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Abstract

Depression is one of the most prevalent but severe of mental disorders, affecting thousands of individuals across the globe. Depression, in its most extreme form, may result in self-harm and an increased likelihood of suicide. Antidepressant drugs are first-line medications to treat mental disorders. Unfortunately, these medications are also prescribed for other in- and off-label conditions, such as deficit/hyperactivity disorders, attention disorders, migraine, smoking cessation, eating disorders, fibromyalgia, pain, and insomnia. This results in an increase in the use of antidepressant medications, leading to clinical and forensic overdose cases that could be either accidental or deliberate. The findings revealed that people who used antidepressants had a 33% greater chance of dying sooner than expected, compared to those who did not take the medications. Analytical techniques for precisely identifying and detecting antidepressants and their metabolic products in a variety of biological matrices are greatly needed to be developed and made available. Hence, this study attempts to discuss various analytical techniques used to identify and determine antidepressants in various biological matrices, which include urine, blood, oral fluid (saliva), and tissues, which are commonly encountered in clinical and forensic science laboratories.

Keywords

antidepressant determination analytical biological matrices chromatography

Introduction

Over 280 million people suffer from depression globally.¹ Depression is one of the most frequently observed disorder which is a common and severe medical condition that can affect anyone, regardless of their economic or social status.^2,3 It is characterised by poor concentration, a depressed mood, loss of interest and desire, sleep issues, exhaustion, self-harming, or suicidal behaviour, as well as by impaired social and professional functioning. According to one approximation, about 3.8% of the population of the world, which would include 5% of adolescents and 5.7% of those over the age of 60, suffer from depression.⁴ It is among the leading causes of morbidity and is connected to a higher mortality rate.³ Antidepressant therapy could potentially reduce the negative health consequences of depression by acting directly on the physiological pathways that lead to greater morbidity and mortality or, indirectly, by improving compliance with the management of comorbid disorders and facilitating a healthier lifestyle.⁵

There are five main classes of antidepressants (ADs) and many more that are less commonly prescribed, such as (1) Selective serotonin reuptake inhibitors (SSRIs), which include escitalopram (Lexapro), citalopram (Celexa), sertraline (Zoloft), paroxetine (Pexeva, Paxil), and fluoxetine (Prozac); (2) Serotonin and norepinephrine reuptake inhibitors (SNRIs), which include venlafaxine (Effexor XR), desvenlafaxine (Pristiq), duloxetine (Cymbalta), and levomilnacipran (Fetzima); (3) Monoamine oxidase inhibitors (MAOIs), including tranylcypromine (Parnate), isocarboxazid (Marplan), and phenelzine (Nardil); (4) Tricyclic antidepressants (TCAs), which include desipramine (Norpramin), doxepin, amitriptyline, nortriptyline (Pamelor), and imipramine (Tofranil); and (5) Atypical, which include vortioxetine (Trintellix), bupropion (Wellbutrin SR, Wellbutrin XL), vilazodone (Viibryd), mirtazapine (Remeron), and trazodone.^6,7

A significant remission of depression has been observed with therapy; even so, as many as 55% of patients were considered treatment-resistant even after more than 12 months of medication.⁸ Previous studies affirmed that people who were on antidepressants had 33% more chances of dying prematurely as compared to those who were not.⁹ Antidepressant users were also found to have 14% more risk of an adverse cardiovascular event, viz. stroke or heart failure.⁹ It has been observed that this class of drug has significant scope for abuse. Especially in cases with a history of simultaneous usage of other drugs or alcohol, both clinical and forensic scenarios come to play, since there may be a possible case of drug abuse or suicide attempt, which can be either voluntary or accidental.¹⁰

The high rate of consumption of ADs might be regarded a public health issue; hence, it is critical to compile the latest literature on the subject (which include analytical techniques used for the detection, identification, characterization, and quantification of ADs) to assist clinical/forensic toxicologists to enhance patient’s quality of life. Hence, the main goal of this research is to summarize various analytical techniques used to identify and determine antidepressants in various biological specimens, such as oral fluid (saliva), blood, hair, urine, and tissues, commonly encountered in emergency department and forensic science laboratories.

The article provides an in-depth, scholarly, and insightful review of analytical techniques adopted for ADs in various matrices with varying infrastructural facilities. Over time, this study illuminates the underlying science and the evolution of the different methods that evaluate the field’s status by putting recent developments into proper perspective and context and speculating on possible future developments. Depending on the infrastructural facilities available with their laboratories, the matrices they have to deal with, the required levels of accuracy, and the speed with which the results are delivered, the analysts/researchers can choose an appropriate method from the different methods given in there for their use. Further, this review helps chemists and toxicologists to improvise and adapt the methodologies for the application of the techniques to a different nature of the sample or matrix.

Identification of Related Studies

For all relevant publications, a comprehensive literature search was conducted by utilizing web-based search engines, ie, ‘PubMed’, ‘Scopus’, using the following keywords: “Antidepressant”, “Biological Matrix”, “Analytical Techniques”, “Detection”, “Analysis”, “Blood”, “Hair”, “Saliva”, “Oral Fluid”, “Intraosseous Fluid”, and “Forensic Toxicology”. The identification of all relevant articles (from 2011 to 2022) was facilitated using broad search terms, with the last search done in July 2022. This systematic review covers about 55 articles that met the search criteria.

Analytical Methodologies Used in Analysing Biological Matrices

Electrophoresis

Świądro et al (2020)¹¹ developed a method that utilizes the combination of Dried Blood Spot (DBS) (assisted by microwave-assisted extraction (MAE)) and the capillary electrophoresis-mass spectrometry (CE–MS) (DBS/MAE/CE-MS) for identifying and determining psychotropic drugs from different groups, including hypnotics (zolpidem), benzodiazepines (tetrazepam), SSRIs (citalopram), and TCAs (imipramine, amitriptyline). Limit of quantification (LOQ) = 5.25-49.0 ng/mL and detection (LOD) = 1.76-14.7 ng/mL, intra-(CV = 3.26-18.52%) and inter-(CV = 1.31-9.43%) day precision of the determinations, matrix effect on ionization of analytes (ME = 98.6-105.5%) and recovery (RE = 85.0-105.4%) were determined. Moreover, the developed technique was selective, and after 7 and after 14 days, it was found that the analytes present in the blood put on DBS cards were stable. Furthermore, the established approach was effectively applied to the analysis of blood and postmortem samples collected from patients who had been treated with the analysed drugs.

