Y.T. Kamal

Rapid RP-HPLC Method for the Quantification of Glabridin in Crude Drug and in Polyherbal Formulation

A simple, economic, selective, precise and robust method has been developed and validated for the analysis of glabridin in crude drugs and polyherbal formulations. Reversed-phase chromatography is performed on a C18 column with water and acetonitrile as mobile phase in gradient elution method at a flow rate of 1 mL/min. Detection is performed at 230 nm and a sharp peak is obtained for glabridin at a retention time of 14.9 ± 0.02 min. Linear regression analysis data for the calibration plot showed a good linear relationship between response and concentration in the range of 1-500 µg/mL; the regression coefficient is 0.9992 and the linear regression equation is y = 26.683x - 142.17. The method is validated for accuracy, precision, reproducibility, robustness and detection and quantification limits, in accordance with International Conference on Harmonization guidelines. Statistical analysis proved that the method is precise, reproducible, selective and accurate for the analysis of glabridin. The proposed, developed and validated high-performance liquid chromatography method for the quantification of glabridin can be used for the quality control and standardization of licorice (Glycyrrhiza glabra Linn.) and different herbal formulations in which licorice is present as a constituent.

Wound healing effects of nanoemulsion containing clove essential oil

Abstract The aim of this study was to investigate the wound healing effects of clove oil (CO) via its encapsulation into nanoemulsion. Optimized nanoemulsion (droplet size of 29.10 nm) was selected for wound healing investigation, collagen determination, and histopathological examination in rats. Optimized nanoemulsion presented significant would healing effects in rats as compared to pure CO. Nanoemulsion also presented significant enhancement in leucine content (0.61 mg/g) as compared to pure CO (0.50 mg/g) and negative control (0.31 mg/g). Histopathology of nanoemulsion treated rats showed no signs of inflammatory cells. These results suggested that nanoemulsion of CO was safe and nontoxic.

Variations in the cytotoxic glycoalkaloids solamargine and solasonine in different parts of the Solanum incanum plant during its growth and development in Oman

Abstract In addition to several important traditional medicine applications of Solanum incanum, the plant is a rich source of important cytotoxic glycoalkaloids, such as solamargine and solasonine. Because S. incanum is a potential source of compounds for steroid synthesis, it is worthwhile to study the content of these important glycoalkaloids during plant developmental stages. Therefore, an attempt has been made to quantitatively estimate solasonine and solamargine content using an optimized isolation process and a newly developed and validated HPTLC method using different parts of S. incanum plants at different developmental stages and comparing changes in the whole-plant GAs profile during the growth and development of S. incanum plants in Oman. Solamargine and solasonine produced well-separated compact bands with Rf values of 0.26 and 0.14, respectively, on silica gel HPTLC plates using chloroform:methanol:5% ammonia (7:3:0.5, v/v/v) after visualization using anisaldehyde sulphuric acid reagent. The chromatograms were scanned at 530 nm wavelength and the simultaneous method was linear (r2 ≥ 0.9962) in concentrations ranging from 50 to 2000 ng/spot for both of the drugs. The validated method was applied to analyse solamargine and solasonine in small, young, immature and mature leaves as well as stem and root parts up to the 40th week of plants’ growth, and showed a rich concentration of glycoalkaloids with large variations at different stages of plant development. Hence, this study highlights the importance of developmental stages of particular organs and the overall age of the plant when harvesting for these GAs from S. incanum plants.

Quantitative analysis of benzyl isothiocyanate in Salvadora persica extract and dental care herbal formulations using reversed phase C18 high-performance liquid chromatography method

Context: Benzyl isothiocyanate is the active antimicrobial agent in Salvadora persica (siwak) widely used in Islamic countries for oral hygiene. Aims: Quantification of benzyl isothiocyanate in the ethanol extract of S. persica and some dental care herbal formulations labeled to contain siwak. Settings and Design: Simple and sensitive high-performance liquid chromatography method was designed. Subjects and Methods: Separation was achieved on reverse phase C18 (250 mm × 4.6 mm, 5 μ ) column with a mobile phase comprising acetonitrile and water (1:1). The detection was carried out at 190 nm using ultra violet-visible detector. The flow rate was kept at 1 mL/min. Results: A sharp and well-defined peak was obtained at the retention time of 9.322 ± 0.3 min. Linear regression analysis data for the calibration plot showed a good linear relationship between response and concentration in the range of 0.5–500 μg/mL with a regression coefficient (r2) of 0.9977. The method was validated for accuracy, precision, robustness, and sensitivity. All the parameters examined met the current recommendations for the International Conference on Harmonization guidelines for method validation. Conclusions: The method was applied for the quantification of benzyl isothiocyanate in siwak extract, dental care powder, mouth wash, and toothpaste claimed to contain siwak. The developed method was found specific, simple, selective, and reliable for routine use in quality control analysis of different commercially available herbal care products. Abbreviations used: RP18: Reversed phase C18, HPLC: High performance liquid chromatography, UV: Ultra violet, r2: regression coefficient, ICH: international conference on harmonization, TLC: Thin layer chromatography, CHCl3: Chloroform, v/v: volume/volume, RSD: Relative standard deviation, LOD: Limit of detection, LOQ: Limit of quantification.

