Chlorpromazine In Plasma Research Articles

A new method for the therapeutic drug monitoring of chlorpromazine in plasma by gas chromatography-mass spectrometry using dispersive liquid-liquid microextraction.

Background: Chlorpromazine is the first antipsychotic drug developed and is included in the list of 'essential drugs' prepared by the WHO. Therapeutic drug monitoring is an important point for psychotropic drugs because of significant genetic variability in their metabolism and poor compliance of the patients with treatment. Method: We developed a novel GC-MS method using dispersive liquid-liquid microextraction for the therapeutic monitoring of chlorpromazine. Results: The method was validated according to the European Medicines Agency guidelines. The developed method's lower limit of quantification was set as 30ng/ml. The calibration curve of chlorpromazine was validated between 30 and 600ng/ml, with correlation coefficients of more than 0.99. Conclusion: The developed method was applied to real human patient plasma.

Determination of Chlorpromazine, Haloperidol, Levomepromazine, Olanzapine, Risperidone, and Sulpiride in Human Plasma by Liquid Chromatography/Tandem Mass Spectrometry (LC-MS/MS).

Background and Objective In this study, turbo-ion spray as an interface of tandem mass spectrometry (MS/MS) was performed for sensitive and accurate quantification of chlorpromazine, haloperidol, levomepromazine, olanzapine, risperidone, and sulpiride in plasma samples. Methods Separation was performed by gradient reversed phase high-performance liquid chromatography using a mobile phase containing ammonium formiate 2 mM, pH 2.7, and acetonitrile flowing through a Restek PFP Propyl C18 analytical column (50 mm×2.1 mm i.d.) with particle size of 5 µm, at a flow rate of 800 µL/min. Positive ion fragments were detected in multiple reaction monitoring (MRM) mode. Sample preparation was achieved by solid phase extraction (SPE) (Oasis HLB). Results Mean extraction recoveries ranged from 82.75% to 100.96%. The standard calibration curves showed an excellent linearity, covering subtherapeutic, therapeutic, and toxic ranges. Intraday and interday validation using quality control (QC) samples were performed. The inaccuracy and imprecision were below 12% at all concentration levels. The limits of detection (LOD) and quantification (LOQ) for all analytes were under therapeutic ranges for all tested analytes. Thus, the proposed method was sensitive enough for the detection and determination of subtherapeutic levels of these antipsychotics in plasma samples. No interference of endogenous or exogenous molecules was observed and no carryover effects were recorded. Conclusion According to the results, the proposed method is simple, specific, linear, accurate, and precise and can be applied for antipsychotic analysis in clinical routine. This method was applied for the determination of the tested antipsychotics in plasma samples taken from 71 individuals.

Magnetic micro-solid-phase extraction based on magnetite-MCM-41 with gas chromatography-mass spectrometry for the determination of antidepressant drugs in biological fluids.

A new facile magnetic micro-solid-phase extraction coupled to gas chromatography and mass spectrometry detection was developed for the extraction and determination of selected antidepressant drugs in biological fluids using magnetite-MCM-41 as adsorbent. The synthesized sorbent was characterized by several spectroscopic techniques. The maximum extraction efficiency for extraction of 500μg/L antidepressant drugs from aqueous solution was obtained with 15mg of magnetite-MCM-41 at pH 12. The analyte was desorbed using 100μL of acetonitrile prior to gas chromatography determination. This method was rapid in which the adsorption procedure was completed in 60s. Under the optimized conditions using 15mL of antidepressant drugs sample, the calibration curve showed good linearity in the range of 0.05-500μg/L (r2 =0.996-0.999). Good limits of detection (0.008-0.010 μg/L) were obtained for the analytes with good relative standard deviations of<8.0% (n=5) for the determination of 0.1, 5.0, and 500.0μg/L of antidepressant drugs. This method was successfully applied to the determination of amitriptyline and chlorpromazine in plasma and urine samples. The recoveries of spiked plasma and urine samples were in the range of 86.1-115.4%. Results indicate that magnetite micro-solid-phase extraction with gas chromatography and mass spectrometry is a convenient, fast, and economical method for the extraction and determination of amitriptyline and chlorpromazine in biological samples.

