Development and Validation of Analytical Method for Clenbuterol Chiral Determination in Animal Feed by Direct Liquid Chr
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Development and Validation of Analytical Method for Clenbuterol Chiral Determination in Animal Feed by Direct Liquid Chromatography Noelia Rosales-Conrado 1 & María Eugenia de León-González 1 & Luis María Polo-Díez 1
Received: 12 December 2014 / Accepted: 3 March 2015 # Springer Science+Business Media New York 2015
Abstract A direct chiral liquid chromatography with UV detection method was developed and validated for enantiomeric determination of clenbuterol β2-agonist in feed samples for poultry. To obtain optimum chromatographic conditions for chiral separation, two commercially available columns packed with vancomycin and teicoplanin chiral stationary phase (150×2.1 mm i.d, 5 μm) were tested. Experimental design and response surface analysis were used to model analytical responses and to get mobile phase composition for acceptable enantioresolution and performance in low analysis time. Different sample treatments based on HCX SPE and on the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method were evaluated and compared to obtain the best extraction recoveries with negligible matrix effect. Special attention was paid to compatibility between sample preparation and chiral separation to get enantioresolution and low detection limits in the high-complex sample. QuEChERS-based extraction allowed obtaining cleaner feed extracts in low sample preparation time, providing a useful and reliable method capable to control the illegal use of clenbuterol enantiomers in feed for growth promotion in food-producing animals. Reproducible recoveries, from 64 to 101 % for clenbuterol enantiomers, with relative standard deviation values within the range 1–6 %, were achieved in feed samples of poultry spiked at low concentration levels of micrograms per gram.
* Noelia Rosales-Conrado [email protected] 1
Analytical Chemistry Department, Faculty of Chemistry, Complutense University of Madrid, Avda. Complutense s/n, 28040 Madrid, Spain
Keywords Clenbuterol . Chiral liquid chromatography . Sample preparation . QuEChERS . Animal feed
Introduction Food safety has become of major concern for consumers, governments, and producers as a result of the global marketplace and an increase in awareness of public health and quality. Trace levels of chemical contaminants in foodstuffs can originate from natural sources, veterinary drug residues, byproducts from food processing, and other intentional and unintentional adulteration (Krska et al. 2012). Clenbuterol (CLB) is a potent chiral β2-adrenergic agonist used as bronchodilatory, tocolytic, and mucolytic agent in the symptomatic treatment of respiratory diseases in both humans and animals. It also promotes the growth of muscular tissue and the reduction in body fat; hence, it is illicitly used as a nutrient repartitioning agent in meat-producing animals. The use of CLB as a growth promoter and its accumulation in foodstuffs cause adverse effects on public health, such as accidental human poisoning associated to consumption of meat products and liver contaminated with CLB r
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