Sensitive assay for catecholamines in pharmaceutical samples and blood plasma using flow injection chemiluminescence analysis.

A rapid and sensitive chemiluminescence (CL) method using flow injection analysis was described for the determination of three catecholamines: dopamine, adrenaline and dobutamine, based on their greatly enhancing effects on the CL reaction of luminol-potassium periodate in basic solutions. Under the optimized conditions, the calibration graphs relating the increase of CL intensity to the concentration of the analytes were linear. The present method allows for the determination of dopamine, adrenaline, and dobutamine over the range of 1.0 x 10(-10) - 1.0 x 10(-7) g/ml. The relative standard deviations for measurements (n=11) of dopamine, adrenaline and dobutamine were 2.9, 2.3 and 1.8% when the concentrations of three catecholamines were at 1.0 x 10(-9) g/ml, respectively. The detection limits of the method were 2.0 x 10(-11) g/ml dopamine, 1.0 x 10(-11) g/ml adrenaline and 4.0 x 10(-11) g/ml dobutamine. The method was successfully applied to the determination of three catecholamines in pharmaceutical samples and blood plasma.     

A multicommutation-based flow system for multi-element analysis in pharmaceutical preparations

A flow system exploiting multicommutation and multidetection is proposed for sequential determinations in pharmaceutical preparations. The feasibilities were demonstrated by the determination of zinc, iron, copper, calcium and magnesium without changing the flow set-up. The gravitational flow of the solutions was exploited for addition of different chromogenic reagents and sample aliquots, thus avoiding the use of a propulsion unit. Transient signals at different wavelengths were measured simultaneously employing a fiber-optic multichannel spectrophotometer. Coefficients of variation of 1.0, 1.5, 1.4, 2.5 and 2.0% were obtained for iron, zinc, copper, calcium and magnesium, respectively. The mean sampling rate for the five species was 60 determinations per hour. In comparison to continuous reagent addition systems, the consumption was up to 160-fold lower. Results for pharmaceutical preparations agreed with those obtained by Flame atomic absorption spectrophotometry (FAAS) at the 95% confidence level.     

Determination of clonazepam in pharmaceutical preparations using simple high-throughput flow injection system

This study was aimed at examining two flow injection-spectrophotometric systems (normal and reverse) for the determination of clonazepam (CLO) at the microgram level in pure and pharmaceutical dosage forms. The estimation of CLO has been developed by conjugating a normal (or reverse) flow injection analysis (nFIA or rFIA) and spectrophotometric detection with phloroglucinol as a coupling reagent. Beer’s law was obeyed over a range of 50–400 and 30–400 μg/mL. The limits of detection were 11 and 8 μg/mL and the sampling rates were 51 and 28 samples per hour for nFIA and rFIA respectively. Both systems were successfully applied for the determination of CLO in its commercially available dosage forms. A comparison between the proposed flow systems was also done. These simple and high throughput methods could be utilized for pharmaceutical analysis of CLO.     

Amperometric detection for capillary flow injection analysis

Ultrasmall-volume measurements of oxidizable compounds have been accomplished by coupling a capillary flow injection system with amperometric detection. Remarkably low (femtomole) mass detection limits result from the combination of nanoliter sample volume and the inherent sensitivity of the wall-jet detector. A substantial economy of reagent consumption and disposal accrues from the operation of the nl/min flow regime. Variables influencing the physical dispersion in the capillary flow injection system, including capillary length, sample volume or flow rate, are explored and optimized.     

Subnanogram determination of aniracetam in pharmaceutical preparations and biofluids by flow injection analysis with chemiluminescence detection based on its enhancement of the myoglobin-luminol reaction

A novel flow injection chemiluminescence method with a myoglobin-luminol system is described for determining aniracetam. Myoglobin-bound aniracetam produced a complex that catalyzed the chemiluminescence reaction between luminol and myoglobin, leading to fast chemiluminescence. The chemiluminescence intensity in the presence of aniracetam was remarkably enhanced compared with that in the absence of aniracetam. Under the optimum reaction conditions the chemiluminescence increment produced was proportional to the concentration of aniracetam in the range of 0.1-1000.0 ng/mL (R2 = 0.9992), with a detection limit of 0.03 ng/mL (3delta). At a flow rate of 2.0 mL/min, the whole process, including sampling and washing, could be completed in 0.5 min, offering a sampling efficiency of 120/h; the RSD was less than 3.0% (n = 5). The method was satisfactory for determination of aniracetam in pharmaceutical preparations and human urine and serum samples. A possible mechanism of the reaction is also discussed.     

