Stir bar sorptive-dispersive microextraction by a poly(methacrylic acid-co-ethylene glycol dimethacrylate)-based magnetic sorbent for the determination of tricyclic antidepressants and their main active metabolites in human urine.

A poly(methacrylic acid-co-ethylene glycol dimethacrylate)-based magnetic sorbent was used for the rapid and sensitive determination of tricyclic antidepressants and their main active metabolites in human urine. This material was characterized by magnetism measurements, zeta potential, scanning electron microscopy, nitrogen adsorption-desorption isotherms, and thermogravimetric analysis. The proposed analytical method is based on stir bar sorptive-dispersive microextraction (SBSDME) followed by liquid chromatography-tandem mass spectrometry. The main parameters involved in the extraction step were optimized by using the response surface methodology as a multivariate optimization method, whereas a univariate approach was employed to study the desorption parameters. Under the optimized conditions, the proposed method was properly validated showing good linearity (at least up to 50 ng mL-1) and enrichment factors (13-22), limits of detection and quantification in the low ng L-1 range (1.4-7.0 ng L-1), and good intra- and inter-day repeatability (relative standard deviations below 15%). Matrix effects were observed for the direct analysis of urine samples, but they were negligible when a 1:1 v/v dilution with deionized water was performed. Finally, the method was successfully applied to human urine samples from three volunteers, one of them consuming a prescribed drug for depression that tested positive for clomipramine and its main active metabolite. Quantitative relative recoveries (80-113%) were obtained by external calibration. The present work expands the applicability of the SBSDME to new analytes and new types of magnetic sorbents.

Application of Bar Adsorptive Microextraction for the Determination of Levels of Tricyclic Antidepressants in Urine Samples.

This work entailed the development, optimization, validation, and application of a novel analytical approach, using the bar adsorptive microextraction technique (BAµE), for the determination of the six most common tricyclic antidepressants (TCAs; amitriptyline, mianserin, trimipramine, imipramine, mirtazapine and dosulepin) in urine matrices. To achieve this goal, we employed, for the first time, new generation microextraction devices coated with convenient sorbent phases, polymers and novel activated carbons prepared from biomaterial waste, in combination with large-volume-injection gas chromatography-mass spectrometry operating in selected-ion monitoring mode (LVI-GC-MS(SIM)). Preliminary assays on sorbent coatings, showed that the polymeric phases present a much more effective performance, as the tested biosorbents exhibited low efficiency for application in microextraction techniques. By using BAµE coated with C18 polymer, under optimized experimental conditions, the detection limits achieved for the six TCAs ranged from 0.2 to 1.6 µg L-1 and, weighted linear regressions resulted in remarkable linearity (r2 > 0.9960) between 10.0 and 1000.0 µg L-1. The developed analytical methodology (BAµE(C18)/LVI-GC-MS(SIM)) provided suitable matrix effects (90.2-112.9%, RSD ≤ 13.9%), high recovery yields (92.3-111.5%, RSD ≤ 12.3%) and a remarkable overall process efficiency (ranging from 84.9% to 124.3%, RSD ≤ 13.9%). The developed and validated methodology was successfully applied for screening the six TCAs in real urine matrices. The proposed analytical methodology proved to be an eco-user-friendly approach to monitor trace levels of TCAs in complex urine matrices and an outstanding analytical alternative in comparison with other microextraction-based techniques.

Gelatin microsphere coated Fe3O4@graphene quantum dots nanoparticles as a novel magnetic sorbent for ultrasound-assisted dispersive magnetic solid-phase extraction of tricyclic antidepressants in biological samples.

