Promazine Detection in Human Hair: Presentation of an LC-MS-MS Method and Pattern of Drug Discontinuation.

Promazine is one of the oldest phenothiazine derivatives that have been proposed for the treatment of various psychiatric disorders. The drug is available as tablets, as syrups and in injectable forms. Despite its prescription to millions of subjects, its detection in human hair has seldom been reported. The aim of the present work is to develop a specific method to identify promazine in human hair by liquid chromatography-tandem mass spectrometry and to apply it to a patient who was self-medicating. The method involves overnight incubation of 20 mg of cut hair in 1 mL of pH 9.5 borate buffer in the presence of amitriptyline-d3 at 40°C. The chromatographic separation was performed using a reverse phase column HSS C18 with a gradient elution for 15 min. Linearity was verified from 0.5 to 500 pg/mg (r2 = 0.9996), after spiking blank hair with the corresponding amounts of promazine. The limit of detection was estimated at 0.1 pg/mg. The precision was lower than 20%. Promazine was detected in the hair of a psychotic subject at 228-270 pg/mg in a 3 × 1 cm segment. Given this was a patient who was self-medicating, her physician requested an immediate drug discontinuation. In a fresh hair specimen collected 3 months later, the proximal segment (0-1 cm) tested positive at 0.9 pg/mg, clearly indicating that the time to obtain a negative result after promazine discontinuation is about 3-4 months.

Use of potentiometric sensors to study (bio)molecular interactions.

Potentiometric sensors were used to study molecular interactions in liquid environments with sensorgram methodology. This is demonstrated with a lipophilic rubber-based and a collagen-based hydrogel sensor coating. The investigated molecules were promazine and tartaric acid, respectively. The sensors were placed in a hydrodynamic wall-jet system for the recording of sensorgrams. Millivolt sensor responses were first converted to a signal, expressing the concentration of adsorbed organic ions. Using a linearization method, a pseudo-first order-kinetic model of adsorption was shown to fit the experimental results perfectly. K(assoc), k(on), and k(off) values were calculated. The technique can be used over 4 decades of concentration, and it is very sensitive to low-MW compounds as well as to multiply charged large biomolecules. This study is the first to demonstrate the application of potentiometric sensors as an alternative and complement to surface plasmon resonance methods.

Validation of derivative spectrophotometry method for determination of active ingredients from neuroleptics in pharmaceutical preparations.

First (DI) and second (D2) order derivative spectrophotometric method with an application of base line to peak technique was used for determination of active pharmaceutical ingredients (API) at two wavelengths: fluphenazine (D1 at lambda = 251 nm and lambda = 265 nm, D2 at lambda = 246 nm and lambda = 269 nm), pernazine (D1 at lambda = 246 nm and lambda = 258 nm, D2 at lambda = 254 nm and lambda = 262 nm), haloperidol (DI at = 235 nm and lambda = 253 nm, D2 at lambda = 230 nm and lambda = 246 nm), and promazine (D1 at lambda = 246 nm and lambda = 251 nm, D2 at lambda = 255 nm and lambda = 262 nm). Linear dependence of derivative values on analyte concentration is maintained in a range 3.12 microg x mL(-1) - 44.80 microg x mL(-1). Detection and determination limits are in the range 0.51 - 3.23 microg x mL(-1) and 1.27 microg x mL(-1) - 9.80 microg x mL(-1), respectively. Determination results of drug constituents are very accurate. Recovery percentage is in a range 95.50% - 103.60%.

Simultaneous chemiluminescence determination of promazine and fluphenazine using support vector regression.

In this work, chemiluminescence (CL) behaviors of two selected phenothiazines, namely promazine and fluphenazine hydrochloride, were investigated for their simultaneous determination using oxidation of Ru(bipy)(3)(2+) by Ce(4+) ions in acidic media. This method is based on the kinetic distinction of the CL reactions of fluphenazine and promazine with Ru(bipy)(3)(2+) and Ce(4+) system in a sulfuric acid medium. Least square support vector regression models were constructed for relating concentrations of both compounds to their CL profiles. The parameters of the model consisting of sigma(2) and gamma were optimized using all possible combinations of sigma(2) and gamma to select the model with the minimum root mean square cross validation. Under optimized conditions, the univariate calibration curve was linear over the concentration ranges of 0.4-30.0 microg mL(-1) and 0.07-5.0 microg mL(-1) with detection limits of 0.1 microg mL(-1) and 0.04 microg mL(-1) for promazine and fluphenazine, respectively. The influence of potential interfering substances on the determination of promazine and fluphenazine were studied. The proposed method was used for simultaneous determination of both compounds in synthetic mixtures and in spiked human plasma.

