A novel and sensitive UPLC-MS/MS method to determine mequitazine in rat plasma and urine: Validation and its application to pharmacokinetic studies.

The aim of this study was to develop an analytical method to determine mequitazine in rat plasma and urine. Mequitazine was separated by UPLC-MS/MS equipped with a Kinetex core-shell C18 column (50 × 2.1 mm, 1.7 µm) using 0.1% (v/v) aqueous formic acid and acetonitrile containing 0.1% (v/v) formic acid as a mobile phase by gradient elution at a flow rate of 0.3 mL/min. Quantitation of this analysis was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique operating in multiple reaction monitoring positive ion mode. Mass transitions were m/z 323.3 → 83.1 for mequitazine and 281.3 → 86.3 for imipramine as internal standard. Liquid-liquid extraction with ethyl acetate and protein precipitation with methanol were used for sample extraction. Chromatograms showed that the method had high resolution, sensitivity and selectivity without interference from plasma constituents. Calibration curves for mequitazine in rat plasma and urine were 0.02-200 ng/mL, showing excellent linearity with correlation coefficients (r2 ) >0.99. Both intra- and inter-day precisions (CV%) were within 4.08% for rat plasma and urine. The accuracies were 99.58-102.03%. The developed analytical method satisfied the criteria of international guidance. It could be successfully applied to pharmacokinetic studies of mequitazine after oral and intravenous administration to rats.

A molecularly imprinted polymer based chemiluminescence array sensor for one-step determination of phenothiazines and benzodiazepines in pig urine.

The residues of phenothiazines and benzodiazepines in foods of animal origin are dangerous to consumers. For inspection of their abuses, this study for the first time reported on the use of a chemiluminescence array sensor for the simultaneous determination of four phenothiazines and five benzodiazepines in pig urine. Two molecularly imprinted polymers were coated in different wells of a conventional 96-well microtiter plate as the recognition reagents. After sample loading, the absorbed analytes were initiated directly by using an imidazole enhanced bis(2,4,6-trichlorophenyl)oxalate-hydrogen peroxide system to emit light. The assay process consisted of only one sample-loading step prior to data acquisition, so one test was finished within 10 min. The limits of detection for the nine drugs in the pig urine were in a range of 0.1 to 0.6 pg/mL, and the recoveries from the fortified blank urine samples were in a range of 80.3 to 95%. Furthermore, the sensor could be reused six times. Therefore, this sensor could be used as a simple, rapid, sensitive and reusable tool for routine screening for residues of phenothiazines and benzodiazepines in pig urine.

Molecularly imprinted polymers as analyte sequesters and selective surfaces for easy ambient sonic-spray ionization.

The use of a molecularly imprinted polymer (MIP) as a selective surface for ambient mass spectrometry is demonstrated. The MIP is used to sequester target analytes from urine and easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is shown to be able to efficiently desorb the analytes from the MIP surface and then transfer them in protonated forms to the gas phase for MS analysis. A set of five phenothiazines (chlorpromazine, perphenazine, triflupromazine, thioridazine and prochlorperazine) were chosen from a proof-of-principle class of drug samples. A chlorpromazine-imprinted methacrylic polymer was synthesized and used to prepare a MIP probe. The MIP-EASI-MS technique using acidified methanol as solvent has been shown to allow quantification of all five drugs in urine with LOQ of ca. 1 micromol L(-1).

Determination of phenothiazine derivatives in human urine by using ionic liquid-based dynamic liquid-phase microextraction coupled with liquid chromatography.

