Monitoring degradation processes of explosives by HPLC analysis with UV- and amperometric detection

 The impact of spilled explosives, their by-products and degradation products on human beings and the environment has been recognised as a serious problem at areas of existing and former ammunition plants. In nature, aerobic and anaerobic degradation processes of explosives and their accompanying compounds yield polar contaminants with relatively high water solubilities. Most are potentially carcinogenic and mutagenic. An HPLC method applying UV-detection for nitroaromatic compounds and amperometric detection for aminoaromatic and phenolic compounds was used for monitoring the degradation of explosives in a polluted groundwater sample under natural conditions. Analysis was performed by direct injection of aliquots of the sample after exposition to daylight for different periods of time. Received: 6 January 1996/Revised: 7 March 1996/Accepted: 13 March 1996     

Determination of clotiazepam by HPLC with spectrophotometric and amperometric detection

Summary Spectrophotometric and amperometric detectors were compared for the determination of clotiazepam by HPLC. Methanol-water (6040) containing 0.01 mol/l ammonium acetate and methanol-water (7030) containing 0.01 mol/l acetate buffer pH 4 were used as mobile phase in the spectrophotometric and amperometric detection, respectively. The minimum detectability for the spectrophotometric detection was 20 ng injected and the method was applied for the determination of clotiazepam in serum, whereas for the amperometric detection a limit of detection of 2.5 g injected was obtained, using this technique for clotizepam determination in tablets.

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Bestimmung von Clotiazepam durch HPLC mit spektralphotometrischer und amperometrischer Detektion

     

Monitoring myoglobin by capillary zone electrophoresis with end-column amperometric detection

Capillary zone electrophoresis was employed for the determination of myoglobin in human urine using end-column amperometric detection with a carbon fiber microelectrode at a constant potential of 1.80 V vs. saturated calomel electrode (SCF). The optimum conditions of separation and detection are: 3.73 x 10-4 mol/L sodium diethyl malonyl urea (barbitone sodium), 1.34 x 10-4 mol/L HCl for the buffer solution, 20 kV for separation voltage, 5 kV and 5 s for injection voltage and injection time, respectively. The limit of detection is 4.4 x 10-8 mol/L or 84 amole signal to noise (S/N = 2). The relative standard deviation is 2.9% for the migration time and 2.5% for the electrophoretic peak current. The method can be used for the determination of myoglobin in human urine. The samples can be directly injected and need no pretreatment. The method is also rapid, less than 2 min, and has a recovery rate of 94-106%.     

Determination of adenine,caffeine, theophylline and theobromine by HPLC with amperometric detection

A relatively simple method for quantifying caffeine, theobromine, theophylline and adenine by HPLC with amperometric detection was developed. A C₁₈-column and an isocratic elution with phosphate buffer pH 3.5/methanol (90 : 10) were employed for the chromatographic separation of the investigated compounds. The optimal detection potential was +1.4 V. The limits of detection were 0.4 ng for adenine, 1 ng for theophylline and 2.5 ng for caffeine and theobromine. The method was applied to the determination of these purine alkaloids in beverages, tea, coffee and cacao. The determination was carried out directly or after solid-phase extraction.     

Speciation of iron coordinated by phytosiderophores by use of HPLC with pulsed amperometric detection and AAS

Phytosiderophores of the mugineic acid family, and the respective iron species, have been separated by anion-exchange chromatography with NaOH gradient elution. Two different detection methods were used in parallel, pulsed amperometry (PAD) for phytosiderophores and atomic absorption spectrometry (AAS) for iron. This combination enables identification of separated iron species. Up to five different iron species were separated and detected within 30 min - two different phytosiderophore species, two amino acid species, and one species which has not yet been identified but which is most probably a decomposition product of phytosiderophores. The detection limit was in the low micro mol L(-1) concentration range, which is sufficiently low for determination in real plant samples, even after dilution. The method has been applied to root washings of iron-deficient wheat and barley plants and to a xylem exudate of non-deficient maize.     

Monitoring human serum transferrin by capillary zone electrophoresis with end-column amperometric detection

Capillary zone electrophoresis was employed for the determination of human serum transferrin using end-column amperometric detection with a carbon fiber microelectrode at a constant potential of 1.9 V vs. saturated calomel electrode (SCE). The optimum conditions of separation and detection are 7.5 x 10(-4) mol/L Tris-3.44 x 10(-4) mol/L HCl for the buffer solution, 20 kV for the separation voltage, 5 kV and 10 s for the injection voltage and the injection time, respectively. The limit of detection is 6.7 x 10(-8) mol/L or 440 amol (S/N = 2). The relative standard deviations are 0.67% for the migration time and 1.5% for the electrophoretic peak current. The method was applied to the determination of transferrin in human serum. The recovery is between 93-104%.     

Analysis of tetracycline antibiotics using HPLC with pulsed amperometric detection.

