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.

Development of a new method for drug detection based on a combination of the dried blood spot method and capillary electrophoresis.

The aim of this study was to develop a new approach to sample preparation of biological material based on a combination of the Dried Blood Spot (DBS) method and capillary electrophoresis coupled with mass spectrometry (CE-MS) for the analysis of blood samples collected in vivo or post-mortem. The proposed approach allowed the identification of typical drugs from different groups, such as tricyclic antidepressants (amitriptyline, imipramine), selective serotonin reuptake inhibitors (citalopram), benzodiazepines (tetrazepam) and hypnotics (zolpidem). In this study, a blood sample was spotted on FTA DMPK C cards, then dried, and 6-mm discs were cut out. The sample preparation procedure involved microwave-assisted extraction (MAE). Various extraction agents, temperatures and durations of extraction were examined in order to achieve the highest efficiency of the process. The method was subjected to a validation procedure. Limits of detection (LOD = 1.76 - 14.7 ng/mL) and quantification (LOQ = 5.25 - 49.0 ng/mL), inter- (CV = 1.31 - 9.43%) and intra- (CV = 3.26 - 18.52%) day precision of the determinations, recovery (RE = 85.0-105.4%) and matrix effect on ionization of analytes (ME = 98.6-105.5%) were determined. Furthermore, the developed DBS/MAE/CM-MS method was selective and analytes present in the blood applied on DBS cards were found to be stable after 7 and after 14 days. Moreover, the developed method was successfully applied to the analysis of both post-mortem samples and blood samples taken from patients treated with the analyzed drugs.

Recent Advances in the HPLC Analysis of Tricyclic Antidepressants in Bio-Samples.

Tricyclic Antidepressants (TCAs) are a group of the main category of antidepressant drugs, which are commonly prescribed to treat major depressive disorder. Determination of TCA drugs is very important for clinical and forensic toxicology, especially for therapeutic drug monitoring in various biofluids. High Performance Liquid Chromatography (HPLC) is a well-established technique for this purpose. A lot of progress has been made in this field since the past 10 years. Novel extraction techniques, and novel materials for sample preparation, novel columns and novel applications of analysis of various biofluids for the determination of TCAs in combination with other drugs are some typical examples. Moreover, advances have been performed in terms of Green Analytical Chemistry principles. Herein, we aim to discuss the developed HPLC methods that were reported in the literature for the time span of 2008-2018.

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.

Magnetic framework composite as sorbent for magnetic solid phase extraction coupled with high performance liquid chromatography for simultaneous extraction and determination of tricyclic antidepressants.

In this study, magnetic framework composites (MFCs) (Fe3O4@TMU-10) microspheres were successfully fabricated and applied as an effective sorbent for preconcentration of the two model tricyclic antidepressants (TCAs) amitriptyline and imipramine from biological samples. MFCs were fabricated by a step-by-step assembly, novel, simple and efficient strategy. The shell thickness of the Metal-organic frameworks (MOFs) could also be easily controlled by tuning the number of assembly cycles. By coupling magnetic solid-phase extraction (MSPE) with high-performance liquid chromatography with UV detector (HPLC-UV), a simple, reliable, fast, sensitive and cost-effective method for simultaneous determination of TCAs was developed. Under optimal conditions, the preconcentration factors and relative recoveries of the studied compounds were obtained in the range of 43-50 and 90.5-99.0% respectively. The calibration curves were obtained in the range of 5-800 µg L-1 with reasonable linearity (R2 > 0.9904) and the limits of detection (LODs) ranged between 2 and 4 µg L-1 (based on S/N = 3). The relative standard deviations of intra- and inter-day tests ranged from 3.1 to 4.6% and from 4.3 to 5.2%, respectively. The results demonstrate that Fe3O4@TMU-10 core-shell magnetic microspheres combine advantages of MOFs and magnetic nanoparticles, and are the promising sorbents for rapid and efficient extraction of target analytes from urine and plasma complex biological samples.

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.

Analysis of tricyclic antidepressants in human plasma using online-restricted access molecularly imprinted solid phase extraction followed by direct mass spectrometry identification/quantification.