Gas Chromatography

Vitreous humor (VH) has emerged as a significant biological matrix in forensic toxicological investigations. Nitoupa et al (2020)¹² simultaneously determined and quantified the 9 most commonly used ADs (venlafaxine, sertraline, mirtazapine, maprotiline, fluoxetine, clomipramine, citalopram, nortriptyline, and amitriptyline) and their 4 metabolites (Odesmethylvenlafaxin, desmethylsertraline, desmethylmirtazapine, desmethylmaprotiline) in VH after solid phase extraction (SPE) and derivatization with heptafluorobutyric anhydride followed by GC/MS. The developed method was applied successfully to VH samples from 43 blood-AD positive cases. Whenever ADs were detected in blood, they were also detected in the corresponding VH samples. The VH/blood concentration ratios were also determined and were found to range from .04 to 7.07. Venlafaxine and its metabolite O-desmethylvenlafaxine, citalopram, mirtazapine, and its metabolite desmethylmirtazapine, were found. The findings indicate that VH might be used in forensic toxicology as an alternate biological material to determine ADs, particularly when urine and/or blood are unavailable.

dos Santos et al (2016)¹³ performed a matrix effect study to verify the analytical compatibility among human and bovine vitreous and saline solution (SS) (NaCl .9%). As model analytes, TCAs [desipramine (DES), imipramine (IMI), nortriptyline (NTR), and amitriptyline (AMI)] were utilised and were extracted from samples utilizing liquid-phase microextraction (LPME) and identified using GC/MS. Human and bovine VH and SS samples were prepared with six different concentrations of ADs (200, 160, 120, 80, 40, and 5 ng/mL) and were analysed. There was no significant matrix effect for NTR or AMI in the three matrices evaluated. However, there was a lot of variation for DES and IMI across all matrices. Once the compatibility of the matrices was established, the approach was completely verified for AMI and NTR in SS. The method was performed on 6 VH samples drawn from real-life cases whose femoral whole blood (FWB) was evaluated using a previously reported methodology. An average ratio (VH/FWB) of ∼.1 was reported for both compounds.

Cabarcos-Fernández et al (2022)¹⁴ developed a dispersive liquid–liquid microextraction (DLLME) method in conjunction with GC–MS for identifying and quantifying antidepressants in pericardial fluid. Validation results met acceptance criteria (U.S. Food and Drug Administration). Thirty-one pericardial fluid samples were analysed using the analytical technique. There were 21 positive samples, ranging in concentration from .19 to 8.48 μg/mL. The most frequently found antidepressants were venlafaxine and olanzapine.

Cartiser et al (2011)¹⁵ developed and validated a 3-step liquid–liquid extraction (LLE) with online derivatization (silylation) prior to GC-MS/MS on rabbit specimens to quantify four benzodiazepines (temazepam, oxazepam, nordazepam, and diazepam) and citalopram in 11 biological matrices (lung, bone marrow, adipose tissue, brain, skeletal muscle, kidney, liver, vitreous humor, bile, urine, and blood). The lower limit of quantitation (LLOQ) was 5 ng/g for citalopram, 1 ng/g for nordazepam, and 10 ng/g for temazepam, diazepam, and oxazepam. The combination of 3-step LLE with MS/MS gave high selectivity in all matrices, even those with the highest lipid content.

Soares et al (2021)¹⁶ employed a combination of dried saliva spots (DSS) extraction and GC-MS/MS method to simultaneously detect and quantify some most commonly prescribed ADs and their metabolites (paroxetine, sertraline, citalopram, O-desmethylvenlafaxine, venlafaxine, and fluoxetine) in 100 μL of oral fluid. The R2 (or coefficients of determination) of at least .99 for all analytes and linearity within the investigated range were considered, with limit of detection (LOD) and limit of quantitation (LOQ) ranging from 10 to 100 ng/mL. The method outlined herein is the first one to employ DSS for the extraction of ADs, and it has proven to be a sensitive, quick, and easy replacement for traditional methods that may be regularly used in forensic and clinical toxicology investigations.

Truta et al (2016)¹⁷ presented a solid phase extraction (SPE) and subsequent analysis by GC–MS/MS technique for determining 14 prevalent antidepressants (venlafaxine, trimipramine, sertraline, paroxetine, nortryptiline, mirtazapine, mianserin, imipramine, fluoxetine, dothiepin, clomipramine, citalopram, and amitriptyline) and 1 metabolite (N-desmethylclomipramine) in whole blood. LOQ and LOD were lower than subtherapeutic and therapeutic concentration limits.¹⁸

Kusano et al (2019)¹⁹ developed an QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe)-dispersive solid-phase extraction (DSPE) technique in combination with GC-MS/MS to quantify 63 pesticides and drugs of abuse in whole blood. The target analytes' LODs in whole blood were in the ng/mL range. Recovery rates ranged from 25% to 118% for pesticides, 28% to 44% for phenethylamines, 15% to 81% for drugs of abuse, 41% to 86% for tri/tetracyclic antidepressants, 36% to 110% for benzodiazepines, and 66% to 84% for barbiturates. The validated results were utilized to develop a systematic, simple-to-operate, and quantitative toxicological GC/MS/MS technique and “Quick-DB Forensic” software.

Papoutsis et al (2012)²⁰ used GC/MS to simultaneously identify and quantify the eleven most frequently prescribed antidepressants and four active metabolites (desmethyl-venlafaxine, venlafaxine, desmethyl-sertraline, sertraline, paroxetine, nortriptyline, desmethyl-mirtazapine, mirtazapine, desmethyl-maprotiline, maprotiline, fluvoxamine, fluoxetine, clomipramine, citalopram, and amitriptyline) in whole blood. The combination of SPE and HFBA derivatization efficiently improved sensitivity and reduced matrix effect of the method. More than 2500 forensic and clinical blood samples were analysed using the developed method, which was shown to be acceptable for routine work.

Fernandez-Lopez et al (2019)²¹ utilized SPE-GC-MS to determine duloxetine, amitriptyline, and venlafaxine in human bone. The assay was validated at concentrations ranging from .3-1 ng/mg (depending on the substance) to 500 ng/mg. The validated approach was then successfully applied to genuine bone specimens from forensic cases where toxicological testing for these drugs in blood had revealed a positive result. In all positive blood results, drugs were identified in the bone, with approximate amounts of 4.6-2 ng/mg for venlafaxine, 19.3-3 ng/mg for duloxetine, and 36.4 ng/mg for amitriptyline.