Gelatin microsphere-coated Fe3O4@graphene quantum dots (Fe3O4@GQD@GM) were designed and synthesized as a novel sorbent via ultrasonic-assisted dispersive magnetic solid-phase extraction (UA-DMSPE) method. The synthesized sorbent was identified and confirmed by FT-IR, XRD, VSM, and SEM techniques. UA-DMSPE was combined with corona discharge ion mobility spectrometry for trace determination of desipramine, sertraline, and citalopram. Effective parameters were considered and optimized. The proposed method, under optimal conditions, showed excellent linearity in different concentration ranges (2-700 ng mL-1, R2 > 0.995), repeatability (RSD < 5.1%), good sensitivity (LODs in the range 0.6-1.5 ng mL-1), high preconcentration factor (PF = 207-218), and acceptable relative recoveries (93.5-101.8%). Eventually, this method was used to determine tricyclic antidepressants in various biological samples. Schematic presentation of the microextraction and monitoring of TCAs by ultrasonic-assisted dispersive magnetic solid phase microextraction-ion mobility spectrometry producer.

Efficacy of DRIVEN-FLOW® M7-II, a new on-site drug screening kit in postmortem urine compared with Triage DOA®.

The efficacy of DRIVEN-FLOW® M7-II(DFM7II) for seven drug groups was compared with Triage DOA® (Triage) using 340 autopsy urine samples taken from bodies within 1 month of death based on mass screening analysis of GC/MS and LC-MS/MS. The sensitivity to benzodiazepines was 0.56 in Triage and 0.53 in DFM7II with few false positives, and their accuracy was 0.88. Triage detected triazolo diazepine derivatives more easily than DFM7II. DFM7II detected diazepam and nitro benzodiazepines more easily than Triage. There were nine amphetamine false-positive cases of more than 10 days after death in Triage, but these were absent in DFM7II during this period. The accuracy of amphetamines for Triage was 0.96 and for DFM7II was 1. Tricyclic antidepressant (TCA) was detected in five cases by mass analysis, while there were four false-positive cases using Triage and eight cases using DFM7II. In the TCA false-positive cases of both kits, tricyclic psychotics such as chlorpromazine, carbamazepine, and quetiapine were included as well as the drug poisoning cases. There were no samples containing cocaine or THC. The accuracy of DFM7II for opiate and barbiturates was 1, but those of Triage was less than 1. Based on the above, DFM7II is a more accurate kit with fewer false-positives for target drug groups, other than TCA, than Triage.

Supramolecular microextraction combined with paper spray ionization mass spectrometry for sensitive determination of tricyclic antidepressants in urine.

This work describes a novel methodology to analyze four tricyclic antidepressants (amitriptyline, doxepin, imipramine and, nortriptyline) in urine samples by combining supramolecular microextraction and paper spray ionization mass spectrometry (PS-MS). The proposed method uses a supramolecular solvent in which reverse micelles of 1-decanol are dispersed in tetrahydrofuran (THF)/water. The extraction of the tricyclic antidepressants at pH 9.0 requires a sample volume of 10.0 mL, short extraction time (1.0 min of extraction and 5 min of centrifugation), low amounts of organic solvent (50 µL of 1-decanol and 200 µL of THF), and provides high preconcentration factors: 96.9 to amitriptyline, 93.6 to doxepin, 71.3 to imipramine, and 146.9 to nortriptyline. The quantification by PS-MS is fast and straightforward because chromatographic separation is not required and all analytes were determined simultaneously. The limits of detection (LOD), quantification (LOQ), and the precision (RSD, %) of the developed method ranged between 5.2 and 8.6 µg L-1, 17.4-28.7 µg L-1 and 1.3-12.9%, respectively. Urine samples of five individuals (three males and two females) were used for accuracy evaluation. The accuracy obtained in these spiked urine samples at µg L-1 levels varied from 95.3 to 112.0%. The method also provided clean mass spectra with a high signal-to-noise ratio, which demonstrates the analytical appeal combination of supramolecular microextraction with determination by paper spray mass spectrometry.

On-disc electromembrane extraction-dispersive liquid-liquid microextraction: A fast and effective method for extraction and determination of ionic target analytes from complex biofluids by GC/MS.

