Two simple, sensitive and specific fluorimetric methods have been developed for the determination of Penicillamine (PNC), a sulphur containing compound. Method (I) involves the reaction of PNC with 2′,7′-bis(acetoxymercuri)-fluorescein (AMF) in the presence of Kolthoff’s buffer, pH 8.2, with subsequent measurement of fluorescence spectra at 520 nm (λEx 497 nm). Method (II) is based on PNC being oxidized into penicillaminic acid using Cerium (IV) in an acidic medium. Method sensitivity has been improved using sodium triphosphate which enhances the luminescence intensity of Ce(III). Fluorescence spectra were then measured at 348 nm (λEx 293 nm). The reaction conditions and the fluorescence spectral properties have been investigated for both methods. Under the described conditions, the proposed methods were applicable over the concentration ranges 0.0048 − 0.0288 µg mL−1 and 0.096 − 0.288 µg mL−1 with mean percentage recoveries 99.95 ± 1.29 and 100.04 ± 1.10 for methods I and II, respectively. The proposed methods were validated in terms of accuracy, precision, LOD and LOQ and robustness and then were successfully applied to the determination of PNC in bulk powder and in capsules as well as in the presence of the related disulphide. The results obtained were determined to be in good agreement with those obtained using a previously reported method.
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