Khraiwesh et al (2011)²² developed an GC/MS technique for determining sertraline and its primary metabolite, desmethyl-sertraline, utilizing SPE and derivatisation with heptafluorobutyric anhydride (HFBA) in whole blood. The LOD and LOQ for desmethyl-sertraline and sertraline were 1.00 and .30 μg/L, respectively. Desmethyl-sertraline extraction efficiency varied from 84.9 (8.2)% to 107.7 (4.4)%, whereas for sertraline, the efficiency ranged from 90.1 (5.8)% to 95.4 (3.0)%. The developed approach might be used in routine clinical and forensic laboratory analysis, pharmacokinetic investigations, therapeutic sertraline monitoring, or examining sertraline-related forensic cases.

Boumba et al (2016)²³ described a rapid method, SPE-GC/MS, for determining the 5 frequently co-administered ADs (citalopram, clomipramine, sertraline, mirtazapine, bupropion) and antipsychotic drug (clozapine) in serum, plasma, and whole blood. The method has been fully validated and used for routine analyses. The calibration curve R2 for the tested drugs in serum, plasma and whole blood ranged from .9977 to .9999. For the analytes and matrices investigated, inter-day and intra-day precisions varied from 3.60-12.91%, and .81-7.85%, respectively.

Oliveira et al (2021)²⁴ developed, validated, and applied a method, combining the bar adsorptive microextraction technique (BAμE) with large-volume-injection gas chromatography-mass spectrometry (LVI-GC-MS) operating in the selected-ion monitoring (SIM) mode, for determining the 6 most common TCAs (dosulepin, mirtazapine, imipramine, trimipramine, mianserin, and amitriptyline) in urine specimens. The developed method (BAμE(C18)/LVI-GC-MS(SIM)) provided suitable matrix effects between 90.2 and 112.9% (RSD ≤ 13.9%), good recovery yields (ranging from 92.3 and 111.5%, RSD ≤ 12.3%), and a remarkable overall process efficiency (ranging from 84.9% to 124.3%, RSD ≤ 13.9%). The six TCAs were satisfactorily screened in real urine matrices using the established technique. In comparison to existing microextraction-based approaches, the suggested analytical methodology was demonstrated to be a superior analytical option for tracking trace levels of TCAs in complicated urine matrices.

Laudani et al (2021)²⁵ developed and validated an LLE-GC-MS (SIM) method for extracting and analysing several various frequently used antipsychotics and antidepressants from spiked pig ribs. The performance of the analytical technique was satisfactory in terms of validation acceptance criteria. Because no matrix interferences were found, this approach appears to have good selectivity. Furthermore, carryover had no effect on the method. Given the encouraging results of the pig rib investigation, this technology might be employed in forensic toxicological protocols (human postmortem cases) as an alternative to traditional procedures.

An alternate matrix for drug testing may be intraosseous fluid (IOF). Rodda et al (2018)²⁶ investigated whether postmortem specimen IOF might provide drug detection capabilities that closely matched the corresponding blood samples. The Arrow EZ-IO Intraosseous Vascular Access System was used to collect IOF from 4 anatomic collection locations of each deceased, the proximal humerus and right and left proximal tibia bone areas of 29 decedents. Standard autopsy samples, such as blood, were also obtained during the autopsy. IOF and blood specimens were immunoassayed using ELISA kits for cannabinoids, benzodiazepines, tricyclic antidepressants, phencyclidine, amphetamines, methamphetamine, cocaine, methadone, oxycodone, fentanyl analogs, and opioids. For drug targets, the correlation between IOF ELISA and cardiac/central blood ELISA findings was nearly perfect. Further blood confirmation analysis using GC/MS revealed a close association to IOF screen findings. There were no significant statistical differences between the IOF locations or body sides. This innovative research confirms the use of IOF as a possible less-compromised tissue in deteriorated or traumatised bodies as an alternative postmortem material for toxicological studies.

Liquid Chromatography

Hamedi and Hadjmohammadi (2016)²⁷ developed a sensitive and quick technique for determining fluoxetine in human plasma and urine samples utilizing alcohol-assisted dispersive liquid-liquid microextraction (AA-DLLME) and high-performance liquid chromatography (HPLC). Plackett-Burman design and Box-Benken design were used, accordingly, to evaluate and optimise the impacts of six parameters on the extraction recovery. In human plasma, the LOD for fluoxetine was 3 ng/mL, with a linearity range of 10-1200 ng/mL. The equivalent results for human urine were 4.2 ng/mL, with a linearity range of 10 to 2000 ng/mL. The method presented here successfully identified fluoxetine in human plasma and urine samples.

Elgawish et al (2020)²⁸ employed solid-phase extraction (SPE) in combination with liquid chromatography-tandem mass spectrometry (LC/MS/MS) technique to determine antidepressant drugs, sertraline, and citalopram levels in plasma samples (rat) after oral administration of a single-dose of 40 mg/kg of sertraline and citalopram. The linear range of this method for citalopram and sertraline was 1-2000 and 1-1000 ng/mL, with lower limits of detection (LLOD) of .35 and .18 ng/mL, respectively.

Breivik et al (2020)²⁹ validated an ultra-high performance liquid-chromatography -tandem mass spectrometry (UPLC-MS/MS) technique in conjunction with LLE for the quantification of mirtazapine, clozapine, and quetiapine in liver tissue, brain tissue, skeletal muscle, and postmortem whole blood. The validated ranges were 13-1060, 16-1960, and 3.8-1534 μg/L for mirtazapine, clozapine, and quetiapine, respectively. Within-run and between-run precision (CV 1.5-8.9%) and accuracy (87.4-122%), matrix effects (95-101%), and recovery (35.7-92%) were validated at 2 concentration levels; 20 and 849 μg/L for mirtazapine, 25 and 1568 μg/L for clozapine, and 5.8 and 1227 μg/L for quetiapine. The methodology was utilized in 3 forensic autopsy instances and involved, respectively, mirtazapine, clozapine, and quetiapine, in supratherapeutic concentrations.

Woźniakiewicz et al (2014)³⁰ validated a microextraction by packed sorbent (MEPS)/Ultrahigh-pressure (or performance) liquid chromatography-mass spectrometry (UHPLC-MS) technique for determining TCAs such as amitriptyline, imipramine, nortriptyline, desipramine, doxepin, and nordoxepin from human oral fluid. The LOQ value of .50 ng/mL obtained makes the MEPS/UHPLC-MS technique a valuable tool in forensic and clinical laboratories, as confirmed by real samples collected from three female patients (55-61 years old), suffering from depression and anxiety, voluntarily participating in the experiment.