The analysis of tetracycline, oxytetracyline, chlortetracycline and doxycycline by high-performance liquid chromatography with pulsed amperometric detection using an anodized boron-doped diamond thin film (BDD) electrode is originally reported. The analyses were carried out using the mobile phase, phosphate buffer (0.01 M, pH 2.5)-acetonitrile (80:20; v/v), on a C18 column (250 x 4.6 mm i.d., 5 microm) at a flow rate of 1.0 mL/min. The optimal PAD waveform parameters at the anodized BDD were 1.5 V (versus Ag/AgCl) detection potential (E(det)) for 290 ms (200 ms delay time and 90 ms integration time), 2.0 V (versus Ag/AgCl) oxidation potential (E(oxd)) for 200 ms oxidation time (t(oxd)) and 0.4 V (versus Ag/AgCI) reduction potential (E(red)) for 200 ms reduction time (t(red)). The proposed method showed the simultaneous determination of tetracycline, oxytetracyline, chlortetracycline and doxycycline with a linear range of 0.1 - 100 microg/mL, detection limits of 0.05 - 0.1 microg/mL and recoveries of 70.8 - 96.0%. The application of this method to real samples was demonstrated and validated using a shrimp sample.     

Determination of sulphur dioxide by flow injection analysis with amperometric detection

Sulphur dioxide can be determined at a sampling rate of 120 h^?1, with amperometric detection after separation in a diffusion cell with a teflon membrane. At 25°C, the calibration graph shows two linear ranges, between 0.06 and 6 mg l^?1 and 12 and 110 ml l^? sulphur dioxide, with a detection limit of 0.03 mg l^?1. At 50°C, the liner range is 04–5 mg l^?1, with a detection limit of mg of l^?1. The procedure has been applied to the determination of sulphur dioxide in wines.     

HPLC analysis of thimerosal and its degradation products in ophthalmic solutions with electrochemical detection

A reverse-phase high-performance liquid chromatographic assay using amperometric detection on a glassy-carbon electrode has been developed for analysis of thimerosal and its main degradation products, thiosalicylic acid and dithiodibenzoic acid, in ophthalmic formulations. A potential value of 0.9 V vs. Ag/AgCl was chosen for simultaneous detection of thimerosal and thiosalicylic acid, obtaining limits of detection of 1.0 and 0.2 ng injected, respectively. A potential value of 1.2V was applied for simultaneous determination of all three compounds studied, obtaining in this case limits of detection of 3,4 and 4 ng injected for thimerosal, thiosalicylic acid and dithiodibenzoic acid, respectively. The results obtained reveal the utility of the HPLC method in quality control of commercial products containing thimerosal with good detectability.     

HPLC with amperometric detection for the determination of pesticides of high polarity

A relatively simple procedure is described for the selective and highly sensitive determination of amitrole (3-amino-1,2,4-triazole) by ionpair HPLC with amperometric detection. The separation was achieved on a polar CN-column using hexane, propanol, water (62:36:2) with LiClO₄ (2.5 g·l^–1) and trichloroacetic acid (2 g·l^–1) as electrolytes. The determination range lies between 500 pg and 500 ng and the limit of detection was found to be 200 pg (s.d.: 11%; N=9). The minimum detectability for the alternative UV-monitoring was 2.5 ng. The developed method may be helpful in production and quality control and for the selective determination of amitrole in commercial formulations.     

Indirect determination of bromide by diffusion flow injection analysis with amperometric detection

Summary A rapid, indirect diffusion flow injection analysis (FIA) method with amperometric detection has been developed for the selective and sensitive determination of Br^–. The method is based on permanganate oxidation of Br^– to bromine. Bromine diffuses through a PTFE membrane and is quantified amperometrically at a platinum working electrode. Calibration graphs were linear up to the maximum concentration of Br^– investigated (10.0 mmol/l). The precision of the technique was better than a relative standard deviation of 0.7% at 10.0 mol/l, with a throughput of 30 samples per hour. The effects of temperature, acidity, working potential, composition of the reagent solution and interferents on the FIA signals were studied. The catalytic effect of Cl^– on the permanganate oxidation of the analyte was utilized to lower the detection limit to 1 mol/l (16 ng Br^–). Similar detection limits were achieved by combining the effects of higher acidity (4.0 mol/l H₂SO₄) and elevated temperatures (40°C). The method was successfully applied to the determination of Br^– in chloride and other reagents, as well as in natural waters.     

Flow injection analysis of warfarin sodium with amperometric detection

Summary A flow-injection method for the determination of warfarin sodium and its dosage forms is reported. The method is based on the oxidizability of the drug at the glassy carbon electrode. Applying an electrode potential of +1.5 V versus Ag/AgCl reference electrode, a calibration curve was found to be linear in the 1–40 g/ml concentration range with minimum detectability of 5 ng/ml (signal-to-noise ratio 2). Although automation was not used in this study, the method could be readily incorporated into automated systems because it employs the technique of continuous analysis in a flowing stream.     

