Determination of nitrite and nitrate in a proposed certified reference material for nutrients in seawater by capillary zone electrophoresis with artificial seawater as the background electrolyte using transient isotachophoresis

We describe a combination of selected ions as a terminating ion which is useful for transient isotachophoresis (ITP) in capillary zone electrophoresis (CZE) for the determination of nitrite and nitrate in seawater. In addition to 150 mM sulfate as the principal terminating ion, 10 mM bromate was added to a sample solution as the additional terminating ion. Artificial seawater containing 3 mM cetyltrimethylammonium chloride (CTAC) was adopted as a background electrolyte (BGE). The limits of detection (LODs) for nitrite and nitrate were 2.2 and 1.0 microg/L (as nitrogen), respectively. The LODs were obtained at a signal to noise ratio (S/N) of 3. The values of the relative standard deviation (RSD) of peak area for these ions were 1.9 and 1.4%. The RSDs of peak height were 1.7 and 1.9%, the RSDs of migration time 0.11%. The proposed method was applied to the determination of nitrite and nitrate in a proposed certified reference material for nutrients in seawater, MOOS-1, distributed by the National Research Council of Canada (NRC). The results almost agreed with the assigned tolerance interval.     

Simultaneous determination of bromide, nitrite and nitrate ions in seawater by capillary zone electrophoresis using artificial seawater as the carrier solution

We describe capillary zone electrophoresis (CZE) for the simultaneous determination of bromide, nitrite and nitrate ions in seawater. Artificial seawater was adopted as the carrier solution to eliminate the interference of high concentrations of salts in seawater. The artificial seawater was free from bromide ion to enable the determination of bromide ion in a sample solution. For the purpose of reversing the electroosmotic flow (EOF), 3 mM cetyltrimethylammonium chloride (CTAC) was added to the carrier solution. A 100 microm ID (inside diameter) capillary was used to extend the optical path length. The limits of detection (LODs) for bromide, nitrite, and nitrate ions were 0.46, 0.072, and 0.042 mg/L (as nitrogen), respectively. The LODs were obtained at a signal to noise ratio (S/N) of 3. The values of the relative standard deviation (RSD) of peak area for these ions were 1.1, 1.5, and 0.97%. The RSDs of migration time for these ions were 0.61, 0.69, and 0.66%. Artificial seawater samples containing various concentrations of bromide, nitrite, and nitrate ions were analyzed by the method. The error was less than +/-12% even if the concentration ratio of bromide ion to nitrite or nitrate ion was 20-240. The proposed method was applied to the determination of bromide, nitrite, and nitrate ions in seawater samples taken from the surface and the seabed. These ions in other environmental waters such as river water and rainwater samples were also determined by ion chromatography (IC) as well as this method.     

Optimum conditions for effective use of the terminating ion in transient isotachophoresis for capillary zone electrophoretic determination of nitrite and nitrate in seawater,with artificial seawater as background electrolyte

We have examined transient isotachophoresis (ITP) conditions, e.g. the nature of the terminating ion, its concentration, and the injection procedure, to improve the limit of detection (LOD) for determination of nitrite and nitrate in seawater by capillary zone electrophoresis (CZE). Artificial seawater containing 3.0 mmol L(-1) cetyltrimethylammonium chloride (CTAC) was used as background electrolyte (BGE). After sample injection 600 mmol L(-1) acetate was separately injected into the capillary as the terminating ion for transient ITP. The LOD for nitrite and nitrate, obtained at a signal-to-noise ratio (S/N) of 3, were 15 and 7.0 microg L(-1) (as nitrogen), respectively. Relative standard deviations (RSD) of peak area for nitrite and nitrate were 7.3 and 0.8%, respectively, and the RSD of peak height were 5.7 and 1.2%, respectively, when the concentrations of nitrite and nitrate were 0.05 and 0.25 mg L(-1). The RSD of migration time for these ions was 0.2%. The proposed method was applied to the determination of nitrite and nitrate in seawater samples. The results for nitrite were nearly in agreement with those obtained by naphthylethylenediamine spectrophotometric analysis (SPA; correlation coefficient 0.9041).     