Ma et al (2021)³¹ employed UPLC/MS analysis and SLE pre-treatment to determine 13 antidepressants (amitriptyline, clozapine, doxepin, trazodone, clomipramine, citalopram, sertraline, maprotiline, paroxetine, norfluoxetine, fluoxetine, desipramine, and imipramine) in blood samples. The LODs ranged between .0003-.003 ng/mL. The linear ranges for norfluoxetine, paroxetine, and doxepin are .01-200 ng/mL, whereas the remainder of the antidepressants has ranges of .001-200 ng/mL. The current approach was applied for antidepressant analysis in actual cases and is expected to be widely utilised for antidepressant analysis in biomedical and forensic cases.

To identify 19 antipsychotics and 22 antidepressants in DBSs and whole blood obtained from postmortem samples, Moretti et al (2019)³² developed an LC-MS/MS method. The LODs for antipsychotics and antidepressants are between .1 and 5.2 ng/mL and between .1 and 3.2 ng/mL. LOQs for both ranged from 5 to 10.0 ng/mL. Sixteen of the 60 cases examined tested positive for 17 different analytes; the technique was fully validated for 14 of them. The concentrations on DBSs were found to be in good agreement with those evaluated in conventional blood samples (taken simultaneously and held at −20°C). During the 3-month period, the percentage of degradation/enhancement for the majority of the compounds was less than 20%. DBSs can be employed for regular sample storage, according to results acquired from actual postmortem cases.

Majda et al (2020)³³ optimized the DI-SPME/LC-TOF-MS technique for detecting and quantifying a large group of psychotropic drugs, including benzodiazepines, SSRIs, SNRIs, TCAs, and sleeping pills “Z” in the postmortem biological matrix (bone marrow aspirate human and blood). Nonbenzodiazepine hypnotics are benzodiazepine derivatives, sometimes known as “Z-Drugs” since the names of many of the first of these drugs to be marketed begin with the letter “z.” There are three major class of z-drugs: imidazopyridine, pyrazolopyrimidine, and cyclopyrrolone. Although structurally unrelated to each other and to benzodiazepines, each has some affinity for the alpha-1 subunit of GABA-A receptors, which are concentrated in the thalamus, cortex, and cerebellum. Commonly prescribed Z-drugs include zolpidem, zopiclone, eszopiclone, and zaleplon, which differ in effectiveness and adverse effects because to differences in their pharmacokinetic profiles.³⁴ The validation parameters were determined, including LOD and LOQ, linearity, within-day and between-days precision, and matrix effect. All of the values obtained were within the acceptable range for toxicological studies. By analysing the postmortem samples, the method’s utility was confirmed. The highly accurate drug concentrations detected in real samples make the future general use of DI-SPME/LC-TOFMS in forensic toxicological investigations.

Johansen (2017)³⁵ developed an LC-MS/MS technique for measuring citalopram and demethylcitalopram enantiomers in human forensic samples' whole blood and postmortem blood. For all 4 enantiomers, the LOQ and LOD were .005 and .001 mg/kg, respectively. Following technique validation, a correlation with the racemic method was evaluated and determined to be satisfactory. The approach was then applied successfully to actual blood samples from forensic investigations (from Eastern Denmark for a period of 3-year), demonstrating that escitalopram was less prevalent in all forensic instances than citalopram.

Nedahl et al (2018)³⁶ established postmortem reference brain values and brain–blood ratios for widely prescribed antidepressants (sertraline, mirtazapine, duloxetine, and citalopram), which might help with postmortem case evaluation. In terms of linearity, precision, process efficiency, and accuracy, the analytical technique was effectively verified in brain tissue. The analytes were quantified using UPLC-MS/MS. Correlations between blood and brain concentrations were obtained, with R2 values ranging from .67 to .91. citalopram’s median brain-blood ratio was 3.71 (range: 1.4-5.9), mirtazapine’s was 1.53 (range: 1.02-4.71), duloxetine’s was 11.0 (range: 5.0-21.6), and sertraline’s was 7.38. (range: 3.2-14.2). The study findings might be used as reference concentrations in the brain in forensic investigations.

Orfanidis et al (2021)³⁷ studied the development of a Mini-QuEChERS based UHPLC–MS-MS technique for detecting and quantifying 84 drugs of abuse (new psychoactive substances (NPS), benzodiazepines, amphetamines, cannabinoids, cocaine, and opiates) and pharmaceuticals (antidepressants, antipsychotics, etc.) in postmortem blood. The method was shown to be sensitive and selective offering LOD ranging from .01 to 9.07 ng/mL. Validation involved assessing the method’s LOQ, matrix effect, carry-over, recovery, precision, and accuracy. The approach satisfied known bioanalytical requirements and was thus used to the study of postmortem blood acquired from chronic drug abusers, providing highly reasonable identification and quantitative measurement of substances in postmortem blood.

Campêlo et al (2021)³⁸ optimized QuEChERS method utilizing design of experiments (DOE) to evaluate the best conditions to obtain the most effective extraction. The antidepressants were identified and quantitated by LC-MS/MS analysis. The technique was verified for all analytes; the calibration curves were linear from 10-1000 ng/mL, with R2 greater than .98 and LOD and LOQ specified as 10 ng/mL. Overall, the optimized method was used to quantify the antidepressants in postmortem real sample analysis. This study established a realistic technique for regular forensic analysis, with rapid and easy sample preparation and an 8-minute total run time for each analysis.

Shi et al (2014)³⁹ have developed a high-throughput automated solid phase extraction coupled with liquid chromatography/quadrupole-time-of-flight mass spectrometry (ASPE-LC-Q-TOF/MS) technique for detecting antidepressants in blood. By correlating the extracting peak area with accurate mass, a quantitative analysis was carried out. Good linearity was detected in the 1-500 μg/L range, with correlation values ranging from .997 6 to .999 7. The LODs ranged from .01 to .5 μg/L. The spiking recoveries ranged from 79.6% to 96.4%, with RSD ranging from 4.1% to 6.4%. The Agilent MassHunter PCDL Manager software was used to create the result screening database, which was then utilised to analyse spiked samples. The findings indicate that all of the spiking antidepressants could be reliably recognised with a modest variance of retention time (<.1 min) and mass (<1 mDa). In summary, the technique is sensitive, rapid, and reliable for antidepressants screening and confirmation in forensic/clinical toxicology laboratories.

Zilfidou et al (2019)⁴⁰ developed an FPSE-HPLC-DAD technique for the extraction and detection of 5 antidepressants, including clomipramine, amitriptyline, fluoxetine, paroxetine, and venlafaxine, in human blood serum. LOD was found at .15 ng/μL, while good absolute recoveries (9.4-88.1%) and low RSD (≤9.2% and ≤ 11.0 for within-day and between-day precision, respectively) were obtained.

