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Ire1 is an endoplasmic reticulum (ER) transmembrane RNase that cleaves substrate mRNAs to help cells adapt to ER stress. Because there are cell types with physiological ER stress, loss of Ire1 results in metabolic and developmental defects in diverse organisms. In Drosophila, Ire1 mutants show developmental defects at early larval stages and in pupal eye photoreceptor differentiation. These Drosophila studies relied on a single Ire1 loss of function allele with a Piggybac insertion in the coding sequence. Here, we report that an Ire1 allele with a specific impairment in the RNase domain, H890A, unmasks previously unrecognized Ire1 phenotypes in Drosophila eye pigmentation. Specifically, we found that the adult eye pigmentation is altered, and the pigment granules are compromised in Ire1 H890A homozygous mosaic eyes. Furthermore, the Ire1 H890A mutant eyes had dramatically reduced Rhodopsin-1 protein levels. Drosophila eye pigment granules are most notably associated with late endosome/lysosomal defects. Our results indicate that the loss of Ire1, which would impair ER homeostasis, also results in altered adult eye pigmentation., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

In this study, a molecularly imprinted polymer based chemiluminescence array capable of simultaneous determining phenothiazines and benzodiazepines was first reported. Two polymers were coated in different wells of the conventional 96-well microtiter plate as the recognition reagents, and the added analytes competed with a horseradish peroxidase-labeled bi-hapten conjugate to bind the recognition reagents. The light signal was induced by using a highly effective luminol-H 2 O 2 -IMP system. The assay procedure consisted of only one sample-loading step prior to data acquisition. Then, the array was used to determine 4 phenothiazines and 5 benzodiazepines in pork simultaneously. The limits of detection for the 9 drugs were in a range of 0.001-0.01 ng/mL, and the recoveries from the fortified blank pork were in a range of 63.5%-94.1%. Furthermore, the array could be reused for 8 times. The detection results for some real pork samples were consistent with an ultra performance liquid chromatography method., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Attempted murder by repeated poisoning is quite rare. The authors describe the case of a 62-year-old man who was admitted to an intensive care unit (ICU) for neurological disturbances complicated by inhalation pneumopathy. He presented a loss of consciousness while his wife was visiting him at the ICU (H0). Forty-eight hours later (H48), police officers apprehended the patient's wife pouring a liquid into his fruit salad at the hospital. Toxicological analyses of a blood sample and the infusion equipment (H0), as well as the fruit salad and its container (H48), confirmed the attempted poisoning with cyamemazine (H0) and hydrochloric acid (H48). In order to evaluate the anteriority of poisonings, hair analysis was requested and the medical records of the 6 previous months were also examined. Two 6-cm brown hair strands were sampled and the victim's medical record was seized in order to determine the treatments he had been given during the previous six months. Segmental hair testing on two 6-cm brown hair was conducted by GC-MS, LC-DAD and LC-MS/MS (0-2/2-4/4-6 cm; pg/mg). Haloperidol (9200/1391/227), amitriptyline (7450/1850/3260), venlafaxine (332/560/260), that had never been part of the victim's treatment were detected, as well as some benzodiazepines (alprazolam, bromazepam, nordazepam); cyamemazine was also detected in all the segments (9960/1610/2367) though only a single dose administration was reported in the medical records. The toxicological analyses performed at H0 and H48 confirmed the homicide attempts in the ICU. In addition, comparison of the results in hair analysis with the medical records confirmed repeated poisoning attempts over the previous six months, and thus explain the origin of the disorders presented by the victim. This case serves to remind us that repeated attempted murder can be difficult to diagnose and that hair analysis can be an effective way to detect such attempts., (Copyright © 2018. Published by Elsevier Ltd.)