In this study, an electromembrane extraction-dispersive liquid-liquid microextraction (EME-DLLME) was performed using a lab-on-a-disc device. It was used for sample microextraction, preconcentration, and quantitative determination of tricyclic antidepressants as model analytes in biofluids. The disc consisted of six extraction units for six parallel extractions. First, 100 µL of a biofluid was used to extract the analytes by the drop-to-drop EME to clean-up the sample. The extraction then was followed by applying the DLLME method to preconcentrate the analytes and make them ready for being analyzed by gas chromatography (GC). Implementing the EME-DLLME method on a chip device brought some significant advantages over the conventional methods, including saving space, cost, and materials as well as low sample and energy consumption. In the designed device, centrifugal force was used to move the fluids in the disc. Both sample preparation methods were performed on the same disc without manual transference of the donor phases for doing the two methods. Scalable centrifugal force made it possible to adjust the injection speed of the organic solvent into the aqueous solution in the DLLME step by changing the spin speed. Spin speed of 100 rpm was used in dispersion step and spin speed of 3500 rpm was used to sediment organic phase in DLLME step. The proposed device provides effective and reproducible extraction using a low volume of the sample solution. After optimization of the effective parameters, an EME-DLLME followed by GC-MS was performed for determination of amitriptyline and imipramine in saliva, urine, and blood plasma samples. The method provides extraction recoveries and preconcentration factors in the range of 43%-70.8% and 21.5-35.5 respectively. The detection limits less than 0.5 µg L-1 with the relative standard deviations of the analysis which were found in the range of 1.9%-3.5% (n = 5). The method is suitable for drug monitoring and analyzing biofluids containing low levels of the model analytes.

Triage DOA® versus INSTANT-VIEW M-1® in Urinary Drug Screening for Acute Drug Poisoning: A Prospective Cross-sectional Study.

Objective In the management of patients with suspected acute drug poisoning, a screening test using the patient's urine is usually performed. The Triage DOA® and INSTANT-VIEW M-1® kits are two commonly used point-of-care screening kits in Japan. However, the relationship between the results of these screening kits and the blood concentration of the poisoning drug is not clear. In this study, we evaluated which kit is more useful for acute drug poisoning screening based on a comparison of their results with the results of a serum drug analysis. Methods This prospective cross-sectional study investigated all patients with acute drug poisoning admitted to a general hospital in Tokyo, Japan, over a nine-month period. The Triage DOA® and INSTANT-VIEW M-1® screening kits were used, and a qualitative serum analysis was conducted simultaneously in all cases. We compared the kits for use in screening patients with acute drug poisoning and evaluated the utility of the kits. Results For the 117 patients enrolled in this study, the 2 kits showed different sensitivities to benzodiazepines (Triage®, 78.6%; INSTANT-VIEW®, 90.5%). Both kits showed high sensitivity to barbiturates (Triage®, 87.0%; INSTANT-VIEW®, 91.3%) but low sensitivity to tricyclic antidepressants (Triage®, 25.0%; INSTANT-VIEW®, 45.8%). Conclusion Because the sensitivity varies depending on the kind of drug, it is difficult to discuss the superiority of these kits. However, this study compared the results of two types of urinary drug screening kits with the results of qualitative analysis of drugs in serum as a gold standard, providing important reference data.

Development of green sodium sulfate-induced solidification of floating organic droplets-dispersive liquid phase microextraction method: Application to extraction of four antidepressants.

Sodium sulfate-induced deep eutectic solvent-based solidification of floating organic droplets-dispersive liquid phase microextraction was developed prior to gas chromatography-mass spectrometry. In this method, a mixture of Na2 SO4 solution (as phase separation agent and disperser) containing menthol-decanoic acid was rapidly injected into an alkaline aqueous solution containing Na2 SO4 . The solution was placed in an ice bath and the menthol-decanoic acid solvent was solidified on the surface of the solution. Under the optimal conditions, the enrichment factors and extraction recoveries were 122-147 and 74-89%, respectively. Finally, an aliquot of the collected organic phase was removed and mixed with acetonitrile and injected into the separation system. The limits of detection and lower limits of quantification were obtained at the ranges of 13-25 and 24-41 ng L-1 , respectively. The relative standard deviations of the proposed method were ≤11% for intra- and inter-day precisions at four concentrations.

