A sensitive RP-HPLC method is presented for the simultaneous quantification of Fluorometholone (FLM) and Tetrahydrozoline hydrochloride (THZ). The method has the advantages of being rapid, accurate, reproducible, ecologically acceptable and sensitive. The separation utilized C8 Xbridge® column and mobile phase mixture of Acetonitrile/phosphate buffer pH 3 ± 0.1 (70:30, v/v) with UV detection at 230 nm. Stepwise optimization and factors affecting separation are properly discussed. Different factors were optimized such as stationary phase, selection of organic solvent and its content, buffer pH and concentration, flow rate, elution type and detection wavelength. The studied drugs were efficiently separated in 3.4 min with high resolution. Also, two univariate spectrophotometric methods have been optimized for the quantification of the studied drugs. Method 1: dual wavelength for THZ and iso-absorptive point for FLM, Method 2: ratio difference (RD) for THZ and first derivative FLM utilizing methanol as a solvent. These methods are accurate, precise with minimal data manipulation. Greenness of the methods was estimated using eco-scale tool where the presented methods were found to be excellent green with eco-score of 83 for HPLC and 80 for spectrophotometry. The methods are validated in conformance with ICH guidelines, with acceptable accuracy, precision, and selectivity. The suggested methods can be employed for the economic analysis of THZ and FLM in their pure form and binary ophthalmic formulation, that can be employed by quality control laboratories.
Different solvent extracts of the aerial parts of Senna italica (Mill.) were investigated for their chemical constituents and biological activities. Moreover, bio-guided fractionation led to isolation and identification of six compounds, namely: physcion (1), emodin (2), 2-methoxy-emodin-6-O-β-d-glucopyranoside (3), 1-hydroxy-2-acetyl-3-methyl-6-hydroxy-8-methoxynaphthalene (tinnevellin) (4), quercetin 3-O-α-l-rhamnopyranosyl-(1→6)-β-d-glucopyranoside (rutin) (5), and 1,6,8-trihydroxy-3-methoxy-9,10-dioxo-9,10-dihydroanthracene (6). The chemical structures of these compounds were established via 1D and 2D 1H- and 13C-NMR spectroscopy. Ethyl acetate and n-butanol extracts as well as compound 3 were evaluated for their anticancer activity against tumor cell lines. The tested extracts showed a moderate to weak activity, while compound 3 showed a moderate activity against human liver cancer (Hep G2) and breast cancer (MCF-7) cell lines with IC50 values of 57.5 and 42.3 μg/mL, respectively. Both ethyl acetate and n-butanol extracts exhibited antimicrobial activities with different strengths, i.e., ethyl acetate exhibited antimicrobial activity against seven test microbes while n-butanol extract showed antimicrobial activity against all tested microbes. This is the first report for the isolation of compound 3 as a new compound from S. italica growing in Egypt.
Eight compounds were isolated and identified from the soil-inhabiting fungus Aspergillus fumigatus 3T-EGY, namely, stearic acid (1), α-linolenic acid (2), physcion (3), di-(2-ethylhexyl) phthalate (4), 2,4,5,17-tetramethoxy pradimicin lactone (5), 3,5-dihydroxy-7-O-α-rhamnopyranoyl-2H-chromen-2-one (6), juglanthraquinone A-5-O-d-rhodosamine-(4′→1″)-2-deoxy-d-glucose (4″→1″′)-cinerulose B (7), and micropeptin (8). Their structures were determined on the basis of one-dimensional (1D-) and two-dimensional nuclear magnetic resonance (2D-NMR) [1H-, 13C-NMR, 1H-1H COSY (COrrelated SpectroscopY), and 1H-13C HMBC (Heteronuclear Multiple Bond Correlation) spectroscopy]. Compound 7 showed moderate in vitro antimicrobial activity against three pathogenic strains with inhibition zones values were ranged from 9.0 to 10.66 mm compared to neomycin as a positive control with inhibition zones values were ranged from 14.0 to 19.0 mm.