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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 C 18 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 ( r 2 > 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.

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.

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 μ 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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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.

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., (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

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.

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.

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.

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., (© 2014 by the Association of Clinical Scientists, Inc.)

Variability in imipramine and desipramine metabolism was evaluated using urinary excretion data from patients with pain. Liquid chromatography-tandem mass spectrometry was used to quantitate concentrations in urine specimens. Interpatient population contained 600 unique imipramine specimens, whereas intrapatient population had 137 patients with two or more specimens. Normal concentration ranges of imipramine, desipramine and the desipramine/imipramine metabolic ratio (MR) were established, and various factors were tested for MR impact. Geometric mean of imipramine urine concentration was 0.46 mg/g of creatinine, and desipramine was 0.67 mg/g of creatinine. Gender, concomitant known CYP2C19 inhibitor use and urine pH did not affect MR. However, proton-pump inhibitor (PPI) users had a significantly lower mean MR than those without a listed PPI. Early age group (18-36 years) had a significantly higher mean MR than middle (37-66 years) and late (67-90 years) age groups. Approximately one-third were positive for one or more of hydrocodone, oxycodone, hydromorphone or oxymorphone. Patients with no opioids reported in the medication list had a significantly lower geometric mean MR than those with prescribed opioids (1.03 vs. 1.54, P = 0.004). Patients with only one prescribed opioid had a lower MR than those with two or more prescribed opioids. Patients with younger age, prescribed opioids and no listed PPI were more likely to have a higher geometric mean urinary desipramine/imipramine MR., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

A sensitive, specific, and rapid high-pressure liquid chromatography/mass spectrometry/mass spectrometry method was developed for the quantitation of 11 tricyclic antidepressants and/or their metabolites; fluoxetine and norfluoxetine; cyclobenzaprine; and trazodone in urine. Samples were alkalinized with 0.2 N NaOH and extracted into 2 ml of hexane: ethyl acetate (1:1), evaporated to dryness, and reconstituted with 100 μl of 20 mM ammonium formate: methanol (20:80). The chromatographic separation was performed using an Allure Biphenyl 100 × 3.2 mm, 5-μ column with a mobile phase consisting of 20 mM ammonium formate: methanol (20:80 v/v) at a flow rate of 0.5 ml/min. The detection was accomplished by multiple-reaction monitoring via electrospray ionization source operating in the positive ionization mode. The calibration curve was linear over the investigated concentration range, 25-2,000 ng/ml, for each analyte using 1.0 ml of urine. The lower limit of quantitation for each analyte was 25 ng/ml. The intra- and inter-day precisions had coefficient of variation less than 15% and the accuracy was within the range from 88% to 109%. The method proved adequate for the tricyclic antidepressants analysis of urine for emergency clinical toxicology and pain management compliance testing., (© 2012 Wiley Periodicals, Inc.)

A rapid method for the simultaneous determination of 42 psychoactive drugs and their metabolites (barbitals, benzodiazepines, tricyclic antidepressants, phenothiazines, etc. ) in human plasma and urine was developed using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). After a simple protein precipitation step, the analysis of the drugs and the metabolites was achieved on an Acquity UPLC BEH C18 column by using the gradient elution with ammonium acetate and methanol-acetonitrile (1: 1, v/v) as the mobile phases, and detected by electrospray ionization-tandem mass spectrometry in the multiple reaction monitoring (MRM) mode operated simultaneously in both positive and negative modes by rapid switching, and quantified by matrix-match standard solution. The average recoveries were 60.2% - 125% and 64.5% - 126% for the drugs in plasma and urine except those of perphenazine, thioridazine and chloropromazine in plasma were 37.6% - 57.5%, 36.3% - 48.3%, 52.4% -67.4%, respectively; trazodone and diazepam in urine were 100% -142% and 108% - 177%, respectively. The relative standard deviations (RSDs) of all the drugs in plasma and urine were within 0.8% - 26% and 2.6%18% (n = 6), respectively. The detection limits of the 4 barbitals ranged from 20 to 100 mg/L and those of the other drugs ranged from 0.05 to 2.0 mg/L. The method is simple, selective and sensitive to detect drugs for both clinical and forensic purposes.

