VALIDATED REVERSE PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR THE SIMULTANEOUS DETERMINATION OF ARTEMETHER AND LUMEFANTRINE IN FIXED COMBINED DOSAGE FORM
Objective: The present aim is to develop simple, precise, and accurate reverse phase high performance liquid chromatographic method (RP-HPLC) for
the simultaneous assay of artemether and lumefantrine in fixed combined dosage form.
Methods: The chromatographic study was carried out on Hypersil C18 column (250Ã—4.6 mm, 5 Î¼) with mobile phase containing a mixture of KH2PO4
buffer (pH-3.5) and acetonitrile in the ratio of 45:55% v/v at a flow rate of 1.0 ml/minute with ultraviolet detection at 218 nm in ambient column
Results: Using the optimized chromatographic conditions artemether and lumefantrine eluted with retention times of 2.207 and 3.733 minutes,
respectively. The method was validated according to ICH guidelines with good reproducibility and linear responses, y=60.813.x+629.53 (rÂ²=0.9982)
for artemether and y=88.3108.x+2370.2 (rÂ²=0.9912). The % relative standard deviations of intra-day precision was ranged 0.378% and 1.26% for
artemether and 0.459% and 1.15% for lumefantrine, respectively. The percentage recoveries were ranged from 99.96% to 100.02% for artemether
and 99.96-99.97% for lumefantrine, respectively.
Conclusions: The developed RP-HPLC method was validated as per ICH guidelines and was found to be best suitable for pharmacokinetic studies of
these mentioned drugs.
Keywords: Artemether, Lumefantrine, ICH guidelines.
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Table 6: Results of robustness studies of artemether and lumefantrine
Flow rate (ml/minute)
Table 7: Stability data of artemether and lumefantrine
% Assay at 0 hr*
% Assay at 24 hrs*
*Average of six determinations
Fig. 4: Calibration curve of artemether
Fig. 5: Calibration curve of lumefantrine
Table 8: Results for analysis in formulations
Sample No. Combither
Artemether 20 mg
Lumefantrine 120 mg
*Average of three determinations. RSD: Relative standard deviation,
SD: Standard deviation
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Rao and Rambabu
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