The use of a new class of hybrid materials, called restricted access molecularly imprinted polymers (RAMIPs) seems to present a good strategy for the sample preparation of complex matrices, since these materials combine good protein elimination capacity with high degree selectivity. Mass spectrometers (MS) have been successfully used for polar drug identification and quantification. In order to combine the advantages of both RAMIPs and mass spectrometry, we proposed a study that joins these properties in a single system, where we could analyse tricyclic antidepressants from human plasma, without offline extraction or chromatographic separation. A RAMIP for amitriptyline was synthesised by the bulk method, using methacrylic acid as a functional monomer and glycidilmethacrylate as a hydrophilic co-monomer. Then, epoxide ring openings were made and the polymer was covered with bovine serum albumin (BSA). A column filled with RAMIP-BSA was coupled to a MS/MS instrument in an online configuration, using water as loading and reconditioning mobile phase and a 0.01% acetic acid aqueous solution: acetonitrile at 30:70 as elution mobile phase. The system was used for on-line extraction and simultaneous quantification of nortriptyline, desipramine, amitriptyline, imipramine, clomipramine and clomipramine-d3 (IS) (from 15.0 to 500.0µgL-1) from plasma samples. The correlation coefficient was higher than 0.99 for all analytes. The CV (coefficient of variation) values ranged from 1.34% to 19.13% for intra assay precision and 1.32-19.77% for inter assay precision. The E% (relative error) values ranged from -19.15% to 19.51% for intra assay accuracy and from -9.04% to 16.22% for inter assay accuracy.

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%.

Ultrasound-assisted combined with nano-sized molecularly imprinted polymer for selective extraction and pre-concentration of amitriptyline in human plasma with gas chromatography-flame detection.

A new process was developed for the selective extraction and pre-concentration of amitriptyline (AT) from human plasma using nano-sized molecularly imprinted polymer (MIP) with ultrasound-assisted extraction (UAE). The nano-sized AT imprinted polymer particles were synthesized using suspension polymerization in silicon oil and characterized by Fourier transform infrared (FT-IR) spectroscopy and scanning electron microscope (SEM) methods. With the application of optimized values, linearity values in the ranges of 20-200µgmL(-1) and 35-200µgmL(-1) were obtained for AT with the correlation of determination values (r(2)) 0.998 and 0.995 in water and plasma, respectively. The limits of detections (S/N=3) for AT were found to be 0.7 and 1.2µgmL(-1) in water and plasma, respectively. The enrichment factors of AT in water and plasma were 52 and 40, respectively. The inter-day precisions (%) were in the range of 5.8-9.2%. Relative recovery rates ranged from 82.4% to 92.3%. The method was successfully applied to determine AT in the human plasma samples.

Development of microextraction by packed sorbent for toxicological analysis of tricyclic antidepressant drugs in human oral fluid.

The aim of this study was to apply microextraction by packed sorbent (MEPS) to the isolation of six tricyclic antidepressants (TCADs): nordoxepin, doxepin, desipramine, nortriptyline, imipramine, and amitriptyline from human oral fluid. Samples were collected from healthy volunteers via free spillage from the oral cavity to disposable test tubes. A method of oral fluid sample pretreatment was developed and optimized in terms of suitability for MEPS extraction and removing of interfering agents (protein, food debris, or air bubbles). Moreover, it was short and simple to perform with limited sample consumption (150µL). Extracts were analysed by UHPLC-MS. The MEPS/UHPLC-MS method was validated at three concentration levels (2.00, 4.00 and 8.00ng/mL) of all analytes in the range 1.25-10.0ng/mL. The following parameters were determined: limit of detection, limit of quantification, precision, and accuracy. For all tested concentration levels, the intra- and inter-day repeatability did not exceeded 8.1% and 12.2%, respectively. Gained LOQ value, 0.50ng/mL, made the MEPS/UHPLC-MS method to be a useful tool in clinical and forensic laboratories, which was demonstrated on the basis of analysis of real samples.

Electro-assisted solid-phase microextraction based on poly(3,4-ethylenedioxythiophen) combined with GC for the quantification of tricyclic antidepressants.

