A new, simple, precise, rapid, selective and stability indicating reversed-phase high performance liquid chromatographic (HPLC) method has been developed and validated for simultaneous quantification of trihexyphenidyl hydrochloride, trifluoperazine hydrochloride and chlorpromazine hydrochloride from combined tablet formulation. The method is based on reverse-phase using C-18 (250×4.6) mm, 5 μm particle size column. The separation is achieved using isocratic elution by methanol and ammonium acetate buffer (1% w/v, pH 6.5) in the ratio of 85:15 v/v, pumped at flow rate 1.0 mL/min and UV detection at 215 nm. The column is maintained at 30 °C through out the analysis. This method gives baseline resolution. The total run time is 15 min. Stability indicating capability is established buy forced degradation experiment. The method is validated for specificity, accuracy, precision and linearity as per International conference of harmonisation (ICH). The method is accurate and linear for quantification of trihexyphenidyl hydrochloride, trifluoperazine hydrochloride and Chlorpromazine hydrochloride between 5 - 15 μg/mL, 12.5- 37.5 μg/mL and 62.5 - 187.5 μg/mL respectively.
Background
Apigenin (4′, 5, 7-trihydroxyflavone), a flavonoid, is present usually in fruits and vegetables possessing numerous biological properties like antioxidant, anti-viral, antibacterial, anti-inflammatory, and chemoprevention activity. So present study was aimed to prepare and characterize nanoliposomes of apigenin and estimate its encapsulation efficiency by stability-assisted reverse-phase (RP)-HPLC method.
Results
The stability indication of the RP-HPLC method developed for apigenin-loaded nanoliposomes was successfully demonstrated and parameters were mainly the retention time which was 4.21 min, limit of detection (LOD) 0.49 μg/mL, limit of quantification (LOQ) 1.48 μg/mL, and %relative standard deviation (RSD) less than 2%. Therefore, the stability indication of the developed reverse-phase HPLC method for apigenin-loaded nanoliposomes was demonstrated successfully and parameters like accuracy, linearity, LOD, LOQ, precision, and %RSD were within the limit range and found to be satisfactory.
Conclusion
The developed RP-HPLC method was found to be suitable for the quantification or estimation of apigenin with its stability in apigenin-loaded nanoliposomes, and this method will be a powerful tool in the future for the estimation of apigenin present in any pharmaceutical preparations.
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