A simple, automated, and sensitive continuous flow-injection analysis/merging zones technique (CFIA/MZ) method was developed for the determination of D-Penicillamine (thiol drug) in pure, pharmaceutical formulations and biological samples. This method involved the reaction of 1,2-Naphthoquinone-4-Sulphonic acid Sodium Salt (NQS) with D-Penicillamine to produce a brown-colored complex that has maximum absorbance at 463 nm. The method was cheap, economic, precise and accurate where the limit of detection was 6.3 µg.mL -1 and the RSD% was 0.34 and Recovery was 99.31%. Various chemical and physical conditions that affected the reaction have been studied. A wide calibration curve was rectilinear within the concentration range (15-1000) μg.mL -1 with a sample throughput of 103 samples.hour -1 . The proposed procedure was applied successfully for the estimation of D-PEN in pharmaceutical and biological samples and the results obtained were favorably compared with those given by a reference method of United States Pharmacopoeia (USP), and there was no significant difference between the obtained results, regarding accuracy and precision at the 95% confidence level.
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