A simple and rapid method for the determination of seven phenothiazines derivatives (chlorpromazine, promethazine, levomepromazine, prochlorperazine, trifluoperazine, fluphenazine and thioridazine) in human urine samples is presented. The analytes are extracted from the sample in 50 microL of the ionic liquid 1-butyl-3-methyl-imidazolium hexafluorophosphate working in an automatic flow system under dynamic conditions. The chemical affinity between the extractant and the analytes allows a good isolation of the drugs from the sample matrix achieving at the same time their preconcentration. The separation and detection of the extracted compounds is accomplished by liquid chromatography and UV detection. The proposed method is a valuable alternative for the analysis of these drugs in urine within the concentration range 0.07-10 microg mL(-1). Limits of detection were in the range from 21 ng mL(-1) (thioridazine) to 60 ng mL(-1) (levomepromazine). The repeatability of the proposed method expressed as RSD (n=5) varied between 2.2% (levomepromazine) and 3.9% (chlorpromazine).

Metabolic fate of phenothiazine in the marmoset (Callithrix jacchus).

The fate of [35S]-phenothiazine, a veterinary anthelmintic, has been investigated in the adult male marmoset (Callithrix jacchus) following oral administration. A near complete recovery of radioactivity (c. 95%) was achieved in 0-3 days, with just over one-third of the dose (c. 37%) being present in the urine and the remainder (c. 58%) being accounted for in the faeces. The majority of the urinary radioactivity (c. 91%) was present as conjugates, tentatively identified as phenothiazine N-glucuronide and leucophenothiazone sulphate. Smaller amounts of phenothiazone, thionol, phenothiazine sulphoxide and unchanged phenothiazine were also identified. The only compound identified in the faeces was unchanged phenothiazine.

[Application of the clinoid dehydration method in forensic medical expert examination of poisoning with narcotic and strong medicine preparations].

The method of clinoid dehydration of biological fluids along with pH measurement is shown to be suitable for the diagnosis of narcotic intoxication and poisoning with narcotic and potent agents. The data obtained by this and conventional methods are compared. Diagnostically significant signs of narcotic intoxication are deduced from the hemogram of a dry blood droplet and serum pH values.

[Extended suicidal poisoning with carbamazepine and phenothiazine derivatives]. / Rozszerzone samobójcze zatrucie karbamazepina i fenotiazynami.

UNLABELLED: Two cases (woman and man) of the extended suicidal poisonings with carbamazepine and phenothiazine derivatives are presented. Drug's blood concentrations during poisoning were monitored. We examine correlation between patient's general status and the drug's blood concentrations, carbamazepine and phenothiazine derivatives interaction due to young, healthy people who received no earlier treatment. MATERIAL AND METHODS: blood samples for toxicological examinations were collected at 0, 12, 24 and 48 hours after admission. Carbamazepine was determined using FPIA method and phenothiazines derivatives by HPLC-DAD. The highest blood concentrations were for carbamazepine: 30.92 mg/l (woman) and 20.95 ng/ml (man); for phenothiazine derivatives: 927 ng/ml (woman) and 733 ng/ ml (man). CONCLUSIONS: In both cases severe central nervous depression was observed due to summed action of the drugs. Sex and individual differences in cytochromes activities should have influence to carbamazepine metabolism and faster elimination time in woman. In the case of phenothiazine derivatives faster elimination time in man was observed. The differences in elimination times between compared drugs confirm their different metabolic routes.

Combination of poly(diallyldimethylammonium chloride) and hydroxypropyl-γ-cyclodextrin for high-speed enantioseparation of phenothiazines by capillary electrophoresis.