Pharmaceuticals do not occur isolated in the environment but in multi-component mixtures and may exhibit antagonist, synergistic or additive behavior. Knowledge on this is still scarce. The situation is even more complicated if effluents or potable water is treated by oxidative processes or such transformations occur in the environment. Thus, determining the fate and effects of parent compounds, metabolites and transformation products (TPs) formed by transformation and degradation processes in the environment is needed. This study investigated the fate and preliminary ecotoxicity of the phenothiazine pharmaceuticals, Promazine (PRO), Promethazine (PRM), Chlorpromazine (CPR), and Thioridazine (THI) as single and as components of the resulting mixtures obtained from their treatment by Fenton process. The Fenton process was carried out at pH7 and by using 0.5-2mgL -1 of [Fe 2+ ] 0 and 1-12.5mgL -1 of [H 2 O 2 ] 0 at the fixed ratio [Fe 2+ ] 0 :[H 2 O 2 ] 0 of 1:10 (w:w). No complete mineralization was achieved. Constitutional isomers and some metabolite-like TPs formed were suggested based on their UHPLC-HRMS n data. A degradation pathway was proposed considering interconnected mechanisms such as sulfoxidation, hydroxylation, N-dealkylation, and dechlorination steps. Aerobic biodegradation tests (OECD 301 D and OECD 301 F) were applied to the parent compounds separately, to the mixture of parent compounds, and for the cocktail of TPs present after the treatment by Fenton process. The samples were not readily biodegradable. However, LC-MS analysis revealed that abiotic transformations, such hydrolysis, and autocatalytic transformations occurred. The initial ecotoxicity tested towards Vibrio fischeri as individual compounds featured a reduction in toxicity of PRM and CPR by the treatment process, whereas PRO showed an increase in acute luminescence inhibition and THI a stable luminescence inhibition. Concerning effects of the mixture components, reduction in toxicity by the Fenton process was predicted by concentration addition and independent action models., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A sensitive electrochemical sensor for determination of phenothiazine (PTZ) was introduced based on molecularly imprinted polymer (MIP) film. A computational study was performed to evaluate the template-monomer geometry and interaction energy in the prepolymerization mixture. The electrode was prepared during electropolymerization of pyrrole (Py) on a pencil graphite electrode (PGE) by cyclic voltammetry (CV) technique. The quantitative measurements were performed using differential pulse voltammetry (DPV) in Britton-Robinson (BR) buffer solutions using 60% (v/v) acetonitrile-water (ACN/H2O) binary solvent. The effect of important parameters like pH, monomer concentration, number of cycles, etc on the efficiency of MIP electrode was optimized and the calibration curve was plotted at optimal conditions. Two dynamic linear ranges of 1-300 µmol L(-1) and 0.5-10 mmol L(-1) were observed. The detection limit (based on S/N=3) of PTZ was obtained 3×10(-7) mol L(-1). The MIP/PGE has been successfully applied as a selective sensor for fast and accurate determination of PTZ in some model and real biological samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

In this study, a novel generic hapten of phenothiazine drugs was synthesized by derivatization of 2-chlorophenothiazine with sodium bromoacetate. Then the hapten was coupled to bovine serum albumin for production of the monoclonal antibody. Results showed that the obtained monoclonal antibody recognized five phenothiazine drugs simultaneously: chlorpromazine, promethazine, acepromazine, perphenazine, and fluphenazine. After evaluation of different coating antigens, a heterologous competitive indirect enzyme-linked immunosorbent assay was developed to determine the residues of the five phenothiazine drugs in swine tissues (muscle, liver, and kidney). The cross-reactivities to the five analytes were in the range of 71 to 98%, and the limits of detection were in the range of 0.2 to 0.4 ng/ml, depending on the drug. Their recoveries from the fortified blank samples were in the range of 73.8 to 96.2%, with coefficients of variation in the range of 4.1 to 14.3%. This is the first study reporting a broad specific immunoassay for multi-determination of the residues of five phenothiazine drugs in animal-derived foods., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Interactions of biomembrane-active compounds with phospholipid monolayers on microfabricated Pt/Hg electrodes in an on-line high throughput flow system are demonstrated by recording capacitance current peak changes as rapid cyclic voltammograms (RCV). Detection limits of the compounds' effects on the layer have been estimated from the data. Compounds studied include steroids, polycyclic aromatic hydrocarbons, tricyclic antidepressants and tricyclic phenothiazines. The results show that the extent and type of interaction depends on the-(a) presence and number of aromatic rings and substituents, (b) presence and composition of side chains and, (c) molecular shape. Interaction is only indirectly related to compound hydrophobicity. For a selection of tricyclic antidepressants and tricyclic phenothiazines the detection limit in water is related to their therapeutic normal threshold. The sensing assay has been tested in the presence of humic acid as a potential interferent and in a tap water matrix. The system can be applied to the screening of putative hazardous substances and pharmaceuticals allowing for early detection thereof in the water supply. The measurements are made in real time which means that potentially toxic compounds are detected rapidly within <10 min per assay. This technology will contribute greatly to environment safety and health., (Copyright © 2014. Published by Elsevier B.V.)

