, stanozolol (Stan) was abused and became the subject of metabolism research. Also in veterinary practice, stanozolol became an important misused anabolic steroid. Like most other anabolic steroids, stanozolol has poor gas chromatographic behavior. It is difficult to detect in urine, because of low urinary excretion and renal clearance. This is due to the rapid metabolization, leading to low concentration levels of the parent compound found in urine. Therefore, most research studies have focused on the detection of its urinary metabolites. For the identification of the metabolites, different methods of extraction and detection are described in the literature. These are reviewed in this article. Most authors use a hydrolysis to free the phase II metabolites. Extraction procedures vary from solid-phase extraction (SPE), liquid-liquid (L-L) extraction to immunoaffinity chromatography (IAC). For the final detection, the use of gas chromatography (GC)-mass spectrometry (MS) can be compared with liquid chromatography (LC)-MSn. Different metabolites are identified depending on the administration of stanozolol in the animal experiment (oral or intramuscular). Analyses for these analytes in other matrices are also briefly discussed.

Evaluation of the bright greenish yellow fluorescence test as a screening technique for aflatoxin-contaminated maize in Malawi

The bright greenish yellow fluorescence (BGYF) test has been used with varying success in screening for aflatoxins in maize. This test was applied to 180 maize samples collected from different markets within 12 districts of Malawi in order to evaluate its performance against high performance liquid chromatographic analysis. The number of BGYF grains in 2.5 kg unground samples ranged from 0 to 35 and about 49% of all tested samples had aflatoxin concentrations ranging from 1 to 382 μg/kg. A total of 65 (36%) of the examined unground samples showed no BGYF. The European Commission recommends a false negative rate of less than 5% for a screening technique to be acceptable. In this study, four BGYF grains per 2.5 kg unground maize sample successfully indicated an aflatoxin contamination of >10 μg/kg (10 μg/kg being the maxium tolerable level proposed by the Common Market for Eastern and Southern Africa), with a 4.4% false negative rate. In this case, the amount of confirmatory analyses would be reduced by 63%...

Aphids transform and detoxify the mycotoxin deoxynivalenol via a type II biotransformation mechanism yet unknown in animals

Biotransformation of mycotoxins in animals comprises phase I and phase II metabolisation reactions. For the trichothecene deoxynivalenol (DON), several phase II biotransformation reactions have been described resulting in DON-glutathiones, DON-glucuronides and DON-sulfates made by glutathione-S-transferases, uridine-diphosphoglucuronyl transferases and sulfotransferases, respectively. These metabolites can be easily excreted and are less toxic than their free compounds. Here, we demonstrate for the first time in the animal kingdom the conversion of DON to DON-3-glucoside (DON-3G) via a model system with plant pathogenic aphids. This phase II biotransformation mechanism has only been reported in plants. As the DON-3G metabolite was less toxic for aphids than DON, this conversion is considered a detoxification reaction. Remarkably, English grain aphids (Sitobion avenae) which co-occur with the DON producer Fusarium graminearum on wheat during the development of fusarium symptoms, tolerate DON much better and convert DON to DON-3G more efficiently than pea aphids (Acyrthosiphon pisum), the latter being known to feed on legumes which are no host for F. graminearum. Using a non-targeted high resolution mass spectrometric approach, we detected DON-diglucosides in aphids probably as a result of sequential glucosylation reactions. Data are discussed in the light of an eventual co-evolutionary adaptation of S. avenae to DON.

Ensuring the quality of results from food control laboratories: laboratory accreditation, method validation and measurement uncertainty

Abstract: Within the framework of control activities by a competent food analysis laboratory, an important basic task is to develop and improve expertise and knowledge continuously in order to report unequivocally reliable results that guarantee safe food distribution and consumption. This scientific message is vital with respect to the whole concept of the general protection of public health. Internationally prescribed and adopted quality regulations are mandatory for quality assurance during production and in quality control before release for consumption. These quality regulatory demands are elaborated within internationally recognized organizations and directorates in order to develop specific aims and activities to assure efficient quality control of traded food. Suitable validated methods of analysis should be able to verify if these integral quality demands for food are fulfilled. In this way the exact composition, safety, falsifications and absence of harmful contaminants should be verified or detected. In this context it is of principal importance that a control laboratory has at its disposal selective, sensitive, rugged, accurate and precise methods of analysis providing highly reliable results with low uncertainty. It is a fundamental duty of a control laboratory to follow actual progress in analysis development and statistical method validation continuously within an accredited quality environment such as prescribed in the ISO 17025 norm and to apply relevant suitable and powerful statistical tools that ensure excellent quality results.