High-speed capillary electrophoresis (CE) enables the simple, rapid, and inexpensive analysis of large sets of chiral samples in the pharmaceutical industry. Hence, we developed a novel method for separating enantiomers of d,L-phenothiazines simply and rapidly, based on using poly(diallyldimethylammonium chloride) (PDDAC) as an additive and hydroxypropyl-γ-cyclodextrin (Hp-γ-CD) as a chiral selector in capillary electrophoresis. Adding 0.9% PDDAC to the background electrolyte generated a stable, high, and reversed electroosmotic flow (EOF). Hp-γ-CD not only worked as a complexing agent to increase the chiral resolution between d,L-phenothiazines but also decreased the effective electrophoretic mobility of these drugs. Combining PDDAC and Hp-γ-CD as buffer additives enabled CE to achieve a high-speed enantioseparation of five pairs of d,L-phenothiazines. A decrease in capillary length and an increase in the intensity of the electric field further shortened the separation time. When the background electrolyte contained 0.9% PDDAC, 5mM Hp-γ-CD, and 75 mM formic acid (pH 3.0), enantioseparation of the d,L-phenothiazines was attained within 230 s by applying a capillary length of 32.5 cm and an electric field of 292 V cm(-1). The limit of detection (LOD) of the d,L-phenothiazines at a signal-to-noise ratio of 3 ranged from 2 to 8 µM. We demonstrated the feasibility of this method by detecting the five pairs of d,L-phenothiazines in urine samples.

A system of screening for the presence of a number of common drugs.

A method is described to provide a rapid screening technique for the presence of barbiturates, glutethimide, carbromal, meprobamate, salicylate, phenothiazine derivatives, bromide, carbon monoxide, and alcohol. Phenothiazines are detected by a spot urine test. The first four drugs are identified, within 60 minutes of blood collection, on thin-layer chromatoplates of microscope slide dimensions. The estimations of bromide, salicylate, carbon monoxide, and of alcohol levels are started in that period so the overall time for the screening is less than two hours, and the amount of blood required is only 10 ml.

Evaluation of a latex agglutination-inhibition slide pregnancy test (Pregnosis).

PIP: A new formulation of a latex agglutination-inhibition slide pregnancy test (Pregnosis) was evaluated over a 21-month period in 2797 unselected specimens of teenagers seen primarily for pregnancy diagnosis. A selected study group was tested of 847 males and nonpregnant females, including 145 patients on moderate to high doses of methadone, 329 patients taking phenothizines or psychotropic drugs, and 25 patients on oral contraceptives. Also tested wer 67 patients who were addicts with moderate to high levels of morphine or codeine, and 100 early or late menopausal protein-free specimens as well as 181 proteinuric specimens. The urines were tested without centrifugation or filtration. The test was 68% accurate for samples 28-40 days from the last menstrual period (LNMP), 93.9% accurate for speciments 41-60 days from LMNP, 97.7% accurate for speciments 61-100 days from LMNP, 98.1% accurate 101-140 days from LMNP and 94.5% accurate on specimens with unspecified LMNPs. Overall accuracy in 1464 uncomplicated pregnancies was 95.4%. False positives were absent on 1287 routine specimens. Test interference from metabolites of methadone, phenothiazines, psychotropic drugs, morphine or codeine, and oral contraceptives.^ieng

Determination of phenothiazines in human body fluids by solid-phase microextraction and liquid chromatography/tandem mass spectrometry.

Eleven phenothiazine derivatives with heavy side-chains were found to be extractable from human whole blood and urine samples by solid-phase microextraction (SPME) with a polyacrylate-coated fiber. The fiber was then injected into the desorption chamber of an SPME-liquid chromatography (LC) interface for LC/tandem mass spectrometry (MS/MS) with positive ion electrospray (ES) ionization. All compounds formed base peaks due to [M + 1](+) ions by LC/ES-MS/MS. By use of LC/ES-MS/MS, the product ions produced from each [M + 1](+) ion showed base peaks due to side-chain liberation. Selected reaction monitoring (SRM) and selected ion monitoring (SIM) were compared for the detection of the 11 phenothiazine derivatives from human whole blood and urine. SRM showed much higher sensitivity than SIM for both types of sample. Therefore, a detailed procedure for the detection of drugs by SRM with SPME-LC/MS/MS was established and carefully validated. The extraction efficiencies of the 11 phenothiazine derivatives spiked into whole blood and urine were 0. 0002-0.12 and 2.6-39.8%, respectively. The regression equations for the 11 phenothiazine derivatives showed excellent linearity with detection limits of 0.2-200 ng ml(-1) for whole blood and 4-22 pg ml(-1) for urine. The intra- and inter-day precisions for whole blood and urine samples were not greater than 15.1%. The data obtained after oral administration of perazine or flupentixol to a male subject are presented.