In this study, a platinum wire coated with poly(3,4-ethylenedioxythiophen) was used as an electro-assisted solid-phase microextraction fiber for the quantification of tricyclic antidepressant drugs in biological samples by coupling to GC employing a flame ionization detector. In this study, an electric field increased the extraction rate and recovery. The fiber used as a solid phase was synthesized by the electropolymerization of 3,4-ethylenedioxythiophen monomers onto a platinum wire. The ability of this fiber to extract imipramine, desipramine, and clomipramine by using the electro-assisted solid-phase microextraction technique was evaluated. The effect of various parameters that influence the extraction efficiency, which include solution temperature, extraction time, stirring rate, ionic strength, time and temperature of desorption, and thickness of the fiber, was optimized. Under optimized conditions, the linear ranges and regression coefficients of calibration curves were in the range of 0.5-250 and 0.990-0.998 ng/mL, respectively. Detection limits were in the range of 0.15-0.45 ng/mL. Finally, this method was applied to the determination of drugs in urine and wastewater samples and recoveries were 4.8-108.9%.

Combination of electromembrane extraction with dispersive liquid-liquid microextraction followed by gas chromatographic analysis as a fast and sensitive technique for determination of tricyclic antidepressants.

For the first time, combination of electromembrane extraction (EME) and dispersive liquid-liquid microextraction (DLLME) followed by gas chromatography-flame ionization detection (GC/FID) was developed for determination of tricyclic antidepressants (TCAs) in untreated human plasma and urine samples. Response surface methodology (RSM) was used for optimization of experimental parameters, so that extraction time of 14min, voltage of 240V, donor phase of 64mM HCl and acceptor phase of 100mM HCl were obtained as optimal extraction conditions. Matrix effect and carry-over were investigated in this work. The results indicated matrix effect for urine and plasma samples in comparison with neat solutions, so match matrix method was used for drawing working calibration curves. However, no carry-over was appeared at the retention time of investigated TCAs (S/N<3). With application of optimized values, good linearity in the range of 2-500µg L(-1) was obtained for TCAs with the correlation of determination values (r(2)) above 0.9968. The limits of detection (S/N=3) for TCAs were found 0.25, 3.0, and 15µg L(-1) in water, urine, and plasma, respectively. The preconcentration factors of TCAs in water, urine, and plasma were from 383 to 1065. The intra- and inter-assay precisions (%) were in the ranges 6.4-11.8% and 6.2-10.8%, respectively, and the intra- and inter-assay accuracies were >86.5%. The results showed that EME-DLLME-GC/FID is a promising combination for analysis of TCAs present at low concentrations in biological matrices.

Micelle to solvent stacking of organic cations in micellar electrokinetic chromatography with sodium dodecyl sulfate.

The on-line sample concentration technique, micelle to solvent stacking (MSS), was studied for small organic cations (quaternary ammonium herbicides, ß-blocker drugs, and tricyclic antidepressant drugs) in reversed migration micellar electrokinetic chromatography. Electrokinetic chromatography was carried out in fused silica capillaries with a background solution of sodium dodecyl sulfate (SDS) in a low pH phosphate buffer. MSS was performed using anionic SDS micelles in the sample solution for analyte transport and methanol or acetonitrile as organic solvent in the background solution for analyte effective electrophoretic mobility reversal. The solvent also allowed for the separation of the analyte test mixtures. A model for focusing and separation was developed and the mobility reversal that involved micelle collapse was experimentally verified. The effect of analyte retention factor was observed by changing the % organic solvent in the background solution or the concentration of SDS in the sample matrix. With an injection length of 31.9 cm (77% of effective capillary length) for the 7 test drugs, the LODs (S/N=3) of 5-14 ng/mL were 101-346-fold better when compared to typical injection. The linearity (R(2), range=0.025-0.8 µg/mL), intraday and interday repeatability (%RSD, n=10) were ≥0.988, <6.0% and <8.5%, respectively. In addition, analysis of spiked urine samples after 10-fold dilution with the sample matrix yielded LODs=0.02-0.10 µg/mL. These LODs are comparable to published electrophoretic methods that required off-line sample concentration. However, the practicality of the technique for more complex samples will rely on dedicated sample preparation schemes.

Optimizing the extraction, separation and quantification of tricyclic antidepressant drugs in human plasma with CE-ESI-TOF-MS using cationic-coated capillaries.

