The manuscript describes validated high performance thin layer chromatography (HPTLC) and first derivative UV spectrophotometric methods for the estimation of diazepam (DZP) and propranolol hydrochloride (PRO) in combined dosage form. The HPTLC separation was achieved on an aluminium-backed layer of silica gel 60F254 using mobile phase ethylacetate-methanol-toluene-triethylamine (1.0 + 3.0 + 6.0 + 0.1, v/v/v/v). Quantification was achieved with UV detection at 235 nm over the concentration range 25 – 250 ng/spot and 200 – 2000 ng/spot for DZP and PRO respectively, with mean recovery of 100.3 ± 0.54 and 100.2 ± 0.35 % for DZP and PRO, respectively by HPTLC method. Derivative spectrophotometric method was based on the estimation of both the drugs at their respective zero crossing point (ZCP). The first-order derivative spectra were obtained at N = 1 (scaling factor), Δλ = 2.0 nm, and the determinations were made at 248 nm (ZCP of PRO) for DZP and 242 nm (ZCP of DZP) for PRO over the concentration range of 2.5–30 μg/mL for both DZP and PRO with mean recovery of 100.2 ± 0.49 and 100.1 ± 0.13 % for DZP and PRO, respectively by first derivative UV spectrophotometric method. These methods were found to be simple, sensitive, accurate, precise, reproducible and economical and applicable for the simultaneous determination of DZP and PRO in combined dosage form.
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