Incorporation of flow injection analysis or capillary electrophoresis with resonance Rayleigh scattering detection for inorganic ion analysis

Resonance Rayleigh scattering (RRS) has been explored as a detection (RRSD) technique for capillary electrophoresis (CE) or flow injection analysis (FIA) of inorganic ions. The detection was achieved through a scattering probe of ion-association complex formed from rhodamine B (Rh B) and iodine. The probe scatters strongly at 630 nm when oxidants such as Cr(2)O(7)(2-), MnO(4)(-) and ClO(-) present in a mixed solution of Rh B and iodide. The scattering disappears once iodine is reduced by reductants. Oxidant or reductant species in a sample can thus be detected by positive or negative RRS signal. To verify the RRSD, FIA-RRSD was first constructed and continuous measurement of testing samples containing Cr(2)O(7)(2-), MnO(4)(-) and/or ClO(-) was performed. The detection limits reached a level of decade nM and a linear range was found between peak height and concentration at the range of 0.255-2.04microM for Cr(2)O(7)(2-), 0.158-3.16microM for MnO(4)(-), and 1.18-9.43microM for ClO(-), with linear regression coefficients of all above 0.99. The run-to-run relative standard deviation of peak height was less than 3% (n=6). CE-RRSD was then set up and studied, using a capillary of 75microm i.d.x33cm filled with a running buffer of 50mM citrate and 25mM Tris (pH 3.32) and worked under -12kV at room temperature. The CE eluent was at-line conducted into a stream of rhodamine B and iodine flowing inner a wide tube by plugging the capillary outlet into the wide tube. Different mixtures prepared from Cr(2)O(7)(2-), MnO(4)(-) and ClO(-) were successfully separated and detected by the CE-RRSD.     

Sequential optimization of a flow injection spectrophotometric method for the assay of chlorpromazine in pharmaceutical preparations

A new simple flow injection spectrophotometric method for the assay of chlorpromazine using cerium(IV) in sulfuric acid media was developed. The oxidized form of the drug was monitored at the maximum absorbance of 526 nm. The optimum conditions were 0.035M sulfuric acid, 3.80 x 10(-3)M cerium(IV), flow rate 4.85 ml/min, coil length 45 cm and sample size 110 mm(3). Optimization was carried out by the modified simplex method. Response surface methodology was employed to investigate the ruggedness of the method. A sampling frequency of 120 hr(-1) was attained. Relative standard deviations for standard sample were usually less than 0.75. The method was applied to the determination of chlorpromazine in proprietary drugs and results were statistically compared with the official British Pharmacopoeia (BP) method.     

Membrane sensors for batch and flow injection potentiometric determination of ethamsylate (cyclonamine) in pharmaceutical preparations

The lipophilic nickel(II) and iron(II) bathophenanthroline derivatives of ethamsylate are used as ion-exchangers with high selectivity characteristics for ethamsylate. Poly(vinyl chloride) membrane sensors incorporating these electroactive materials display fast linear response for 1 × 10^–1–1 × 10^–4 M ethamsylate under static and hydrodynamic modes of operation. In an acetate buffer of pH 4, the calibration slope is 51–53 mV/concentration decade and the lower limit of detection is 5.3 g/ml. Except for salicylate and nitrate, most common anions, organic sulfonates, carboxylates, phenolates and various pharmaceutical excipients and diluents do not interfere. Determination of ethamsylate in various dosage forms shows an average recovery of 98.9% of the nominal and a mean standard deviation of 0.7%.     

Novel lipoate-selective membrane sensor for the flow injection determination of alpha-lipoic acid in pharmaceutical preparations and urine

A potentiometric lipoate-selective sensor based on mercuric lipoate ion-pair as a membrane carrier is reported. The electrode was prepared by coating the membrane solution containing PVC, plasticizer, and carrier on the surface of graphite electrode. Influences of the membrane composition, pH, and possible interfering anions were investigated on the response properties of the electrode. The sensor exhibits significantly enhanced response toward lipoate ions over the concentration range 1 × 10⁻⁷ mol L⁻¹ to 1 × 10⁻² mol L⁻¹ with a lower detection limit of (LDL) of 9 × 10⁻⁸ mol L⁻¹ and a slope of −29.4 mV decade⁻¹, with S.D. of the slope is 0.214 mV. Fast and stable response, good reproducibility, long-term stability, applicability over a pH range of 8.0–9.5 is demonstrated. The sensor has a response time of ≤12 s and can be used for at least 6 weeks without any considerable divergence in its potential response. The proposed electrode shows good discrimination of lipoate from several inorganic and organic anions. The CGE was used in flow injection potentiometry (FIP) and resulted in well defined peaks for lipoate ions with stable baseline, excellent reproducibility and reasonable sampling rate of 30 injections per hour. The proposed sensor has been applied for the direct and FI potentiometric determination of LA in pharmaceutical preparations and urine; and has been also utilized as an indicator electrode for the potentiometric titration of LA.     

