Hassan F. Askal

Use of charge-transfer complexation in the spectrophotometric analysis of certain cephalosporins.

Three simple, rapid and sensitive spectrophotometric procedures were developed for the analysis of cephapirin sodium (1), cefazoline sodium (2), cephalexin monohydrate (3), cefadroxil monohydrate (4), cefotaxime sodium (5), cefoperazone sodium (6) and ceftazidime pentahydrate (7) in pure form as well as in their pharmaceutical formulations. The methods are based on the reaction of these drugs as n-electron donors with the sigma-acceptor iodine, and the pi-acceptors: 2,3-dichloro-5,6-dicyano-p-benzo-quinone (DDQ) and 7,7,8,8-tetracyanoquinodimethane (TCNQ). Depending on the solvent polarity, different coloured charge-transfer complexes and radicals were developed. Different variables and parameters affecting the reactions were studied and optimized. The obtained charge-transfer complexes were measured at 364 nm for iodine (in 1,2-dichloroethane), 460 nm for DDQ (in methanol) and 843 nm for TCNQ (in acetonitrile). Ultraviolet-visible, infrared and (1)H-nuclear magnetic resonance techniques were used to study the formed complexes. Due to the rapid development of colours at ambient temperature, the obtained results were used on thin-layer chromatograms for the detection of the investigated drugs. Beers plots were obeyed in a general concentration range of 6-50, 40-300 and 4-24 mug ml(-1) with iodine, DDQ and TCNQ, respectively, with correlation coefficients not less than 0.9989. The proposed procedures could be applied successfully to the determination of the investigated drugs in vials, capsules, tablets and suspensions with good recovery; percent ranged from 96.47 (+/-1.14) to 98.72 (+/-1.02) in the iodine method, 96.35 (+/-1.62) to 98.51 (+/-1.30) in the DDQ method, and 95.98 (+/-0.78) to 98.40 (+/-0.87) in the TCNQ method. The association constants and standard free energy changes using Benesi-Hildebrand plots were studied. The binding of cephalosporins to proteins in relation to their molar absorptivities was studied.

Colourimetric and AAS determination of cephalosporins using Reineck's salt.

Two simple, accurate, sensitive and selective procedures for the determination of eight cephalosporins are described. These procedures are based on the formation of ion-pair complexes between the drugs and ammonium reineckate, the formed precipitates are quantitatively determined either colourimetrically or by atomic absorption spectrometrically. The methods consist of reacting drugs with Reineckes salt in an acidic medium at 25+/-2 degrees. The first colourimetric procedure (procedure I) is based on dissolving the formed precipitate with acetone, the volume was completed quantitatively and the absorbance of the solution was measured at 525 nm against pure solvent blank. Also, the formed precipitates on the atomic absorption spectrometric procedure (procedure II) are quantitatively determined directly or indirectly through the chromium precipitate formed or the residual unreacted chromium in the filtrate at 358.6 nm. The optimum conditions for precipitation have been carefully studied. Beers law is obeyed for the studied drugs in the range 5-35 microg ml(-1) with correlation coefficients not less-than 0.9989. Both procedures I and II hold well accuracy and precision when applied to the analysis of the cited cephalosporins in different dosage forms with good recovery percent ranged from 98.7+/-0.90 to 100.1+/-0.74 without interference from additives.

Selective spectrophotometric method for the determination of ascorbic acid in pharmaceutical preparations and fresh fruit juices

A simple and selective method for the determination of ascorbic acid in pharmaceutical preparations and fresh fruit juices is described. The procedure is based on the reaction of ascorbic acid with the zinc chloride salt of diazotized 1-aminoanthraquinone (Fast Red AL salt) in an acidic medium, followed by development of a blue colour (λmax 630 nm) in alkaline solution. Different variables affecting the colour development were studied and optimized. The method was used to determine between 5 and 25 µg ml–1 of ascorbic acid in the final measured solution. The simplicity of the method permits rapid analysis, suitable for routine control. The method is highly specific for the determination of ascorbic acid in the presence of dehydroascorbic acid and all other vitamins normally encountered with it in pharmaceutical dosage forms. Moreover, the proposed method can also be applied to the determination of ascorbic acid in some fresh fruit juices without interference from coloured and other substances present in the fruit extracts. The reliability of the method was established by parallel determinations against the official British pharmacopoeial method.

