Electrochemically controlled fiber-in-tube solid-phase microextraction method for the determination of trace amounts of antipsychotic drugs in biological samples.

In this study, a novel 'fiber-in-tube' configuration was applied to electrochemically controlled fiber-in-tube solid-phase microextraction of antipsychotic drugs (perphenazine and chlorpromazine) from biological samples. To prepare an electrochemically controlled fiber-in-tube solid-phase microextraction column, first eight stainless-steel wires were placed into the stainless-steel column. Then, a nanostructured Cu-Cr-Al ternary layered double hydroxide/polythiophene coating was prepared on the inner surface of the stainless-steel tube and on the surfaces of the stainless-steel wires by a facile in situ electrodeposition method. The nanostructured coating exhibited enhanced long lifetime, good mechanical stability, high porosity, and large specific surface area compared with polythiophene and Cu-Cr-Al layered double hydroxide coatings. Under the optimal conditions, the limits of detection were in the range of 0.07-0.8 µg/L. This method showed good linearity for perphenazine and chlorpromazine in the ranges of 0.3-300 and 0.2-300 µg/L, respectively, with coefficients of determination more than 0.9982. The inter- and intra-assay precisions (RSD%, n = 3) were in the ranges of 3.0-5.1 and 2.5-4.5% at three concentration levels of 5, 25 and 50 µg/L, respectively. Finally, the method was applied for the analysis of the drugs in human urine and plasma samples.

Facile synthesis of magnetic molecularly imprinted polymer: Perphenazine template and its application in urine and plasma analysis.

Synthesis of magnetic iron oxide nanoparticles and its surface modification with methacrylic acid (MAA) was performed simultaneously by adding Fe(2+)/Fe(3+) to an alkaline MAA solution under nitrogen atmosphere. MAA coated magnetite (Fe3O4@MAA) has abundant reactive double bonds on the surface that can initiate polymerization. Magnetic molecularly imprinted polymers (MMIPs) were synthesized through distillation-precipitation polymerization of MAA as monomer, perphenazine (PPZ) as template, and ethylene glycol di-methacrylate (EGDMA) as cross linker on Fe3O4@MAA, with concise control of experimental conditions in about 90min. The produced super paramagnetic MMIPs can be separated from the solution in the presence of external magnetic field in less than 1min. Characterizations of the synthesized particles were performed by electron microscopes, thermo-gravimetric analysis (TGA), vibrating sample magnetometer (VSM), Fourier transform infrared (FT-IR) spectroscopy, and BET. The data showed that Fe3O4@MAA was well encapsulated in the polymer shell. The MMIPs showed high porosity. Moreover, MMIPs were used for rapid pre-concentration and separation of PPZ in human plasma and urine without any dilution and pretreatments using high performance liquid chromatography equipped with a photo diode array detector (HPLC-PDA). The calibration curve in urine and plasma has shown the same slope as the external calibration curve. Linear range of 20-5000ngmL(-1), and a detection limit of 5.3ngmL(-1) was obtained. The results showed 97.92% recovery along with the relative standard deviation of 6.07% (n=6) for 1µgmL(-1) PPZ. Pre-concentration factor was 13. The MMIPs adsorbed PPZ in 1min and then desorbed it by MeOH:HOAc in 2min.

Analysis of perphenazine and fluphenazine by capillary electrophoresis coupled with tris (2,2'-bipyridyl) ruthenium (II) electrochemiluminescence detection.

The coupling of end-column tris (2,2'-bipyridyl) ruthenium (II) electrochemiluminescence (ECL) detection with capillary electrophoresis (CE) was developed for the analysis of two antipsychotic drugs, perphenazine (PPH) and fluphenazine (FPH). The parameters related to CE separation and ECL detection, including the detection potential, the buffer pH value and concentration, the separation voltage, and Ru(bpy)3(2+) concentration, were investigated in detail. Under optimum conditions, PPH and FPH were well separated and detected within 11 min. The linear ranges were 0.1-5 µM for PPH, and 0.1-7.5 µM for FPH, respectively. The limits of detection of PPH and FPH were 5 and 10 nM (S/N=3). The relative standard deviations (n=3) of the ECL intensity and the migration time were less than 2.5 and 0.65% in a day, and less than 3.4 and 1.7% in different three days. The proposed method was successfully applied to determine PPH and FPH in spiked urine samples with satisfactory results.