A novel promising approach to the improvement of analytical properties of horseradish peroxidase based on its inclusion into self-assembled structures of chitosan is discussed. It is shown that the reasonable choice of a polyelectrolyte, a detailed investigation of its interaction with the enzyme and the conditions of the {peroxidase-polyelectrolyte} complex formation allow for stabilizing the biocatalyst in aqueous and aqueous-organic media without a substantial loss in its activity and developing corresponding analytical procedures and biosensors. The latter provides highly selective determination of a number of organic compounds and sensitive determination of heavy metal ions that becomes possible due to the specific interactions of the analytes with the polymer matrix. Besides, the application of the proposed analytical systems and biosensors provides the expansion of the range of the compounds, and poorly water soluble and slowly oxidized substrates of peroxidase as well, which could be determined and real samples which could be analyzed by enzymatic methods. Analytical performance of the developed spectrophotometric indicator procedures and biosensors based on the self-assembled complex {peroxidase-chitosan} is demonstrated in the determination of metal ions (Hg(II), Cd(II), and Pb(II)), phenothiazines (promazine, chloropromazine, and trifluoroperazine), phenolic compounds (phenol, hydroquinone, catechol, pyrogallol, quercetin, rutin, and esculetin), organic peroxides (tert-butyl peroxide, 2-butanone peroxide, and benzoyl peroxide) in various samples, including water-insoluble matrices., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A rapid, sensitive and fully automated chemiluminometric method is described for determination of five phenothiazine derivatives, namely, trifluoperazine, fluphenazine, perphenazine, thioridazine and chlorpromazine. The method is based on the chemiluminescence (CL) induced by the oxidation of drugs with Ce(IV) in nitric acid. A flow manifold based on the association of multi-commutation and multi-pumping flow methodologies is proposed. The active operated solenoid devices consisted of a micro-pump (propelling 50μL per stroke) and a six ports solenoid valve. Reconfiguration of the flow manifold was performed by using software settings, without physical alteration of the instrument manifold. It permits the design of flexible miniaturized networks for flow analysis based on a time-pulse-counting strategy. The proposed method allows the determination of the drugs at the ngmL(-1) level with a sample throughput of 38h(-1) (chlorpromazine) and 40h(-1) (trifluoperazine, fluphenazine, perphenazine, and thioridazine). The method was successfully applied to the determination of the phenothiazine derivatives in pharmaceuticals formulations., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The occurrence of residues of malachite green and its leuco-metabolite in tissues of farmed fish for human consumption have long been of concern and there is extensive literature on methods of analysis and surveillance for these compounds. Recently, concern has been expressed that the use of other related compounds in place of malachite green may go undetected. This paper describes a new method for extending the range of triarylmethane and related phenothiazine dyes that can be detected in fish. In this procedure 13 parent compounds are monitored, with any potential leuco-forms being oxidized back to the parent prior to determination. The method utilizes a buffer-acetonitrile extraction followed by liquid-liquid extraction. Oxidant is added and the extracts further purified by cation exchange chromatography. Final determination is carried out using LC-MS/MS. The method has been validated to the standards of Commission Decision 2002/657/EC.

