Katarzyna Filip

Preparation of selectively protected protoescigenin derivatives for synthesis of escin analogs and neoglycoconjugates

Protoescigenin, the main aglycone of horse chestnut saponin mixture known as escin, was selected as substrate for exploratory chemistry towards selective protection, followed by propargyl ether formation and subsequent condensation with azido-monosaccharides, to obtain novel triazole linked conjugates of the triterpene.

Process-related impurities of eplerenone: determination and characterisation by HPLC methods and Raman spectroscopy

Graphical abstract Figure. No caption available. HighlightsDevelopment and validation of RP and HPLC methods for impurity profiling of eplerenone API and its starting material.Application of Raman spectroscopy as a fast method for the diagnostics and characterization of the isomers during the synthesis.The bands assignment supported by the quantum chemical calculations. ABSTRACT Two novel high‐performance liquid chromatography methods for the determination of process‐related impurities of eplerenone drug substance and the designated starting material were developed and validated. Process impurities, including stereoisomers of eplerenone and the intermediate, were controlled using a Kromasil C18 column (250 mm x 4.6 mm; particle size 5 &mgr;m), under gradient conditions. Simple mobile phases: water and acetonitrile, as well as a PDA detector set at 240 nm were used. In order to control the stereochemical purity of the starting material (SM) in the eplerenone synthesis the polysaccharide‐based Kromasil 5‐AmyCoat chiral stationary phase was applied. To confirm the identity of the process‐related impurities (nine compounds) Raman Spectroscopy (RS), as a fast and convenient method, was applied. Differences in the wavenumbers of C=C and C=O stretching vibrations were the most distinctive features for the identification by means of RS. The bands assignment was supported by quantum mechanical computations. For one pair of the epimers containing the hydroxyl group the O‐H…O bond geometry was correlated with the wavenumbers of stretching vibrations of this group. Wherever possible, experimental results were compared with literature data.

N‐terminal guanidinylation of the cyclic 1,4‐ureido‐deltorphin analogues: the synthesis, receptor binding studies, and resistance to proteolytic digestion

The synthesis of a series of N‐guanidinylated cyclic ureidopeptides, analogues of 1,4‐ureido‐deltorphin/dermorphine tetrapeptide is described. The δ‐ and μ‐opioid receptor affinity of new guanidinylated analogues and their non‐guanidinylated precursors was determined by the displacement radioligand binding experiments. Our results indicate that the guanidinylation of cyclic 1,4‐ureidodeltorphin peptide analogues does not exhibit a uniform influence on the opioid receptor binding properties, similarly as reported earlier for some linear peptides. All analogues were also tested for their in vitro resistance to proteolysis during incubation with large excess of chymotrypsin, pepsin, and papain by means of mass spectroscopy. Guanidinylated ureidopeptides 1G–4G showed mixed μ agonist/δ agonist properties and high enzymatic stability indicating their potential as therapeutic agents for treatment of pain. Copyright

TLC-densitometric analysis of α-escin in bulk drug substance and in pharmaceutical dosage forms

Abstract A quite simple and rapid TLC-densitometric method for the identification of α-escin (Aescin) in bulk drug substances was developed. In so doing, different chromatographic conditions, including various mobile and stationary phases, were tested. A TLC densitometric determination of the examined compound was performed without using visualizing reagent, yet with the use of appropriate dipping reagents, in order to obtain reliable UV-densitometric measurements of α-escin - a substance which has weak chromophore groups. Herein, the application of a mobile phase containing n-butanolacetic acid-water in volume composition 30:7:13, the use of silica gel 60F254 plates with concentrating zone, and subsequent application of 10% sulphuric acid in ethanol or 5% vanillin in methanol/sulphuric acid, respectively, provided the best results in a TLCdensitometric study of α-escin. The described method was successfully employed to identify α-escin in commercial samples that were in an oral dosage form (tablets) and also in the form of gel containing 20 mg of α-escin.

A Validated TLC-Densitometric Method for the Determination of Mesterolone in Bulk Material and in Tablets

Mesterolone is a synthetic androgenic steroid indicating a weak anabolic activity. A new, simple in use, and economical TLC-densitometric method in normal phase system (NP-TLC) has been developed and validated for the identification and quantitative determination of mesterolone in bulk drug and in tablet formulation. NP-TLC analysis was performed on aluminium plates precoated with silica gel 60F254 as the stationary phase using chloroform-acetone (40 : 10, v/v) as mobile phase. Densitometric analysis was carried out at λ = 745 nm after staining with phosphomolybdic acid. These conditions were found to give visible (dark blue) spot and sharp peak, respectively, for mesterolone at R F  0.75 ± 0.02 and enabled satisfactory separation of mesterolone from its related substance (potential impurity). The proposed NP-TLC-densitometric method was validated for specificity, linearity, precision, accuracy, robustness, and sensitivity according to ICH guideline and other validation requirements. The limit of detection (LOD) and limit of quantification (LOQ) were 61.0 ng·spot−1 and 184.0 ng·spot−1, respectively. The percent content of mesterolone in marketed tablet formulation was found to be 99.40% of label claim. The developed TLC-densitometric method can be successfully used in quality control of mesterolone in bulk material and also tablet formulation.