Catalytic solid substrate-room temperature phosphorimetry for the determination of residual perphenazine based on the electronic effect of rhodamine 6G.

The rhodamine 6G(+) -perphenazine (Rhod 6G(+) -PPH) compound is formed in the ester-exchange reaction between -OH of PPH and -COOC2 H5 of Rhod 6G(+) . PPH was oxidized to a red compound (PPH') in the presence of K2 S2 O8 . Interestingly, the room temperature phosphorescence (RTP) of Rhod 6G(+) was quenched because the -OH of PPH' reacted with -COOC2 H5 of Rhod 6G(+) -PPH to form Rhod 6G(+) -PPH' and PPH, which decreased the π-electron density (δ) of the carbon atom in the Rhod 6G(+) -PPH' conjugated system and enhanced the nonradiation energy loss of the excited Rhod 6G(+) of the triplet state. The PPH content was directly proportional to the ΔIp of the system. Thus, a new catalytic solid-substrate room temperature phosphorimetry (SSRTP) method was established for the determination of PPH. The method had high sensitivity (the limit of detection was 0.019 fg/spot, corresponding to a concentration of 4.8 × 10(-14) g/mL; the sampling quantity was 0.40 µL/spot), good selectivity, convenience and speed. The analytical results were in accordance with those of high-performance liquid chromatography (HPLC). The structures of Rhod 6G(+) , PPH and Rhod 6G(+) -PPH were characterized by infrared spectra. The reaction mechanism by which PPH was determined is discussed.

Chemiluminescence sensor for the determination of perphenazine based on a molecular imprinted polymer.

A luminol-potassium ferricyanide-perphenazine CL reaction with high sensitivity for the determination of perphenazine was found. A perphenazine molecular imprinted polymer (MIP) was synthesized. Using the perphenazine MIP as a recognition material, a novel molecular imprinting-chemiluminescence (MI-CL) sensor for the determination of perphenazine was made based on the luminol-potassium ferricyanide-perphenazine CL reaction. The sensor displayed good selectivity and high sensitivity. The linear-response range of the sensor was 5.0 x 10(-8) - 1.0 x 10(-5) g/ml with a linear correlation coefficient of 0.9961. The detection limit was 2 x 10(-8) g/ml perphenazine and the relative standard deviation for 1.0 x 10(-6) g/ml perphenazine solution was 3.7% (n = 11). The sensor was applied to the determination of perphenazine in urine samples with satisfactory results.

Stability and characterization of perphenazine aerosols generated using the capillary aerosol generator.

Perphenazine (a potent antiemetic) was aerosolized using capillary aerosol generator to generate respirable condensation aerosols from drug in propylene glycol (PG) solutions, by pumping the liquids through a heated capillary tube. The study characterized the stability of perphenazine during and following aerosol generation. The stability-indicating HPLC method (C-8 column with a mobile phase of 52% 0.01 M pH 3.0 acetate buffer+48% acetonitrile) also enabled the study of perphenazine stability in solution under acidic, basic, oxidizing and photolysing conditions. An LC-MS (ESI+) method was used to characterize the degradation products. Perphenazine was found to be stable in acidic and basic conditions, while perphenazine sulfoxide was the major product formed in dilute peroxide solutions. Two photo-degradation products were formed in PG that were tentatively identified by LC-MS; one of these was synthesized and confirmed to be 2-[4-(3-phenothiazin-10-yl-propyl)-piperazino]-ethanol. Both photolysis products showed that aromatic dechlorination had occurred and one appeared to also result from interaction with the solvent. Within an aerosolization energy window of 84-95 J, fine particle aerosols were generated from perphenazine PG formulations with no significant degradation. Small amounts of degradation products were produced in all samples during aerosolization at elevated (non-optimal) energies. These were largely consistent with those seen to result from oxidation and photolysis in solution, showing that oxidation and dehalogenation appeared to be the main degradation pathways followed when the CAG system was overheated.