A new restricted access molecularly imprinted polymer capped with albumin for direct extraction of drugs from biological matrices: the case of chlorpromazine in human plasma

A new restricted access molecularly imprinted polymer coated with bovine serum albumin (RAMIP-BSA) was developed, characterized, and used for direct analysis of chlorpromazine in human plasma samples. The RAMIP-BSA was synthesized using chlorpromazine, methacrylic acid, and ethylene glycol dimethacrylate as template, functional monomer, and cross-linker, respectively. Glycerol dimethacrylate and hydroxy methyl methacrylate were used to promote a hydrophilic surface (high density of hydroxyl groups). Afterward, the polymer was coated with BSA using glutaraldehyde as cross-linker, resulting in a protein chemical shield around it. The material was able to eliminate ca. 99% of protein when a 44-mg mL(-1) BSA aqueous solution was passed through it. The RAMIP-BSA was packed in a column and used for direct analysis of chlorpromazine in human plasma samples in an online column switching high-performance liquid chromatography system. The analytical calibration curve was prepared in a pool of human plasma samples with chlorpromazine concentrations ranging from 30 to 350 μg L(-1). The correlation coefficient obtained was 0.995 and the limit of quantification was 30 μg L(-1). Intra-day and inter-day precision and accuracy presented variation coefficients and relative errors lower than 15% and within -15 and 15%, respectively. The sample throughput was 3 h(-1) (sample preparation and chromatographic analysis steps) and the same RAMIP-BSA column was efficiently used for about 90 cycles.

Highly sensitive determination of chlorpromazine by electrochemically treated pencil graphite fiber as both solid-phase microextraction fiber and working electrode for use in voltammetry method

This paper describes the electrochemical modification of a pencil graphite fiber used as both a solid phase microextraction fiber and a working electrode in differential pulse voltammetry (DPV). The combination of solid phase microextraction and DPV was applied as a highly selective and sensitive method for the determination of chlorpromazine. The pencil graphite fibers were electrochemically treated in phosphoric acid and sodium hydroxide solutions. Experimental parameters influencing the extraction efficiency of SPME, such as NaOH concentration, ionic strength of the sample, stirring rate, and extraction time were studied and optimized. Under the optimum conditions, analytical parameters such as linearity, precision and limit of detection were evaluated. The linear dynamic range was from 0.35 to 35 μg L−1 and the limit of detection was 0.2 μg L−1. Intra- and inter-day precision of the method was satisfactory with a relative standard deviation of 2.2 and 4.8%, respectively. The method was successfully applied for the determination of chlorpromazine in human plasma and urine.

Drug screening in human plasma by cloud-point extraction and HPLC

Abstract Cloud-point extraction (CPE) with RP-HPLC/DAD detection was used to develop a screen for six model basic drugs (paracetamol, promazine, amitriptyline, nortriptyline, clomipramine and chlorpromazine) in human plasma. These drugs’ varied hydrophobicities entail different affinities for the micelle-rich phase and CPE extraction efficiencies. Extraction recovery (except paracetamol) was above 80% and reproducibility (RSD%) ranged from 2.88 to 10.26 intraday and from 3.12 to 12.33 interday. The limits of detection were: 0.125 µg mL−1 (promazine and chlorpromazine), 0.25 µg mL−1 (amitriptyline and nortriptyline) and 0.5 µg mL−1 (paracetamol and clomipramine). The method was linear over the ranges: 0.125–1.0 µg mL−1 (promazine and chlorpromazine), 0.25–1.0 µg mL−1 (amitriptyline and nortriptyline), 0.5–1.0 µg mL−1 (clomipramine) and 0.5–10 µg mL−1 (paracetamol). The procedure is a good alternative to the SPE or LLE sample preparation usually used.