Densitometric high performance thin-layer chromatography identification and quantitative analysis of psychotropic drugs.

A thin-layer chromatography (TLC)-densitometry method has been developed to identify and quantify haloperidol, amitriptyline, sulpiride, promazine, fluphenazine, doxepin, diazepam, trifluoperazine, clonazepam, and chlorpromazine in selected psychotropic drugs. Separation was performed on precoated silica gel 60 F254 TLC plates. Chromatograms were developed in various mobile phases, and 8 of 30 tested phases were selected based on spot location and developing time. The identification and quantification were carried out based on ultraviolet densitometric measurements at chosen wavelengths. In addition to retention coefficients, the absorption spectra recorded directly from chromatograms were also used in qualitative analysis. Under established experimental conditions, high sensitivity of the method was achieved. The limit of detection ranged from 0.009 to 0.260 microg, depending on the wavelength selected for measuring. A satisfactory recovery, ranging from 92.99 to 104.70%, was achieved for individual constituents.

Analytical studies and application of reaction of promazine and thioridazine hydrochlorides with some oxidants.

The run and analytical application of oxidation reaction of promazine (PM) and thioridazine (TR) hydrochlorides with iron(III) and ferrocyanide(III) are described. The coloured products of oxidation absorb at 512 nm for promazine and 634 nm for thioridazine. This property is a basis of a rapid, accurate and sensitive spectrophotometric method for determination of these phenothiazines. The elaborated methods allow to determine 3-28 microgPM/ml in Fe(III)-PM and 2-11 microgPM/ml in [Fe(CN)6](3-)-PM systems, 2.5-22 microgTR/ml in the Fe(III)-TR and 4-16 microgTR/ml in [Fe(CN)6](3-)-TR systems.

Application of reinecke salt and alizarin S for the determination of promazine.

Reinecke salt and alizarin S have been tested as reagents for the determination of promazine. They react in neutral and acidic media with promazine forming a reddish and brown crystalline compounds. The compounds are sparingly soluble in water but fairly soluble in acetone and methanol. The quantitative extraction of compound of promazine with alizarin S has been used for the extractive spectrophotometric determination of promazine. Formation of promazine ion-association complex with Reinecke salt has been applied for indirect determination of low concentrations of promazine by AAS measurement of the chromium content of the reineckate counter - anion.

Examination of iron (III) and hexacyanoferrate (III) ions as reagents for the spectrophotometric determination of promazine and perazine.

Iron (III) chloride and potassium hexacyanoferrate (III) have been tested as reagents for the determination of promazine hydrochloride and perazine. The methods are based on the oxidation of phenothiazines by FeCl3 and K3[Fe(CN)6] in perchloric acid medium. The optimal conditions for the formation of oxidation products of promazine and perazine were examined. The absorption spectra in the UV-VIS region were recorded.

Simultaneous quantification of promazine hydrochloride and its sulfoxide in pharmaceutical preparations.

The use of derivative UV-spectrophotometry is proposed for the simultaneous quantification of promazine hydrochloride in the presence of sulfoxide, and vice versa. For this purpose, mathematical parameters were established for generating derivative spectra of analytes. The determination of promazine was made using the first-order derivative (deltalambda = 10 nm, second polynomial degree) at 268 nm. The quantification of sulfoxide was achieved by applying third-derivative spectra (deltalambda = 14 nm, sixth polynomial degree) based on measurements of the amplitude at 342 - 344 nm. An elaborated method was successfully used to determine analytes in commercial promazine pharmaceuticals. The obtained results agreed well with those obtained by the HPLC method.

Determination of propionylpromazine hydrochloride in formulation matrixes using reversed-phase ion-pair small bore liquid chromatography.

Propionylpromazine hydrochloride (PPZHCl) has been investigated for use with leghold traps to reduce the amount of self-inflicted trauma experienced by animals restrained by these traps. Three types of PPZHCl formulations made with Karo dark syrup, K-Y Jelly, and Vaseline were used in 2 types of tranquilizer trap devices (TTDs). A reversed-phase ion-pair liquid chromatography (LC) method using a small bore C18 column was used to: (1) determine the purity of the PPZHCl material used in these formulations, and (2) to determine the resulting PPZHCl content of each formulation. Analyte quantitation was done using UV absorption at 280 nm. Regression analysis of calibration standard solutions indicated a linear and directly proportional relationship between analyte response and PPZHCl concentration over the range evaluated. Recovery data from: (1) Vaseline formulations containing 38.8, 16.2, and 8.78% PPZHCl were 104, 92.9, and 90.2%, respectively, (2) Karo dark syrup formulations containing 26.5, 18.1, and 10.3% PPZHCl were 97.7, 99.3, and 106%, respectively, and (3) K-Y Jelly formulations containing 33.0, 23.5, and 13.4% PPZHCl were 100, 99.4, and 88.7%, respectively. The relative standard deviation (RSD) values from triplicate analysis of these formulations ranged from 0.7 to 6.7%. The PPZHCl content from 9 manufactured TTDs, 3 for each formulation type, were analyzed in triplicate and produced RSD values ranging from 0.7-6.8%. These results indicate that the formulation extraction presented could be used to evaluate the PPZHCl content in TTDs prior to field use. The use of a small bore LC column reduced the amount of solvents consumed and hazardous waste generated, compared to sample analysis that uses a more conventional analytical LC column.