For the detection and quantification of 38 ADs, antipsychotics, and related metabolites in small amounts (200 μL) of human whole blood, Zhu et al (2021)⁴¹ validated an LLE-LC-MS-MS methodology. The validated method was shown to be extremely sensitive, with a LOD range of .0005 to 1 ng/mL and an LLOQ range of .002 to 2 ng/mL. For all analytes, bias and precision within- and between runs were within 14.7%. To make sure that the target compounds' therapeutic and toxic blood concentration ranges were completely covered, dilution integrity was assessed. Furthermore, this methodology was employed to analyse real whole blood samples acquired from 2 forensic cases, demonstrating its practical value in giving accurate and thorough information on the deceased’s past medication.

Vignali et al (2020)⁴² validated an LC-MS/MS technique to detect and quantify quetiapine and its 2 main metabolites (7-hydroxyquetiapine and N-desalkylquetiapine) in tissues, biological fluids, and blood. LODs of .9, .3, and .3 ng/mL were reported for quetiapine, 7-hydroxyquetiapine, and N-desalkylquetiapine respectively; with a LOQ of 10.0 ng/mL defined for all 3 analytes. The experiment includes 13 postmortem actual examples. The findings indicate that 7-hydroxyquetiapine is less impacted by this factor, but quetiapine and N-desalkylquetiapine may experience considerable postmortem redistribution. It is usually good to analyse both analytes since N-desalkylquetiapine might be detected in blood at substantially higher proportions than quetiapine.

Di Rago et al (2021)⁴³ developed and validated a quick approach for detecting and semi-quantifying a wide range of drugs, including various antidepressants, synthetic cannabinoids (SCs), amphetamines, anaesthetics, anti-psychotics, anti-convulsants, beta-blockers, benzodiazepines, stimulants, opioid and non-opioid analgesics, Tetrahydrocannabinol (THC), and other NPS, in blood. To achieve quick turnaround times, this approach incorporates a small sample volume, a one-step liquid extraction, and automated data processing. The extracts were chromatographed for 5 min on a Kinetex C18 50 × 4.6 mm 2.6 m solid-core analytical column before being analysed on a Sciex 3200 Q-TRAP MS-MS (+ESI, MRM mode). The methodology has been employed in over 6000 coronial investigations utilising both clinical and postmortem blood specimens in standard practice at the Victorian Institute of Forensic Medicine’s forensic toxicology service. This approach has significantly boosted throughput, shortened turnaround times, and enabled same-day examination of findings when required. The approach is widely used in overnight testing, with results delivered to pathologists within 4 hours after data collection. To offer an effective death investigation procedure, this quick toxicological approach is employed in concert with other investigative techniques such as full-body CT imaging, assessment of case circumstances, and medical histories.

Musshoff et al (2020)⁴⁴ summarized the concentration distributions of over 100 legal and illicit drugs, including benzodiazepines, antipsychotics, antidepressants, opioids, narcotic drugs, and major metabolites in Caucasian hair samples. Following extraction of finely chopped specimens by ultrasonic in methanol, the proximal sections of 1-6 cm in length were examined by LC-MS/MS, depending on their availability. Other laboratories may find the data a helpful starting point for a preliminary analysis of their findings. However, the facts of the specific case and the analytical techniques employed should be considered when using this statistical data.

Rodrigues et al (2021)⁴⁵ developed an effective method to quantify 16 ADs, 7 antipsychotics and 3 metabolites, as well as semi-quantify norsertraline and norfluoxetine in postmortem blood utilizing a modified micro-QuEChERS method coupled to LC/MS-MS. The calibration curve was found to be linear in the range of 1-500 ng/mL, with R2>.99, and all standards quantified within 15% of the target except 20% at the LOQ of 1 ng/mL for 26 compounds. This approach has the benefits of a low sample volume of 100 μL, straightforward but efficient sample preparation, and a short run time of 8.5 min. One hundred genuine postmortem samples were analysed using the approved analytical approach.

Pouliopoulos et al (2018)⁴⁶ developed QuEChERS in conjunction with the UHPLC/MS-MS method for detecting and quantifying a variety of commonly prescribed antidepressant and antipsychotic drugs for toxicological reasons. The technique was validated and offered satisfactory precision with CV ranging from 1.2 to 13.2% and accuracy with recoveries ranging from 85 to 113%. For the 15 drugs, LODs ranged between .0003 to .017 μg/mL, whereas LOQs ranged between .001 to .05 μg/mL. The approach was shown to be appropriate for analysing clinical samples for Therapeutic Drug Monitoring (TDM) and postmortem or ante-mortem whole blood samples from forensic cases. A variety of clinical and forensic samples were successfully evaluated, proving QuEChERS’s efficiency in this sector.

Orfanidis et al (2018)⁴⁷ developed a UHPLC-MS/MS approach for detecting and quantifying 27 drugs (benzodiazepines, amphetamines, cannabinoids, cocaine, and opiates) and pharmaceuticals (antidepressants and antipsychotics) in human bones. The method was found to be sensitive and selective, with LOD ranging from .03 to 1.35 ng/g of bone. The technique satisfied standard bioanalytical parameters and was thus used to the analysis of bone marrow and bone collected postmortem from chronic drug users, enabling precise identification and quantitative drug determination in medicolegal cases when blood analysis was not possible.

Chen et al (2019)⁴⁸ developed a rapid and sensitive LC-MS/MS technique for determining duloxetine and quetiapine in rat plasma. Good linearity was obtained over the range of 1.00-200 ng/mL (R2 = .9982) for duloxetine and .50-100 ng/mL (R2 = .9972) for quetiapine utilizing 50 μL of rat plasma, respectively. The approach was used in a pharmacokinetic interaction study, and the findings showed that quetiapine had a substantial influence on the increased plasma exposure of duloxetine in rats when used in combination. This study might be easily implemented in TDM of patients with major depressive disorder (MDD).

Remane et al (2011)⁴⁹ validated a LLE-UHPLC-MS/MS technique for detecting and quantifying 34 antidepressants in plasma. LOQ and LOD indicated that the method was selective, sensitive, precise, and accurate for 28 of the 34 tested drugs. Analysing real plasma samples and external quality control samples satisfactorily demonstrated the applicability.

Wietecha-Posłuszny et al (2012)⁵⁰ optimized an MEPS/HPLC-DAD technique for extracting 6 tricyclic TCAs (nordoxepin, doxepin, desipramine, imipramine, nortriptyline, amitriptyline) from human serum. The LOD (.02-.05 μg/mL), interday (4.4-11.6%) and intraday (2.7-8.8%) precision (RSD), and the accuracy of the assay (94.5-108.8%) were estimated at 3 concentration levels-.2, .5, and .8 μg/mL. The method’s accuracy was determined by analysing certified reference material. Furthermore, the SPE method and the verified approach were contrasted. The suitability of MEPS as a tool for forensic/clinical methods for serum sample preparation was evaluated at the end. Quetiapine was analyzed by Johansen⁵¹ (2017) by UHPLC-MS/MS in a reported a drug-facilitated sexual assault (DFSA).