A method for the simultaneous determination of six commonly prescribed cyclic antidepressants and their major metabolites in urine is presented. This method can be used for quantitation of amitriptyline, nortriptyline, imipramine, desipramine, doxepin, desmethyldoxepin, and maprotiline in human urine, in addition to the qualitative determination of their hydroxylated metabolites. This method is suitable for confirmation of drug abuse in health care professionals and overdose cases where the identity of the abused cyclic antidepressant may not be known. Samples are spiked with internal standard and hydrolyzed with beta-glucuronidase from Escherichia coli. Hydrolysis is found to be essential to the extraction procedure as the tertiary cyclic antidepressants are found to be extensively conjugated in urine. The secondary cyclic antidepressants, on the contrary, are found to be minimally conjugated. Drugs are extracted from alkalinized urine into solvent and derivatized with MSTFA/ammonium iodide/ethanethiol reagent. This reagent produces more stable derivatives compared to reagents previously employed. Gas chromatographic (GC)-mass spectrometric analysis is performed in electron ionization mode by selective ion monitoring, using hydrogen as a carrier gas, a short narrow bore GC capillary column, and fast temperature program, allowing for a rapid analytical cycle. While maintaining specificity for these drugs, concentrations in human urine ranging from 50 to 20,000 ng/mL can be measured with intraday and interday precisions, expressed as variation coefficient, of less than 2.8% for all analytes.

Patients taking tricyclic antidepressants (TCA) can experience toxicity or severe side effects. As a rapid and less technically demanding alternative to quantitative serum analysis, most laboratories offer qualitative immunoassays to assist in the evaluation of a suspected TCA overdose. However, the relationship between quantitative serum and qualitative urine levels of TCA-related compounds and their metabolites has not been comprehensively studied. Serum high-performance liquid chromatography results were compared to the qualitative urine results using the Syva Rapid Test and the Biosite Triage. Serum concentrations of amitriptyline, desipramine, doxepin, imipramine, and nortriptyline ranging from subtherapeutic to toxic triggered a positive response on both urine immunoassay devices. On the other hand, neither immunoassay uniformly detected clomipramine, even at serum levels greater than the therapeutic range. False positives due to cyclobenzaprine were more common with the Biosite assay. For virtually all positive urine TCA findings, it was not possible to determine whether the positive results corresponded to subtherapeutic, therapeutic, supratherapeutic, or toxic serum concentrations. Because urine immunoassays are the only option for many laboratories analyzing specimens for TCAs (especially in an emergency setting), clinicians must understand the limitations and interpret results in conjunction with clinical findings and/or quantitation of serum levels.

Improvements have been made in recent years in on-site or point-of-care (POC) immunoassay testing devices. We evaluated the DrugCheck 9 cup, a new qualitative visually read, competitive binding, immunoassay cup that measures nine analytes. The study was performed according to the recent National Laboratory Certification Program (NLCP) guidelines for validating a laboratory-based immunoassay. The study included a linearity challenge with 5 replicates at concentrations 0, 25%, 50%, 75%, 100%, 125%, and 150% of the cutoff and also determination of the limit of detection. Interference (specificity) studies were included to evaluate the cross-reactivity of common structural analogues or others purported to interfere with the assay. The ability to differentiate positive and negative samples was evaluated by positive and negative quality control specimens and also parallel comparisons of laboratory verified positive and negative donor specimens previously analyzed by immunoassay and gas chromatography-mass spectrometry. The results indicated a high degree of correlation (97.6%) with the laboratory immunoassay and a high degree of specificity and sensitivity with extended linearity below the cutoff.

In six cases of suspected opipramol overdose, commercially available immunoassays for tricyclic antidepressants (TCA) EMIT tox serum Assay and ADxR serum TCA Assay indicated arbitrarily high or toxic TCA concentrations. However, opipramol concentrations determined by high-performance liquid chromatography (HPLC) analysis were in the high-normal or low-toxic range. This finding prompted us to study opipramol metabolism by mass spectral techniques and to determine the cross-reactivity of opipramol and its metabolites in immunoassays. Three previously unknown metabolites (I, II, V) included an oxidation product of the hydroxyethyl moiety to an acetic acid group at the piperazine side chain (1), a decarboxylation product of the latter metabolite (II), and opipramol-N-oxide (V). In addition, two previously reported metabolites were identified, which included a deshydroxyethyl metabolite (III) and dibenzazepine (IV). One of the major metabolites of opipramol is the acetic acid metabolite (I), which may exceed the opipramol plasma concentration immensely and contribute to an arbitrarily high concentration in commercially available immunoassays. The cross-reactivities of the metabolite (I) were determined to be 64 and 66% with EMIT and ADx, respectively.