In this study, the extraction and CE-ESI-TOF-MS analysis of tricyclic antidepressant (TCA) drugs imipramine, desipramine, clomipramine and norclomipramine in human plasma has been optimized. The CE capillaries were modified with ω-iodo-alkyl ammonium salt (M7C4I coating) to reduce analyte adsorption to the silica wall. The use of a strong cation exchange (SCX) solid-phase extraction (SPE) column specifically designed for the extraction of basic drug species from biofluids gave very clean extracts with high and reproducible recoveries. The extraction recoveries were ranging between 87 and 91% with % RSD values of 0.5-1.7% (n=3). The obtained strong cation exchange-SPE extracts of the TCA in human plasma only contained the analytes of interest. The optimized CE separation conditions were obtained by adding ACN and acetic acid to the sample while using an aqueous BGE. The CE-ESI-TOF-MS analysis was performed within 6 min for all TCA analytes under the optimized condition with peak efficiencies up to 1.4 x 105 plates/m and an average % RSD of the migration times of the analytes of 0.3% (n=5). The presented method can readily be used for the extraction and quantification of basic drug species in human biological fluids and in pharmaceutical formulations.

Microwave-assisted hydrolysis and extraction of tricyclic antidepressants from human hair.

The objective of this research was to develop, optimize, and validate a modern, rapid method of preparation of human hair samples, using microwave irradiation, for analysis of eight tricyclic antidepressants (TCADs): nordoxepin, nortriptyline, imipramine, amitriptyline, doxepin, desipramine, clomipramine, and norclomipramine. It was based on simultaneous alkaline hair microwave-assisted hydrolysis and microwave-assisted extraction (MAH-MAE). Extracts were analyzed by high-performance liquid chromatography with diode-array detection (HPLC-DAD). A mixture of n-hexane and isoamyl alcohol (99:1, v/v) was used as extraction solvent and the process was performed at 60°C. Application of 1.0 mol L(-1) NaOH and microwave irradiation for 40 min were found to be optimum for hair samples. Limits of detection ranged from 0.3 to 1.2 µg g(-1) and LOQ from 0.9 to 4.0 µg g(-1) for the different drugs. This enabled us to quantify them in hair samples within average therapeutic concentration ranges.

Preparation of a poly(N-isopropylacrylamide-co-ethylene dimethacrylate) monolithic capillary and its application for in-tube solid-phase microextraction coupled to high-performance liquid chromatography.

A poly(N-isopropylacrylamide-co-ethylene dimethacrylate) (poly(NIPAAm-co-EDMA)) monolith was in situ prepared in the capillary and was investigated for in-tube solid-phase microextraction (SPME). NIPAAm, an acrylamide monomer with isopropyl group, was crosslinked with EDMA. PEG of 400-20,000 Da molecular weight and methanol were selected as the binary porogens. The porous structures of the resulting monoliths have been assessed by SEM, nitrogen adsorption-desorption, and pressure drop measurements. To investigate the extraction mechanism, several groups of model analytes (including neutral, acidic, and basic) were examined. The result showed that this monolithic material exhibited high extraction efficiencies for compounds under highly acidic and basic conditions, which was due to the hydrophobic interactions and excellent pH stability of the monolith. The equilibrium extraction time profiles were also monitored for model compounds to evaluate the extraction capacity of monolithic capillary. Finally, the developed monolith in-tube SPME-HPLC method was applied to the determination of three tricyclic antidepressants from urine samples.

Selectivity differences between alkyl and polar-modified alkyl phases in reversed phase high performance liquid chromatography.

Compared to conventional C18 phases, polar-modified phases have distinct differences with regards to chromatographic behavior. In the present study, ODS phases and polar-modified phases were synthesized. The columns containing these new packings demonstrated satisfactory stability under both acidic (pH 1.5) and basic (pH 10) conditions. We evaluated the selectivity differences between alkyl and polar-modified alkyl RP columns by using a range of neutral analytes. The polar-modified alkyl phases showed excellent peak shapes for almost all compounds. We also compared the selectivity differences between them for separating nucleotides by using 100% aqueous mobile phase and tricyclic antidepressants in the intermediate pH mobile phases. The results demonstrated that polar-modified phases display a significantly reduced hydrophobic nature and a significantly reduced silanol activity compared to the conventional C18 phases.

Capillary electrophoretic separation of tricyclic antidepressants using a polymer-coated capillary and beta-cyclodextrin as an electrolyte additive.