Quantification of glycyrrhizin biomarker in Glycyrrhiza glabra rhizome and baby herbal formulations by validated RP-HPTLC methods

Background: A simple and sensitive thin-layer chromatographic method has been established for quantification of glycyrrhizin in Glycyrrhiza glabra rhizome and baby herbal formulations by validated Reverse Phase HPTLC method. Materials and Methods: RP-HPTLC Method was carried out using glass coated with RP-18 silica gel 60 F254S HPTLC plates using methanol-water (7: 3 v/v) as mobile phase. Results: The developed plate was scanned and quantified densitometrically at 256 nm. Glycyrrhizin peaks from Glycyrrhiza glabra rhizome and baby herbal formulations were identified by comparing their single spot at Rf = 0.63 ± 0.01. Linear regression analysis revealed a good linear relationship between peak area and amount of glycyrrhizin in the range of 2000-7000 ng/band. Conclusion: The method was validated, in accordance with ICH guidelines for precision, accuracy, and robustness. The proposed method will be useful to enumerate the therapeutic dose of glycyrrhizin in herbal formulations as well as in bulk drug.

Reversed-phase high-performance thin-layer chromatographic method for the simultaneous determination of glycyrrhizin and glabridin biomarkers in Glycyrrhiza glabra roots, rhizomes and herbal formulations

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 RF = 0.63 ± 0.02 and RF = 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.

Investigation of antioxidant compounds in commercial pomegranate molasses products using matrix-solid phase dispersion extraction coupled with HPLC

Pomegranate is a well known fruit for its unique flavor, taste and health benefits. The medicinal properties of this fruits directly associated with the phenolic content present, with great anti-oxidant potential. The research is intended to develop matrix solid phase dispersion method (MSPD) and HPLC quantification of four major anti-oxidant marker constituents (vitamin C, gallic acid, rutin & ellagic acid) in pomegranate molasses samples. The effects of several important experimental parameters like type of dispersant, sample-dispersant ratio, solvents and its volume, time of extraction were investigated. A C18 column with the specification (5 µm, 250 × 4.0 mm) was used for the separation. A gradient flow of mobile phase was selected after many trials containing 0.1%, v/v solution of orthophosphoric acid and acetonitrile. The flow rate was 1.0 mL/min; and the chromatograms were recorded at 254 nm. The validation parameters, like linearity (r2 = 0.9985, 0.9965, 0.9925 & 0.9986), accuracy (100.3, 99.5, 100.9 & 101.9%), intra-day precision (%RSD = 1.09, 1.02, 1.26 & 0.97), inter-day precision (%RSD = 1.32, 0.83, 1.07, & 1.15) LOD (0.07, 4.50, 0.45 & 0.40 µg/mL), LOQ (0.095, 9.50, 0.85 & 9.5 µg/mL) and robustness (% RSD = 0.92, 0.76, 0.81 & 0.83) respectively for vitamin C, gallic acid, rutin & ellagic acid, were found satisfactory as per ICH guidelines.

Matrix solid-phase dispersion method coupled with high-performance liquid chromatography for the simultaneous quantification of selected compounds from freeze dry pomegranate juice

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 (vit...

Simultaneous quantification of piperlongumine and piperine in traditional polyherbal formulation using validated HPLC method

The alternative system of medicines like Unani and Ayurveda is preferred worldwide nowadays due to its therapeutic efficacy, lower side effects, holistic approach, psychological dimensions, and qualitative action of weather and seasonal requirement. A simple procedure is described for the simultaneous extraction and estimation of piperlongumine and piperine in a well-known Unani polyherbal formulation using reversed-phase high-performance liquid chromatography (HPLC). The chromatography was carried out on reversed-phase C18 (250 × 4.6 mm) column with a mobile phase containing acetonitrile—water (50:50 v/v). Detection was accomplished with ultraviolet (UV) detection at λ = 325 nm. The flow rate was kept as 1.0 mL−1. The proposed method was validated according to International Conference on Harmonization (ICH) guidelines for accuracy (94.4–105.0%), precision (0.37–2.17% RSD), and robustness (0.14–2.11% RSD). The limit of detection (LOD) values were found as 30 and 10 ng mL−1, while limit of quantification (...