A.A. Al-Majed

Methods of analysis of 4-quinolone antibacterials.

A comprehensive review with 270 references for the analysis of the members of an important class of drugs, 4-quinolone antibacterials, is presented. The review covers most of the methods described for the analysis of these drugs either per se, in dosage forms or in biological fluids.

Spectrophotometric estimation of d-penicillamine in bulk and dosage forms using 2,6-dichloroquinone-4-chlorimide (DCQ)

A simple colorimetric method for the determination of D-penicillamine in pure form and pharmaceutical formulations is described. The method is based on coupling between D-penicillamine and 2,6-dichloroquinone-4-chlorimide (DCQ) in dimethylsulphoxide. The optimum conditions for the reaction were investigated and incorporated into the procedure. The reaction forms a yellow 1:1 complex with maximum absorbance at 431 nm (epsilon = 3,700). Regression analysis of Beers plot showed good correlation (r = 0.9998) and the calibration graph was rectilinear over the range 4-20 microg/ml(-1) with a detection limit of 0.15 microg/ml(-1) . The average recovery for the commercial capsules was 101.66% with an RSD of 1.57%. The results obtained were sufficiently accurate, reproducible and in accordance with those given by the official method. The method is specific for the intact drug, and can be adopted in the presence of some interfering substances.

The voltammetric study and determination of ramipril in dosage forms and biological fluids

The voltammetric behavior of ramipril was studied using cyclic voltammetry, direct current polarography (DCt), differential pulse polarography (DPP) and alternating current polarography (ACt). Ramipril developed well-defined cathodic waves in Britton-Robinson buffers over the pH range 6-12. The waves were characterized as being diffusion-controlled, irreversible and partially affected by adsorption phenomenon. The diffusion-current constant (Id) was 1.24 +/- 0.02. The current-concentration plots were rectilinear over the range 10-50, 4-40 and 0.16-12 micrograms/ml in the DCt, DPP and ACt modes, respectively, with a minimum detectability (S/N = 2) of 0.02 microgram/ml (4.8 x 10(-8) M) using the latter mode. The proposed method was successfully applied to the determination of ramipril in commercial tablets. Hydrochlorothiazide, which is frequently co-formulated with ramipril, did not interfere with the assay. Furthermore, the proposed method was applied to the determination of ramipril in urine and plasma adopting the ACt technique. The percentage recoveries were 97.12 +/- 0.56 and 94.97 +/- 0.62%, respectively. A pathway for the electrode reaction was proposed.

SPECTROPHOTOMETRIC DETERMINATION OF RAMIPRIL (A NOVEL ACE INHIBITOR) IN DOSAGE FORMS

A simple and sensitive spectrophotometric method has been developed for the determination of ramipril in its dosage forms. The method is based on reacting the drug with potassium permanganate in alkaline medium whereby a bluish green colour peaking at 610 nm is produced. The absorbance-concentration plot is rectilinear over the range 0.1 − 7.5 μg · mlΣ 1 (r = 0.9992) with minimum detectability of 0.05 μg · ml−1 (1.2 × 10−7 M). The molar absorptivity was 2.42 × 104 L · Mol−1 · cm−1. The different experimental parameters affecting the development and stability of the colour were carefully studied and optimized. The proposed method was further applied to the determination of ramipril in its dosage forms, whether alone or in combination with hydrochlorothiazide. The results obtained were in good agreement with those obtained by a reference HPLC method. A proposal of the reaction pathway was postulated.

Voltammetric determination of N-nitrosoderivatives of atenolol and propranolol in simulated gastric juice.

A highly sensitive and simple voltammetric method is proposed for the determination of N-nitrosoatenolol (NA) and N-nitrosopropranolol (NP) in simulated gastric juice. The method is based on measuring the differential-pulse polarographic peak produced by NA and NP in Britton-Robinson buffers of pH 3 and 4 for NA and NP, respectively. Both compounds yielded diffusion-controlled current with diffusion-current constants of 7.23 +/- 0.03 and 9.46 +/- 0.06 for NA and NP, respectively. The current-concentration plots were rectilinear over the range 0.16-9.6 micrograms ml-1 with minimum detectability (S/N = 2) of 0.015 microgram ml-1 (5 x 10(-8) M) for NA; for NP the range was 0.08-8.0 micrograms ml-1 with minimum detectability (S/N = 2) of 0.009 microgram ml-1 (3 x 10(-8) M). The proposed method was successfully applied to study the possible in vivo production of the nitroso-derivatives under the standard nitrosation reaction conditions recommended by WHO. The method is characterized by simplicity and higher sensitivity as compared with the reported HPLC method.

Use of 7-fluoro-4-nitrobenzo-2-oxo-1,3-diazole (NBD-F) for the determination of ramipril in tablets and spiked human plasma

Ramipril, as a secondary amine compound, reacts with 7-fluoro-4-nitrobenzo-2-oxo-1,3-diazole (NBD-F) producing the corresponding fluorescent NBD-ramipril. According to this fact, spectrophotometric and fluorimetric methods for the determination of ramipril were developed. The effect of these parameters on the reaction product were carefully studied to optimize reaction conditions. The relationship between the absorbance at 465 nm and the concentration was found to be linear over the range 1-10 microg/ml. Moreover, the fluorescence intensity was also found to be directly proportional at the concentration over the range of 20-100 ng/ml at 530 nm after excitation at 465 nm. The proposed procedure was successfully applied to the determination of ramipril in both tablet dosage form and in plasma. Spectrophotometric determination of ramipril tablets yielded a percentage recovery of 98.66+/-0.38, while the percentage recovery of spectrofluorimetric determination of ramipril in spiked human plasma was 99.08+/-1.11%. The results obtained are in good agreement with those obtained by the reference method. No interference could be observed from the co-administered drug (hydrochlorothiazines).