Snamina et al (2019)⁵² validated a new MAE/UHPLC-TOFMS technique to determine the antidepressant drugs (paroxetine, sertraline, fluoxetine, citalopram, and venlafaxine) in bone marrow aspirates in real forensic investigations. Due to a dearth of data on therapeutic and harmful levels of considered drugs in bone marrow, serum data has been utilised as a reference. According to this supposition, the devised approach enables the measurement of all drugs indicated at therapeutic concentrations. Additionally, this approach has been applied in actual instances. Lastly, 4 analytes were detected in 3 case samples: citalopram (68 ng/mL), paroxetine (about 3.6 μg/mL), fluoxetine (84 ng/mL), and venlafaxine (104 ng/mL).

Mohebbi et al (2018)⁵³ developed an effective sample pretreatment method by combining DLLM, DSPE, and salt induced-homogenous liquid-liquid extraction (SI-HLLME) based on the solidification of floating organic droplets for the extraction of commonly prescribed TCAs (imipramine, clomipramine, desipramine, amitriptyline, and nortriptyline) in human urine prior to their determination by HPLC-UV. The obtained LODs and LOQ were ranging between .22-.31 and .71-1.1 μg/L, respectively. The recommended method was used to identify TCA drugs in urine samples from various individuals. The technique may be used in future forensic and pharmacokinetic investigations of TCAs.

Fuentes et al (2019)⁵⁴ developed a method for identifying and determining 6 antidepressants (including venlafaxine, sertraline, mirtazapine, fluvoxamine, fluoxetine, and escitalopram) in human urine by utilizing microextraction by packed sorbent (MEPS) accompanied by ultra-performance liquid chromatography with photodiode array detection (MEPS-UPLC-PDA). The ensuing analytical technique was validated in terms of linearity (25-1000 ng/mL), LOD (<7.1 ng/mL), LOQ (25 ng/mL), precision (4.7-15.1% as RSD), and accuracy (80.4-126.1% as mean recovery for 4 replicate determinations). The suggested approach allowed the concentrations of the 6 target antidepressants to be assessed at therapeutic to toxic levels. The application to modest amounts of urine (300 μL) allowed for quick analyte extraction and findings comparable to existing clinical and forensic options.

Lioupi et al (2019)⁵⁵ validated an FPSE-HPLC-DAD technique for the simultaneous extraction and analysis of 5 widely known ADs (clomipramine, amitriptyline, fluoxetine, paroxetine, venlafaxine) in human urine samples. For the first time, FPSE was assessed in terms of extracting the tested antidepressants from human urine, and it turned out to be an efficient, and rapid sample preparation method. Among other polarities, the sol-gel graphene sorbent coated on cellulose FPSE media was the most efficient. The absolute recovery values were 29.0% for clomipramine, 43.0% for amitriptyline, 67.0% for fluoxetine, 33.9% for paroxetine, and 25.5% for venlafaxine, while relative recoveries were more than 90%. The described approach yields an acceptable LOD of .15 ng/L.

López-Rabuñal et al (2020)⁵⁶ developed and validated an SPE-LC-MS-MS technique for determining 15 antidepressants (desmethylcitalopram, citalopram, desmethylvenlafaxine, venlafaxine, paroxetine, norsertraline, sertraline, norfluoxetine, fluoxetine, norclomipramine, clomipramine, desipramine, imipramine, nortriptyline, and amitriptyline) and 14 benzodiazepines (zolpidem, midazolam, triazolam, 7-aminoflunitrazepam, flunitrazepam, oxazepam, lormetazepam, lorazepam, nordiazepam, diazepam, bromazepam, clonazepam, α-hydroxyalprazolam, and alprazolam) in meconium. Method validation included the following: linearity [n = 5, LOQ to 400 ng/g], LOD (n = 6, 1-20 ng/g), LOQ (n = 9, 5-20 ng/g), imprecision (n = 15, 0-14.6%), accuracy (n = 15, 90.6-111.5%), selectivity (no exogenous or endogenous interferences), matrix effect (n = 10, −73 to 194.9%). Meconium samples (n = 4) were evaluated with and without hydrolysis (alkaline and enzymatic) using this approach. Norclomipramine, clomipramine, norfluoxetine, fluoxetine, nordiazepam, diazepam, clonazepam, α-hydroxyalprazolam, and alprazolam were all detected in the authentic meconium samples. In light of this, the current LC-MS-MS approach enables a high throughput identification of the most prevalent antidepressants and benzodiazepines in meconium, which may be helpful in clinical and forensic contexts.

Steuer et al (2015)⁵⁷ developed and validated robust methods for quantifying 40 neuroleptic and antidepressant drugs in whole blood based on fast and simple protein precipitation, and secondly evaluated the utility of microflow liquid chromatography coupled to mass spectrometry (MFLC–MS/MS) in comparison to conventional LC–MS/MS in terms of method validation parameters, particularly LODs, matrix effects, imprecision, and bias. With a few exceptions, both approaches have comparable LOQs, LODs, and calibration models. Low mobile phase consumption, a short runtime, and better peak separation were benefits of the MFLC system.

Ogawa et al (2022)⁵⁸ developed an extraction method utilizing an ISOLUTE PLD⁺ protein and phospholipid removal column to quantify 20 psychoactive drugs, like amphetamines, sedative-hypnotics, antipsychotics, and antidepressants, in postmortem whole blood samples by LC-MS/MS. When compared to traditional protein precipitation and the QuEChERS, the method showed improvement in extract cleanliness. For all analytes, the technique was validated; the calibration curves showed good linearity (R2 >.991). This approach has been used successfully to quantify psychoactive substances in postmortem forensic samples.

After almost 2 years of decomposition, Desrosiers et al (2012)⁵⁹ examined the distribution of desmethylcitalopram, citalopram, nortriptyline, and amitriptyline in 13 different porcine skeletal tissues. Following a simple wash operation, bones were pulverised, and drugs were extracted using passive incubation in methanol followed by SPE. UHPLC was used to analyse the samples and confirmed with GC/MS. The Kruskall-Wallis test revealed that bone type had a significant impact on drug levels for all analytes, with values ranging from 33 to 166-fold. Less erratic were ratios of drug levels to those of the equivalent metabolite, which varied by around a factor of one to eight. This implies that the interpretative value of drug measurements in bone has limitations and those relative levels of drug and metabolite concentrations should be further examined in terms of their forensic relevance.