Residue analysis of ethylenethiourea in vegetables by high-performance liquid chromatography with amperometric detection

Summary A method for the residue analysis of ethylenethiourea (ETU) in vegetables by HPLC with amperometric detection was developed. ETU was separated with Nucleosil C₁₈ as stationary phase and methanol-water (5:95, v/v) containing 0.05 mol/l ammonium acetate as eluent. The voltage of the working electrode (glassy carbon) was set at +1150 mV vs. Ag/AgCl. The limit of detection for ETU was found to be 0.3 ng. The method was applied to the determination of trace amounts of ETU in tomatoes and cucumbers. The minimum quantitation level was 0.01 ppm. The method can be performed easily; it is selective and highly sensitive.     

Selective detection of metal species in HPLC and FIA by means of pulsed amperometric detection (PAD)

Triple-step pulsed amperometry is used to detect selectively metal complex species after HPLC separation in the presence of coeluting matrix compounds. Optimized pulse parameters enable accurate and selective detection of species even in the presence of electroactive compounds such as phenols or sulfurcontaining peptides. Complexes of tin(IV) with rutin and of platinum(II) with glutathione are presented as examples for these classes of compounds. It is demonstrated that by changing the pulse waveform direct and indirect detection modes can be realized. The method is applied to detect platinum species in a grass extract and iron(II)-lactate in fruit juice. Furthermore it is shown, that pulsed amperometry can be used as a detector in FIA for determining the fraction of metal complex formed after mixing of metal and excess ligand without separation.     

Characterization of low-molecular-weight metal species in plant extracts by using HPLC with pulsed amperometric detection and cyclic voltammetry

An analytical separation scheme is presented for the isolation of low-molecular-weight metal species (< 10 kDa) in plants. After ultrafiltration of the aqueous plant extracts isotachophoresis or gel chromatography is used for pre-separation and HPLC at a cyclodextrin-phase for further separation of metal containing fractions. Trace metals are detected off-line by using either adsorptive stripping voltammetry for platinum or AAS for zinc and magnesium. It is shown that platinum and zinc detection closely correlate with pulsed amperometric detection (PAD), while magnesium behaves differently. PAD is carried out after post-column addition of sodium hydroxide, thus enabling a sensitive and selective detection of carbohydrates. It is demonstrated that cyclic voltammetry is a useful tool for further characterization of the PAD-detectable metal species. By comparison of the cyclic voltammograms of carbohydrate standards, including carbohydrate-metal species, with the respective voltammograms obtained from HPLC fractions, carbohydrate species isolated from the samples can be considered as being either partly oxidized sugars (sugar alcohols or sugar acids) or glycosidically bound sugars.     

Internal standard in flow injection analysis with amperometric detection

In this work, we describe for the first time the use of the internal standard method in flow injection analysis (FIA) with amperometric detection. The method is based on the application of sequential potential pulses to the working electrode in an electrochemical flow cell. The sequence of potential pulses is selected in such a way that the analyte and internal standard compound are detected and monitored individually and independently at the same working electrode. This approach compensates for random errors associated with variations of flow rate, injection volume, ionic strength difference between standards and samples, and accidental insertion or formation of air bubbles in the carrier stream. In addition, this method can overcome the major drawback of amperometric detection using solid electrodes, which is gradual electrode passivation. To illustrate the potential of this method, the flow-injection amperometric detection of uric acid using [Fe(CN)₆]^3? as an internal standard (IS) is presented as an example.     

Characterization of low-molecular-weight metal species in plant extracts by using HPLC with pulsed amperometric detection and cyclic voltammetry

An analytical separation scheme is presented for the isolation of low-molecular-weight metal species (< 10 kDa) in plants. After ultrafiltration of the aqueous plant extracts isotachophoresis or gel chromatography is used for pre-separation and HPLC at a cyclodextrin-phase for further separation of metal containing fractions. Trace metals are detected off-line by using either adsorptive stripping voltammetry for platinum or AAS for zinc and magnesium. It is shown that platinum and zinc detection closely correlate with pulsed amperometric detection (PAD), while magnesium behaves differently. PAD is carried out after post-column addition of sodium hydroxide, thus enabling a sensitive and selective detection of carbohydrates. It is demonstrated that cyclic voltammetry is a useful tool for further characterization of the PAD-detectable metal species. By comparison of the cyclic voltammograms of carbohydrate standards, including carbohydrate-metal species, with the respective voltammograms obtained from HPLC fractions, carbohydrate species isolated from the samples can be considered as being either partly oxidized sugars (sugar alcohols or sugar acids) or glycosidically bound sugars. Received: 26 January 1998 / Revised: 19 June 1998 / Accepted: 24 June 1998