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Binding of therapeutic agents to plasma proteins, particularly to serum albumin, provides valuable information in the drug development. This study was designed to evaluate the binding interaction of neratinib with bovine serum albumin (BSA). Neratinib blocks HER2 signaling and is effective in trastuzumab-resistant breast cancer treatment. Spectrofluorometric, UV spectrophotometric, and fourier transform infrared (FT-IR) and molecular docking experiments were performed to study this interaction. The fluorescence of BSA is attributed to the presence of tryptophan (Trp) residues. The fluorescence of BSA in presence of neratinib was studied using the excitation wavelength of 280 nm and the emission was measured at 300-500 nm at three different temperatures. Neratinib quenched the BSA intrinsic fluorescence by static mechanism. A complex formation occurred due to the interaction leading to BSA absorption shift. The fluorescence, UV- absorption, three dimensional fluorescence and FT-IR data showed conformational changes occurred in BSA after interaction with neratinib. The binding constant values decreased as the temperature increased suggesting an instable complex formation at high temperature. Site I (sub-domain IIA) was observed as the principal binding site for neratinib. Hydrogen bonding and Van der Waals forces were suggested to be involved in the BSA-neratinib interaction due to the negative values of entropy and enthalpy changes.

High performance liquid chromatography (HPLC) and second-order derivative spectrophotometry have been used for simultaneous determination of pravastatin (PS) and fenofibrate (FF) in pharmaceutical formulations. HPLC separation was performed on a phenyl HYPERSIL C18 column (125 mm × 4.6 mm i.d., 5 μm particle diameter) in the isocratic mode using a mobile phase acetonitrile/0.1 % diethyl amine (50:50, V/V, pH 4.5) pumped at a flow rate of 1.0 mL min-1. Measurement was made at 240 nm. Both drugs were well resolved on the stationary phase, with retention times of 2.15 and 5.79 min for PS and FF, respectively. Calibration curves were linear (R = 0.999 for PS and 0.996 for FF) in the concentration range of 5-50 and 20-200 µg mL-1 for PS and FF, respectively. Pravastatin and fenofibrate were quantitated in combined preparations also using the second-order derivative response at 237.6 and 295.1 nm for PS and FF, respectively. Calibration curves were linear, with the correlation coefficient R = 0.999 for pravastatin and fenofibrate, in the concentration range of 5-20 and 3-20 µg mL-1 for PS and FF, respectively. Both methods were fully validated and compared, the results confirmed that they were highly suitable for their intended purpose.

Background: The construction and electrochemical response characteristics of Poly (vinyl chloride) membrane sensors for moxifloxacin HCl (MOX) are described. The sensing membranes incorporate ion association complexes of moxifloxacin cation and sodium tetraphenyl borate (NaTPB) (sensor 1), phosphomolybdic acid (PMA) (sensor 2) or phosphotungstic acid (PTA) (sensor 3) as electroactive materials., Results: The sensors display a fast, stable and near-Nernstian response over a relative wide moxifloxacin concentration range (1 × 10(-2) - 4.0 × 10(-6), 1 × 10(-2) - 5.0 × 10(-6), 1 × 10(-2) - 5.0 × 10(-6) M), with detection limits of 3 × 10(-6), 4 × 10(-6) and 4.0 × 10(-6) M for sensor 1, 2 and 3, respectively over a pH range of 6.0 - 9.0. The sensors show good discrimination of moxifloxacin from several inorganic and organic compounds. The direct determination of 400 μg/ml of moxifloxacin show an average recovery of 98.5, 99.1 and 98.6% and a mean relative standard deviation of 1.8, 1.6 and 1.8% for sensors 1, 2 and 3 respectively., Conclusions: The proposed sensors have been applied for direct determination of moxifloxacin in some pharmaceutical preparations. The results obtained by determination of moxifloxacin in tablets using the proposed sensors are comparable favorably with those obtained using the US Pharmacopeia method. The sensors have been used as indicator electrodes for potentiometric titration of moxifloxacin.