Multi-residue analysis of tranquillizers in meat: confirmatory assays using mass spectrometry.

A rapid and sensitive multi-residue method was developed to attempt to confirm the presence of the beta-blocker carazolol and the tranquillizers acepromazine, azaperone, chlorpromazine, propionylpromazine and xylazine in pig muscle tissues. The procedure involves determination by liquid chromatography coupled with tandem mass spectrometry. The liquid chromatographic separation was performed on a Symmetry C18 column with gradient elution. A mixture of aqueous buffer, containing 0.01% m/v trifluoroacetic acid (pH 3.5), and acetonitrile at a flow rate of 0.4 ml min-1 was used as the mobile phase. The abundant parent ions [M+ H+] produced by positive electrospray ionisation were selected for collisional dissociation with argon. Fragment ions were recorded with daughter ion scan and multiple reaction monitoring. The analytes were identified unambiguously by assessing retention times and diagnostic ions in meat samples spiked from 50 micrograms kg-1 [maximum residue limit (MRL) for azaperone and azaperol] to 5 micrograms kg-1 (MRL for carazolol).

Flow injection spectrophotometric determination of promazine.

A method for the spectrophotometric determination of promazine by flow injection analysis is reported. The procedure is based on the reaction of promazine with molybdophosphoric acid. Concentrations of promazine in the range 1-25 ppm have been determined with a relative standard deviation of 1.8% at 10 ppm (n = 20). The method was applied to the determination of promazine in injections and compared favourably with an independent reference method based on spectrophotometry.

Spectrophotometric determination of promazine with an oxidative column in FIA manifolds.

A simple flow-injection spectrophotometric method for the determination of promazine is described. The two proposed procedures are based on the oxidation of analyte with a manganese dioxide column. Concentrations of promazine in the ranges 2-20 and 1-6 are determined with a relative standard deviation of 1.0%. The injection rates are 62 and 80 samples h-1, respectively. The influence of foreign species and the determination of promazine in a pharmaceutical formulation are also reported.

[Study on the multiwavelength coefficient spectrophotometry and its application].

This paper reveals a general principle of multiwave length spectrophotometry and puts forward a multiwave length coefficient method to determine mixed components system. The determination of the coefficients and the connection between the coefficients with the method are discussed in the paper. Using several groups of coefficients, according to the method, a mixed components of chlorpromazine hydrochloride and promazine hydrochloride were assayed at the same time. The laboratory results of recoveries indicates that the precision and the accuracy are good, and, different coefficients do not make a notable impact on the method.

Thin-layer chromatographic screening method for the tranquillizers azaperone, propiopromazine and carazolol in pig tissues.

A procedure is described for the detection of azaperone, propiopromazine and carazolol in pig muscle, liver and kidney tissue. The method comprises extraction from an alkaline tissue homogenate with diethyl ether, followed by cleaning up and concentration of the extract on a silica gel solid-phase extraction column. Two-dimensional thin-layer chromatography on a silica plate was used for the detection of the tranquillizers. Detection levels were 25 micrograms kg-1 for propiopromazine, 50 micrograms kg-1 for azaperone (or its metabolite azaperol) and 125 micrograms kg-1 for carazolol. In pigs treated with the usual doses the presence of propiopromazine and azaperol could be established in kidney tissue 8 h after administration, whilst in injection sites all three tranquillizers could be detected.

Liquid chromatographic determination of identity, content, and content uniformity of desipramine, fluphenazine, and promazine.

A liquid chromatographic procedure has been developed for the assay, content uniformity, and identification of single active ingredient formulations of desipramine, fluphenazine, and promazine. The drugs are extracted from formulations with methanol or dilute hydrochloric acid and quantitated against an internal standard (norephedrine). The drugs are identified by comparison of retention times with those of the reference standards.