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  • Author or Editor: Prawez Alam x
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High-performance thin-layer chromatographic (HPTLC) densitometric method for analysis of arbutin in commercial whitening creams was developed and validated. Aluminum-backed silica gel 60 F254 plates were used as stationary phase while methanol-chloroform-acetic acid 3.5:6:0.5 (%, v/v/v) mixture was used as mobile phase. Under these chromatographic conditions, arbutin was well separated from other ingredients. This system was found to give a well defined, sharp, and compact spot of arbutin at retention factor (R F) value of 0.40 ± 0.02. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 42.25 and 112.45 ng per spot respectively. The proposed method with high degree of precision and accuracy was employed for the analysis of arbutin both qualitatively and quantitatively in commercial whitening creams. Due to the efficiency of the method in separating arbutin from other ingredients including its degradation products, it can be applied as a stability-indicating method.

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An attempt was made to develop and validate a simple, accurate high-performance thin-layer chromatographic (HPTLC) method for the simultaneous determination of diosmin, hesperidin, and ascorbic acid in pharmaceutical preparations. Analysis was performed on 10 cm × 20 cm aluminum-backed plates coated with 0.2 mm layers of silica gel 60 F254 (Merck, Germany). CAMAG TLC Scanner III was used for UV densitometric scanning. The used HPTLC system was found to give sharp, symmetrical, and well-resolved peaks at R F values of 0.34 ± 0.02, 0.40 ± 0.04, and 0.56 ± 0.03, and linearity was found in the ranges 100–800 ng/spot (r 2 = 0.9985), 100–800 ng/spot (r 2 = 0.9966), and 50–400 ng/spot (r 2 = 0.9951) for diosmin, hesperidin, and ascorbic acid, respectively.

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Novel, simple, and sensitive high-performance thin-layer chromatography (HPTLC) with fluorescence detection method has been successfully established and validated for the simultaneous determination of vulgarin and epivulgarin in different collections of Artemisia judaica. HPTLC method was carried out using glass plates coated with silica gel 60 F254 using petroleum ether‒acetone (7:3, v/v) as the mobile phase. After development, the plates were scanned and quantified densitometrically at 224 nm for both vulgarin and epivulgarin. The two compounds’ peaks from A. judaica were identified by comparing their single spot at R F = 0.30 ± 0.02 and R F = 0.36 ± 0.01, respectively, with those of vulgarin and epivulgarin. Linear regression analysis revealed a good linear relationship between peak area and amount of vulgarin and epivulgarin in the range of 100–700 ng band−1 for both compounds. The method was validated, in accordance with the International Conference on Harmonization (ICH) guidelines, for precision, accuracy, and robustness. The proposed method will be useful to measure the therapeutic dose of vulgarin and epivulgarin in A. judaica extracts.

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A simple and sensitive high-performance thin-layer chromatographic (HPTLC) method was developed for the evaluation of biomarker lupeol in the leaves of five different species of genus Ficus (Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata, and Ficus vest) belonging to family Moraceae. Chromatography was performed on glass-backed silica gel 60 F254 precoated HPTLC plates with solvents toluene-methanol (9:1, v/v) as the mobile phase. After development, the HPTLC plate was derivatized with p-anisaldehyde reagent, scanned, and quantified at 540 nm. The system was found to give compact spot for lupeol at R F = 0.32 ± 0.01. The precisions and accuracy (n = 6) for lupeol were found to be 1.47–1.64% and 1.63–1.86%, and 99–100% and 99.4–99.7%, respectively, for inter-day and intra-day. Lupeol was found to be present in four species, i.e., F. carica (0.4%, w/w), F. nitida (1.4%, w/w), F. palmata (0.33%, w/w), and F. vest (0.59%, w/w), while it was absent in F. ingens. The statistical analysis proved that the developed method is reproducible and selective. The developed method can be used as an important tool to assure the therapeutic dose of active ingredients in herbal formulations as well as for standardization and quality control of bulk drugs and in-process formulations. This method can also be employed for the further study of degradation kinetics and determination of lupeol in plasma and other biological fluids.

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A novel, simple, and sensitive high-performance thin-layer chromatography (HPTLC) method has been successfully developed and validated for the quantification of benzyl isothiocyanate in Salvadora persica (Miswak or Siwak) extract and some dental care herbal products labeled to contain Siwak extract. A modified HPTLC method was carried out using glass-coated silica gel 60 F254 plates and mobile phase consisting of n-hexane—ethyl acetate (9:1) furnished compact spots at R F 0.61 ± 0.01. The developed plate was scanned and quantified densitometrically at 191 nm. Linear regression analysis revealed a good linear relationship between peak area and amount of benzyl isothiocyanate in the range of 100–600 ng band−1 (r 2 = 0.9991). The method was validated in accordance with the International Conference on Harmonization (ICH) guidelines for precision, accuracy, and robustness. The proposed method will be useful to evaluate the therapeutic efficacy of Siwak extracts and dental care herbal formulations containing Siwak.

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A densitometric high-performance thin-layer chromatographic (HPTLC) method for analysis of hydroquinone has been developed and validated. Chromatography was performed on aluminum foilbacked silica gel 60 F254 plates with chloroform-methanol 85:15 (% v/v) as mobile phase. This system furnished a compact band for hydroquinone at R F 0.51. Hydroquinone was quantified densitometrically at 289 nm. The limits of detection (LOD) and quantification (LOQ) were 38.50 and 115.50 ng per band, respectively. High precision and accuracy were achieved. The method was used for both qualitative and quantitative analysis of hydroquinone in commercial formulations. Because the method can effectively separate the hydroquinone in the presence of its degradation products, it can be used as a stability-indicating method.