A new analytical method, based on liquid chromatography with coulometric detection, has been developed and applied to the determination of selected phenothiazines (chlorpromazine, promazine, fluphenazine and levomepromazine) in human plasma. The drugs were separated on a Discovery pentafluorophenylpropyl column, using a mobile phase composed of acetonitrile (32%) and a pH 1.9 phosphate buffer (68%). Promethazine was used as the internal standard. Detection was carried out at an oxidation potential of +0.500 V. A novel clean-up procedure was developed by means of solid-phase extraction, using cyanopropyl cartridges, which gave good extraction yield for all the analytes, with absolute recovery values higher than 91.0%. The detector response was linear over a plasma concentration range of 0.5-250.0 ng mL(-1) for chlorpromazine, promazine and levomepromazine and of 0.2-4.0 ng mL(-1) for fluphenazine. Precision results, expressed by the intra-day and the inter-day relative standard deviation values, were good, being lower than 3.9%. Accuracy data were satisfactory as well. The method has been successfully applied to the analysis of drug plasma levels of psychiatric patients undergoing therapy with selected phenothiazines.

Chromatographic behavior of two main classes of neuroleptics, derivatives of phenothiazine and thioxanthene in RP systems modified by anionic additives: sodium perchlorate and sodium hexafluorophosphate possessing chaotropic properties, was examined. Influence of the method of pH lowering (by addition of acids: trifluoroacetic or perchloric or by adding the appropriate concentration of phosphate buffer) and the kind of organic modifier in the mobile phase (methanol, acetonitrile) were estimated. Stability of complexes created between protonated drugs and anions of added salts was evaluated by comparison of their desolvation parameters (K), limiting retention factors for unsolved molecules calculated on the basis of chromatographic data. Experimentally obtained parameters were used in QSAR studies. It appeared that chosen parameters reflect not only physico-chemical properties of analytes but also contain information about the strength of their antipsychotic activity. Multidimensional cluster analysis has been performed. On the basis of the results obtained, it could be concluded that chaotropic systems can generate useful parameters for further QSAR studies.

The amount of DNA was measured by using thioridazine, which would be attached to the DNA, as an electrochemical indicator. An indicator (thioridazine) solution, a test solution (DNA solution), and a poly-l-lysine solution were successively placed on a glassy carbon electrode, and the electrode was allowed to dry; DNA was immobilized on an electrode surface by the electrostatic binding between DNA and poly-l-lysine. The electrode was immersed into a buffer solution for 15 min, and then differential pulse voltammetry (DPV) was carried out: the oxidation current peak of thioridazine was observed, and its magnitude depended on the amount of DNA in the solution which was used for preparing the electrode. It could be estimated between 0.2 microg DNA (corresponds to 630 pmol nucleotides) to 20 microg DNA (63 nmol nucleotides) from the oxidation peak current of DPV.

A spectrophotometric method is described for the determination of four commonly used psychotropic phenothiazine drugs. The method is based on the measurement of the intensely coloured charge-transfer complex formed by the interaction of these drugs as n-electron donors with chloranilic acid as the pi-acceptor. The coloured species measured exhibits maximum absorption at 535 nm. The molar-combining ratio and optimum assay conditions are reported. Beer's law is obeyed over the range, 25-250 microg ml(-1) with apparent molar absorptivity in the range, 0.93 x 10(3)-1.45 x 10(3) l mol(-1) cm(-1). The limits of detection and quantification are reported. The proposed method was applied to the determination of these psychotropics in pharmaceutical formulations and the results demonstrated that the method is equally accurate, precise and reproducible as the official methods. The validity of method was established by recovery studies via standard-addition technique with satisfactory results.

Four simple, selective, accurate and reproducible procedures are described for the assay of methdilazine in bulk form and in formulations. One titrimetric and three spectrophotometric methods are based on the oxidation of the drug with potassium iodate, and determination of either excess iodate or iodine released in the reaction. In the titrimetric method (Method A) the drug is reacted with a known excess of iodate in sulphuric acid medium followed by the iodometric determination of residual oxidant. The residual oxidant is determined by reacting it with variamine blue and measuring the absorbance of the oxidised dye at 540 nm (Method B). The second spectrophotometric method (Method C) is based on the oxidation of the drug in sulphuric acid medium in the presence of chloride ions by a large excess of iodate and the iodate being reduced to iodine. The ICl(2)(-) generated in this reaction is used to iodinate 2',7'-dichlorofluorescein dye, and the red colour of the iodinated dye is measured at 525 nm. The other spectrophotometric method (Method D) also involves the oxidation of the drug in acid medium by a large excess of iodate with the liberation of iodine and its subsequent extraction with carbon tetrachloride followed by measuring the absorbance 520 nm. The methods were successfully applied to the determination of methdilazine in tablets and syrup and the results obtained in agreement with the label claim.