Capillary zone electrophoretic determination of iodide in seawater using transient isotachophoresis with artificial seawater as the background electrolyte

We describe an application of capillary zone electrophoresis (CZE) with transient isotachophoresis (ITP) as the on-line concentration procedure for the determination of iodide in seawater. The effective mobility of iodide was decreased by the addition of 10 mM cetyltrimethylammonium chloride (CTAC) to an artificial seawater background electrolyte (BGE) so that transient ITP functioned and iodide was separated from other coexisting anions such as bromide, nitrite, and nitrate in seawater samples. After sample injection, 600 mM acetate was separately injected into the capillary as the terminating ion to generate transient ITP. The limit of detection (LOD) for iodide was 3.0 microg/L. The LOD was obtained at a signal-to-noise ratio (S/N) of 3. The values of the relative standard deviation (RSD) of peak area, peak height, and migration time for iodide were 2.9, 2.1, and 0.6%. The proposed method was applied to the determination of iodide in seawater collected around the Osaka Bay. The results obtained by use of the calibration graph were agreed with those obtained by the addition of the standard solutions for iodide.     

Two-dimensional nano-liquid chromatography-mass spectrometry system for applications in proteomics

Transient isotachophoresis-capillary zone electrophoresis with artificial seawater as the background electrolyte (BGE) was improved to further lower the limit of detection (LOD) for determination of nitrite and nitrate in seawater. By lowering the pH of BGE, the difference between effective mobility of nitrite and that of nitrate increased, thereby permitting increased sample volumes to be tolerated and their LOD values to decrease. Artificial seawater with pH adjusted to 3.0 using phosphate buffer was adopted as the BGE. To reverse electroosmotic flow (EOF), a capillary was flushed with 0.1 mM dilauryldimethylammonium bromide for 3 min before the capillary was filled with the BGE. Limits of detection (LODs) for nitrite and nitrate were 2.7 and 3.0 microg/l (as nitrogen), respectively. The LODs were obtained at a signal-to-noise ratio of 3. Values of the relative standard deviation (RSD) of peak area for these ions were 2.0 and 0.75%, respectively, when nitrite concentration was 0.05 mg/l and that of nitrate was 0.5 mg/l. The RSDs of peak height were 4.4 and 2.3%. The RSD values of migration time for these ions were 0.19 and 0.17%. The proposed method was applied to determination of nitrite and nitrate in a proposed certified reference material for nutrients in seawater, MOOS-1, distributed by the National Research Council of Canada. Results agreed with the assigned tolerance interval. This method was also applied to determination of these ions in seawater collected around Osaka Bay. Results nearly agreed with those obtained by a conventional spectrophotometric method.     

Determination of nitrate in seawater by capillary zone electrophoresis with chloride-induced sample self-stacking

A capillary zone electrophoresis (CZE) method was established to determine low concentration nitrate which was online preconcentrated with chloride-induced leading-type sample self-stacking for seawater samples. The sample self-stacking was based on transient isotachophoresis in which chloride served as leading ion, and dihydrogenphosphate in the background electrolyte (0.1 M phosphate) as the terminating one. Due to the small mobility difference between nitrate and chloride, the isotachophoresis time was so long that nitrate could not separate from the rear sharp boundary between chloride and the background electrolyte (BGE) when it migrated to the detection window. A zwitterionic surfactant, 3-(N,N-dimethyldodecylammonio)propane sulfonate was added to the BGE to enlarge the mobility difference for its selective interaction with anions. Thus, a highly conductive sample could be injected in a large volume with about fourfold sensitivity enhancement compared to that of field amplification sample stacking in which nitrate was dissolved in pure water. The relative standard deviations (n=5) of migration time, peak area, peak height were 0.1, 3.0, 1.5%, respectively. The limit of detection (S/N=3) for nitrate was 35 microg/l in seawater samples with relatively low concentration BGE (0.1 M sodium phosphate, pH 6.2). The overall procedure consisting of online preconcentration and separation was as simple as routine CZE except for a slightly longer sample injection time (3-4 min).     