Enantioselective binding analysis of verapamil to plasma lipoproteins by capillary electrophoresis–frontal analysis

Capillary electrophoresis coupled with frontal analysis was applied to the study of enantioselective binding of verapamil (VER) to plasma lipoproteins. The drug-lipoprotein mixed solution, which had been in the binding equilibrium, was hydrodynamically introduced into a non-coated fused-silica capillary. Since VER is positively charged in the neutral run buffer (pH 7.4), the unbound VER enantiomers migrated toward the cathodic end much faster than negatively charged lipoproteins and their bound forms. Once unbound VER migrated apart from lipoprotein, the bound VER was quickly released from the protein to maintain the binding equilibrium. Thus, VER migrated as a zone through the capillary and gave a trapezoidal peak with a plateau region on the electropherogram. The VER concentration in this plateau region was equal to the unbound VER concentration in the initial sample solution. It was found that the bindings of VER to high-density lipoprotein (HDL), low-density lipoprotein (LDL) and oxidized LDL were not site-specific and not enantioselective. Partition-like binding to lipid part of these lipoproteins seemed to be dominant. The total binding affinities of LDL to VER were about seven-times stronger than those of HDL, and the oxidation of LDL by copper ion enhanced the binding affinities significantly.

Utility of certain π-acceptors for the spectrophotometric determination of some penicillins

Two simple and sensitive spectrophotometric methods are described for the determination of six penicillin derivatives. The methods are based on the reaction of these drugs as n-electron donors with either 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ) or 7,7,8,8-tetracyanoquinodimethane (TCNQ) as pi-acceptors, to give a highly coloured radical anion. The coloured products are quantified spectrophotometrically at 460 and 842 nm for DDQ and TCNQ, respectively. The optimization of the different experimental conditions is described. The interference from streptomycin sulphate and common degradation products was also studied. The proposed methods were applied successfully to the determination of the different penicillins investigated, either in pure or dosage forms, with good accuracy and precision. The results were compared with those given by the official United States Pharmacopeial XXI method.

Frontal analysis of drug-plasma lipoprotein binding using capillary electrophoresis.

High performance frontal analysis coupled with capillary electrophoresis (HPFA/CE) was applied to the ultramicroanalysis of enantioselective binding of drug to plasma lipoproteins. A small volume (ca. 80 nl) of (R)- or (S)-propranolol (PRO, 25-150 microM) and human high-density lipoprotein (HDL, 2.63 g/l) or human low-density lipoprotein (LDL, 4.37 g/l) mixed solution, which was in the state of binding equilibrium, was introduced hydrodynamically into a non-coated fused silica capillary. Positively charged unbound PRO enantiomers migrated toward cathodic end much faster than negatively charged lipoproteins and the bound form. Once unbound PRO migrated apart from lipoprotein, the bound PRO was quickly released from the lipoprotein to maintain the binding equilibrium. Thus, PRO migrated as a zone in the capillary, giving a peak with a plateau region, where the concentration is the same as the unbound PRO concentration in the original sample solution. The unbound PRO concentration calculated form the plateau height agreed with that determined by a conventional ultrafiltration method used as a reference method. It was found that the bindings of PRO to HDL and PRO to LDL were not enantioselective, while the total binding affinity of PRO to LDL (4.01 x 10(5) per M) was 17 times higher than that of PRO-HDL binding (2.38 x 10(4) per M).