Voltammetric determination of josamycin (a macrolide antibiotic) in dosage forms and spiked human urine.

The voltammetric behaviour of josamycin (a macrolide antibiotic) has been studied using direct current (DC(t)) alternating current (AC(t)) and differential pulse polarography (DPP). In Britton-Robinson buffers, josamycin developed cathodic waves over the pH range 7-12. At pH 10, a well-defined cathodic wave with diffusion current constant of 1.06 +/- 0.19 (n = 5) was obtained. The wave was characterized as being diffusion-controlled; and partially affected by adsorption phenomenon. The current-concentrations plots are rectilinear over the range 10-60 and 6-50 microg/ml using DC(t) mode and DPP mode, respectively. The minimum detectability limit was 1.2 microg/ml (1.9 x 10(-6) M) adopting the DPP mode. A method was proposed for the determination of josamycin in its tablets adopting both DC(t) and DPP modes. The results obtained were in good agreement with those given by the manufacturer. The method was extended to the in-vitro determination of the drug in spiked human urine; the % recovery was 98.06 +/- 1.76% (n = 5). The number of electrons involved in the reduction process was accomplished and a proposal of the electrode reaction was presented.

Voltammetric determination of isradipine in dosage forms and spiked human plasma and urine

The voltammetric behavior of isradipine was studied using direct current (DC(t)), differential-pulse (DPP) and alternating current (AC(t)) polarography. Isradipine exhibited well-defined cathodic waves over the whole pH range in Britton-Robinson buffer (BRb). At pH 5, the analytical pH, the diffusion-current constant (Id) was 8.27+/-0.52. The current-concentration plots were rectilinear over the range 1-20 and 0.1-18 microg/ml using the DC(t) and DPP modes, respectively, with minimum detectability of 0.01 microg/ml (2.7 x 10(-8) M) using the latter technique. The current has been characterized as being diffusion-controlled, although adsorption phenomenon played a limited role in the electrode process. The proposed method was applied to commercial tablets and capsules. The percentage recoveries were in good agreement with those given by the manufacturer. The method was further extended to the in-vitro determination of the drug in spiked human urine and plasma, the percentage recoveries were (n = 4) 100.12+/-1.42 and 103.88+/-5.13, respectively. The number of electrons involved in the reduction process was accomplished and a proposal of the electrode reaction was presented.

A stability-indicating spectrophotometric method for the determination of fenoterol in pharmaceutical preparations

ABSTRACT A simple and highly sensitive spectrophotometric method was developed for the determination of fenoterol hydrobromide in pharmaceutical formulations. The method is based on measurement of the orange-yellow color peaking at 436 nm, produced when the drug is coupled with diazotized benzocaine in triethylamine medium. The method is applicable over the range of 0.5 – 10 μg/ml, with minimum detectability of 3.6 ng/ml (≈ 9.4×10−9 M). The molar absorptivity is 4.61×104 L. mol−1. cm−1. with relative standard error of 0.196%. The proposed method was successfully applied to the determination of fenoterol in its dosage forms; the mean recoveries of 99.61 ± 1.59 and 98.97 ± 1.52 for tablets and aerosols, respectively, were obtained. The results of the proposed method were favorably compared to those obtained by the official and reference methods. No i8543168 was encountered from the co-formulated drug ipratropium bromide, common excipients and alkaline induced degradation product. A proposal of the reaction pathway is presented.

Generic simple enzyme immunoassay approach to avert small molecule immobilization problems on solid phases: Application to the determination of tobramycin in serum

Generic simple and sensitive universal enzyme immunoassay approach for the determination of small analytes has been developed to avert the problems associated with small molecule immobilization onto solid phases. The developed assay employed a heterogeneous non-competitive binding format. The assay used anti-analyte antibody coupled to polyacrylamide beads as a solid-phase and beta-d-galactosidase enzyme-labeled analyte as a label. In this assay, the analyte in a sample was firstly incubated to react with an excess of the antibody-coupled beads, and then the unoccupied antibody binding sites were allowed to react with the enzyme-labeled analyte. Analyte bound to the antibody-coupled beads was separated by centrifugation, and the enzyme activity of the supernatant was measured spectrophotometrically at 420nm, after reaction with 4-nitrophenyl-beta-d-galactopyranoside as a substrate for the enzyme. The signal was directly proportional to the concentration of analyte in the sample. The optimum conditions for the developed assay were established and applied to the determination of tobramycin, as a representative example of the small analytes, in serum samples. The assay limit of detection was 10ngmL(-1) and the effective working range at relative standard deviation of </=10% was 40-800ngmL(-1). The assay precisions were acceptable; the relative standard deviations were 4.36-5.17 and 5.62-7.40% for intra- and inter-assay precision, respectively. Analytical recovery of tobramycin spiked in serum ranged from 95.89+/-4.25 to 103.45+/-4.60%. The assay results correlated well with those obtained by high-performance liquid chromatography (r=0.992). The assay described herein has great practical value in determination of small analytes because it is sensitive, rapid, and easy to perform in any laboratory. Although the assay was validated for tobramycin, however, it is also anticipated that the same methodology could be used for essentially any analyte for which a selective antibody exists, and an appropriate enzyme conjugate can be made.