Capillary electrophoresis using high ionic strength background electrolytes containing zwitterionic and non-ionic surfactants and its application to direct determination of bromide and nitrate in seawater

In capillary electrophoresis, it is commonly considered that even a moderately high ionic concentration in the background electrolyte (BGE) leads to high currents, resulting in Joule heating and serious peak distortion. As a new approach to overcome this problem, zwitterionic (Zwittergent-3-14) and/or non-ionic (Tween 20) surfactants have been added to BGEs containing high salt concentrations (e.g. 0.3 M NaCl) and have been shown to result in acceptable separation currents (<200 microA). In turn, these BGEs could be applied to the separation of samples containing high salt concentrations (such as undiluted seawater) without the occurrence of any significant peak broadening due to electrodispersion of the sample. For example, a BGE comprising 10 mM Zwittergent-3-14, 50 mM Tween 20, 0.3 M NaCl and 5 mM phosphate (ph 7) could be used for the determination of UV-absorbing anions in seawater, giving good peak shapes and detection limits of 0.8 microM and 0.6 microM for nitrate and bromide, respectively. The beneficial effects of the non-ionic surfactant on the separation were attributed largely to suppression of the electro-osmotic flow. On the other hand, the zwitterionic surfactant was found to be capable of the incorporation of some anions in accordance with the behaviour of these same surfactants in electrostatic ion chromatography. This incorporation resulted in a decreased conductivity of the BGE and also a change in the separation selectivity of the system.     

Simultaneous determination of nitrite and nitrate in human plasma by on-capillary preconcentration with field-amplified sample stacking

A simple method for the determination of nitrite and nitrate in human plasma has been developed using CZE with minimal sample preparation. Field‐amplified sample stacking (FASS) was used to achieve submicromolar detection by dilution of the plasma sample with deionized water. In CZE, the separation of nitrite and nitrate was achieved within 10 min without adding EOF modifier. The optimal condition was achieved with 50 mM phosphate buffer at pH 9.3. The ninefold diluted plasma samples were injected hydrodynamically for 40 s into a 60 cm×75 μm id uncoated fused‐silica capillary. The separation voltage was 20 kV (negative potential) and UV detection was performed at 214 nm. The linearity curves for nitrite and nitrate were obtained by the standard addition method. The estimated LODs for nitrite and nitrate in ninefold diluted plasma sample were 0.05 and 0.07 μM, respectively. The LODs for nitrite and nitrate in original plasma samples were 0.45 and 0.63 μM. The intra‐ and inter‐day precisions for both analytes were <2.6% and the recovery ranged between 92.3 and 113.3%. It was found that nitrite was more stable than nitrate in the plasma after the sample preparation. This proposed method was applied to a number of human plasma samples and the measured nitrite and nitrate concentrations in human plasma were consistent with the literature ranges.     

DNA fragment separations by on‐line combination of capillary isotachophoresis–capillary zone electrophoresis with UV detection

A high‐speed DNA fragment separation system based on an on‐line combination of capillary ITP with CZE (CITP‐CZE) and using UV detection at 260 nm was developed. A novel CITP‐CZE buffer system of pH 6.1 was designed for the separation of ten DNA fragments with sizes ranging from 100 to 1000 bp. An effect of underivatized α‐, β‐ and γ‐cyclodextrins on the resolution of DNA fragments in the CZE step of the CITP‐CZE combination was systematically investigated. Methylhydroxyethylcellulose present in the BGE was used to eliminate the EOF. DNA ladder fragments were separated within 10 min with LODs in the range of 1–5 ng/μL (S/N = 3). The RSDs of the migration time and peak area of individual DNA fragments were in the range of 1–3 and 3–9%, respectively. The developed CITP‐CZE system was further applied to the analysis of digest plasmid DNA samples.     

Analysis of sub-ppb levels of Fe(II), Co(II), and Ni(II) by electrokinetic supercharging preconcentration, CZE separation, and in-capillary derivatization

The analysis of sub-ppb levels of Fe(II), Co(II), and Ni(II) in heat exchanger fluids of nuclear power plants is needed to monitor corrosion. A method involving preconcentration with electrokinetic supercharging (electrokinetic injection with transient ITP), CZE separation, and in-capillary derivatization with ortho-phenanthroline (o-Phe) for direct UV detection was thus developed. First, a multizone BGE was loaded into the capillary by successive hydrodynamic introduction of zones of (i) o-Phe-containing BGE, (ii) BGE for the zonal separation, and (iii) ammonium-based leading electrolyte. Metal cations were electrokinetically injected and stacked at the capillary inlet behind this last leading zone. Finally, a terminating electrolyte zone was hydrodynamically introduced. When a constant voltage was applied, metal ions kept on concentrating isotachophoretically, then separated in CZE mode, were complexed by migrating through an o-Phe zone, and finally detected by direct absorbance. To detect extremely thin peaks, it was attempted for the first time to focus the derivatization reagent by inducing a second transient ITP, before labeling analytes, already separated in CZE mode. With this arrangement, LODs were about 30 ppt in pure water. In heat exchanger fluid matrices containing 1000 ppm bore and 2 ppm lithium, only Fe(II) cation was detected among the three cations of interest at the 1 ppb level using the present method, and its LOD was about ten times higher, due to the lower loading rate during electrokinetic injection.     

