Title:Development and Validation of RP-HPLC Method for the Simultaneous Determination of Levamisole HCl and Oxyclozanide and its Application in the Assay of Veterinary Bolus Formulations
VOLUME: 17 ISSUE: 5
Author(s):Kemal Hussien Seid, Tarekegn Berhanu, Kaleab Asres and Ayenew Ashenef*
Affiliation:Department of Pharmaceutical Chemistry and Pharmacognosy, School of Pharmacy, College of Health Sciences, Addis Ababa University, P.O.Box. 1176, Addis Ababa, Previously Pesticides Residue Analysis Laboratory, Federal Ministry of Agriculture, Addis Ababa, Ethiopia.Currently Academic Vice President, Department of Industrial Chemistry, Addis Ababa Science and Technology University, P.O. Box.16417, Addis Ababa, Department of Pharmaceutical Chemistry and Pharmacognosy, School of Pharmacy, College of Health Sciences, Addis Ababa University, P.O.Box. 1176, Addis Ababa, Department of Pharmaceutical Chemistry and Pharmacognosy, School of Pharmacy, College of Health Sciences, Addis Ababa University, P.O.Box. 1176, Addis Ababa
Keywords:Analytical method development, bolus formulations, RP-HPLC, levamisole, oxyclozanide, imidazothiazole.
Abstract:
Introduction: A reverse-phase high-performance liquid chromatography (RP-HPLC) method
was developed and validated for the simultaneous analysis of two drugs, levamisole hydrochloride
(LH) and oxyclozanide (OX), in co-formulation for veterinary use.
Materials and Methods: The new HPLC method was validated as per the ICH and other guidelines. A
C18 column was used with a gradient program; eluent A was an equal mixture of methanol and acetonitrile,
and eluent B included a 25 mM phosphate buffer at pH 7.0 containing 30 mM sodium decanesulfonate
and triethylamine (50:50:1 v/v) with pH adjusted to 7.0 using H3PO4 [51:49 v/v] .The detection
wavelength was set at 220 nm. For the final gradient program, the retention times were 8.2(for
LH) and 13.6(for OX) minutes, respectively, at a flow rate of 1 ml/min over 20 minutes run time.
Results: The method was precise, specific and robust. The correlation coefficient, R2 was 0.9998 and
0.9999 for LH and OX, respectively in the range of 5 – 280 μg / mL. The percent y-intercepts and percent
residual standard deviations were 1.6%/0.4% and 1.4%/1.0% for LH and OX, respectively. The
LOD and LOQ of the method were 0.21 μg / mL and 0.62 μg / mL for LH and 0.06 μg / mL and 0.18
μg / mL for OX. The method has an average accuracy of 100.5% for LH and 101.1% for OX when
tested on veterinary bolus formulations, and the samples could be stored under typical lab conditions
for about 7 days without significant degradation.
Conclusion: This HPLC method is suitable for assaying levamisole hydrochloride and oxyclozanide
simultaneously from veterinary formulations.
