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  • Author or Editor: Saleh Alqasoumi 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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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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Extensive research on Ficus species has shown their excellent cytotoxic potential which motivated the authors for further evaluation of its other species. In this article, the β-sitosterol content in the chloroform extract of the leaves of five Ficus species (Ficus carica [FCCE], Ficus nitida [FNCE], Ficus ingens [FICE], Ficus palmata [FPCE], and Ficus vasta [FVCE]) was estimated by a validated high-performance thin-layer chromatography (HPTLC) method along with cytotoxic activity. The chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with hexane and ethyl acetate (8:2, v/v) as the mobile phase. The developed plate was derivatized with p-anisaldehyde reagent, scanned, and quantified at λ = 550 nm. It furnished a compact and intense peak of β-sitosterol at R F = 0.17 ± 0.001. The contents of β-sitosterol (μg mg−1 of the dried weight of the extract) in the selected Ficus species were found as: FCCE (1.047 μg mg−1) > FVCE (0.771 μg mg−1) > FNCE (0.372 μg mg−1) > FPCE (0.309 μg mg−1), while it was absent in F. ingens. Methylthiazol tetrazolium (MTT) assay was used to compare the cytotoxic potential of all Ficus species against HepG2 (liver), HEK-293 (kidney), MCF-7 (breast), and MDA-MB 231 (breast) cell lines. The FCCE exhibited good cytotoxic property against HepG2, HEK-293, and MDA-MB-231 cells (IC50: 32.5, 41.4, and 47.3 μg mL−1, respectively), while FICE showed against HepG2 and MDA-MB-231 cells (IC50: 31.4 and 41.2 μg mL−1, respectively). The remaining Ficus extracts were found to be very less effective or insignificant. The cytotoxic property of FCCE is also supported by the HPTLC estimation of β-sitosterol which is reported to exhibit anticancer properties by interfering with multiple cell signaling pathways, including cell cycle, apoptosis, and proliferation. Our data suggest that the developed HPTLC method can be further employed in the analysis of marketed herbal formulations, and the active Ficus species can be further subjected to isolation of cytotoxic phytoconstituents.

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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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JPC - Journal of Planar Chromatography - Modern TLC
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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