Application of two chemometric methods for the determination of imipramine, amitriptyline and perphenazine in content uniformity and drug dissolution studies.

Double-divisor spectra derivative and partial least squares methods were developed for content uniformity and dissolution tests in binary or ternary mixtures. The simultaneous determinations of perphenazine (PER) combined with amitriptyline hydrochloride (AMI) and/or imipramine hydrochloride (IMI) have been accomplished using the information of the absorption spectra of appropriate solutions. The double-divisor method is based on the use of the first derivative of the ratio spectrum obtained by dividing the absorption spectrum of the ternary mixture PER-AMI-IMI by a standard spectrum resulted from the addition of two of the three analytes in equal concentrations. The concentration of each component is then determined from their respective calibration graphs established by measuring the ratio derivative analytical signal at a specific wavelength. In this method, the linear determination ranges were of 3.65-18.24 microg/mL for PER, 4.32-21.60 microg/mL for AMI, and 4.83-24.19 microg/mL for IMI. The results were compared with those obtained by partial least squares multivariate calibration (PLS) method pre-treated by a wavelet compression-orthogonal signal correction (W-OSC) filter in zero-order derivative spectra. The calibration model was evaluated by internal validation (cross-validation) and by external validation over synthetic mixtures, content uniformity and dissolution tests. According to the dissolution profile test more than 95% of the three substances were dissolved within 10 min. The results from both techniques were statistically compared with each other and can be satisfactorily used for quantitative analysis and dissolution tests of multicomponent tablets.

Kinetic spectrophotometric method for determination of perphenazine based on monitoring the oxidation intermediate by applying a stopped-flow technique.

A novel and high-sensitive stopped-flow kinetic spectrophotometic method for the determination of perphenazine based on monitoring the variation of absorbance of the intermediate within a few seconds has been developed. The optimum conditions for various parameters on which the forming of the intermediate depends, were investigated. It was found that the initial reaction rate increased linearly with an increase in the perphenazine concentration in the range from 1.0 x 10(-5) to 1.6 x 10(-4)M. The detection limit was calculated to be 5.3 x 10(-6)M. The kinetics of the reaction was established by the aid of single-mixing or double-mixing stopped-flow techniques, a successive reaction model was proposed to analyze and simulate the reaction. The influence of both ascorbic acid and NADH on the time courses was also investigated.

Simultaneous determination of promethazine, chlorpromazine, and perphenazine by multivariate calibration methods and derivative spectrophotometry.

Partial least-squares (PLS) regression, singular value decomposition-based PLS, and an artificial neural network (ANN) were tested as calibration procedures for the simultaneous determination of promethazine, chlorpromazine, and perphenazine by both conventional and derivative spectrophotometry. Comparison of the results revealed that the application of the ANN to the derivative spectra is superior to the application of the 2 PLS methods used. Different binary and ternary synthetic mixtures of the phenothiazine drugs in pure form and in tablets were analyzed by the proposed method, and acceptable results were obtained.

Extractive-spectrophotometric determination of some phenothiazines with dipicrylamine and picric acid.

Dipicrylamine and picric acid have been tested as reagents for the determination of promethazine and perphenazine. They react in neutral media with these drugs forming the coloured compounds. The compounds are sparingly soluble in water and quantitatively extracted into organic solvents. The extracts are intensely coloured and very stable. These properties have been exploited for the extractive spectrophotometric determination of promethazine and perphenazine in pure solutions and pharmaceuticals. Linear calibration graphs were obtained in the concentration range 4-40, 3-30 microg ml(-1) of promethazine and 4-80, 8-60 microg ml(-1) of perphenazine for picric acid and dipicrylamine, respectively. The relative standard deviation (RSD) is less than 0.8%.

Capillary zone electrophoresis determination of perphenazine in pharmaceutical preparations and in human serum.

Perphenazine belongs to the group of phenothiazine-based neuroleptic drugs frequently used in the long-term treatment of psychotic disorders. In this work, a new capillary zone electrophoresis (CZE) method for the rapid determination of perphenazine (PPZ) in pharmaceutical preparations and human blood serum was developed. Using solid-phase extraction (SPE) as a preclean/purification and preconcentration step before CZE analysis, a detection limit of 3 ng/mL for the monitoring of PPZ in blood serum (for a signal-to-noise ratio of 3) was reached.