Combination of dispersive solid phase extraction and deep eutectic solvent-based air-assisted liquid-liquid microextraction followed by gas chromatography-mass spectrometry as an efficient analytical method for the quantification of some tricyclic antidepressant drugs in biological fluids.

A dispersive solid phase extraction coupled with deep eutectic solvent-based air-assisted liquid-liquid microextraction has been developed and applied to the extraction and preconcentration of some tricyclic antidepressant drugs in the human urine and plasma samples prior to their determination by gas chromatography-mass spectrometry. In this method, a sorbent (C18) is first added into an alkaline aqueous sample and dispersed by vortexing. By this action, the analytes are adsorbed onto the sorbent. Then, the sorbent particles are isolated from the aqueous solution by centrifugation. Afterward, a deep eutectic solvent, prepared from choline chloride and 4-chlorophenol is used to desorb the analytes from the sorbent. Subsequently, the supernatant solution is removed and added into an alkaline deionized water placed into a test tube with a conical bottom. The resulting mixture is rapidly sucked into a glass syringe and then injected into the tube. This procedure is repeated for several times and a cloudy solution consisting of fine droplets of deep eutectic solvent dispersed into the aqueous phase is formed. After centrifuging the obtained cloudy solution, the tiny droplets of the extractant, containing the extracted analytes, settle at the bottom of the tube. Finally, an aliquot of the extractant is taken and injected into the separation system for quantitative analysis. Several significant factors affecting the performance of the proposed method are evaluated and optimized. Under optimum extraction conditions, the method shows low limits of detection in the ranges of 5-10, 8-15 and 32-60 ng L-1 in deionized water, urine, and plasma, respectively. Enrichment factors are observed to be between 325 to 385 in deionized water, 155 to 185 in urine, and 64 to 72 in plasma. Extraction recoveries are in the range of 65-77 (in deionized water), 62-74 (in urine), and 64-72% (in plasma). The relative standard deviations of the proposed method are ≤ 6% for intra- (n = 6) and inter-day (n = 4) precisions at a concentration of 200 ng L-1 of each analyte. Finally, the applicability of the introduced method is investigated by analyzing the selected drugs in different biological fluids. In the proposed method, for the first time, a deep eutectic solvent composed of safe, cheap, and biodegradable compounds was synthesized and used (at µL-level) as an elution and extraction solvent, simultaneously which led to omit the consumption of toxic organic solvents. This represents a significant advantage in the era of green chemistry. In addition, the introduced method is sensitive, simple in operation, rapid, and efficient.

A new carbon paste electrode modified with MWCNTs and nano-structured molecularly imprinted polymer for ultratrace determination of trimipramine: The crucial effect of electrode components mixing on its performance.

A novel carbon nanocomposite paste electrode was prepared and used as a voltammetric sensor for ultratrace determination of trimipramine (TRI) which currently used for the treatment of psychiatric disorders. For this aim, nanoparticles of molecularly imprinted polymer (MIP), synthesized by precipitation polymerization method, and multi-walled carbon nanotubes (MWCNTs) were embedded in a nanocomposite paste electrode. The nanocomposite mixing style demonstrated a significant influence on the final electrode performance. The sensor exhibited linear response range of 1.0 × 10-10-2.5 × 10-8 mol L-1 and very high sensitivity of 2131 µA µâ€¯mol L-1. The lower detection limit of the sensor was calculated to be 4.5 × 10-11 mol L-1 (S/N = 3). This sensor was applied successfully for highly selective determination of TRI in pharmaceutical formulations, urine and serum samples without applying any sample pretreatment.

Pharmacokinetic determination and analysis of nortriptyline based on GC-MS coupled with hollow-fiber drop-to-drop solvent microextraction technique.

AIM: A simple, sensitive and robust technique of hollow-fiber drop-to-drop solvent microextraction coupled with GC-MS has been successfully developed for the detection of antidepressant drug nortriptyline in human blood and urine samples. The recoveries of the drug from the spiked samples are found to be well within the range and appropriate to support the method. RESULTS: The LOD for the drug was obtained to be 0.007, 0.009 and 0.021 µg ml-1 in deionized water, urine and blood samples of human subjects, respectively. Linearity was obtained over the concentration range of 0.5-5.0 mg l-1 in deionized water with correlation coefficient 0.99672.

