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One of the alternative methods for the measurement of radiochemical purity (RCP) of 99mTechnetium-sestamibi uses Whatman 3MM paper as the stationary phase and ethyl acetate as the mobile phase. However, alternative method shows peak splitting and shouldering of the radiopharmaceutical. This could be because of higher levels of analyte. We aimed to assess the effect of sample dilution (1 : 1) on this method and to compare these RCP values with the reference method. The diluted samples showed a unique peak in the radiopharmaceutical zone, but the RCP values were significantly different (P<0.05) and correlated poorly with the reference method.