[A systematic screening and identification method for 29 central nervous system drugs in body fluid by high performance capillary electrophoresis].

A systematic screening method has been developed for the detection of 29 central nervous system (CNS) drugs in human plasma, urine and gastric juice by high performance capillary electrophoresis (HPCE). The first step is sample preparation. The patient's or normal human plasma (0.5 ml) spiked with CNS drugs was extracted with 2 x 4 ml dichloromethane, while 2 ml of patient's or spiked urine was extracted with 2 x 6 ml chloroform. The combined extract from plasma or urine was evaporated to dryness in a rotation evaporator at 35 degrees C. The residue was dissolved in 100 microliters methanol and subsequently 400 microliters of redistilled water was added. The patient gastric juice (3 ml) was centrifuged at 2,000 r.min-1 for 5 min. The supernatant was filtered through 0.45 micron microporous membrane for injection onto capillary columns. The second step was to perform CZE separation in acidic buffer composed of 30 mmol.L-1(NH4)3PO4(pH 2.50) and 10% acetonitrile (condition A). Most of the benzodiazepines (diazepam, nitrazepam, chlordiazepoxide, flurazepam, extazolam, alprazolam) and methaqualone were baseline separated and detected at 5-13 min, while thiodiphenylamines showed group peaks at 3-5 min and barbiturates migrate with electroosmotic fluid (EOF) together. The third step is to separate the drugs in basic buffer constituted of 70 mmol.L-1 Na2HPO4(pH 8.60) and 30% acetonitrile (condition B). The thiodiphenylamines and some other basic drugs could be well separated, which include thihexyphenidyl, imipramine, amitriptyline, diphenhydramine, chlorpromazine, doxepin, chlorprothixene, promethazine and flurazepam, while the rest of the CNS drugs did not interfere with the separation. The last step was to separate the drugs by micellar electrokinetic chromatography (MEKC) in such a buffer as 70 mmol.L-1 SDS plus 15 mmol.L-1 Na2HPO4 (pH 7.55) and 5% methanol (condition C). Barbiturates (barbital, phenobarbital, methylphenobarbital, amobarbital, thiopental, pentobarbital, secobarbital) and some hydrophobic drugs (glutethimide, alprazolam, clonazepam, carbamazepine, trifluoperazine, oxazepam) could be well separated. These drugs might be identified by both the relative migration time (rtm = tdrug/tEOF) and the ratios of peak heights (rh) monitored at different wavelength, since the ratios are characteristic of the spectrum of a drug. This method has been used in several real clinical samples of intoxication. For example, perphenazine and doxepin were detected in the gastric juice and phenobarbital in blood and gastric juice of an intoxicated patient.

[Stripping voltammetric determination of perphenazine and some antipsychotic drugs at carbon paste electrodes].

A sensitive electrochemical method for the determination of four drugs: perphenazine, fluphenazine chlorpromazine and trifluperazine is reported. These compounds exhibited accumulation at carbon paste electrodes in an open circuit. Differential pulse voltammetric determination gave linear calibration curves in the range of 0.02-1 microgram/ml. Detection limits found were 5-7 ng/ml. The drugs in tablets and body fluids were determined directly without separation. The preconcentration mechanism and determination conditions in a new kind of carbon paste piston electrode were investigated. Adsorption and extraction played important roles in accumulation process and the ratio of these two parts was measured by chronocoulometry.

Liquid chromatographic method for perphenazine and its sulfoxide in pharmaceutical dosage forms for determination of stability.

A liquid chromatographic method is described for determination of perphenazine and perphenazine sulfoxide in representative dosage forms. Sulfoxide levels were nondetectable or less than 1% in tablets and in an injectable product. Sulfoxide levels increase with time in some syrup formulations and may be as high as 11% in syrup formulations before their expiration date.

High-performance liquid chromatographic determination of perphenazine and amitriptyline hydrochloride in two-component tablet formulations.

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.