A new method for the CE separation of nine tricyclic antidepressants (TCAs), viz. amitriptyline, clomipramine, desipramine, doxepin, fluphenazine, imipramine, nortriptyline, promazine, and thioridazine, is described. The capillary was statically coated with a layer of poly(N,N-dimethylacrylamide) (PDMA) to suppress the EOF, and beta-CD was used as an additive in the BGE solution. The optimal resolution of nine TCAs was obtained by using a 1% v/v PDMA-coated capillary and a BGE solution of 50 mM sodium phosphate buffer (pH 3.0) containing 0.5 mM beta-CD. Efficiencies were typically >10(5 )plates/m. Complete separation of nine TCAs could be achieved in about 28 min; the two diastereomers of doxepin and the two enantiomers of thioridazine could also be separated. The RSD values of migration time and peak area of the TCAs were in the ranges 0.5-0.8 and 3.3-4.9% (n = 10), respectively. In combination with a suitable sample clean-up technique, such as hollow fiber-based liquid phase microextraction (HF-LPME), the polymer-coated capillary can be employed for the CE-UV analysis of TCAs in human plasma.

Separation and determination of amitriptyline and nortriptyline by dispersive liquid-liquid microextraction combined with gas chromatography flame ionization detection.

Dispersive liquid-liquid microextraction (DLLME) coupled with gas chromatography-flame ionization detection (GC-FID) was applied for the determination of two tricyclic antidepressant drugs (TCAs), amitriptyline and nortriptyline, from water samples. This method is a very simple and rapid method for the extraction and preconcentration of these drugs from environmental sample solutions. In this method, the appropriate mixture of extraction solvent (18 microL Carbon tetrachloride) and disperser solvent (1 mL methanol) are injected rapidly into the aqueous sample (5.0 mL) by syringe. Therefore, cloudy solution is formed. In fact, it is consisted of fine particles of extraction solvent which is dispersed entirely into aqueous phase. The mixture was centrifuged and the extraction solvent is sedimented on the bottom of the conical test tube. 2.0 microL of the sedimented phase is injected into the GC for separation and determination of TCAs. Some important parameters, such as kind of extraction and disperser solvent and volume of them, extraction time, pH and ionic strength of the aqueous feed solution were optimized. Under the optimal conditions, the enrichment factors and extraction recoveries were between 740.04-1000.25 and 54.76-74.02%, respectively. The linear range was (0.005-16 microg mL(-1)) and limits of detection were between 0.005 and 0.01 microg mL(-1) for each of the analytes. The relative standard deviations (R.S.D.) for 4 microg mL(-1) of TCAs in water were in the range of 5.6-6.4 (n=6). The performance of the proposed technique was evaluated for determination of TCAs in blood plasma.

CE of tricyclic antidepressant clomipramine and metabolites: electromigration and wall adsorption.

CE of tricyclic antidepressants clomipramine and its metabolites demethylclomipramine, didemethylclomipramine and 8-hydroxyclomipramine resulted in partly extremely tailing peaks in bare fused-silica capillaries. Especially at high pH of the BGE this behavior was not unexpected as adsorption of the cationic analytes onto the negatively charged wall due to electrostatic attraction can be supposed. Less expected was the observation that peak tailing could not be overcome neither by using a capillary with dynamic coating with cationic CTAB added to the BGE, nor by the usage of a capillary permanently coated with polyvinyl alcohol (PVA), both operated at acidic pH. As this tailing was even more pronounced than with bare fused silica, and was suppressed upon addition of MeCN to the BGE, another source of adsorption than pure ion-ion interaction seems plausible. In the bare silica capillary the mobility, mu, of the analytes followed roughly the pH dependence of a monoacidic base, but two deviations from the sigmoid theoretical curve were evident: (i) even at low pH the mobilities were not constant; they decreased in contrary with pH over the entire range; (ii) the apparent pK(a) values of two analytes, derived at the pH with halve the mobility at low pH, are significantly smaller than the thermodynamic pK(a). Upon modifying the expression for mu = f(pH), and considering the pH dependence of the negative charge density at the wall by an additional term which takes chromatographic retention into account, an equation was derived which enables the description of the observed electromigration of the analytes as function of pH, pK(a) of analytes and surface silanol groups, actual mobility of analytes, distribution coefficient (or retention factor) due to adsorption including its pH dependence. The interplay of electrophoretic movement and residual adsorptive retention allowed to resolve the analytes finally in an uncoated capillary, namely at pH 7.65 (30 mM ionic strength), whereas at the cost of the robustness of the separation system.