A densitometric HPTLC method has been established for analysis in alcohol extracts of the seeds and leaves of Achyranthes aspera L. The extracts were separated on aluminum foil-backed silica gel 60 F 254 plates. Detection was by measurement of absorbance at 490 nm, in absorbance/reflectance mode, after derivatization with 10% H 2 SO 4 in 95.5% (v/v) ethanol. The R F of oleanolic acid was 0.22. The oleanolic acid content of both extracts was compared statistically and the amount in the seed extract found to be higher (0.970%) than that in the leaf extract (0.666%).
A simple, precise, rapid, selective, and economic reversed phase high-performance liquid chromatography (HPTLC) method has been established for simultaneous analysis of Metformin Hydrochloride and Repaglinide. HPTLC method was developed using on precoated silica gel G60 F254 plates as stationary phase, using methanol:ammonium sulphate (0.25%) (pH-5.7) (2.5:7.5, v/v) as mobile phase. The plates were scanned at approximately 243 and 236 nm for HPLC and HPTLC both respectively. In HPTLC method both the drugs were resolved using proposed mobile phase and R f value was found to be 0.34 for MET and R f 0.60 for REPA. The method was found to linear in the range 500-2500 ng/band for MET, and 100-500 ng/band based for REPA respectively. This HPTLC procedure is economic, sensitive, and less time consuming than other chromatographic procedures. It is a user-friendly and importance tool for analysis of combined dosage form.
The present paper describes stability indicating high-performance thin-layer chromatography (HPTLC) assay method for nitazoxanide in bulk drugs. The method employed TLC aluminium plates precoated with silica gel 60F-254 as the stationary phase. The solvent system consisted of carbon tetra chloride: ethyl acetate (7:3 v/v). The system was found to give compact spot for nitazoxanide (R f value of ± 0.02). Densitometric analysis of nitazoxanide was carried out in the absorbance mode at 345 nm. The linear regression analysis data for the calibration plots showed good linear relationship with r 2 = 0.9992 ± 0.0001 with respect to peak area in the concentration range 200-1200 ng spot-1. The mean value ± S.D. of slope and intercept were 9.3726 ± 0.023 and 3795.46 ± 13.940 with respect to peak area. The developed HPTLC method was validated with respect to accuracy, precision, recovery and robustness. Also to determine related substance and assay determination of nitazoxanide that can be used to evaluate the quality of regular production samples. The developed method can also be conveniently used for the assay determination of nitazoxanide in pharmaceutical formulations. The limits of detection and quantitation were 17.12 and 51.88 ng spot-1 , respectively. Nitazoxanide was subjected to acid and alkali hydrolysis, oxidation, photochemical and thermal degradation. The drug undergoes degradation under acidic, basic, oxidation and heat conditions. This indicates that the drug is susceptible to acid, base hydrolysis, oxidation and heat. Statistical analysis proves that the method is repeatable, selective and accurate for the estimation of said drug. The proposed developed HPTLC method can be applied for identification and quantitative determination of nitazoxanide in bulk drug and tablet formulation.
A simple, precise, rapid, selective, and economic reversed phase high-performance liquid chromatography (RP-HPLC) method has been established for simultaneous analysis of A Phenomenex C 18 (250×4.6 mm i.d) chromatographic column equilibrated with mobile phase 0.02 M Potassium dihydrogen o-phosphate/acetonitrile (55/45, v/v) adjusted to pH 6.5 with Triehtylamine (1% v/v) was used. Mobile phase flow rate was maintained at 1 ml/min and effluents were monitored at 278 nm. The sample was injected using a 20 µl fixed loop, and the total run time was 10 min. Experimental conditions such as pH of mobile phase, column saturation time, selection of wavelength, etc. were critically studied and the optimum conditions were selected. The retention time for PCM, DMP and TMD were 3.76 min, 5.18 min and 4.28 min, respectively. The calibration curve for DMP, PCM and TMD was found to be linear in the range of 0.2-1 µg/ml, 6.5-32.5 µg/ml and 0.75-3.75 µg/ml with a correlation coefficient of 0.9998, 0.9976 and 0.9974. The detection limits for PCM, DMP and TMD were 20 ng/ml, 1.06 ng/ml and 2 ng/ml, respectively, while quantitation limits were 60 ng/ml, 3.23 ng/ml and 6 ng/ml, respectively. This HPLC procedure is economic, sensitive, and less time consuming than other chromatographic procedures. It is a user-friendly and importance tool for analysis of combined tablet dosage forms.
A simple, rapid and accurate thin-layer chromatography (TLC)-densitometric method has been established and validated for the simultaneous determination of nebivolol hydrochloride and hydrochlorothiazide in tablets. The method is based on TLC separation of the two drugs followed by densitometric measurements of their spots at 281 nm. The separation was carried out on Merck TLC aluminium plate of silica gel 60F 254 using toluene:ethyl acetate:methanol:ammonia (3:2.7:1.7:0.1 v/v/v) as a mobile phase. Nebivolol hydrochloride and hydrochlorothiazide gave sharp and well defined peak at R f 0.68 and 0.38, respectively. Calibration curves for nebivolol hydrochloride and hydrochlorothiazide were linear in range 500-3000 ng/spot and 1000-6000 ng/spot, respectively. Method was successively applied to tablet formulation. No chromatographic interferences from the tablet excipients were found. The method was validated in accordance with the requirements of ICH guidelines.
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