Patel et al (2018)⁶⁰ developed a LLE-LC–MS/MS technique for determining doxepin and its metabolite, nordoxepin in human plasma. A linear dynamic range of 5.00-1300 pg/mL for nordoxepin and 15.0-3900 pg/mL for doxepin was established with R2 of .9993 and .9991, respectively. The extraction recovery ranged between 88.0%–99.1% and 86.6%–90.4% for nordoxepin and doxepin, respectively. The approach was successfully used in a bioequivalence study using a 6 mg oral disintegrating tablet of doxepin hydrochloride in forty-one healthy Indian participants under fasting and feeding conditions.

Maciel et al (2020)⁶¹ presented an automated multidimensional technique based on LC-MS/MS utilizing a lab-made, graphene-based capillary extraction column coupled to a C8 analytical column to determine four ADs, such as desipramine, clomipramine, citalopram, and carbamazepine, and 1 anticonvulsant AC, sertraline, in urine. The study reports good linearity (R2 > .98), and LODs were in the range of .5 to 20 μg/L. The procedure was then used to the direct examination of 10 untreated urine samples, with citalopram traces discovered in one of them. The results imply that the proposed technique might be a viable option for offering direct and completely automated analysis of ADs in biological specimens.

In clinical and forensic settings, antidepressants detection in Oral fluid (OF) is crucial. OF testing provides a simple and non-intrusive sample collecting method. Shin et al (2020)⁶² validated an SPE-LC-MS/MS technique for quantifying 18 antidepressants (venlafaxine, trimipramine, trazodone, sertraline, paroxetine, nortriptyline, mirtazapine, imipramine, fluoxetine, duloxetine, doxepin, desvenlafaxine, desipramine, cyclobenzaprine, clomipramine, citalopram, bupropion, and amitriptyline) and metabolites in OF collected by Quantisal^® oral collection devices. The linear ranges were 10 to 1000 ng/mL for all analytes with a LOD of 10 ng/mL. The real patient sample revealed a percent variation of 86.3-111% from the concentration that had previously been estimated. This technique enables the quick identification of 18 antidepressants and metabolites in OF and may be simply used in a routine laboratory.

Fisher et al (2013)⁶³ validated an LC-MS/MS technique for the analysis of sulpiride, risperidone and 9-hydroxyrisperidone, quetiapine, olanzapine, clozapine and norclozapine, aripiprazole and dehydroaripiprazole, and amisulpride in small (200 μL) volumes of serum or plasma for TDM purposes. The method’s applicability to haemolysed whole blood and to OF were also studied. The calibration of the assay was linear across the concentration ranges investigated. There was no evidence of ion suppression/enhancement or interference from certain recognised metabolites of the antipsychotics investigated.

Vignali et al (2021)⁶⁴ employed GC/MS, LC/MS/MS and LC/Q-TOF to identify fluvoxamine a SSRI in biological fluids (cardiac and femoral blood, bile and urine) collected from a corpse. Fluvoxamine (2.20 mg/L) was quantified in blood using a validated LC–MS-MS method. The blood level of fluvoxamine was roughly 10 times higher than the .23 mg/L upper limit for therapeutic blood levels. Nevertheless, it is impossible to rule out contributing factors in deaths brought on by the use of multiple drugs. Fluvoxamine distribution in all biological fluids was assessed, and a postmortem redistribution effect was detected (C/P blood ratio: 1.86). An liquid chromatography quadrupole time-of-flight (LC-QTOF) analytical technique was used to detect fluvoxamine acid metabolites in cardiac blood, bile, and urine.

Micellar Liquid Chromatography

Agarwal et al (2014)⁶⁵ developed, micellar liquid chromatographic (MLC) method for determining paroxetine in serum, urine and physiological samples, utilizing C18 column and electrochemical detection (ECD). The proposed methodology was validated in terms of specificity, accuracy (88-97.5%, recovery), linearity (r(2) > .9999; .5-5 μg/mL range), intermediate precision (RSD < .54%), repeatability (RSD < .54%), and LOD and LOQ (.001 and .005 μg/mL, respectively). The method developed was effectively applied to real urine and blood samples, and also spiked urine and serum samples. The developed method was quick, accurate, and inexpensive, making it appropriate for routine paroxetine analysis in monitored samples.

Wietecha-Posłuszny et al (2017)⁶⁶ developed and validated an effective MAE/UHPLC–MS-TOF technique for simultaneous identification and quantification of psychoactive substances, such as piperazines (n = 3), imidazopyridines (n = 1), anticonvulsants (n = 1), SNRI (n = 1), SSRI (n = 4), TCAs (n = 5), and benzodiazepines (n = 16), in human bone marrow for the first time. Following the method’s international criteria-based validation, it was used to analyse 3 case samples, where paroxetine, doxepin, citalopram, nordiazepam, and diazepam were successfully detected. Lastly, the method presented here is an excellent demonstration of a method that might be utilized in forensics.

Touch Spray Mass Spectrometry

Morato et al (2019)⁶⁷ used Mitra^® microsampling for sample collection, which is based on the volumetric absorptive microsampling approach, and analyzed directly using touch spray mass spectrometry (TS-MS) to simultaneous determine 30 common drugs of abuse, which include antipsychotic, antidepressants, cathinones, substituted methylenedioxyphenethylamines, cocaine, methamphetamines, fentanyl derivatives, benzodiazepines, and opioids, in oral fluid (OF). Using TS-MS provided high selectivity, specificity, and sensitivity using a small sample volume (10 μL) that can be collected easily and non-invasively. Furthermore, direct analysis from swabs might be employed for point-of-care testing in clinical or forensic contexts when linked with miniature mass spectrometers, allowing for the rapid acquisition of crucial toxicological data in situ.

Surface-Enhanced Raman Spectroscopy

Boroujerdi et al (2022)⁶⁸ presented for the first time a surface-enhanced raman spectroscopy (SERS) protocol for the detection of amitriptyline, the common and widely used TCA on dried saliva spots and dried blood spots using a test substrate modified with silver nanoparticles. The validated protocol is rapid and non-destructive, with a detection limit of 95 ppb, and linear range between 100 ppb and 1.75 ppm on the SERS substrate, which covers the therapeutic window of amitriptyline (150 ppb–300 ppb)¹¹ in biological fluids.

