Simultaneous determination of some dihydroxybenzenesulphonic acid derivatives and their degradation product and main impurity (hydroquinone) by ion-pair liquid chromatography.

A simple and sensitive high-performance liquid chromatography method was developed and validated for the determination of calcium dobesilate (DOB) or ethamsylate (ETM) in the presence of their degradation product, hydroquinone (HQ). The analyses were carried out on Promosil C18 column (4.6 mM × 250 mM, 5 µm particle size) using an ion-pair mobile phase consisting of methanol-1.5 mM tetra-butyl ammonium bromide in 0.06 M phosphate buffer (25 : 75, v/v) at pH 6.0 with fluorescence detection at 286/333 nm. Pindolol was used as an internal standard. The proposed method was found to be rectilinear over the concentration ranges of 0.05-0.5 µg/mL for DOB, 0.1-0.8 µg/mL for ETM and 0.005-0.1 µg/mL for HQ. The method was applied for the determination of the studied drugs in different dosage forms and biological fluids. The results of the proposed method were statistically compared with those obtained by the comparison methods revealing no significance differences in the performance of the methods regarding accuracy and precision. Moreover, applying a time-programmed fluorescence technique was valuable for the detection of trace amounts of HQ as an impurity and allowed purity testing of ETM or DOB within the BP pharmacopeial limit (0.1%).

Successive spectrophotometric resolution as a novel technique for the analysis of ternary mixtures of pharmaceuticals.

A novel spectrophotometric technique was developed for the simultaneous determination of ternary mixtures, without prior separation steps. This technique was called successive spectrophotometric resolution technique. The technique was based on either the successive ratio subtraction or successive derivative subtraction. The mathematical explanation of the procedure was illustrated. In order to evaluate the applicability of the methods a model data as well as an experimental data were tested. The results from experimental data related to the simultaneous spectrophotometric determination of lidocaine hydrochloride (LH), calcium dobesilate (CD) and dexamethasone acetate (DA); in the presence of hydroquinone (HQ), the degradation product of calcium dobesilate were discussed. The proposed drugs were determined at their maxima 202 nm, 305 nm, 239 nm and 225 nm for LH, CD, DA and HQ respectively; by successive ratio subtraction coupled with constant multiplication method to obtain the zero order absorption spectra, while by applying successive derivative subtraction they were determined at their first derivative spectra at 210 nm for LH, 320 nm or P(292-320) for CD, 256 nm or P(225-252) for DA and P(220-233) for HQ respectively. The calibration curves were linear over the concentration range of 2-20 µg/mL for both LH and DA, 6-50 µg/mL for CD, and 3-40 µg/mL for HQ. The proposed methods were checked using laboratory-prepared mixtures and were successfully applied for the analysis of pharmaceutical formulation containing the cited drugs with no interference from other dosage form additives. The proposed methods were validated according to the ICH guidelines. The obtained results were statistically compared with those of the official BP methods for LH, DA, and CD, and with the official USP method for HQ; using student t-test, F-test, and one way ANOVA, showing no significant difference with respect to accuracy and precision.

Selective electrochemical sensing of calcium dobesilate based on the nano-Pd/CNTs modified pyrolytic graphite electrode.

A new palladium nanoparticle functionalized multi-wall carbon nanotubes (nano-Pd/CNTs) modified pyrolytic graphite electrode (PGE) has been fabricated for electrochemical sensing of calcium dobesilate (CD) in pharmaceutical capsules. The nano-Pd/CNTs were characterized by transmission electron microscopy (TEM) and X-ray diffraction (XRD). The nano-Pd/CNTs composite showed a strong electrocatalytic property for CD. The anodic peak current is 6-fold than that obtained in bare PGE and the oxidation potential has an obvious shift to negative. The anodic peak current is proportional to the concentration of CD in the range of 1.0 x 10(-7) to 7.0 x 10(-4)mol L(-1), with a linear relative coefficient r=0.999 and a detection limit 4.0 x 10(-8)mol L(-1) (S/N=3). This kind of electrode shows good stability, sensitivity, reproducibility, large linear range and low detection limit towards electrochemical determination of CD. The proposed method provides a selective and sensitive electrochemical sensor of calcium dobesilate.

An electrochemical sensor for determination of calcium dobesilate based on PoPD/MWNTs composite film modified glassy carbon electrode.