Binding analysis of nilvadipine to plasma lipoproteins by capillary electrophoresis-frontal analysis

Capillary electrophoresis coupled with frontal analysis (HPCE/FA) was applied to the ultramicro analysis of enantioselective binding of nilvadipine (NV), a calcium channel blocker, to plasma lipoproteins. The drug lipoprotein mixed solution was hydrodynamically introduced into a non-coated fused silica capillary for capillary electrophoresis. Since NV has no electric charge in the run buffer (pH 7.4), the unbound NV moved towards the cathodic end by electroosmotic flow, which was faster than the electrophoretic migrations of negatively charged lipoproteins and the bound NV. Once unbound NV migrated apart from lipoprotein, and bound NV was quickly released from the protein to maintain the binding equilibrium. Thus, NV migrated as a zone with a plateau region. The concentration of NV in this plateau region appearing on the electrophorogram was the same as the unbound NV concentration in the initial sample solution. It was found that the binding of NV to high-density lipoprotein (HDL), low-density lipoprotein (LDL) and oxidized LDL was non-specific and not enantioselective. Partition-like binding to the lipid part of these lipoproteins seemed to occur dominantly. The total binding affinities of NV to LDL were about seven times stronger than those to HDL, and the oxidation of LDL enhanced the binding affinity significantly.

New spectrophotometric methods for determination of captopril bulk drug and tablets

Three simple, rapid and sensitive methods for the assay of captopril which is an effective alternative to digitalis were developed. These methods are based on the oxidation reaction in aqueous solution of captopril with either ferric chloride or iodine. The indirect quantitation of the product was carried out at 523, 351 and 620 nm for ferro-bipyridyl, residual iodine and residual iodine-starch complex, respectively. All variables were studied to optimize the reaction conditions. Regression analysis of Beers plot showed good correlation in a general concentration range of 0.25-25 mug captopril/ml. No interference was observed from hydrochlorothiazide diuretic which was recently introduced in combination with captopril or other common pharmaceutical adjuvants. The validity of the methods was tested by analysing capoten and capozide tablets. Recoveries were 99.1-102.8%.

Spectrophotometric study of the charge transfer complexes of some pharmaceutical butyrophenones

The molecular interactions between haloperidol and droperidol as electron donors and each of iodine; 7,7,8,8-tetracyanoquinodimethane (TCNQ); 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ); tetracyanoethylene (TCNE); 2,4,7-trinitro-9-fluorenon (TNF); and 2-3-5-6-tetrabromo-1,4-benzoquinone (Bromanil) as acceptors have been investigated spectrophotometrically. Different variables affecting the reaction were studies and optimized. Beers law was obeyed in a concentration limit of 2.5-2500 mug ml(-1) for the studied drugs with various acceptors used. Electron affinities (E(A)) of the acceptors were found to correlate with both the time required for maximum colour formation and the molar absorptivities of haloperidol and droperidol. A Jobs plot of the absorbance versus the molar ratio of the drugs to iodine indicated 1:1 ratio. The proposed methods were found to be rapid and sensitive and may be applied for estimation of named drugs in pharmaceutical dosage forms without interferences from the common additives encountered. Percentage recoveries ranged from 99.1-102.2%.

Selective spectrophotometric determination of ascorbic acid in drugs and foods

A new analytical method was developed for the determination of ascorbic acid. The method is based on the reaction of ascorbic acid with 4-chloro-7-nitrobenzofurazane (NBD-Cl) in the presence of 0.2M sodium hydroxide, where a bluish green colour (lambda(max) 582 nm) is developed after dilution with 50% (v/v) aqueous acetone solution. Beers law was obeyed in a concentration range of 5-20 microg ascorbic acid/ml with a good correlation coefficient (r = 0.9990). The method was found to be highly specific for the determination of ascorbic acid in the presence of dehydro-ascorbic acid, all other vitamins and minerals possibly present in multivitamin preparations, rutin, salicylamide, acetyl salicylic acid, paracetamol, caffeine, phenylephrine hydrochloride and dipyrone. Moreover, the proposed procedure was also successfully applied for the determination of ascorbic acid in some canned and fresh fruit juices, some vegetables and infant milk products without interference from coloured and other substances present in the plant extracts.