VALIDATED CAPILLARY ZONE ELECTROPHORESIS METHOD FOR THE DETERMINATION OF ACUTE NEUROTOXIN AZIDE IN AN ANTIRETROVIRAL DRUG
Appalacharyulu Salapaka*, Hemanth Kumar Sharma, Kishore Babu Bonige, Raghu Babu Korupolu, Uttam Kumar Ray
ABSTRACT
A rapid and sensitive capillary zone electrophoresis method was developed, optimized and validated for determination of acute neurotoxin azide, in an antiretroviral drug - Zidovudine. Electrophoresis was carried out in an aqueous run electrolyte containing 7.5 mM of sodium chromate, 5 mM of boric acid and 0.1 mM of cetyltrimethylammonium bromide. Separation of azide was achieved using a bare fused silica capillary had an effective length of 56 cm and an internal diameter of 50 μm with an extended light path detection window. Capillary was thermostated at 25° C and -12 μA current applied across on it. Nanolitre volumes of standard and sample solutions injected hydrodynamically by applying 50 mbar pressure for 45 sec. Analyte was monitored at a wavelength, signal: 460 nm and reference: 275 nm. The detector response found to be linear for azide in the concentration range of 0.09 μg/ml to 1.5 μg/ml. LOD and LOQ values of azide are found to be 0.09 μg/mL and 0.28 μg/mL respectively. RSD for replicate (n=6) injections of standard solution containing 1.0 μg/mL of azide was < 5% and for precision in intra-day (n=6) and inter-day (n=6) experiments was within 5%. Accuracy for the azide is 94.3%. The proposed method is specific, accurate, sensitive, rugged and precise for routine analysis of azide in quality control laboratories.
Keywords: Antiretroviral, Azide, Capillary zone electrophoresis, Indirect UV detection, Zidovudine.
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