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- Author or Editor: Arti Katiyar x
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Asensitive, selective, and precise high-performance thin-layer chromatography (HPTLC) method was developed for simultaneous analysis of six bioactive phenolic compounds, i.e., juglone, quercetin, myricetin, rutin, caffeic acid, and gallic acid in methanol extract and its fractions (hexane, chloroform, ethyl acetate, and butanol fractions) from bark of Juglans regia. Good separation was achieved on RP-18 F254S TLC plate using methanol-water-formic acid-acetic acid (48.8:46.4:2.4:2.4, ν/ν). The densitometric determination of the compounds was carried out at 254 nm in reflectance/absorbance mode. The method was validated in terms of linearity, sensitivity, accuracy, precision, robustness, and specificity. The linear regression data for the calibration plots of the reference compounds showed a good linear relationship with higher correlation coefficient (r 2 ≥ 0.997). Accuracy of the method was evaluated in terms of average percent recovery, which ranged from 98.63 to 101.06%. HPTLC results revealed qualitative and quantitative differences in the phenolic compounds in the extract and fractions. The ethyl acetate fraction contained gallic acid followed by myricetin, rutin, quercetin, and caffeic acid in higher amount in comparison to the extract and fractions. The present method can be used for routine quality control of J. regia extracts.
A new quantitative method using thin-layer chromatography silica gel 60F254 plates as the stationary phase and the mobile phase consisting of toluene-ethyl acetate-acetic acid (15.0:7.5:0.5, v/v) was employed for simultaneous determination of lignans and flavonoid in Podophyllum hexandrum. Densitometric determination of the constituents was performed at 254 nm in reflectance/absorbance mode. The method was validated for precision, accuracy, specificity, robustness, and recovery. The linear regression analysis data for the calibration plots showed a good linear relationship (r 2 = 0.9903–0.9976) in the calibration range of 1–8 μg per band for 4′-O-demethylpodophyllotoxin (1), podophyllotoxin (2), and podophyllotoxone (4), and 2–10 μg per band for kaempferol (3) and deoxypodophyllotoxin (5) with respect to peak area. Limits of detection and quantitation were in the range of 250–617 ng per band and 856–1974 ng per band. The average recovery for 4′-O-demethylpodophyllotoxin, podophyllotoxin, kaempferol, podophyllotoxone, and deoxypodophyllotoxin was 96.38 ± 1.92 to 101.84 ± 1.05%, indicating the excellent reproducibility. Podophyllotoxin was found in highest content (9.92 μg mg−1) and podophyllotoxone in the lowest content (0.94 μg mg−1). The proposed method is rapid, simple, precise, specific, sensitive, accurate, and robust.
A reverse phase high-performance thin-layer chromatography (RPHPTLC) method was developed for determination of vitexin, hyperoside, vitexin-2″-O-rhamnoside, quercetin, and apigenin in the Crataegus oxyacantha extract. The method employed precoated plate of RP-18 silica gel 60F254 as the stationary phase with acetonitrile-methanol-water-formic acid (10:10:20:0.05, ν/ν) as mobile phase, and densitometric determination was carried out at wavelength 254 nm in reflection/absorption mode. The linear regression analysis data for the calibration plots showed linear relationship (r) from 0.9985 to 0.9992. The method was validated for accuracy, precision, and robustness. The limits of detection and quantification were in the range of 100–300 ng and 310–960 ng, respectively, for analytes. The method is reproducible and convenient for quantitative analysis of these flavonoids in the leaves of C. oxyacantha. This is the first report of simultaneous densitometry quantification of major bioactive constituents in C. oxyacantha by developed and validated RP-HPTLC method. The change in the content of bioactive constituents with growth of the plant was also examined in leaf samples collected for 2 years.