Facile immobilization of cholesterol oxidase on Pt,Ru-C nanocomposite and ionic liquid-modified carbon paste electrode for an efficient amperometric free cholesterol biosensing.

In present work, the enzyme cholesterol oxidase (ChOx) was immobilized by Nafion® (Naf) on Pt,Ru-C nanocomposite and an ionic liquid (IL)-modified carbon paste electrode (CPE) in order to create cholesterol biosensor (Naf/ChOx/Pt,Ru-C/IL-CPE). The prepared working electrodes were characterized using scanning electron microscopy-energy-dispersive spectrometry, while their electrochemical performance was evaluated using electrochemical impedance spectroscopic, cyclic voltammetric, and amperometric techniques. Excellent synergism between IL 1-allyl-3-methylimidazolium dicyanamide ([AMIM][DCA]), Pt,Ru-C, and ChOx, as modifiers of CPE, offers the most pronounced analytical performance for improved cholesterol amperometric determination in phosphate buffer solution pH 7.50 at a working potential of 0.60 V. Under optimized experimental conditions, a linear relationship between oxidation current and cholesterol concentration was found for the range from 0.31 to 2.46 µM, with an estimated detection limit of 0.13 µM and relative standard deviation (RSD) below 5.5%. The optimized amperometric method in combination with the developed Naf/ChOx/Pt,Ru-C/IL-CPE biosensor showed good repeatability and high selectivity towards cholesterol biosensing. The proposed biosensor was successfully applied to determine free cholesterol in a human blood serum sample via its enzymatic reaction product hydrogen peroxide despite the presence of possible interferences. The percentage recovery ranged from 99.08 to 102.81%, while RSD was below 2.0% for the unspiked as well as the spiked human blood serum sample. The obtained results indicated excellent accuracy and precision of the method, concluding that the developed biosensor can be a promising alternative to existing commercial cholesterol tests used in medical practice.

Chemometrics of anisotropic lipophilicity of anticancer androstane derivatives determined by reversed-phase ultra high performance liquid chromatography with polar aprotic and protic modifiers.

The research of the use of steroid compounds in the treatment of various types of cancer has a history of more than 50 years. Numerous steroid derivatives have expressed significant anticancer activity, however the thorough analysis of their physicochemical and toxicological properties is required prior to their clinical application. The present study is focused on the characterization of physicochemical properties of a series of previously synthesized new androstane derivatives (16E-hydroxyimino derivatives, D-homo lactones and D-seco dinitriles) with anticancer activity applying reversed-phase ultra high performance liquid chromatography (RP-UHPLC) system under isocratic conditions and chemometric tools, including in silico determination of their absorption, distribution, metabolism, excretion and toxicity (ADMET) properties. The chromatographic analysis was carried out applying two two-component and one ternary mobile phases with aprotic (acetonitrile) and protic (water and methanol) solvents. The conducted quantitative structure-retention relationship modeling resulted in two univariate and nine multivariate linear mathematical models that successfully correlate the retention parameters (logk) with descriptors of molecular polarity/lipophilicity (tPSA, REF, Average LogP, ClogP, ALOGP, LogS-SILICOS-IT, AClogS), descriptors that mainly depend on intermolecular interactions (BP, CP, CT, DE, HL) and ADMET descriptors (SWISSLogKpSP, GPCR, HIA, EI). The quality of the models was proved by the internal and external validation, while the robustness of the models was confirmed by Y-randomization test. Considering the established meaningful relationships between physicochemical/ADMET properties and retention parameters, the determined logk parameters of the analyzed series of steroid derivatives can be considered to be a biopharmaceutical property from the perspective of lipophilicity.

Electroanalytical performance of a ß-cyclodextrin and ionic liquid modified carbon paste electrode for the determination of verapamil in urine and pharmaceutical formulation.

The analytical performance of sensitive and cost-effective electrochemical sensors based on ionic liquids (ILs) with the bis(trifluoromethylsulfonyl)imide anion, [NTf2]-, and the imidazolium cation with different alkyl chain lengths for electrochemical oxidation of verapamil (VER) was investigated. 1-Ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([EMIM][NTf2]), 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([BMIM][NTf2]) and 1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([HMIM][NTf2]) were studied as possible materials for modification of a carbon paste electrode (CPE) for trace-level determination of VER. The experimental parameters including selection of the working electrode, the pH of working media, and the amount of CPE modifiers were investigated. Among them, the [EMIM][NTf2]-CPE with 4.3 wt% of IL was selected as the most appropriate for the square wave voltammetric (SWV) determination of VER at pH 5.0. Cyclic voltammetric studies showed that the electrochemical oxidation of VER was adsorption controlled. Consequently, the square wave adsorptive stripping voltammetric (SW-AdSV) parameters were optimized with Eacc = -0.4 V and tacc = 180 s as the most suitable for accumulation of VER on the electrode surface. The electroanalytical performance of the [EMIM][NTf2]-CPE was further improved by its in situ electrochemical modification with ß-cyclodextrin (ß-CD) and the linear concentration range of VER was from 0.006 to 0.129 µg mL-1; the relative standard deviation did not exceed 0.7%, and the evaluated limit of detection in model solution was 0.002 µg mL-1. The ß-CD/[EMIM][NTf2]-CPE showed adequate selectivity towards VER in the presence of inorganic ions and interferents usually found in human urine. The proposed sensor was successfully applied for VER determination in a spiked human urine sample and pharmaceutical formulation with good repeatability and recovery.