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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

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2013

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Vol. 25, no. 1

159--169

EN

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.

2

Development and validation of a TLC-densitometric method for the simultaneous quantitation of icariin and L-arginine from commercial formulations

Gupta P. K, Kuber V. V, Ghosh V. K, Bhope S. G, Sharma V., Purohit S

Acta Chromatographica

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2011

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Vol. 23, no. 3

447--458

EN

A simple, rapid, and specific thin layer chromatographic (TLC) method has been developed and validated for the simultaneous estimation of icariin and L-arginine from commercial polyherbal formulations for sexual dysfunction. The separation of the methanol extract of these formulations was achieved on silica gel 60 F254 aluminum backed TLC plates by using ethyl acetate-acetone-glacial acetic acid-formic acid-water 12:2:1:2:2 (υ/υ) as mobile phase. Densitometric analysis of icariin and L-arginine was monitored in absorbance mode at 270 and 195 nm, respectively. The linear regression analysis data for the calibration plots for icariin and L-arginine showed good linear relationship with r2 = 0.9984 +- 0.01 and 0.9968 +- 0.02, in the concentration ranges of 250–750 and 500–1500 ng/spot, respectively. The method was validated for precision, robustness, and recovery. The average percentage recovery was found to be 98.26% for icariin and 99.63% for L-arginine. The limits of detection and quantitation were 72, 116 and 238, 383 ng/spot, respectively, for icariin and L-arginine. Statistical analysis proves that the method is repeatable and selective for the estimation of the targeted drugs. Since the proposed mobile phase effectively resolves the icariin and L-arginine, this method can be applied for the identification and quantitation of these components in herbal extracts and marketed formulations.

3

Validated HPTLC method for simultaneous quantification of sennoside A, sennoside B, and kaempferol in Cassia fistula Linn.

Bhope S. G, Kuber V. V, Nagore D. H

Acta Chromatographica

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2010

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Vol. 22, no. 3

481--489

EN

This paper describes a simple, precise, and accurate HPTLC method for simultaneous quantification of sennoside A, sennoside B, and kaempferol in Cassia fistula whole plant extract. Chromatographic separation of the sample extract was performed on aluminium foil plates coated with silica gel 60 F 254 as stationary phase. The mobile phase was toluene-ethyl acetate-methanol-formic acid 8:10:5:2 ( υ/υ ). Densitometric evaluation of the separated bands was performed at 270 nm. Sennosides A and B and kaempferol were satisfactorily resolved at R F 0.22 ± 0.05, 0.19 ± 0.05, and 0.81 ± 0.05, respectively. Recovery of sennosides A and B and kaempferol from Cassia fistula extract was 98.03, 98.74, and 99.08%, respectively. The method was validated for specificity, accuracy, linearity (100–400 ng per band), and precision (instrument precision in the range 1.03–1.33 and method precision in the range of 1.31–1.75) in accordance with ICH guidelines.