A poly-o-phenylenediamine and multi-wall carbon nanotubes composite (PoPD/MWNTs) modified glassy carbon electrode (GCE) was prepared by in situ electropolymerization using an ionic surfactant as the supporting electrolyte. The morphology of the resulting PoPD/MWNTs composite was characterized by TEM and the electrochemical properties of the modified electrode were characterized by cyclic voltammetry. The electrochemical behavior of calcium dobesilate on PoPD/MWNTs modified electrode was also investigated. The large current response of calcium dobesilate on PoPD/MWNTs modified electrode is probably caused by the synergistic effect of the electrocatalytic property of PoPD and MWNTs. The reductive peak current increased linearly with the concentration of calcium dobesilate in the range of 0.1-1.0 micromol/L and 4.0-400 micromol/L by square wave adsorptive stripping voltammetry, respectively. The detection limit (three times the signal blank/slope) was 0.035 micromol/L. The modified electrode could eliminate the interference of dopamine, norepinephrine and epinephrine at 100-, 90- and 70-fold concentration of 1.0 micromol/L calcium dobesilate, respectively. The proposed modified electrode provides a new promising and alternative way to detect calcium dobesilate.

Validation of liquid chromatographic method for analysis of lidocaine hydrochloride, dexamethasone acetate, calcium dobesilate, buthylhydroxyanisol and degradation product hydroquinone in suppositories and ointment.

In this paper, there was developed a sensitive, precise and accurate reversed-phase liquid chromatographic (RP-HPLC) method and validated for simultaneous determination of lidocaine hydrochloride, dexamethasone acetate (DA) and calcium dobesilate (CD) in suppositories and ointment. Also there was achieved a parallel analysis of buthylhydroxyanisol, as a preservative, and hydroquinone, as a degradation product of calcium dobesilate, present in these dosage forms. The relative standard deviation (RSD) values for all five compounds indicated a good precision and accuracy of the RP-HPLC method. Method is selective, sensitive and reproducible with good recovery values and can be applied in simultaneous determination of all mentioned compounds.

Flow injection chemiluninescence for detecting picogram amounts of dobesilate in human urine.

A sensitive method for the determination of dobesilate in pharmaceutical preparations and human urine is described by using controlled-reagent-release technology. The method entailed the use of luminol and periodate, which are immobilized on anion exchange resin and react in alkaline medium, giving chemiluminescence (CL) at 425 nm. Dobesilate was detected by measuring the decrease of CL intensity, and which was observed linear over the dobesilate concentration range of 10-600 pg ml(-1), and the limit of detection was 3.5 pg ml(-1) (3sigma) and a relative standard deviation of less than 3.0%. At a flow rate of 2.0 ml min(-1), the determination of dobesilate, including sampling and washing, could be performed in 0.5 min, giving a throughput of about 120 times per hour. The proposed method has an extremely low limit of detection down to 3.5 pg ml(-1), thus it can be applied directly in the assay of human urine without any pre-treatment. It was also found that the dobesilate concentration reached its maximum after orally administrated for 3.5h, and the excretion ratio in 24h was 58.8% in the body of volunteers.

Flow-injection biamperometric direct determination of calcium dobesilate in irreversible couple system.

A flow-injection biamperometric method for direct determination of calcium dobesilate had been proposed based on biamperometric detection for irreversible couple. The detection was realized by coupling the oxidation of dobesilate at one platinum wire electrode with the reduction of MnO(4)(-) at another one with the applied potential difference of 0 V between two platinum wire electrodes. Dobesilate was determined in the range of 4.0 x 10(-6) to 1.0 x 10(-4) M with the detection limit of 8.0 x 10(-7) M (S/N=3). The relative standard derivation of 1.7% was obtained for 24 successive determinations of 4.0 x 10(-5) M dobesilate. The proposed method had been shown to be sensitive, simple and rapid.

[Spectrodensitometric determination of lidocaine-chloride in antihemorrhoidal agents]. / Spektrodenzitometrijsko odredivanje lidokain-hlorida u nekim antihemoroidalijama.

Spectrodensitometric method was used in determination of lidocaine-chloride in some antihemorrhoidal agents after thermomicrodetection and its direct thin layer coating. This procedure is based on transference of the studied substance into the gaseous state, its quantitative transference on the basic layer of silicagel GF254, isolation in the benzene-methanol mixture and its identification. Determination was performed by spectrodensitometric determination of absorption. The results obtained are precise and reproducible and the procedure is suitable for determination of lidocaine-chloride in micrograms.