Two new simple, accurate and precise titrimetric micro-procedures are described for the analysis of phenothiazines in pure sample, tablets, injections and syrup using periodate as the oxidant. The first method is based on the oxidation of phenothiazines with periodate in acid medium and the iodate formed in the reaction is determined by reacting it with iodide and titrating the liberated iodine with thiosulfate after masking the excess of periodate with molybdate. In the second procedure, the unreacted (excess) periodate is determined iodometrically under basic conditions. The reaction conditions have been optimised and the stoichiometry of the reaction has been evaluated. A linear relationship exists between the amount of the drug and the titration end-point as shown by the values of correlation coefficient, r (0.9991-0.9999). The slope of the linear relationship has been calculated and found to be in the range, 0.1457-0.3120. The methods were applied to the analysis of dosage forms with results comparable to those given by the official methods. Both the methods are indirect visual titration methods, and are simpler than, and superior to, many existing methods for the assay of phenothiazines.

In this study, enantioseparations of five phenothiazines, including promethazine, ethopropazine, trimeprazine, methotrimeprazine, and thioridazine, in cyclodextrin (CD)-modified capillary zone electrophoresis were investigated using a phosphate buffer (40 mM) at pH 3.0. We focussed on the separation of phenothiazines with the use of CDs at low concentrations. Three different CDs, including beta-CD, hydroxypropyl-beta-CD (HP-beta-CD) and gamma-CD, were chosen as chiral selectors. The results indicate that effective enantioseparation of phenothiazines, except for methotrimeprazine, is simultaneously achievable with addition of gamma-CD at a concentration of 2.5-6.0 mM. The enantiomers of ethopropazine and trimeprazine are effectively separated with addition of HP-beta-CD at low concentrations, in the range 0.4-6.0 mM, whereas those of promethazine and trimeprazine are baseline resolved with beta-CD at much lower concentrations (0.02-3.0 mM) than with HP-beta-CD. The results also confirm that the separation window is greatly enlarged at low CD concentrations. Moreover, the drastic variations of the electrophoretic mobility of phenothiazines as a function of CD concentration reveal that phenothiazines interact very strongly with CDs in the order gamma-CD

A spectrophotometric procedure for the determination of phenothiazines in pure form and in a number of their pharmaceutical preparations has been developed that offers the advantages of simplicity, accuracy, precision and sensitivity over many other methods. The method is based on the oxidation of phenothiazines by a known excess amount of potassium dichromate followed by the estimation of unreacted amount of dichromate by reacting with excess of iron(II) and measuring the iron(III) formed by complexing with thiocyanate. The reacted oxidant corresponds to the drug content. Different variables affecting the reaction between drugs and dichromate were studied and optimized. At the maximum absorption of 480 nm, Beer's law is obeyed in the range 2.5-29.75 microg/ml. The molar absorptivity and Sandell sensitivity of the procedure were calculated in addition to detection limit. Statistical treatment of the experimental results indicates that the procedure is precise and accurate. Excipients used as additives in pharmaceutical formulations did not interfere in the proposed procedure. The reliability of the method was established by parallel determination against the official BP methods. The procedure described was successfully applied to the determination of the bulk drugs and in pharmaceutical formulations.