Acute Toxicity From Intravenous Use of the Tricyclic Antidepressant Tianeptine.

Tianeptine (7-[([3-chloro-6,11]-dihydro-6-methyldibenzo[c,f][1,2]thiazepin-11-yl) amino] heptanoic acid S, S dioxide) is a tricyclic compound prescribed as an antidepressant in European countries, but is not currently approved for use in the United States. There are few published case reports of tianeptine intoxication. Presented is the first case of acute toxicity associated with the intravenous use of tianeptine. A 36-year-old male intentionally injected tianeptine powder intravenously to "help him see into the future". He became unresponsive and a bystander called emergency medical services. Upon arrival to the Emergency Department, excessive constriction of the pupils, sedation, and a respiratory rate of 6 respirations per minute (rpm) were noted. Blood and urine were collected ~2 h post admission. The patient's serum ethanol concentration was 133 mg/dL. His toxicity was successfully reversed with two doses of naloxone 0.4 mg IV. He was started on a naloxone infusion at 0.2 mg/h and discharged 13 h after admittance awake, alert and oriented. The patient's urine sample screened negative for common drugs of abuse and total tricyclic antidepressants. A high performance liquid chromatography tandem mass spectrometry method was developed and validated to quantify tianeptine in urine. The calibration range was 1-100 ng/mL with linear regression correlation (r2) of 0.9996 or greater. The limit of quantitation was administratively set at 1 ng/mL. The bias of the assay was determined to be within ±20% of the target value for each quality control specimen. The intra-day and inter-day precision did not exceed 15% coefficient of variation for each quality control specimen. Matrix effects, absolute recovery, carryover and specificity were also evaluated. The patient's tianeptine urine concentration was determined to be 2 ng/mL.

Development of a novel ultrasonic-assisted magnetic dispersive solid-phase microextraction method coupled with high performance liquid chromatography for determination of mirtazapine and its metabolites in human urine and water samples employing experimental design.

Ultrasonic-assisted magnetic dispersive solid-phase microextraction coupled with high performance liquid chromatography has been developed for extraction and determination of mirtazapine, N-desmethyl mirtazapine, and 8-hydroxy mirtazapine in human urine and water samples. Magnetic graphene oxide-polyaniline nanocomposite (MGOPA) as a novel SPME sorbent was synthesized and used for the microextraction process. The analytical performance of MGOPA was compared with magnetic graphene oxide nanocomposite and indicated that the new sorbent was quite effective for extraction of mirtazapine, N-desmethyl mirtazapine, and 8-hydroxy mirtazapine. A two-stage experimental design approach, Plackett-Burman screening design and Box-Behnken optimization design, was used for screening and optimizing of significant variables in the microextraction process. The practical applicability of the proposed method was assessed by studying the linearity, intra-day and inter-day accuracy, enrichment factor, and precision. This method can be satisfactorily applied to the determination of mirtazapine and its metabolites in human urine and environmental water samples. Graphical Abstract Magnetic graphene oxide-polyaniline nanocomposite.

Metabolism and bioactivation of the tricyclic antidepressant amitriptyline in human liver microsomes and human urine.

AIM: Amitriptyline is a widely used tricyclic antidepressant, but the metabolic studies were conducted almost 20 years ago using high-performance liquid chromatography coupled with ultraviolet detector or radiolabeled methods. RESULTS: First, multiple ion monitoring (MIM)- enhanced product ion (EPI) scan was used to obtain the diagnostic ions or neutral losses in human liver microsome incubations with amitriptyline. Subsequently, predicted multiple reaction monitoring (MRM)-EPI scan was used to identify the metabolites in human urine with the diagnostic ions or neutral losses. Finally, product ion filtering and neutral loss filtering were used as the data mining tools to screen metabolites. Consequently, a total of 28 metabolites were identified in human urine after an oral administration using LC-MS/MS. CONCLUSION: An integrated workflow using LC-MS/MS was developed to comprehensively profile the metabolites of amitriptyline in human urine, in which five N-acetyl-l-cysteine conjugates were characterized as tentative biomarkers for idiosyncratic toxicity.