Simultaneous determination of iodide and iodate in seawater by transient isotachophoresis-capillary zone electrophoresis with artificial seawater as the background electrolyte

We developed capillary zone electrophoresis with transient isotachophoresis (ITP) as an on-line concentration procedure for simultaneous determination of iodide and iodate in seawater. The effective mobility of iodide was decreased by addition of 20 mM cetyltrimethylammonium chloride to an artificial seawater background electrolyte so that transient ITP functioned for both iodide and iodate. Limits of detection for iodide and iodate were 4.0 and 5.0 microg/l (as iodine) at a signal-to-noise ratio of 3. Values of the relative standard deviation of peak area, peak height, and migration times for iodide and iodate were 2.9, 1.3, 1.0 and 2.3, 2.1, 1.0%, respectively. The proposed method was applied to simultaneous determination of iodide and iodate in seawater collected at a pond at our university.     

Determination of phosphate in seawater by CZE with on-line transient ITP

We developed CZE with indirect UV detection for the determination of phosphate in seawater using transient ITP as an on-line concentration procedure. The following optimum conditions were established: BGE, 5 mM 2,6-pyridinedicarboxylic acid (PDC) containing 0.01% hydroxypropylmethylcellulose (HPMC) adjusted to pH 3.5; detection wavelength, 200 nm; vacuum injection period of sample, 3 s (45 nL); terminating ion solution, 500 mM MES adjusted to pH 4.0; vacuum injection period of the terminating ion solution, 30 s (450 nL); applied voltage, 30 kV with the sample inlet side as the cathode. The LOD for phosphate was 16 microg/L (PO(3-)(4) -P) at S/N of 3. The respective values of the RSD of the peak area, peak height, and migration time for phosphate were 2.6, 2.3, and 0.34%. The proposed method was applied to the determination of phosphate in a seawater certified reference material for nutrients, MOOS-1, distributed by the National Research Council of Canada (NRC). The results were very similar to certified values. The method was also applied to the determination of phosphate in coastal seawaters. The results agreed with those obtained using a conventional spectrophotometric method.     

CZE study on adsorption processes of aliphatic and aromatic amines on PMMA chip

Adsorption processes on a PMMA chip linked with CZE separations of a group of 13 aliphatic and aromatic mono‐ and di‐amines were studied. Due to the lack of chromophores within aliphatic amines, contact conductivity detection implemented directly onto the chip was used for monitoring of cationic CZE separations. To prevent an adsorption of studied amines to the chip channels, the surface of PMMA chip was modified by dynamic coating. Different surface modifiers, such as aliphatic oligoamines (diethylenetriamine and triethylenetetramine), were added to the BGE solutions filling the chip channels. The effect of various concentrations of surface modifiers on peak profiles and separation parameters of amines was monitored. Of these, mainly, aliphatic di‐amines and aromatic mono‐amines adversely affected the CZE resolution of a whole group of analytes by their strong adsorption to the chip channels. A propionate BGE with pH 3.2 containing 100 μM triethylenetetramine and 25 mM 18‐crown‐6‐ether was found suitable for CZE resolution of 12 from a total of 13 amines studied. Simple dynamic modification of the surface of PMMA chip enabled fast (analysis time lasted 9 min), sensitive (sub‐μM LODs reached) and reproducible (1–3% RSD of the peak areas) CZE analysis of the aliphatic and aromatic amines.     

Onboard determination of submicromolar nitrate in seawater by anion-exchange chromatography with lithium chloride eluent.

