ABSTRACT
Simple and rapid determinations of some psychotropic drugs in some pharmaceutical wastewater and human plasma samples were successfully accomplished via the tandem dispersive liquid-liquid microextraction combined with high performance liquid chromatography-ultraviolet detection (TDLLME-HPLC-UV). TDLLME of the three psychotropic drugs clozapine, chlorpromazine, and thioridazine was easily performed through two consecutive dispersive liquid-liquid microextractions. By performing this convenient method, proper sample preconcentrations and clean-ups were achieved in just about 7min. In order to achieve the best extraction efficiency, the effective parameters involved were optimized. The optimal experimental conditions consisted of 100µL of CCl4 (as the extraction organic solvent), and the pH values of 13 and 2 for the donor and acceptor phases, respectively. Under these optimum experimental conditions, the proposed TDLLME-HPLC-UV technique provided a good linearity in the range of 5-3000ngmL-1 for the three psychotropic drugs with the correlation of determinations (R2s) higher than 0.996. The limits of quantification (LOQs) and limits of detection (LODs) obtained were 5.0ngmL-1 and 1.0-1.5ngmL-1, respectively. Also the proper enrichment factors (EFs) of 96, 99, and 88 for clozapine, chlorpromazine, and thioridazine, respectively, and good extraction repeatabilities (relative standard deviations below 9.3%, n=5) were obtained.
Subject(s)
Chromatography, High Pressure Liquid/methods , Liquid Phase Microextraction/methods , Psychotropic Drugs/analysis , Psychotropic Drugs/blood , Wastewater/analysis , Chlorpromazine/analysis , Chlorpromazine/blood , Chlorpromazine/isolation & purification , Chromatography, High Pressure Liquid/economics , Clozapine/analysis , Clozapine/blood , Clozapine/isolation & purification , Humans , Limit of Detection , Liquid Phase Microextraction/economics , Psychotropic Drugs/isolation & purification , Thioridazine/analysis , Thioridazine/blood , Thioridazine/isolation & purification , Time FactorsABSTRACT
Staphylococcus aureus biofilms are involved in a wide range of infections that are extremely difficult to treat with conventional antibiotic therapy. We aimed to identify potentiators of antibiotics against mature biofilms of S. aureus Mu50, a methicillin-resistant and vancomycin-intermediate-resistant strain. Over 700 off-patent drugs from a repurposing library were screened in combination with vancomycin in a microtitre plate (MTP)-based biofilm model system. This led to the identification of 25 hit compounds, including four phenothiazines among which thioridazine was the most potent. Their activity was evaluated in combination with other antibiotics both against planktonic and biofilm-grown S. aureus cells. The most promising combinations were subsequently tested in an in vitro chronic wound biofilm infection model. Although no synergistic activity was observed against planktonic cells, thioridazine potentiated the activity of tobramycin, linezolid and flucloxacillin against S. aureus biofilm cells. However, this effect was only observed in a general biofilm model and not in a chronic wound model of biofilm infection. Several drug compounds were identified that potentiated the activity of vancomycin against biofilms formed in a MTP-based biofilm model. A selected hit compound lost its potentiating activity in a model that mimics specific aspects of wound biofilms. This study provides a platform for discovering and evaluating potentiators against bacterial biofilms and highlights the necessity of using relevant in vitro biofilm model systems.
Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Drug Evaluation, Preclinical , Drug Repositioning , Drug Synergism , Methicillin-Resistant Staphylococcus aureus/drug effects , Thioridazine/pharmacology , Methicillin-Resistant Staphylococcus aureus/physiology , Models, Theoretical , Thioridazine/isolation & purification , Treatment Outcome , Wound Infection/drug therapyABSTRACT
The purpose of this study was to develop a method for the stereoselective analysis of thioridazine-2-sulfoxide (THD-2-SO) and thioridazine-5-sulfoxide (THD-5-SO) in culture medium and to study the biotransformation of rac-thioridazine (THD) by some endophytic fungi. The simultaneous resolution of THD-2-SO and THD-5-SO diastereoisomers was performed on a CHIRALPAK AS column using a mobile phase of hexane:ethanol:methanol (92:6:2, v/v/v)+0.5% diethylamine; UV detection was carried out at 262 nm. Diethyl ether was used as extractor solvent. The validated method was used to evaluate the biotransformation of THD by 12 endophytic fungi isolated from Tithonia diversifolia, Viguiera arenaria and Viguiera robusta. Among the 12 fungi evaluated, 4 of them deserve prominence for presenting an evidenced stereoselective biotransformation potential: Phomopsis sp. (TD2) presented greater mono-2-sulfoxidation to the form (S)-(SE) (12.1%); Glomerella cingulata (VA1) presented greater mono-5-sulfoxidation to the forms (S)-(SE)+(R)-(FE) (10.5%); Diaporthe phaseolorum (VR4) presented greater mono-2-sulfoxidation to the forms (S)-(SE) and (R)-(FE) (84.4% and 82.5%, respectively) and Aspergillus fumigatus (VR12) presented greater mono-2-sulfoxidation to the forms (S)-(SE) and (R)-(SE) (31.5% and 34.4%, respectively).