Discussion

The preparation of samples is an important phase in the analytical process since it takes up more than 80% of the time used for analysis.⁶⁹ Eliminating interfering species, extracting analytes, and increasing the concentration of the analyte to improve the final signal are the main objectives of the pre-treatment procedure.⁷⁰ We carried out an in-depth review of the different techniques, particularly recent trends, employed in laboratories for antidepressant extraction from various biological matrices.

As mentioned above, biospecimens where antidepressants are usually detected include urine,^{24,51,53-55,61,64,65} whole blood,^{11,17,22,23,29,31,35,36,39,41,57,58} dried blood spots,^11,32 plasma,^{23,26,28,48,48,60,63} serum,^{23,40,46,50,63,65} hair,^44,51 OF,^30,62,67 pericardial fluid,¹⁴ bone marrow,^33,52,66 bones,^{21,25,34,47,59} brain tissue,³⁶ intraosseous fluid,²⁶ meconium,⁵⁶ and VH^12,13 with each matrix requiring specific treatment depending on the type of antidepressant under investigation.

Protein precipitation,^26,48,58,59 LLE,^{15,25,29,41,46,53,60} SPE,^{12,17,20-23,28,32,34,36,42,51,56,59,62,64} and some miniaturized techniques—such as SPME,³³ MEPS,^30,50,54 DLLME,¹⁴ AA-DLLME,²⁷ BAμE,²⁴ and LPME¹³ are the most utilized procedures for extracting antidepressants from the biological samples.

QuEChERS has strengthened its position for analysis of antidepressants residues in human biospecimens.^{19,37,38,45,46} The approach provides quality, safety, authenticity, and traceability of the analytes in the various matrices by requiring just two steps, extraction and clean-up. The QuEChERS protocol ensures a high recovery rate and better analytical performance by controlling various factors such as the extraction solvent, sample quantity, sample/solvent ratio, pH, salts, and sorbent phases, making QuEChERS the procedure of excellence in the pretreatment phase of samples destined for analysis by chromatography or spectrometry.

The accuracy, precision, and robustness of analytical procedures are often impacted by sample treatment, and preconcentration²⁷ of analytes enhances sensitivity and enables lower LODs.

Sample preparation is an important step in a forensic toxicological analysis. Technological advancements and the availability of mass spectrometers with increasing sensitivities and the need to remove potential interferences such as matrix components or nonrelevant analytes to the analysis are essential to optimizing an analytical method. Forensic toxicological analysis involves diverse analytes with a variety of chemical properties that need to be considered when selecting an appropriate sample preparation technique. Different extraction techniques are employed to remove interferences from the biological samples, such as blood, tissues, and hair, reducing matrix effects and concentrating the target analytes, among others. The push for broad standards and best practice documents for forensic toxicology demonstrates the need for laboratories to ensure they meet the required capabilities to detect specific classes of drugs commonly encountered in forensic casework and ensure method validity.⁷¹

Analytical Methods

Several analytical methods range from basic wet analytical techniques to sophisticated instrumental analytical methods for identifying and quantifying ADs in biological samples. There are more complex techniques with specialized implementations, and there are also applications and combinations of the overall techniques within each major analytical technique. The analytical techniques reported in this review vary from simple to highly sophisticated complex techniques, depending on several factors such as the nature of the sample, matrices, admixtures, accuracy needed, speed of analysis, instrumentation available and cost, etc.

The majority of these techniques rely on LC approaches, utilizing MS/MS detection modes,^60,62,63 TOF/MS,^33,52 and U(H)PLC with MS/MS.^{29,46,47,51,61} Additionally, several studies employed GC coupled to MS detection^{12-14,20,22,24-26,34,62} or tandem MS detection modes.^15-17,19 Most frequently, these methods have shown low detection limits, selectivity, sensitivity, and acceptable results, enabling precise quantification of ADs. Additional methods for determining TCAs in biological matrices have also been explored, such as capillary electrophoresis,¹¹ TS-MS,⁶⁷ HPLC-UV,⁵³ and HPLC-DAD.^40,50,55

As explained earlier, forensic and toxicological analysis is associated with so much variance in the nature of sample, the objective of the analysis, and infrastructural facilities that the earlier work reference will be a great guiding factor. The analyst/researcher will be at liberty to use a method of choice suitable for analysis under a specified set of conditions and requirements. Many a time, the analyst has to carry out research work to analyze a new type of sample by his ingenuity, experience, knowledge, and the resources at his disposal, giving birth to a new alternative procedure of analysis.

Conclusion

A review of literature from 2011 to 2022 for the analysis of antidepressants and their metabolites in biological samples, including blood, urine, tissues, saliva, hair, and vitreous humour, etc., are presented with their relative merits.

Most recent methods rely on LC-MS-MS, which has excellent sensitivity, selectivity, precision, and accuracy. Some papers deal with GC-MS techniques for thermally stable antidepressants and their metabolites. Some researchers have used CE-MS and LC with UV and DAD detectors. LC-UV approach has the benefit of being simpler, faster, more reliable, and more extensively applicable to antidepressant quantification. For some antidepressants, MLC, an RP-HPLC technique, was utilized, which has advantages over conventional HPLC in terms of sample preparation, selectivity, and adaptability. Its key benefit is that samples can be injected directly into the chromatographic system with no previous preparation step. The TSMS technique has the potential to provide quick and non-invasive point-of-care drug testing in clinical or forensic settings.

The analysis of the antidepressants in various biological matrices can be successfully carried out using a wide variety of methods reported in this review, depending on the nature of the analyte/s and the matrix. Several other non-antidepressant drugs and their metabolites, likely to be present in the matrices, can also be analyzed simultaneously by the techniques presented.

Supplemental Material

Supplemental Material - Recent Advances in Analytical Techniques for Antidepressants Determination in Complex Biological Matrices: A Review

Supplemental Material for Recent Advances in Analytical Techniques for Antidepressants Determination in Complex Biological Matrices: A Review by Sachil Kumar, Siddaraj Darshan, and Tulsidas R. Baggi in International Journal of Toxicology

Footnotes

Author Contributions

Kumar, S. contributed to conception and design, contributed toacquisition, drafted manuscript, and critically revised manuscript; Baggi, T.contributed to design, contributed to analysis, and critically revisedmanuscript; Darshan, S. contributed to acquisition, analysis, andinterpretation, drafted manuscript, and critically revised manuscript. Allauthors gave final approval and agree to be accountable for all aspects of workensuring integrity and accuracy.

Declaration of Conflicting Interests

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Funding

The author(s) received no financial support for the research, authorship, and/or publication of this article.

ORCID iD

Sachil Kumar

Supplemental Material

Supplemental material for this article is available online.

Abbreviations

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