Chlorpromazine quantification in human plasma by UPLC–electrospray ionization tandem mass spectrometry. Application to a comparative pharmacokinetic study

In the present study a method to quantify chlorpromazine in human plasma using cyclobenzaprine as the internal standard (IS) is described. The analyte and the IS were extracted from human plasma by a liquid–liquid extraction with diethyl ether/dichloromethane (70/30, v/v) and analyzed by an ultra performance liquid chromatography (UPLC) coupled to an electrospray tandem triple quadrupole mass spectrometer in positive mode (UPLC–ES +-MS/MS). Chromatography was performed isocratically on an Aquity UPLC BEH C18 1.7 μm (50 mm × 2.1 mm i.d.) operating at 40 °C. The mobile phase was a mixture of 65% water + 1% formic acid and 35% of acetonitrile at a flow-rate of 0.5 mL/min. The lowest concentration quantified was 0.5 ng/mL and a linear calibration curve over the range 0.5–200 ng/mL was obtained, showing intra-assay precisions from 2.4 to 5.8%, and inter-assay precisions from 3.6 to 9.9%. The intra-assay accuracies ranged from 96.9 to 102.5%, while the inter-assay accuracies ranged from 94.1 to 100.3%. This analytical method was applied in a relative bioavailability study in order to compare a test chlorpromazine 100 mg simple dose formulation versus a reference in 57 volunteers of both sexes. The study was conducted in an open randomized two-period crossover design and with a fourteen days washout period. Plasma samples were obtained over a 144-h interval. Since the 90% CI for both C max, AUC last and AUC 0–inf were within the 80–125% interval proposed by the Food and Drug Administration and ANVISA, it was concluded that chlorpromazine 100 mg/dose was bioequivalent to the reference formulation, according to both the rate and extent of absorption.

Determination of selected phenothiazines in human plasma by solid-phase extraction and liquid chromatography with coulometric detection

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.

Sensitive liquid chromatography/tandem mass spectrometry method for the determination of the lipophilic antipsychotic drug chlorpromazine in rat plasma and brain tissue

A simple, sensitive and robust liquid chromatography/electrospray ionization tandem mass spectrometry (LC–ESI-MS/MS) method was developed and validated for quantification of chlorpromazine in rat plasma and brain tissue. Chlorpromazine was extracted from rat plasma and brain homogenate using liquid–liquid extraction. The compounds were separated on a Waters Atlantis™ dC-18 (30 mm × 2.1 mm i.d., 3 μm) column using a mobile phase of acetonitrile/20 mM ammonium formate (pH 4.25 adjusted with formic acid) with gradient elution. Chlorpromazine was detected in positive ion mode using multiple reaction monitoring (MRM). The method was validated and the specificity, linearity, lower limit of quantitation (LLOQ), precision, accuracy, recoveries and stability were determined. The LLOQ was 0.2 ng/ml for plasma and 0.833 ng/g for brain tissue. The method was linear over the concentration range from 0.2 to 200.0 ng/ml for plasma and from 0.833 to 833.3 ng/g for brain tissue. The correlation coefficient ( R 2) values were more than 0.998 for both plasma and brain homogenate. The precision and accuracy for intra-day and inter-day were better than 7.54%. The relative and absolute recovery was above 84.9% and matrix effects were lower than 5.6%. This validated method has been successfully used to quantify the rat plasma and brain tissue concentration of chlorpromazine after chronic treatment.

Radioimmunoassay for the N-Oxide Metabolite of Chlorpromazine in Human Plasma and Its Application to a Pharmacokinetic Study in Healthy Humans

Antibodies specific to chlorpromazine N-oxide (CPZNO) were produced in rabbits immunized with a hapten–bovine serum albumin conjugate, which was prepared by linking the 7-position of the phenothiazine ring of the metabolite to the protein via a 4-carbon bridge. An extraction radioimmunoassay (RIA) was developed using this antiserum and shown to have adequate sensitivity and specificity for determination of plasma concentrations of CPZNO in the presence of chlorpromazine (CPZ) and its major metabolites. It was used together with the previously developed RIAs for CPZ, chlorpromazine sulfoxide (CPZSO), and 7-hydroxychlorpromazine (7-OHCPZ) to study the pharmacokinetics of CPZ and these metabolites in five healthy volunteers after they received a single 50-mg oral dose of CPZ. It is interesting to note that peak plasma concentrations of CPZNO were considerably higher than CPZ, and the apparent elimination half-lives of this metabolite were shorter than those of CPZ.

Therapeutic monitoring of chlorpromazine I: Pitfalls in plasma analysis.

