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Research Journal of Pharmacy and Technology
Year : 2017, Volume : 10, Issue : 7
First page : ( 2249) Last page : ( 2254)
Print ISSN : 0974-3618. Online ISSN : 0974-360X.
Article DOI : 10.5958/0974-360X.2017.00398.5

Development and Validation of a Bioanalytical method for the Determination of Levamisole Residue in Backyard Poultry Egg

M Hossain d Kamal1,*, Nahar Kamrun1, M Mazumder ohammed Ehsanul Hoque1, M Wahiduzzaman d1, Alqahtani Ali S.2, Dr Gestier Tony1, Hamid Kaiser1,*

1Vetafarm Pty Ltd. R and D Centre, Wagga Wagga, NSW, 2650, Australia.

2Faculty of Pharmacy, King Saud University, Saudi Arabia

*Corresponding author e-mail: kamal@vetafarm.com.au

Online published on 17 October, 2017.


Development of a bioanalytical method is often tricky and labour intensive due to its nature or the criteria set by regulatory bodies. In the present study, a simple, reliable and sensitive analytical method was developed for the determination of levamisole residue in chicken eggs. Levamisole and ronidazole (internal standard, IS) were separated on a C18 reversed-phase HPLC column. Following liquid-liquid extraction, chromatographic separation was accomplished with a mobile phase consisting of acetonitrile: 0.05M KH2PO4 at ratio of 20: 80, and the drug was detected at 235nm using a UV detector at a flow rate of 1.0ml/min at the ambient temperature. Linearity was obtained over the range 0.3125–10.0μg/ml for levamisole hydrochloride with lower limit of quantitation of 0.3125μg/ml. For each level of quality control samples, inter-and intra-day precision (%CV) was within ±15%. Absolutes extraction recovery of drug from plasma was ≥;60%. The residue limit of levamisole gradually increased from day 1 to day 3 and then declined from day4 to day 7. The highest level of residue after oral administration of 80mg/kg levamisole to chickens was on day 3 (0.599μg/g). This level was lower than the recommended maximum residue limit (MRL), thus, no withholding period was required for egg consumption following the administration of the levamisole solution.



Levamisole, Bioanalytical, Residue, Withholding period, Backyard poultry.


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