Spectrophotometric determination of magnesium in pharmaceutical preparations by cost-effective sequential injection analysis

A simple and rapid, inexpensive spectrophotometric method was proposed for magnesium assay in pharmaceutical preparations by sequential injection analysis (SIA). The method is based on the reaction between o-cresolphthalein complexone (CPC) and Mg(II) in alkaline media, yielding a pink colored complex with absorption maximum at 570 nm. Since the formation constant between Ca-CPC and Mg-CPC is similar, initially a sample/standard solution was aspirated into the holding coil followed by a mixture of masking-buffer solutions. This was done because masking of calcium should be accomplished before Mg-CPC complexation. Then the reagent was introduced into the reaction coil to produce a colored complex, which is measured spectrophotometrically at 570 nm. In this way the interference of calcium was reduced. Furthermore, all the parameters that affect the reaction were evaluated. The calibration curve is linear over a range of 0-20 mg l⁻¹ of Mg(II) with a detection limit of 0.24 mg l⁻¹. A sample throughput of 80 samples per hour and relative standard deviation <2.0% were achieved. The proposed method was successfully applied for the assay of magnesium in three different compositions of pharmaceutical preparations (tablets). The results were found to be in good agreement with the manual flame atomic absorption spectrophotometry (FAAS) and UV-Vis spectrophotometry methods and with the claimed values by the manufactures. The t-test shows no significant difference at 95% confidence level.     

Cost-effective flow injection spectrophotometric assay of iron content in pharmaceutical preparations using salicylate reagent

A new flow injection procedure for an assay of Fe(III) by using salicylate obtained from antipyretic powder, which is a cheap and easily available reagent, is proposed. A red complex was continuously monitored by a laboratory-made green LED colorimeter. A linear calibration was obtained in the range of 1–20 mg Fe l⁻¹ with a detection limit of 0.5 mg Fe l⁻¹ and R.S.D.s of 1.4–5.4% (n=3, for 1–20 mg Fe l⁻¹). The new procedure was applied to assay iron contents in pharmaceutical preparations. The results were in good agreement with those of the USP standard method.     

Potassium permanganate-glyoxal chemiluminescence system for flow injection analysis of cephalosporin antibiotics: cefalexin, cefadroxil, and cefazolin sodium in pharmaceutical preparations

A sensitive flow injection chemiluminescence (FL-CL) method for the determination of cephalosporin antibiotics, was developed. The method was based on that cephalosporin antibiotics could enhance the CL reaction of glyoxal and KMnO₄ in sulfuric acid. Method development included the optimization of reagent concentrations and flow-rate. Under the optimized conditions, three cephalosporin antibiotics: cefalexin, cefadroxil, and cefazolin sodium, were determined. The detection limits of the method are 10 ng ml⁻¹ cefalexin, 2 ng ml⁻¹ cefadroxil, and 2 ng ml⁻¹ cefazolin sodium. The method was successfully applied to the determination of three cephalosporin antibiotics in pharmaceutical preparations.     

Chemiluminescence detection flow cells for flow injection analysis and high-performance liquid chromatography

We have examined a range of new and previously described flow cells for chemiluminescence detection. The reactions of acidic potassium permanganate with morphine and amoxicillin were used as model systems representing the many fast chemiluminescence reactions between oxidising agents and organic analytes, and the preliminary partial reduction of the reagent was exploited to further increase the rates of reaction. The comparison was then extended to high-performance liquid chromatography separations of α- and β-adrenergic agonists, with permanganate chemiluminescence detection. Flow cells constructed by machining novel channel designs into white polymer materials (sealed with transparent films or plates) have enabled improvements in mixing efficiency and overall transmission of light to the photodetector.     

Diazocoupling reaction for the spectrophotometric determination of physiologically active catecholamines in bulk and pharmaceutical preparations

A simple and sensitive spectrophotometric method for the determination of four catecholamines viz., adrenaline bitartarate (ABT), methyldopa (MDP), dopamine hydrochloride (DPH), and levodopa (LDP), in both pure form and in pharmaceutical formulations is described. The method is based on the reaction of diazotized sulphanillic acid (DSA) with catecholamines in a basic medium to yield orange-red colored products having absorption maxima at 507 nm for MDP and at 475 nm for others analytes. The colored species obeyed Beer’s law in the range of 1–27, 0.5–17, 0.6–15, and 1.5–19.2 μg/mL for ABT, MDP, DPH, and LDP, respectively. The molar absorptivity values as obtained from Beer’s law data were found to be 0.812 × 10⁴, 0.947 × 10⁴, 0.927 × 10⁴, and 0.709 × 10⁴ L/(mol cm), while Sandell’s sensitivity values were observed to be 412.03, 25.15, 20.44, and 27.81 ng/cm² for ABT, MDP, DPH, and LDP, respectively. Common excipients did not interfere with the proposed method. The results of the proposed methods compare favorably with those of official methods. The proposed method offers a simplicity, reliability, rapidity, and accuracy compared to the existing methods. The text was submitted by the author in English.     