Electromembrane extraction through a virtually rotating supported liquid membrane.

Electromembrane extraction (EME) of model analytes was carried out using a virtually rotating supported liquid membrane (SLM). The virtual (nonmechanical) rotating of the SLM was achieved using a novel electrode assembly including a central electrode immersed inside the lumen of the SLM and five counter electrodes surrounding the SLM. A particular electronic circuit was designed to distribute the potential among five counter electrodes in a rotating pattern. The effect of the experimental parameters on the recovery of the extraction was investigated for verapamil (VPL), trimipramine (TRP), and clomipramine (CLP) as the model analytes and 2-ethyl hexanol as the SLM solvent. The results showed that the recovery of the extraction is a function of the angular velocity of the virtual rotation. The best results were obtained at an angular velocity of 1.83 RadS(-1) (or a rotation frequency of 0.29 Hz).The optimization of the parameters gave higher recoveries up to 50% greater than those of a conventional EME method. The rotating also allowed the extraction to be carried out at shorter time (15 min) and lower voltage (200 V) with respect to the conventional extraction. The model analytes were successfully extracted from wastewater and human urine samples with recoveries ranging from 38 to 85%. The RSD of the determinations was in the range of 12.6 to 14.8%.

Buyer Beware: Pitfalls in Toxicology Laboratory Testing.

Urine drug screens are commonly used in various clinical settings and situations. Immunoassays are the most commonly available method of testing for urine drug screens in hospitals. Although convenient, immunoassays are prone to false positive and false negative results. It is important for the health care provider to understand the principles of the laboratory methods involved with urine drug screens as this will help guide appropriate result interpretation and therefore improve clinical care.

[The determination of amitriptyline and its metabolite, nortriptyline, in the biological objects of the corpse by the high-performance liquid chromatography technique].

Tricyclic antidepressants are among the preparations that most frequently cause intoxication in adults and children; moreover, poisoning with these substances not infrequently has a fatal outcome. Medications belonging to this group, such as amitriptyline, are extensively used to manage manifestations of depression, anxiety, migraine, neuropathic pain, and hyperactivity syndrome. Amitriptyline overdosage causes non-specific symptoms of intoxication, and its clinical picture does not allow to identify the nature of a psychotropic xenobiotic. Of primary importance in connection with this is to establish the cause of intoxication or death by the clinical toxicological and forensic medical methods based on the results of the fast identification and quantitation of amitriptyline in biological materials including blood, urine, hepatic tissues, etc. The authors describe the method for the determination of amitriptyline and its principal physiological metabolite nortriptyline in biological objects with the help of high performance liquid chromatography (HPLC).

Method validation of a tricyclic antidepressant drug panel in urine by UPLC-MS/MS.

UNLABELLED: Tricyclic antidepressant (TCA) drugs are also used as adjunctive therapy to treat chronic pain. To monitor patient compliance to therapy, urine specimens may be preferred since collection is non-invasive, and the specimen can provide a longer detection window. TCA drugs are frequently monitored by immunoassay; however, poor antibody specificity may compromise results. The purpose for this study was to develop a confirmation method for determining TCA in urine specimens by ultra-pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Our method can quantify 9 TCA drugs in less than 5 min. Method validation experiments were performed, and the coefficient variation for inter- and intra-day precision was within 12% for each analyte at five different concentrations. Accuracy studies had good agreement with another laboratory that performs testing by GC/MS. HIGHLIGHTS: 1. Tricyclic antidepressants are commonly used to treat depression, anxiety, and neuropathic pain. 2. A confirmation method by ultra-pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) to determine TCA in urine specimens was developed. 3. Urine testing for TCA is preferred for monitoring adherence to therapy due to ease of collection.