Mechanism of action of narcotics in the production of menstrual dysfunction in women.

The ability of morphine to block ovulation in animals prompted investigation of the frequency and mechanisms of menstrual abnormalities in women addicted to narcotic analgesics. Menstrual histories obtained from 76 former heroin addicts receiving daily methadone maintenance revealed that more than one-half of these women had experienced menstrual abnormalities while taking heroin or methadone. In order to determine the specific physiologic effects of narcotic analgesics on reproductive function, detailed endocrinologic studies were carried out in seven of these patients who complained of amenorrhea or irregular menses while receiving methadone. Four of the seven women manifested abnormalities of the control of gonadotropin secretion. Three of these four failed to exhibit cyclic gonadotropin release, as evidenced by an absence of increased levels of follicular phase follicle-stimulating hormone, midcycle gonadotropin peaks or luteal phase progesterone increments. In the fourth patient a prolonged follicular phase (30 days) of the menstrual cycle was detected. One of these four patients also had low basal gonadotropin levels and failed to exhibit luteinizing hormone increments greater than control levels in response to ethinyl estradiol (positive feedback). The remaining three women exhibited normal patterns of gonadotropin secretion during the observation period. In these women, menstrual bleeding occurred in response to withdrawal from luteal phase (10 to 20 ng/ml) progesterone levels and to exogenous ethinyl estradiol, suggesting normal uterine responsivity to progesterone and estrogen. Although not documented, it is likely that oligo-ovulation was the cause of the irregular menses in these three patients. Amenorrhea is commonly associated with methadone ingestion or heroin addiction and appears to be related to an alteration of the hypothalamic mechanisms controlling gonadotropin secretion. Tolerance to these effects of methadone may develop after chronic ingestion.

Positive- and negative-ion mass spectrometry and rapid clean-up of 19 phenothiazines.

Positive-ion electron impact (PIEI), positive-ion chemical ionization (PICI) and negative-ion chemical ionization (NICI) mass spectra of 19 phenothiazines are presented. In the PIEI mode, peaks due to M, M minus side chain (M - R1), M - R1 + H, and side chain itself (R1) appeared for most compounds. The M - R1 and R1 ions were very useful for drug screening. In the PICI mode, most spectra showed base or intense peaks due to M + H, and small peaks due to M + C2H5; peaks due to M - R1 + 2H and R1 also appeared in many compounds. In the NICI mode, fragmentation modes were different in different compound groups; molecular or [M - H]- quasi-molecular anions appeared in many compounds with aliphatic side chains. Anions at m/z 98 and 115 were characteristic for compounds with (N-methylpiperazinyl)propyl side chains. Selected ion monitoring in the PIEI mode generally gave much higher sensitivity than in the PICI and NICI modes. Phenothiazines present in urine or plasma could be rapidly isolated by use of Sep-Pak C18 cartridges. Thirteen of 19 phenothiazines could be detected by HP-17 wide-bore capillary gas chromatography with satisfactory separation from impurities in their underivatized forms.

Study of the metabolism of phenothiazines: determination of N-demethylated phenothiazines in urine.

When phenothiazine drugs (chlorpromazine, promazine, promethazine, propiomazine, propionylpromazine, trifluoperazine and trimeprazine) were reacted with m-chloroperbenzoic acid, N-demethylated phenothiazines were obtained in moderate yield. The mass spectra of the N-demethylated phenothiazines showed either m/z 44 and 72 or m/z 58 as characteristic ions depending on their side chain. The N-demethylated propiomazine was identified as a metabolite of propiomazine from the urine of a rat which was given propiomazine orally.

Use of the EMITtox serum tricyclic antidepressant assay for the analysis of urine samples.

