Validated Stability Indicating HPTLC, UHPLC and UV-Spectrophotometric Techniques for the Determination of Bepotastine Besilate in Presence of Its Oxidative Degradate

Abstract

The study is aimed at developing methods which have a complete validation as stipulated in the ICH guidelines and to be applied for the determination of Bepotastine besilate (BB) in pure form and in pharmaceutical formulations in the presence of its oxidative degradation product. High performance thin layer chromatography (HPTLC), Ultra high performance liquid chromatography (UHPLC) and different spectrophotometric methods (first derivative, first derivative of ratio spectra and ratio difference are developed for simultaneous determination of bepotastine besilate in laboratory-prepared mixtures of bepotastine besilate with its oxidative degradate and in pharmaceutical formulations were used in the study design. Firstly, HPTLC was performed and separation occurred on silica gel 60 F254 plates, with butanol: ammonia (8:2, v/v) as a developing system. UHPLC in which separation occurred on a Kinetex C 18 column using methanol- 0.1% O-phosphoric acid - acetonitrile (70:20:10, by volume) as mobile phase, followed. And lastly was UV/Vis spectrophotometry which included first derivative determination of the drug at 252.6 nm, first derivative of ratio of peak amplitudes at 233.4, 250 and 275.6 nm and the ratio difference with the amplitude difference between (240 nm and 260 nm). Result showed that HPTLC method was applicable over the concentration range of 0.5-5 μg / band, while UHPLC method was linear over the concentration 2- 12 μg/mL and spectrophotometric methods were linear over the concentration range 20-120 μg/mL for bepotastine besilate. The proposed three techniques are quite accurate and precise. They can be used for routine analysis of bepotastine besilate in pharmaceutical formulation and stability indicating methods.

Chemical Stability Study of H1 Antihistaminic Drugs from the First and the Second Generations, Diphenhydramine, Azelastine and Bepotastine, in Pure APIs and in the Presence of Two Excipients, Citric Acid and Polyvinyl Alcohol

“…These two products were detected after stressing BEPO at extreme pH values, as under thermolytic as photolytic stress. A representative chromatogram showing the disappearance of the peak corresponding to unmodified BEPO and two additional peaks from the decomposition products B1 and B2 is presented in Figure 7 C. One study from the literature allowed the detection of two degradants of BEPO after storing in acidic or oxidative conditions [ 24 , 25 ]. In the next studies from the literature [ 26 ], irradiation under alkaline conditions led to the formation of five degradants ( Figure 4 ).…”

“…According to the literature, oxidative and acidic degradation of BEPO could be due to the oxidation or cleavage of butanoic acid from the structure [ 24 , 25 ]. Photodegradation of BEPO was proposed by the cleavage of its ether bond, especially in strong pH conditions.…”

“…As far as BEPO is concerned, two reports were found in the literature presenting stability-indicating HPLC methods together with stress degradation experiments [ 23 , 24 ]. In addition, the sensitivity of BEPO to oxidation leading to one degradation product (DP1) has been confirmed using HPTLC, UHPLC and spectrophotometric methods [ 25 ].…”