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A new rapid, simple, economical, and environment-friendly reversed- phase high-performance thin-layer chromatography (RPHPTLC) method has been established for the simultaneous determination of glycyrrhizin and glabridin in Glycyrrhiza glabra roots, rhizomes and selected herbal formulations. The method was carried out using RP-18 silica gel 60 F254S HPTLC glass plates and methanol–water (7:3 v/v) as the mobile phase. The developed plates were scanned and quantified densitometrically at 256 and 233 nm for glycyrrhizin and glabridin, respectively. Glycyrrhizin and glabridin peaks from G. glabra roots and rhizomes and herbal formulations were identified by comparing their single spots at R F = 0.63 ± 0.02 and R F = 0.28 ± 0.01, respectively. Linear regression analysis revealed a good linear relationship between the peak areas and the amounts of glycyrrhizin and glabridin in the ranges of 1000–7000 and 100–700 ng band−1, respectively. The method was validated, in accordance with the International Conference on Harmonization (ICH) guidelines for precision, accuracy, and robustness. The proposed method will be useful to determine the therapeutic doses of glycyrrhizin and glabridin in herbal formulations as well as in bulk drug.

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In this research, a novel method was developed for the matrix solid phase dispersion (MSPD) followed by high-performance liquid chromatography (HPLC) quantification of four marker constituents (vitamin C, gallic acid, rutin, and ellagic acid) in the freeze-dried pomegranate fruit juice. Various MSPD parameters like type of dispersant, sample–dispersant ratio, solvents, its volume, and time of extraction have been optimized after many trials. Furthermore, HPLC method has been developed and optimized for the analysis of all four components. The HPLC separation was achieved using a 250 × 4.6 mm column, particle size of 5 μm, C18 reverse phase column, with a mobile phase consisting of acetonitrile and 0.05% H3PO4, in gradient elution mode with a mobile phase flow rate of 1 mL/min, using ultraviolet (UV)–visible detection at 254 nm. All calibration curves showed good linear regression (r 2 ≥ 0.9925) within test ranges. The extraction recoveries of the marker constituents analyzed by MSPD methods were found as ranging from 97.5% to 103.5%. From comparing the chromatograms, validation data and other parameters like time, labor, and feasibility, we found that MSPD technique was most suitable for the analysis as compared to conventional liquid–liquid extraction technique.

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Authors: Omer A. Basudan, Perwez Alam, Nasir A. Siddiqui, Mohamed F. Alajmi, Adnan J. Alrehaily, Saleh I. Alqasoumi, Maged S. Abdel-Kader, Prawez Alam and Abd El Raheim M. Donia

A simple and sensitive high-performance thin-layer chromatographic (HPTLC) method was developed for the evaluation of biomarker β-amyrin in the leaves of fve different species of genus Ficus (Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata, and Ficus vasta) grown in the Kingdom of Saudi Arabia. Chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with solvents toluene–methanol (9:1, v/v) as the mobile phase. After development, the HPTLC plate was derivatized with p-anisalde-hydereagent to give well-resolved and compact spot of β-amyrin. Scanning and quantifcation were done at 550 nm. The system was found to give compact spot for β-amyrin at R F = 0.58. The linear regression analysis data for the calibration plots showed good linear relationship with r 2 = 0.998 with respect to area in the concentration range of 100–900 ng. The regression equation for β-amyrin standard was found to be Y = 5.835X + 87. The precisions (n = 6) for β-amyrin were found to be 1.64–1.77% and 1.68–1.84%, respectively, for intra-day and inter-day batches, and the recovery values were found to be 97.6–98.3%. β-Amyrin was found to be present in three species, i.e., F. carica (0.29%, w/w), F. nitida (0. 5 4% w/w), and F. p almata (0.31%, w/w), while it was absent in F. vasta and F. ingens. The statistical analysis proves that the developed method for the quantifcation of β-amyrin is reproducible; hence, it can beemployed for the determination of β-amyrin in plasma and other biological fuids as well as in fnished products avai lable in the market.

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Authors: Ahmed I. Foudah, Prawez Alam, Abdulaziz I. Al Furaih, Mohammad Ayman A. Salkini and Maged S. Abdel-Kader

Fenugreek is one of the oldest medicinal plants known by mankind. The aim of this study is to evaluate the effect of environmental conditions on the saponin contents of the plant. The amount of diosgenin in different samples of fenugreek was estimated by high-performance thin-layer chromatography (HPTLC). The developed method was validated, in accordance with the International Conference on Harmonization (ICH) guidelines for precision, accuracy, and robustness. HPTLC was carried out using hexane‒acetone (8:2%, v/v) on 20 cm × 10 cm glass coated silica gel 60 F254 plates. The developed plates were scanned and quantified densitometrically at λ = 430 nm. Linear regression analysis revealed a good linear relationship between the area under the peak and the amount of diosgenin in the range of 50–400 ng per band. The amount of diosgenin which reflects the total saponin contents varied according to the country of origin. The sample obtained from Yemen showed the highest amount of diosgenin followed by the Saudi sample. Both locations represent areas with the highest altitude (Yemen) and the highest temperature (Saudi).

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