A first post-column chemical derivatization method for the liquid chromatographic determination of phenothiazines is presented. Peroxyacetic acid is introduced as a derivatizing agent for phenothiazines, yielding the colored radical cations or fluorescent sulfoxides, depending on reaction conditions. Both reaction products were successfully employed for the detection of the phenothiazines after their liquid chromatographic separation. The fluorescence spectroscopic detection of the sulfoxides proved to be the more robust and sensitive method. Limits of detection ranged from 4 nM for triflupromazine and trimeprazine to 300 nM for phenothiazine for the fluorescence spectroscopic detection of the sulfoxide and from 0.3 microM for phenothiazine and triflupromazine to 2 microM for trifluperazine for the UV-Vis spectroscopic detection of the radical cation. The calibration functions for the fluorimetric sulfoxide determination ranged from two to more than three decades, starting at the limit of quantification.

A new spectrophotometric method for the assay of phenothiazines in pure form as well as in pharmaceutical formulations with the chromium(VI)-metol-sulfanilic acid system has been developed. The method is based on the oxidation of the drugs by a known excess of chromium(VI) and subsequent determination of the unreacted oxidant by interacting with metol and sulfanilic acid. The reacted oxidant corresponds to the drug content. The coloured species exhibits maximum absorbance at 530 nm. Beer's law is obeyed over the concentration range 5-60 micrograms ml-1 and the relative standard deviation is found to be less than 2%. The apparent molar absorptivities are in the range 3.77 x 10(3)-3.98 x 10(3) l mol-1 cm-1, the detection limits being in the range 0.6133-1.1349 micrograms ml-1. The method was successfully applied to the determination of the studied drugs in their formulations and the mean percentage recoveries were found to be 97.32-102.80%.

A simple and rapid isocratic high-performance liquid chromatographic method for the analysis of methylene violet, a neutral phenothiazine dye, in the presence of DNA has been developed. These chromatographic conditions are also applicable to its N-demethylated, bromo and iodo analogs. The method utilizes an internal surface reversed-phase column and a mobile phase consisting of 20% acetonitrile in 50 mM phosphate buffer (pH 7.0) and detection at 280 nm. Under these conditions all five dyes are well resolved from one another and from the faster migrating DNA. The effects of organic modifiers, ionic strength and pH of the buffer on the capacity factors of the dyes have been investigated. The method has successfully been applied to detect the photoproducts of methylene violet and its bromo analog in the presence of DNA without removing the biopolymer from the reaction mixture.

A simple and sensitive high-performance liquid chromatographic (HPLC) method for the determination of phenothiazine (PHE) is described. PHE is converted to diphenylamine (DIP) by desulfurization with Raney nickel catalyst. DIP is highly sensitive to electrochemical detection. The calibration graph for PHE quantification after desulfurization was linear between 0.1 and 2.0 ng per injection. The detection limit (signal-to-noise ratio = 3) of PHE after desulfurization was 10 pg, which is twenty times higher than that of the parent compound PHE. The proposed desulfurization technique was applied to other PHE-related compounds. The structural confirmation of the desulfurized product of PHE was carried out by LC-MS using atmospheric pressure chemical ionization.

Fenoverine is a spasmolytic, non-anticholinergic phenothiazine derivative that inhibits calcium channel currents. We describe a high-performance liquid chromatographic method for the determination of fenoverine in striated muscle, smooth muscle, myocardium, and liver tissue. Reversed-phase liquid chromatography was performed on a 5 microns Nucleosil C18 column with acetonitrile-0.015 M phosphate buffer (28:72, v/v) as a mobile phase and detection with ultraviolet at 214 nm. The limit of quantitation of fenoverine in tissue samples was 25 ng injected. This method is well suited for the determination of fenoverine in various organs in animal experiments.

Fenoverine is a novel, potent, musculotropic, spasmolytic agent that affects primarily the gastrointestinal tract, bile duct, and female genital organs. A simple, specific, and accurate HPLC method was developed for the determination of fenoverine in capsules and plasma. This method has been successfully applied to stability studies of fenoverine capsules and to a pilot study in a normal, healthy volunteer following oral administration of fenoverine. For the determination of fenoverine in capsules, a Nucleosil 5-micron CN column, with acetonitrile:0.1 M ammonium acetate (60:40) as mobile phase and detection at 254 nm, was employed. The mean correlation coefficient of the calibration curve (n = 6) for the assay was 0.9999 over a concentration range of 24.6 to 147.6 micrograms/mL of fenoverine standard solutions. Fenoverine did not decompose significantly at 4, 45, 55, and 65 degrees C for 3 months. The mean correlation coefficients of within-day and between-day calibration curves were 0.9995 and 0.9999, respectively, over a range of 10 to 1000 ng/mL of fenoverine in plasma. The limit of detection was 10 ng in plasma.

