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3 Acta Chromatographica

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Stability-indicating TLC—image analysis method for determination of andrographolide in bulk drug and Andrographis paniculata formulations

Phattanawasin P., Burana-Osot J., Sotanaphun U., Kumsum A.

Acta Chromatographica

2016

Vol. 28, no. 4

525--540

A simple and rapid thin-layer chromatographic (TLC)–image analysis method was developed for stability-indicating studies and determination of andrographolide in bulk drug and in Andrographis paniculata formulations. Good chromatographic separation of andrographolide and the degradation products under various stress conditions was achieved on a silica gel G60 F254 TLC plate with the use of two mobile phases, dichloromethane—toluene—ethanol (6:3:1, v/v/v) and toluene—ethyl acetate—formic acid (5:3.5:1.5, v/v/v) and p-anisaldehyde as visualization reagent. Image analysis of the scanned TLC plate was performed by Sorbfil TLC Videodensitometer, UN-SCAN-IT, JustTLC, and ImageJ software to measure the quantity of the dark bluecolored band of andrographolide on a TLC plate. The TLC—image analysis method was validated in terms of specificity, linearity, sensitivity, precision, accuracy, and robustness and was also applied for determination of the amount of andrographolide in A. paniculata formulations and the content of andrographolide remained in the samples under forced degradations. The analytical results determined by the TLC—image analysis method, TLC—densitometry, and high-performance liquid chromatography (HPLC) methods were compared and found to be not significantly different.

Simultaneous estimation of anti-cancer terpenoids in pharmaceutical nanoformulation by RP-HPLC and HPTLC

Parveen R., Ahmad F. J., Iqbal Z., Singh M., Kamal Y., Ahmad S.

Acta Chromatographica

2014

Vol. 26, no. 2

391--400

Andrographolide and betulinic acid are the terpenoids having potential anti-cancer activity. The cytotoxicity activity of both the drugs was carried out separately and in combination on liver cancer HepG2 cell lines. High-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) methods were developed and validated for simultaneous estimation of these two terpenoids as per the International Conference on Harmonization (ICH) guidelines, which was applied for quantification in nanoformulation. The retention time by HPLC and retardation factor by HPTLC for andrographolide and betulinic acid were found to be 2.2 and 6.6 min, and 0.24 ± 0.01 and 0.66 ± 0.01, respectively. Both the methods were validated for accuracy, precision, repeatability, robustness, limit of detection (LOD), and limit of quantitation (LOQ). The content of andrographolide and betulinic acid in nanoformulation was found to be 96.0% and 98.0% by HPLC and 96.59% and 98.33% by HPTLC, respectively, of labelled claim.

Development and validation of RP-HPLC method for simultaneous analysis of andrographolide, phyllanthin, and hypophyllanthin from herbal hepatoprotective formulation

Bhope S. G., Kuber V. V., Nagore D. H., Gaikwad P. S., Patil M. J.

Acta Chromatographica

2013

Vol. 25, no. 1

159--169

A simple, sensitive, and accurate liquid chromatographic method with photodiode array detector was developed for the determination of andrographolide, phyllanthin, and hypophyllanthin. The separation was carried out on a reverse-phase 250 mm × 4.6 mm, 5μ symmetry C8 column (Waters). The gradient was prepared from 0.1% orthophosphoric acid (solvent A) and (1:1) acetonitrile:methanol (solvent B) as mobile phase delivered at a flow rate of 1 mL min -1. A linear behavior was observed between observed peak area response, and concentration of analytes was investigated, with good correlation coefficient. The method established was successfully applied to quantify andrographolide, phyllanthin, and hypophyllanthin from the herbal hepatoprotective formulation.