Pooled plasma from healthy volunteers was spiked with pure, synthetic chlorpromazine (CPZ), chlorpromazine sulfoxide (CPZSO), or chlorpromazine N-oxide (CPZNO), and then made alkaline with either sodium hydroxide or sodium carbonate. The samples were allowed to stand at room temperature for various timed intervals before extraction with organic solvent. It was found that CPZNO was reduced to CPZ in plasma made alkaline with sodium hydroxide, but not in protein-free buffer solution at high pH nor in plasma made alkaline with sodium carbonate. The reaction appears to take place through reducing equivalents generated by the action of sodium hydroxide on plasma proteins. Thus, apparent concentrations of CPZ in plasma from patients were elevated by as much as 343% when sodium hydroxide was used compared with concentrations in aliquots of the same plasma samples alkalinized with sodium carbonate. The amount of CPZ produced from CPZNO depends on the type of extraction procedure employed as well as on the quantity of sodium hydroxide added to the plasma. By contrast, no interconversion between CPZ and CPZSO or CPZNO and CPZSO was observed in plasma alkalinized and extracted under any of the conditions tested.

The effect of sample storage on the stability of chlorpromazine in plasma and whole blood

The stability of chlorpromazine in heparinized whole blood and plasma was investigated at three different concentrations (5, 20, and 50 ng/ml) after storage at −20°C for 0, 7, 14, 28, 56, and 84 days. A statistical analysis was done to determine whether there was any significant difference between the concentration of chlorpromazine in these body fluids as determined on day one versus each of the later dates of analysis. The results indicate that no significant difference was found over the course of these investigations.

Quantitative determination of chlorpromazine and thioridazine by high-performance thin layer chromatography.

An assay for quantitation of chlorpromazine and thioridazine in patient plasma is developed. The procedure utilizes high-performance thin layer chromatography for separation and in situ absorption densitometry for quantitation. Depending upon the HPTLC plate size, 30 to 60 samples may be processed in less than 6 hr, with an intra-assay coefficient of variation of less than 4%. The procedure can be used to measure as little as 10 ng/ml of drug in plasma, well below expected concentrations in patients receiving these medications. Investigations of extraction procedures, sample application procedures, chromatographic conditions, sample derivatization, and in situ absorption densitometry are described.

Subnanogram Quantitation of Chlorpromazine in Plasma by High-Performance Liquid Chromatography with Electrochemical Detection

A specific and sensitive high-performance liquid chromatographic (HPLC) method for the quantitative determination of subnanogram levels of chlorpromazine in plasma is described. Following extraction of chlorpromazine and the internal standard, prochlorperazine, HPLC analysis is carried out on a cyano column with a mobile phase consisting of 0.1 M ammonium acetate in acetonitrile (10:90 v/v). The use of oxidative thin-layer amperometric detection allowed the quantitation of 0.25ng of chlorpromazine/ml of plasma with a coefficient of variation of 5.1%. The HPLC method has adequate sensitivity to follow plasma concentration–time profiles up to 24hr following low single oral doses of chlorpromazine in healthy volunteers.

Recent advances in the analysis of phenothilazine drugs and their metabolites using high performance liquid chromatography

1. 1. HPLC procedures with electrochemical or ultraviolet detection were developed in order to analyze plasma samples from patients and volunteers who had received antipsychotic drugs. 2. 2. Procedures were described for the determination of chlorpromazine and trimeprazine in human plasma. 3. 3. The described procedures for chlorpromazine and trimeprazine demonstrated sufficient sensitivity for their use in pharmacokinetic studies following low single oral doses of these agents. 4. 4. The HPLC-EC procedures for chlorpromazine and trimeprazine were compared with GC-MS and radiolmmunoassay procedures, respectively.

Gas chromatographic—mass spectrometric procedure for the quantitation of chlorpromazine in plasma and its comparison with a new high-performance liquid chromatographic assay with electrochemical detection

A gas chromatographic—mass spectrometric assay using selected ion monitoring is compared with a high-performance liquid chromatographic assay using an electrochemical detector for single-dose studies of the psychotherapeutic phenothiazine drug chlorpromazine. Measurements were made after extraction of chlorpromazine and the internal standard, prochlorperazine, from basified plasma with an isopropanol—pentane solvent mixture. Following evaporation of the organic solvents the residue was reconstituted in a small volume of methanol and subjected to gas chromatographic—mass spectrometric selected ion detection. The residual sample was then evaporated and made up in a larger volume of acetonitrile and analyzed by high-performance liquid chromatography using an electrochemical detector. These specific methods display excellent correlation for plasma concentration determinations in the range of 0.25–10 ng ml −1 and will allow for the study of the pharmacokinetics of chlorpromazine following single low doses of the drug.