Ion-exchange chromatography using a high-capacity anion exchanger with UV detection was applied to the determination of nitrate in seawater. Major ions in seawater samples did not affect the peak shape and the retention time of the nitrate when an alkaline metal cation-chloride solution was used as an eluent at high concentrations (0.5-2 mol/l). At a wavelength of 220 nm, the peak of bromide was very small because of low absorption, while its separation from the nitrate peak was good at high concentrations. Among the eluents tested, lithium chloride gave the best separation of nitrate from bromide. It was estimated that the lithium ion had the least potential for ion-pair formation with nitrate, and its retention time was prolonged compared with the retention times when using other cations; with bromide and nitrite, such an effect was not observed. The results of shipboard seawater nitrate determination by our method in the South Pacific Ocean and Antarctic Sea showed good agreement with those by the conventional photometric method using continuous flow.     

Simultaneous determination of weakly ionizable analytes in urine and plasma samples by transient pseudo-isotachophoresis in capillary zone electrophoresis

A rapid method for the simultaneous determination of several non-steroidal anti-inflammatory drugs (NSAIDs) in human plasma and urine was developed using transient pseudo-isotachophoresis (ITP) in capillary zone electrophoresis (CZE). The influence of different parameters on resolution and preconcentration efficiency, such as background electrolyte (BGE) composition, sample injection, sample matrix composition, and pH, were studied to optimize the transient pseudo-ITP performance. Optimized conditions were a BGE consisting of 100 mM Na₂B₄O₇ in 10% aqueous MeOH solution and hydrodynamic injection of the sample at 50 mbar for 90 s. The sample was prepared in a solution mixture of 1% NaCl/ethanol (30:70 v/v) at pH 10. Our results show that this simple strategy offers improved sensitivity compared to conventional CZE analysis, reaching a 45-fold preconcentration factor. The detection limits (LODs) were as low as 0.07 mg/L for standard samples with good repeatability (values of relative standard deviation, %RSD < 11%). The method was applied to the analysis of NSAIDs in biological samples. Validation for human plasma and urine samples demonstrated good linearity, low detection limits, and satisfactory repeatability values.     

Determination of trace cationic impurities in butylmethylimidazolium-based ionic liquids: from transient to comprehensive single-capillary counterflow isotachophoresis-zone electrophoresis

Determination of impurities in ionic liquids (ILs) remains a difficult task. In this work, the hyphenation of isotachophoretic (ITP) preconcentration to zone electrophoresis (ZE) has been explored for the trace analysis of the cationic impurities Na(+), Li(+), and methylimidazolium (MI(+)) in butylmethylimidazolium (BMI(+))-based ILs. Simultaneous detection of UV-transparent and UV-absorbing impurities was ensured by a BGE composed of creatinine-acetate buffer. To induce ITP, three different strategies were evaluated: (i) Sample self-stacking ensured by the addition of ammonium acetate (NH(4)Ac) to 25-50-fold diluted IL solution (transient ITP). (ii) Complete ITP-ZE separation performed in a single capillary: ITP was realized in discontinuous electrolytes comprising an 80 mM NH(4)Ac, 40 mM acetic acid, 30 mM alpha-CD, pH 5.05, leading electrolyte (LE) and a 10 mM creatinine, 10 mM acetic acid, pH 4.9, terminating electrolyte (TE). To create the ZE stage, the ITP stack of analytes was moved back toward the capillary inlet by pressure and simultaneously the capillary was filled with the BGE. This protocol made it possible to accommodate a 2.5-times diluted IL sample. (iii) Complete counterflow ITP-ZE with continuous electrokinetic sample supply: the ITP stage was performed in a capillary filled with a 150 mM NH(4)Ac, 75 mM acetic acid, 30 mM alpha-CD, pH 5.0 LE, with 40-times diluted IL at the capillary inlet. BMI(+) from IL acts as the terminating ion. The LODs reached in this latter case were at the 10 and 1 ppb levels for MI(+) and Li(+) in diluted IL matrix, respectively.     

Measurement of nitrate and chlorate in swimming pool water by capillary zone electrophoresis

Capillary zone electrophoresis (CZE) of nitrate and chlorate in swimming pool water are described. Nitrate and chlorate were determined simultaneously with an indirect detection method in an electrolyte containing 10 mM chromate and 0.1 mM cetyltrimethylammonium bromide (CTAB). Where chloride concentration was so high that nitrate could not be determined satisfactorily because of interference, a direct detection technology was developed in which 10 mM sulfate and 0.1 mM CTAB were used as the buffer. The wavelength for indirect detection was 254 nm and 214 nm for direct detection. Relative standard deviations of the quantification of nitrate and chlorate in real samples were below 6%. The detection limits were 7 mug ml(-1) for chlorate, and 4 mug ml(-1) (indirect detection) and 0.4 mug ml(-1) (direct detection) for nitrate.     