Subject(s)
Antipsychotic Agents/isolation & purification , Asteraceae/microbiology , Chromatography, High Pressure Liquid/methods , Fungi/metabolism , Thioridazine/analogs & derivatives , Thioridazine/isolation & purification , Amylose/analogs & derivatives , Amylose/chemistry , Antipsychotic Agents/chemistry , Antipsychotic Agents/metabolism , Aspergillus fumigatus/isolation & purification , Aspergillus fumigatus/metabolism , Biotransformation , Buffers , Carbamates/chemistry , Chromatography, High Pressure Liquid/standards , Culture Media/chemistry , Diethylamines/chemistry , Ethanol/chemistry , Ether/chemistry , Fungi/isolation & purification , Hexanes/chemistry , Methanol/chemistry , Phyllachorales/isolation & purification , Phyllachorales/metabolism , Reproducibility of Results , Solvents/chemistry , Spectrophotometry, Ultraviolet , Stereoisomerism , Thioridazine/chemistry , Thioridazine/metabolismABSTRACT
Enantioseparations of five chiral phenothiazines in CD-modified CZE using the single isomer sulfate-substituted beta-CD (heptakis(2,3-dihydroxy-6-O-sulfo)-beta-CD, SI-S-beta-CD) and dual CD systems consisting of SI-S-beta-CD and a neutral CD as chiral selectors in a citrate buffer at pH 3.0 were investigated. The results indicate that SI-S-beta-CD is an excellent chiral selector for enantioseparation of promethazine. The enantiomers of trimeprazine were well separated, while those of ethopropazine could also be baseline-resolved with SI-S-beta-CD. With dual CD systems, especially with hydroxypropyl-beta-CD (HP-beta-CD) as neutral CD, the enantioselectivity of thioridazine and ethopropazine was considerably enhanced. Effective enantioseparation of phenothiazines, except for methotrimeprazine, could thus be favorably and simultaneously achieved. Moreover, reversal of the enantiomer migration order of ethopropazine and thioridazine occurred by varying the concentration of gamma-CD in the presence of SI-S-beta-CD. These phenomena may be attributable to the opposite effects of sulfated beta-CD and gamma-CD on the mobility of the enantiomers of ethopropazine and of thioridazine. Comparative studies on the enantioseparations of phenothiazines with single CD and dual CD systems containing SI-S-beta-CD and randomly sulfate-substituted beta-CD (MI-S-beta-CD) were made.
Subject(s)
Electrophoresis, Capillary/instrumentation , Electrophoresis, Capillary/methods , Phenothiazines/isolation & purification , beta-Cyclodextrins/chemistry , gamma-Cyclodextrins/chemistry , Electrolytes , Hydrogen-Ion Concentration , Indicators and Reagents , Molecular Structure , Phenothiazines/chemistry , Promethazine/chemistry , Promethazine/isolation & purification , Stereoisomerism , Sulfates/chemistry , Thioridazine/chemistry , Thioridazine/isolation & purification , Trimeprazine/chemistry , Trimeprazine/isolation & purificationABSTRACT
Resolution of racemic thioridazine obtained from Thioril tablets (Cipla Ltd., Goa, India) into its enantiomers has been achieved by HPLC using a beta-cyclodextrin (CD)-bonded stationary phase. Thioridazine was isolated from commercial formulations and was purified using preparative TLC. The purity was ascertained by RP-HPLC. For the resolution of rac-thioridazine using cyclodextrin based CSP and mobile phase of 0.05 M phosphate buffer (pH 6.5)-acetonitrile (50:50) was found to be successful. The optimum conditions of resolution were established by systematically studying the effect of organic modifier, concentration of buffer, pH and flow rate of mobile phase. The detection limit was found to be 10 microg (5 microg of each enantiomer). The enantiomeric purity of each of the resolved isomers was verified by optical rotation.