Simultaneous determination of chlorpromazine and levomepromazine in human plasma and urine by high-performance liquid chromatography using electrochemical detection

A rapid, selective and sensitive method for the simultaneous determination of chlorpromazine and levomepromazine in human plasma and urine has been developed using high-performance liquid chromatography with electrochemical detection. The unchanged drugs and internal standard extracted from plasma and urine were separated by reversed-phase high-performance liquid chromatography. The influence of acetonitrile concentration and of the pH of the mobile phase were investigated. The detection limits were 100 pg for chlorpromazine and for levomepromazine. In comparison with three other detection systems this was found to be the most sensitive method. This method was successfully applied to the simultaneous determination of chlorpromazine and levomepromazine in human plasma and urine for pharmacokinetic studies.

Simultaneous Determination of Chlorpromazine and Levomepromazine in Human Plasma by High-Performance Liquid Chromatography Using Electrochemical Detection

Simultaneous Determination of Chlorpromazine and Levomepromazine in Human Plasma by High-Performance Liquid Chromatography Using Electrochemical Detection

Thin-layer differential pulse voltammetric determination of chlorpromazine in biological fluids

The combination of a thin-layer electrochemical cell with differential pulse voltammetry can be used to determine chlorpromazine in plasma and urine. The thin-layer cell (23 μl capacity) has a wax-impregnated graphite electrode. Direct determination of chlorpromazine in urine gave a linear calibration curve for the range 4.8 × 10 -3–2.4 × 10 -4 M with 97% recovery. No interference from glutethimide, dextropropoxyphene, meprobamate, diazepam, and methaqualone-HCl was detected. Direct measurement of chlorpromazine in plasma gave a linear calibration curve for the range 2.4 × 10 -5–4.8 × 10 -4 M with 89% recovery. The procedure for plasma and urine requires only 2 min per determination. Detection levels are below that required for monitoring therapeutic levels of chlorpromazine in urine.

Analysis of chlorpromazine in plasma: Effect of specimen storage

Two analytical methods for the analysis of chlorpromazine (CPZ), a radioimmunoassay (RIA) and a GLC-MS method, were compared in a bioequivalence study of two CPZ tablet formulations (Thorazine: film coated and sugar coated). Thirty-six nonsmoking, healthy, male volunteers completed the study. Each subject ingested single doses (2 × 25 mg) of the test (T) and the reference (R) formulations in a two-way crossover design with a two-week drug-free interval between doses. Following each administration, plasma concentrations of CPZ were monitored over a period of 24 h by both RIA and GLC-MS methods. Plasma concentrations and pharmacokinetic parameters determined by either analytical method showed wide intersubject variation, with the GLC-MS data showing relatively higher magnitude of intersubject variation than the RIA data. In general, plasma concentrations measured by RIA were significantly different from those measured by GLC-MS (paired t tests: p < 0.0001). As indicated by the regression analysis, concentrations determined by RIA were 1.3–1.4 times higher than those determined by GLC-MS. There were strong and significant correlations between the two methods for both T and R (r > 0.75: p < 0.0001). Similar statistical relationships were found between the plasma concentrations of CPZ determined by the two methods at each sampling time and the bioequivalence parameters area under the plasma level versus time curves up to the last measurable concentration (AUC0t) and the maximum plasma concentration (Cmax). Bioequivalence parameters AUC0t, AUC0∞, and (Cmax). determined separately from the data obtained by the two methods for the two formulations were examined by analyses of variance (ANOVA) and other criteria and tests for bioequivalence. Results of ANOVA, confidence intervals of test/reference ratios of AUC0t, AUC0∞, and Cmax, and statistical tests for bioequivalence indicated that, except for Cmax by GLC-MS, data generated by both methods indicated bioequivalence of the two formulations (i.e., test/reference ratio was within 100 ± 20%). The experiments suggest that despite significant differences between the absolute plasma concentration values obtained, both methods yielded data for the two formulations which demonstrated that they were bioequivalent.