On-line coupling of capillary isotachophoresis and capillary zone electrophoresis for the determination of flavonoids in methanolic extracts of Hypericum perforatum leaves or flowers

Five flavonoids (hyperoside, isoquercitrin, quercitrin, quercetin and rutin) were separated and determined in extracts of Hypericum perforatum leaves or flowers by capillary zone electrophoresis (CZE) with isotachophoretic (ITP) sample pre-treatment using on-line column coupling configuration. The background electrolyte (BGE) used in the CZE step was different from the leading and terminating ITP electrolytes but all the electrolytes contained 20% (v/v) of methanol. The optimal leading electrolyte was 10 mM HCl of pH* approximately 7.2 (adjusted with Tris) and the terminating electrolyte was 50 mM H3BO3 of pH* approximately 8.2 (adjusted with barium hydroxide). This operational system allowed to concentrate and pre-separate selectively the flavonoid fraction from other plant constituents before the introduction of the flavonoids into the CZE capillary. The BGE for the CZE step was 50 mM Tris buffer of pH* approximately 8.75 containing 25 mM N-[tris(hydroxymethyl)methyl]-3-aminopropanesulfonic acid as co-ion and 55 mM H3BO3 as complex-forming agent. The ITP-CZE method with spectrophotometric detection at 254 nm was suitable for the quantitation of the flavonoids in real natural samples; kaempferol was used as internal standard. The limit of detection for quercetin-3-O-glycosides was 100 ng ml(-1) and calibration curves were rectilinear in the range 1-10 microg ml (-1) for most of the analytes. The RSD values ranged between 0.9 and 2.7% (n=3) when determining approximately 0.07-1.2% of the individual flavonoids in dried medicinal plants.     

Dermal nitrate: an old marker of firearm discharge revisited with capillary electrophoresis

The present work describes a capillary electrophoretic method for nitrite and nitrate determination to be used as a screening tool for investigating the residues of firearm discharge. The use of capillary electrophoresis allowed the rapid determination of nitrite and nitrate, which are major inorganic components of gunshot residues, offering a quantitative and selective alternative to the traditional paraffin test (dermal nitrate test). The method is simpler, cheaper, and faster than the modern approaches to gunshot residue analysis based on the determination of barium, lead and antimony by using flameless absorption spectrometry, inductively coupled plasma-mass spectrometry (ICP-MS), or scanning electron microscopy. The analysis was carried out in a bare fused-silica capillary (75 microm inner diameter) with a 100 mM borate buffer (pH 9.24). The detection was by UV absorption at 214 nm. Separation took place under reversed voltage of 15 kV. Bromide was used as the internal standard. Sensitivity was about 1 mM for both nitrite and nitrate. Reproducibility (intraday and day-to-day) was also good with relative standard deviations (RSDs) < 1.0% for relative migration times and < 4.5% for peak areas in both standard solutions and real matrix. Hair and skin samples from a victim shot in the head were successfully analyzed for the presence of nitrite and nitrate.     

Rapid separation of tetracycline derivatives and their main degradation products by capillary zone electrophoresis.

A mixture of five tetracycline (TC) derivatives: minocycline (MC), demeclocycline (DMCTC), doxycycline (DC), and sancycline (SC), as well as each TC derivative from its main degradation product were separated by capillary zone electrophoresis (CZE). The influence of the pH and the concentration and nature of the background electrolyte (BGE) on the separations was investigated. Ethylenediaminetetraacetic acid (EDTA; 1 mM) was used as additive in a 25 mM phosphate buffer (pH 2.3) because this BGE enabled the rapid separation of the TC derivatives and of each TC derivative from its respective degradation product in less than 6 min. After optimization of the separation conditions, the analytical characteristics of the method were investigated. The parameters involved were linearity, precision (repeatability and reproducibility), and limits of detection (LODs). LODs obtained for the five TC derivatives studied were about 3 microg/mL. Finally, the CZE method developed was applied to study the stability of TC derivatives and to analyze the TC derivative content in three different pharmaceutical preparations.