The EMITtox serum tricyclic antidepressant (TCA) assay is intended for use with serum or plasma. Currently, there are no commercially available immunoassay kits available for the detection of TCAs in urine. The proposed method utilizes the EMITtox serum TCA assay for the direct analysis of urine samples. The minimum detectable concentration of nortriptyline in urine is 25 ng/mL. The within-run CV was determined to be 0.6% and the between-run CV was 2.6%. The TCA assay cross-reacts with phenothiazines and antihistamines. The proposed methodology should be applicable to other EMIT serum assays to allow their use with urine.

Serum levels and urinary excretion of mequitazine after a single oral dose.

Pharmacokinetics of mequitazine, a recently introduced peripheral H1-histamine receptor antagonist of phenothiazine type, was followed up to 72 h after the single oral dose of 5 mg of the drug to eight fasted healthy volunteers. Each subject was treated thrice with a dosing interval of 15 days or more. Thus all the results were triplicated. Serum mequitazine was measured by mass fragmentography using a gas-liquid chromatograph/mass spectrometer set in the electron impact mode. Urine phenothiazines were determined fluorometrically before and after cleaving phenothiazines from their glucuronide conjugates. Peak concentration of mequitazine in serum was 3.19 +/- 1.70 (s.d.) ng.ml-1, time to peak concentration 5.67 +/- 1.68 h, elimination half-life 45 +/- 26 h, and elimination rate constant 0.018 +/- 0.007 h-1. Only 10.9 +/- 3.3% of the dose appeared in urine in unconjugated plus the glucuronidated form during the first 72 h. About 46% of the urinary phenothiazines were glucuronide conjugates. The results suggested that after the oral administration only low mequitazine concentrations appeared in serum, most of the drug seemed to be deactivated by the extrarenal route, and the kinetic properties of the drug resembled those of several phenothiazines used for psychiatric therapy.

[Evaluation of an automated liquid chromatograph permitting toxicologic screening of biological fluids]. / Evaluation d'un automate de chromatographie liquide permettant un screening toxicologique dans les liquides biologiques.

This study evaluated an automated high performance liquid chromatography algorithm that allows identification of 330 substances and metabolites in biological fluids. We assessed its ability to detect in urine and gastric lavage fluids the main psychotropic drugs ingested with suicidal intent. Antidepressants and most of the phenothiazines were recognised; identification of benzodiazepines was more uncertain. Compared to EMIT techniques, chromatography is less sensitive, but allows precise identification of the offending poison, quantifies the amount, and allows broad toxicological screening of pharmacological classes inaccessible to the EMIT technique.

Hollow fiber-liquid phase microextraction combined with gas chromatography for the determination of phenothiazine drugs in urine.

A simple method of hollow fiber-liquid phase microextraction (HF-LPME) combined with gas chromatography (GC) was developed for the analysis of four phenothiazine drugs (promethazine, promazine, chlorpromazine and trifluoperazine) in human urine samples. All variables affecting the extraction of target analytes including organic solvent type, stirring rate, extraction time, extraction temperature, pH of sample solution and ionic strength were carefully studied and optimized. Under the optimal conditions, the analytical performance of HF-LPME-GC-flame photometric detector (FPD) and HF-LPME-GC-flame ionization detector (FID) were evaluated and compared. The results showed that the HF-LPME-GC-FID was more sensitive than HF-LPME-GC-FPD for the determination of four target phenothiazine drugs, while the signal peak shape and resolution obtained by HF-LPME-GC-FPD was better than that obtained by HF-LPME-GC-FID. HF-LPME-GC-FPD/FID was successfully applied for the assay of the interested phenothiazine drugs in urine sample, and the excretion of the drugs was also investigated by monitoring the variation of the concentration of chlorpromazine in urine of a psychopath within 8 h after drug-taking. The proposed method provided an effective and fast way for the therapeutic drug monitoring (TDM) of phenothiazine.