Subject(s)
Chromatography, High Pressure Liquid/methods , Cyclodextrins/chemistry , Dosage Forms , Pharmaceutical Preparations/chemistry , Thioridazine/isolation & purification , Buffers , Chromatography, Thin Layer , Hydrogen-Ion Concentration , Sensitivity and Specificity , Stereoisomerism , Thioridazine/chemistryABSTRACT
Promethazine and thioridazine were separated and detected by capillary electrophoresis with end-column amperometric detection. The influence of pH value on oxidation potential, the peak current and the resolution were studied and the following conditions was selected: 0.03 M Na2HPO4 and 0.015 M citric acid at pH 3.0, detection potential at 1.10 V. The detection limits of these two substances were in the range of 10(-8) mol/l. The linear range spanned two to three orders of magnitude. This method was applied to the detection of promethazine and thioridazine spiked in urine.
Subject(s)
Antipsychotic Agents/isolation & purification , Histamine H1 Antagonists/isolation & purification , Promethazine/isolation & purification , Thioridazine/isolation & purification , Antipsychotic Agents/urine , Electrochemistry , Electrodes , Electrophoresis, Capillary , Histamine H1 Antagonists/urine , Humans , Hydrogen-Ion Concentration , Oxidation-Reduction , Promethazine/urine , Reproducibility of Results , Thioridazine/urineABSTRACT
In this paper, we describe the direct, preparative enantioselective chromatography of racemic (rac)-propranolol hydrochloride (HCI) and rac-thioridazine.HCl using Chiralpak AD chiral stationary phase and mobile phase systems containing carbon dioxide and methanol without the use of basic or acidic additives. Isolated fractions of propranolol.HCl were positively identified by mass spectrometry, Beilstein flame test, melting point, and chemical analysis to be HCI enantiomers of propranolol-HCl salts exhibited characteristic mass spectra peaks at 36 and 38 mass-to-charge ratio in the expected 3:1 isotopic ratio for the solute that were absent in the mass spectra for the free-base forms. To our knowledge, the direct, preparative enantioselective isolation of HCI enantiomeric salts of rac-propranolol and of rac-thioridazine have not been previously demonstrated and published.
Subject(s)
Adrenergic beta-Antagonists/isolation & purification , Antipsychotic Agents/isolation & purification , Chromatography/methods , Propranolol/isolation & purification , Thioridazine/isolation & purification , Adrenergic beta-Antagonists/chemistry , Antipsychotic Agents/chemistry , Carbon Dioxide , Mass Spectrometry , Methanol , Propranolol/chemistry , Stereoisomerism , Thioridazine/chemistryABSTRACT
(+/-)-Thioridazine and (+/-)-desmethylthioridazine have been oxidized to produce a number of chiral sulphoxide and amine oxide compounds. Diastereoisomeric isomers were separated by thin-layer and high performance liquid chromatography. Thioridazine-5-sulphoxide, N-desmethylthioridazine-5-sulphoxide and thioridazine-N-oxide diastereoisomers were found to be thioridazine metabolites following dosing in rats or after in-vitro incubation with rat liver homogenate.
Subject(s)
Thioridazine/analogs & derivatives , Thioridazine/isolation & purification , Animals , Biotransformation , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , In Vitro Techniques , Liver/metabolism , Magnetic Resonance Spectroscopy , Male , Rats , Rats, Inbred Strains , Stereoisomerism , Thioridazine/metabolismABSTRACT
A selective oxidation of thioridazine to give exclusively its ring sulfoxides and a separation of the resulting products as diastereoisomeric pairs of enantiomers (DL, LD and DD, LL) are reported. These pairs were characterized by TLC, high-performance liquid chromatographic, IR, UV, 1H-NMR, 13C-NMR, GC-MS, and elemental analyses, and by reduction to thioridazine by lithium aluminum hydride. Structural data for the separated diastereoisomeric pairs or their nitric acid salts were obtained from NMR and IR studies. Gram quantities of each of the two diastereoisomeric pairs of enantiomers were isolated in better than 99% purity.