Determination of ephedrine and related compounds in pharmaceutical preparations by ion chromatography with direct conductivity detection

An ion chromatographic method with conductivity detection for the simultaneous determination of ephedrine, pseudoephedrine and norephedrine was developed. A mixture of 2.0 mmol/L HNO3 and 2% (v/v) acetonitrile was used as eluent. The three ephedrine-like compounds were separated and determined within 20 min. The linear ranges were 0.08-50 microg/mL for ephedrine, 0.08-40 microg/mL for pseudoephedrine and 0.06-40 microg/mL for norephedrine. The detection limits were 0.03 microg/mL for ephedrine and pseudoephedrine, and 0.02 microg/mL for norephedrine. The method has been applied successfully to the determination of these sympathomimetics in pharmaceutical preparations and in Ephedra herbs.     

流动注射化学发光法测定甲硝唑

在碱性条件下 ,铁氰化钾氧化鲁米诺产生化学发光 ,甲硝唑对该体系有显著的增强作用 (亚铁氰化钾存在时 )。基于此 ,建立了流动注射化学发光测定痕量甲硝唑的新方法。甲硝唑浓度在 2 .0× 1 0 -6～ 4 .0× 1 0 -4 mol L范围内与发光强度呈良好的线性关系 ;检出限 (3σ)为 1 .5× 1 0 -7mol L。相对标准偏差 (c =1 .0× 1 0 -5mol L ,n=1 1 )为 3.6 %。方法已用于制剂中甲硝唑含量测定     

Indirect chemiluminescence-based detection of mefenamic acid in pharmaceutical formulations by flow injection analysis and effect of gold nanocatalysts

A highly sensitive flow injection-chemiluminescence detection (FI-CL) method based on periodate oxidation of two popular luminescent compounds for the determination of mefenamic acid (MFA) is presented. The method is an indirect CL detection method based on the CL emission generated during the oxidation of Pyrogallol (Pg) or Luminol (Lu) with the excess of periodate that remains after oxidation of MFA within the time period of 15 min. The MFA calibration curves obtained with either luminescent compounds were linear over a wide concentration range, depending on the system employed, offering detection limits in the range of low to ultra-low μg L⁻¹ levels. Gold nanoparticles (Au-NPs) were also assessed as means for enhancing the CL signal. Pg-periodate was not affected by the presence of gold nanocatalysts as opposed to Lu-periodate CL signal which exhibited a significant increase in the presence of citrate synthesized Au-NPs. The reproducibility of the method, expressed by the relative standard deviation (R.S.D.), was very satisfactory and always below 5% as evidenced by repeated measurements (n ≥ 10) of standard solutions at two concentration levels (2 and 20 μg L⁻¹).     

Simultaneous multiwavelength detection in flow injection analysis

The potential of the use of the diode-array detector in conjunction with flow injection analysis is outlined. Methods for multicomponent resolution can be based on the formation of complexes absorbing at different wavelengths. The example given is the determination of mixtures of copper(II) and iron(III) with a mixed 1:10-phenanthroline/neocuproine reagent. Amplification and dilution methods are based on the sum of the absorbances at several wavelengths and on monitoring absorbances at wavelengths away from the absorption maximum, respectively. The determination of nitrite via the Griess reaction is used as an example; the viable determination range is extended to 0.002/2-60.0 μg ml^?1 nitrite. Software for the implementation of the suggested methods is outlined.     

Improvement of detection limits in flow injection analysis

Summary When using a conventional UV/VIS-Spectrophotometer with a large sample compartment as a detector for FIA, flow cells with longer pathlengths can be applied. Despite of a unavoidable band broadening a significant improvement of detection limit can be achieved compared to standard cells. Band broadening is compensated by slight modifications of the manifold, so that the dispersion is in the same range for long path cells and standard flow cells. With a 5 cm flow cell phosphate can be detected down to 0.005 mg/l P.     

Determination of ceftriaxone sodium in pharmaceutical formulations by flow injection analysis with acid potassium permanganate chemiluminescence detection.

Based on the chemiluminescence (CL) emission generated from the oxidation of ceftriaxone sodium alkali hydrolysate by potassium permanganate in polyphosphoric acid (PPA), a novel determination method for ceftriaxone sodium was developed by using a flow-injection technique. The calibration curve appears to be linear in the range between 0.05 and 100 microg mL(-1) with a detection limit (3sigma) of 25 ng mL(-1), and a relative standard deviation (RSD) of 0.6% for eleven replicate determinations of 5.0 microg mL(-1) ceftriaxone sodium. The proposed method has been successfully utilized for the determination of ceftriaxone sodium in pharmaceutical formulations, while the chemiluminescence reaction mechanisms were investigated.