Authors:Daniil Olennikov and Vyacheslav Partilkhaev
Athin-layer chromatographic (TLC) method was used for the quantification of four dominant flavonoids (rutin, narcissin, nicotiflorin, isoquercitrin) in the shoots of Cargana spinosa (L.) DC. (Fabaceae). Chromatographic separation was performed on silica gel HPTLC plates with double development of ethylacetate-1,2-dichloroethane- acetic acid-85% formic acid-water 10:2.5:1:1:0.8 (v/v) as mobile phase. The plates were scanned densitometrically at 360 and 387 nm. The method was validated for precision, repeatability, and accuracy. It was found to be precise: intraday and interday RSD were 1.48–1.87% and 1.59–1.97%, respectively. Instrumental precision and repeatability for the method were found to be 0.51–0.64 and 1.18–1.32 (CV (%)), respectively. Accuracy was checked by measuring the recovery at four levels; the average recovery was 98.19–101.36% for rutin, 98.04–101.14% for narcissin, 98.16–100.54% for nicotiflorin, and 98.11–101.98% for isoquercitrin. The method was used for the analysis of the flavonoids mentioned in C. spinosa shoots samples. Nicotiflorin was detected in Caragana genus for the first time.
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 (r2 ≥ 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.
Authors:Supriya Jirge, Pratima Tatke, and Satish Gabhe
Standardization has become mandatory for global acceptance of herbal oriental medicines which lack validated methods of analysis due to their complex nature. Thus, a simple and fast method was developed for the simultaneous quantitative determination of three biologically active flavonoid compounds, i.e., kaempferol, rutin, and quercetin in extracts and marketed formulations containing Phyllanthus niruri (Bhuiamla) and Emblica officinalis (Amla). The separation was performed on thin-layer chromatography (TLC) aluminum plates precoated with silica gel 60 F254. Good separation was achieved in the mobile phase composed of chloroform-methanol-formic acid and detected at wavelength 254 nm in reflectance-absorbance mode. The developed method was well validated by different parameters mentioned in International Conference on Harmonization (ICH) guidelines such as linearity, accuracy, precision, etc. The linear regression data for the calibration plots showed a good linear relationship with r = 0.9997, 0.9998, and 0.9998 for kaempferol, rutin, and quercetin, respectively. The validated method was successfully applied to determine the contents of three biomarkers in a single run in Bhuiamla Vati and capsule (Ayurvedic and modern solid dosage form, respectively) and Amla Vati and Amla tincture (homeopathic preparation). The present high-performance thin-layer chromatographic (HPTLC) method, which was used for the standardization of various marketed traditional medicines of two different medicinal plants, is being reported the first time with these three biomarkers.
Authors:Anshul Shakya, Neelutpal Gogoi, Sushil Kumar Chaudhary, Hans Raj Bhat, and Surajit Kumar Ghosh
Background: The fruits of Benincasa hispida (Thunb.) Cogniaux, a ‘rasayana’ in ‘Ayurveda’, are enriched with several secondary metabolites, and rutin is one of them. Fruits are used for their anabolic, brain tonic, carminative, diuretic, memory enhancer, refrigerant, and vitalizer properties. Objective: In view of the fact that herbal medicines and/or products are facing challenges towards global acceptance due to the lack of universally accepted standardization method (s), the aim of the current investigation was to develop and validate a high-performance thin-layer chromatography (HPTLC)-densitometry method for the quantification of rutin in the hydroalcoholic extracts of the fruit pulp of B. hispida (HABH). Materials and methods: The separation was achieved in a solvent system consisting of ethyl acetate-formic acid-acetic acid-water at a ratio of 7.2:0.7:0.7:1.4 by volume on a TLC aluminum plate pre-coated with silica gel 60 F254. Quantifications were performed by densitometric scanning under a deuterium lamp at a wavelength of 268 nm in the absorbance mode. The precision, accuracy, and reproducibility of the HPTLC method were validated by the International Conference on Harmonization (ICH) guidelines. Results: The mobile phase employed for HPTLC/TLC resulted in good separation for rutin (RF = 0.357). The limit of detection and limit of quantification of the analysis were found to be 0.1 and 0.3 µg per band, respectively. The rutin concentration in the HABH was found to be 178.28 ± 3.62 µg in 10 mg of the extract. Conclusion: The method developed here is simple, fast, reliable, and sensitive and can be implemented in the analysis and routine quality control of B. hispida formulations containing rutin.
Authors:Sayan Biswas, Ranjit K. Harwansh, Amit Kar, and Pulok K. Mukherjee
A simple, rapid, quantitative, and validated high-performance thin-layer chromatographic (HPTLC) method has been developed for the simultaneous estimation of quercetin (QU), rutin (RU), and gallic acid (GA) in the methanolic extract of Amaranthus tricolor L. aerial parts. Densitometric analysis of QU, RU, and GA were carried out in the absorbance mode at 254 nm. The method gave spot at RF 0.49, 0.14, and 0.28 corresponding to quercetin, rutin, and gallic acid, respectively. The limit of detection (22.31, 14.12, and 16.24 ng per spot) and limit of quantification (67.80, 55.32, and 52.54 ng per spot), respectively, were confirmed with the mobile phase toluene–ethyl acetate–formic acid in a ratio of 7:5:1 (v/v). Linear regression analysis data for the calibration plot for QU, RU, and GA showed a good linear relationship with a correlation coefficient r > 0.9955 in the concentration range of 200–800 ng per spot, 200–500 ng per spot, and 200–600 ng per spot, respectively. The method was validated for linearity, accuracy, precision, detection, and quantification limits, specificity, and robustness as per the International Conference on Harmonization (ICH) guidelines. The proposed validated high-performance thin-layer chromatographic method provides a novel approach for the quality control and standardization of A. tricolor L.
Authors:Y.T. Kamal, Sayeed Ahmad, Nanjaian Mahadevan, Prawez Alam, Shahana Salam, Yahya I Asiri, Abdullatif Bin Muhsinah, and Abdulrhman Alsayari
free flavonoids like rutin and quercetin forms the basis of bioactivity of these Itrifal formulations [ 5 ]. The anthraquinone glycosides like sennoside A and B were selected as a specific marker for C. angustifolia , which is present in a significant