Ahmed I. Foudah

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.

HiGh-performance thin-layer chromatography method for the simultaneous determination of vulgarin and epivulgarin biomarkers in Artemisia judaica L.

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 RF = 0.30 ± 0.02 and RF = 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.

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.

GC quantitative analysis of benzyl isothiocyanate in Salvadora persica roots extract and dental care herbal products

An accurate, sensitive, precise and simple method was developed utilizing Gas Chromatography for the quantitative analysis of benzyl isothicyanate in Siwak extract and dental care herbal products claimed to contain Siwak. Rtx (30.0 m × 0.25 mm ID, 25 µm thickness) column was used and helium as carrier gas at a flow rate of 0.74 mL/min. The retention time of standard benzyl isothicyanate was 13.470 min under the described conditions. Linear regression data analysis indicated a good linear relationship between peak height measurement and concentration of benzyl isothiocyanate in the range of 10–50 µg/ml (R2 = 0.9971). The regression equation was y = 11,471x. The developed GC method was subjected to validation requirements set by the ICH for precision, accuracy, and robustness. The entitled GC analyses expected to be valuable for the determination of benzyl isothiocyanate in Siwak extracts and other formulations containing Siwak extract. The amount of benzyl isothiocyanate reflects the efficacy of the products.

Wound Healing Study of Eucalyptus Essential Oil Containing Nanoemulsion in Rat Model

The objective of this investigation was to develop nanoemulsion formulations of Eucalyptus essential oil (EEO) and to evaluate its wound healing effects in comparison with standard gentamycin in rat model. Various nanoemulsionns of EEO were prepared using aqueous phase titration method and the zones of nanoemulsion were identified by the construction of phase diagrams. EEO nanoemulsions were investigated in terms of physical stability, self-nanoemulsification efficiency and physicochemical characterization. Optimized nanoemulsion of EEO was selected for wound healing study, collagen estimation and histopathological evaluation in rats in comparison with pure EEO and standard gentamycin. Optimized nanoemulsion presented significant would healing activity in rats as compared with pure EEO upon oral administration. The wound healing activity of optimized nanoemulsion was comparable with standard gentamycin. Optimized EEO nanoemulsion also presented significant enhancement in collagen content as compared with pure EEO and negative control. However, the collagen contents of optimized nanoemulsion treated animals were comparable with standard gentamycin-treated animals. Histopathological studies of optimized nanoemulsion treated rats showed no signs of inflammatory cells which suggested the safety and non-toxicity of EEO nanoemulsion. This study suggested the potential of nanoemulsion in enhancing the wound healing activity of EEO upon oral administration.

Protective Effect of Arnebia hispidissima Against Carbon Tetrachloride-induced Heart and Kidney Injury in Rats

Background and Objective: Arnebia hispidissima (F. Boraginaceae) has been found to have cardiac and febrifuge properties. It has long been used in traditional system of medicine for the treatment of heart ailments, headache and fever. This study aimed to investigate the protective effect of Arnebia hispidissima extract (AHE) on carbon tetrachloride (CCl4)-induced cardiotoxicity and nephrotoxicity. Materials and Methods: Adult rats were divided into five groups and medicated as follows: (1) The control group received the vehicles (olive oil; 1 mL kgG1, i.p.+3% Tween 80; 1 mL kgG1, p.o.). (2) CCl4 group of animals was administered with 20% CCl4 in olive oil (1 mL kgG1, p.o.). (3) Silymarin group was co-administered with CCl4 plus silymarin (50 mg kgG1, p.o.). (4) CCl4 plus AHE (200 mg kgG1, p.o.). (5) CCl4 plus AHE (400 mg kgG1, p.o). Rats received vehicle, CCl4, silymarin or AHE twice a week for 8 weeks. Serum AST (aspartate transaminase), LDH (lactate dehydrogenase), CK (creatine kinase), CK-MB (creatine kinase-MB isoenzyme) and cTnI (cardiac troponin) were measured to assess the heart damage markers. Serum levels of creatinine, urea, uric acid and BUN (blood urea nitrogen) were measured as markers of the renal function. Markers of oxidative stress in the cardiac and renal tissues were estimated by determining the levels of SOD (superoxide dismutase), GPx (glutathione peroxidase), CAT (catalase), GSH (reduced glutathione) and MDA (malondialdehyde). Heart and kidney tissues were investigated for histopathological changes. Results: Administration of CCl4 significantly increased the levels of cardiac and renal damage markers. Co-administration of CCl4 plus AHE significantly relieved the adverse effect of CCl4 in rat and reduced the increased serum levels of cardiac and renal damage markers. AHE compensated the deficits in the antioxidant defense mechanisms (SOD, GPx and CAT) and suppressed LPO (lipid peroxidation) in rat heart and kidney resulting from CCl4 administration. Moreover, histopathological changes induced with CCl4 in heart and kidney tissues of rat were also reduced with the co-administration of AHE. Conclusion: In this study, we have found that oral administration of AHE prevented CCl4-induced cardio- and nephrotoxicity by accelerating heart and kidney antioxidant defense mechanisms and down regulating the LPO near to the normal levels.

Standardization and Antioxidant Studies of Arnebia hispidissima

Background and Objective: Arnebia hispidissima (A. hispidissima) a member of the family Boraginaceae, is dye yielding and medicinally important plant. It is widely used in the cosmetic industries. This study has been made for the preliminary standardization of A. hispidissima plant. Methodology: The standardization evaluation comprises of powder microscopy and fluorescence analysis and TLC profiling. In addition, preliminary phytochemical screening, determination of total phenol and in vitro free radical scavenging activity (DPPH radicals scavenging assay and reducing power activity) were preformed utilizing the methanol extract. Results: A microscopic study of powder of whole plant of A. hispidissima showed different types of trichomes, vessel and fibres. Fluorescence analysis showed different colours under visible light, low UV and high UV. TLC of the hexane extract developed 8, 6, 9 and 10 spots with visible light, low UV, high UV and ninhydrin-H2SO4 spray, respectively. The phytochemical analysis of the methanol extract gave a positive indication for the presence of active compounds including alkaloids, carbohydrates, glycosides, steroids, triterpenoids saponins, phenols, tannins, flavonoids and proteins. The quantitative analysis showed the presence of a significant quantity of total phenol and the in vitro antioxidant activity clearly showed the terrific antioxidant property. Conclusion: The data generated from the present study can be utilized for the identification and quality control of A. hispidissima plant.