A reversed-phase thin-layer chromatographic method has been developed for the determination of partition coefficients. A support phase has been chosen, following investigation of the lack of adsorptive properties, which has a minimal effect on the pH of the buffer system. A stationary phase has been chosen to give deltaRm values of the same magnitude as Hansch pi values for a series of phenothiazines. The method can be applied to molecules of a wide range of lipophilicity following preliminary investigations of suitable phase-volume ratios and of the pH and composition of the binary mobile phase, providing adsorption on the support phase is excluded.

Experimental dipole moments measured in anhydrous benzene and dioxane are compared to the calculated values of some 2-substituted phenothiazine derivatives. The dipole moments and the solvent effects support the existence of a conformational mixture of these compounds as solutes.

The response and elemental selectivity of the electrolytic conductivity detector (E1CD) to nitrogen-, chlorine-, and sulfur-containing compounds has been investigated as a function of chemical structure, furnace chemistry conditions (i.e., reaction gas, furnace temperature, catalyst, and post-furnace chemical abstractors), and solubilization-and-ionization processes (electrolytic chemistry and electrolytic conductance). The "profiling" of detector response for the gas-liquid chromatographic-E1CD analysis of selected compounds under defined furnace chemistry reaction conditions and electrolytic chemistry included the determination of detector signal to noise ratio, peak tailing, and elemental selectivity. Detector response and conductivity phenomena are discussed in terms of gas-phase furnace chemistry reactions, post-furnace reaction or abstraction processes, and solution-phase ionization and neutralization processes occurring in the conductivity cell.

The UV and visible spectra of radical cations of several phenothiazine derivatives were studied using different solvents. The establishment of a relationship between these bands and the R2 and R10 substituents was attempted. The influence of the disolvents on the bands also was studied. The characteristic charge transfer band was observed in the solid state using diffuse reflectance spectroscopy. The R2 substituent did not appear to influence the band, while the R10 substituent influenced the band considerably, probably due to steric effects.

A reversed-phase thin-layer chromatographic technique was used for the characterization of 26 phenothiazine drugs. With two chromatographic systems having the sam stationary phase and phase volume ratio, but mobile phases of different pH, all but two of the compounds could be identified. Rf values in the different systems were standardized by applying a reference compound to the plates next to each compound under investigation; the corrected Rf values were calculated from the differences in the Rm values of the compounds and the reference compound, and the theoretical Rm value of the reference. It was shown that Rf values for different chromatographic systems with the same stationary phase could be predicted with reasonable accuracy. The pH of the mobile phase, for which a maximum difference in Rf values was obtained for pairs of compounds, could also be calculated and corresponded well with the observed values.

The use of ternary solvent systems as mobile phases in combination with highly efficient chromatographic columns is of particular interest for the analysis of pharmaceutical and biological compounds. The solute-silanol group interactions decrease when the eluent is blended with water. Good selectivities are found with such systems.

A thin-layer chromatographic-second derivative spectrofluorimetric procedure has been developed for low levels of sulphoxide (down to 0.01%, w/w) in phenothiazine drug substances and formulations. The method has been applied to prochlorperazine maleate, chlorpromazine hydrochloride and promethazine hydrochloride. It has also been applied to pharmaceutical formulations of promethazine hydrochloride and comparison of the results with those obtained by a published difference spectrophotometric procedure for promethazine sulphoxide showed that there was good agreement. The method is simple, rapid, accurate and precise and seems to have general application to the determination of low levels of sulphoxide in phenothiazine drug substances or formulations.

Contrary to earlier literature reports, the impurities in toluidine blue O were shown by column chromatography, TLC, and mass spectrometry to be N-methyl homologs of 2-methylthionine rather than N-methyl homologs of thionine. Small amounts of 2-methyl-3-amino-7-methylaminophenothiazine and 2-methyl-3,7-diaminophenothiazine were identified in commercial samples of toluidine blue O. However, sample handling and a warm alkaline environment can cause rapid demethylation of the dye.

Operational difficulties have been encountered with many of the official phenothiazine assay procedures in this laboratory. Although some difficulties were corrected, the procedures still lack specificity. Accordingly, the general scope of differential UV and differential fluorescence methods was investigated. These specific methods are facile, reproducible, and applicable to both composite and single-tablet assays.

Commercially available phenothiazine derivatives were used for the study of cyclodextrin complex formation by cationic isotachophoresis with alpha-, beta- and gamma-cyclodextrin and methylated analogues of beta-cyclodextrin as leading electrolyte additives. The relationships between the type of solute substituent in the 10- and/or 2-position and the stability of the created cyclodextrin complex were studied and the results were utilized for the optimization of isotachophoretic conditions suitable for the resolution of the studied phenothiazine derivatives. Successful resolution of three racemic solutes was achieved.

A simple method is reported for the preparation of gram quantities of phenothiazine sulfoxides by aqueous nitrous acid oxidation of phenothiazines at room temperature. The chiral levomepromazine gave rise to diastereoisomeric products analogous to those reported for thioridazine sulfoxidation.

The usefulness of amperometric detection in pharmaceutical analyses was investigated for different groups of drugs. The UV response at 254 nm and that at the absorption maximum of the solute were compared with the electrochemical signal obtained. The minimum detectable concentration (nanograms on-column) of each substance is reported for the three different detection systems. This comparison was performed for 72 drugs (local anaesthetics, antipyretics, tricyclic antidepressants, sulphonamides, sex hormones, beta-adrenoceptor blocking agents, phenothiazines, alkaloids, diuretics and penicillins). The median limit of detection of the amperometric detector (see definition in the text) is 1.0 ng on-column and the median gain in sensitivity, compared with UV detection is 22.5.

A rapid, precise, and accurate high-performance liquid chromatographic procedure is presented for the simultaneous determination of perphenazine and amitriptyline hydrochloride in two-component tablet formulations. An aliquot of a methanolic extract of the tablet, containing trifluoperazine hydrochloride as an internal standard, is chromatographed on a nitrile bonded phase microparticulate column using a 0.005 M ammonium acetate-methanol (20:80) mobile phase. Quantitation is by peak area. The relative standard deviations for the procedure are 0.34 and 0.54% for the simultaneous determination of perphenazine and amitriptyline, respectively. Eight commercial tablet formulations were analyzed and found to contain 96.5-101.5 and 96.5-103.3% of the labeled amounts of perphenazine and amitriptyline hydrochloride, respectively.

The chemistry of the red color formed during perchloric acid titration of chlorpromazine hydrochloride in acetic acid in the presence of mercuric acetate is discussed. Addition of ascorbic acid prevents the color formation and allows titration using a crystal violet end-point. Ascorbic acid addition also sharpens the potentiometric end-point. Ascrobic acid and its oxidation product, dehydroascorbic acid, being neutral to perchloric acid, do not interfere with the titration.

The degradation kinetics of 10-acetylphenothiazine were studied after isolation and identification of its degradation products, phenothiazine, phenothiazine-5-oxide, 3H-phenothiazine-3-one, and 7-(10'-phenothiazinyl)-3H-phenothiazine-3-one. The acetyl group was not present in any degradation product. The degradation rate was independent of oxygen but highly pH dependent, indicating a specific hydrogen-ion-catalyzed hydrolysis of 10-acetylphenothiazine to give phenothiazine, which was oxidized. The experimental values for the phenothiazine concentration were in good agreement with values calculated on the basis of this assumption, both in the presence and absence of oxygen.

A procedure is described for TLC detection and differentiation of the butyrophenone-diphenylbutylpiperidine group and phenothiazine derivatives at the microgram level. A two-dimensional TLC method to separate butyrophenone and diphenylbutylpiperidine compounds is reported. A variety of possible detection reagents were examined. The solvent systems and spray reagents described should be useful for the identification of these drugs in various dosage forms.