A highly selective, sensitive, and rapid high-performance liquid chromatography (HPLC) method has been developed and validated for the quantification of darifenacin in mouse plasma. Bisoprolol was used as an internal standard (IS). Darifenacin and the IS were extracted using the deproteinisation technique, followed by injection of an aliquot of the supernatant into the chromatographic system. The chromatographic separation was achieved on a reversed phase C18 column with a mobile phase of acetonitrile: 0.1% diethyl amine (pH 3.5) (60:40, v/v) pumped at a flow rate of 1.0 mL min(-1). The analytes were detected at 210 and 314 nm for excitation and emission, respectively. The assay exhibited a linear range of 100-3000 ng mL(-1), with a lower detection limit of 35 ng mL(-1). The method was statistically validated for linearity, accuracy, precision, selectivity and stability according to the FDA guidelines. The intra- and inter-assay coefficients of variation did not exceed 13.5% from the nominal concentration. The accuracy for darifenacin was within ±15% of the theoretical value. The assay was successfully applied in a pharmacokinetic study., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Enantiomeric resolution of atenolol was achieved on the HPLC vancomycin macrocyclic antibiotic chiral stationary phase Chirobiotic V. The polar ionic mobile phase consisted of methanol-glacial acetic acidtriethylamine (100 + 0.025 + 0.75, v/v/v) at a flow rate of 0.8 mL/min. Fluorescence detection at 2751305 nm for excitation and emission, respectively, was used. Plasma samples were purified using SPE on Oasis HLB cartridges. The calibration curves in plasma were linear over the range of 5-400 ng/mL (r = 0.999) for each enantiomer with an LOD of 1.0 ng/mL. The proposed method was validated in compliance with International Conference of Harmonization guidelines in terms of linearity, accuracy, precision, LOD, LOQ, and selectivity. The overall recoveries for S-(-)- and R-(+)-atenolol enantiomers from plasma were 95.0-99.5%; RSD ranged from 2.5 to 3.3%. The developed method was applied for the trace analysis of atenolol enantiomers in plasma and for the pharmacokinetic investigation of atenolol enantiomers in mouse plasma.

A novel poly (vinyl chloride) PVC membrane sensor for Fe(2+) ions is described. The sensor is based on the use of newly synthesized chiral 2,6-bis-(carboxamide methyl ester)pyridine derivative as neutral ionophore in plasticized PVC membrane. The sensor display a fast, stable and near-Nernstian response over a relative wide ferrous concentration range (1 × 10(-3) to 6 × 10(-6) M), with cationic slope of 31.5 ± 0.5, mV per concentration decade over a pH range of 5.0-9.0. The direct determination of 0.25-56.0 µg/ml of ferrous in aqueous solution shows an average recovery of 98.5% and a mean relative standard deviation of 1.5% at 20.0 µg/ml. The sensor displays long life-span, long-term stability, high reproducibility, and short response time. Selectivity coefficients for Fe(II) relative to a number of interfering substances were investigated. The sensor shows high significantly for Fe(2+) over Fe,(3+) Cu,(2+) Zn,(2+) Cd,(2+) Hg,(2+) Pb,(2+) Ni,(2+) Co,(2+) Mn,(2+) Al,(3+) alkaline earth and alkali metal ions. The sensor is successfully applied for measurement of ferrous in drug formulations. The results obtained for the determination of ferrous using the proposed sensor are comparable favourably with those obtained using the spectrophotometric method. Copyright © 2010 John Wiley & Sons, Ltd., (Copyright © 2010 John Wiley & Sons, Ltd.)

The photostability of selected benzocycloheptane antihistaminic agents, namely, loratadine (I), pizotifen (II), ketotifen fumarate (III) and cyproheptatidine (IV), was investigated. Both I and II were photolabile while III and IV were photostable. To perform stability studies on the photolabile compounds (I and II), specific stability-indicating high performance liquid chromatographic (HPLC) methods were established. The accuracy, precision and reliability of the developed HPLC methods for the assay of I and II in their pharmaceutical dosage forms were reported. Assay results for both drugs were within R.S.D. values <2%. The stability-indicating power of the developed methods was validated through study of UV-degraded solutions of I and II contained in quartz cells. The photostability of both drugs was studied under UV-irradiation at 254 nm. The photodegradation kinetics of both drugs, studied in different solvents, are also reported. TLC fractionation of photodegraded solutions of both drugs, revealed two fluorescent photodegradates of drug I. The use of UV-absorbers (ascorbic acid and p-aminobenzoic acid (PABA)) enhanced the photostability of both drugs possibly through a spectral-overlay effect.

The degradation of the amoebicide diloxanide furoate in alkaline medium at different temperatures was investigated using both a spectrophotometric and a developed HPLC method. In solutions, the drug was found to undergo decomposition, i.e., temperature and pH dependent. The pH-rate profile at pH between 7.6 and 9.6 indicated a first-order dependence of Kobs on [-OH]. Arrhenius plot obtained at pH 8 was linear between 40 and 63 degrees C. The estimated activation energy of hydrolysis was found to be 18.25 kcal degree.mol(-1). The effect of simulated gastric and intestinal fluids on the drug was also investigated. A new thin-layer chromatographic (TLC) procedure for the fractionation of the drug and its alkaline hydrolysis products has been developed and was found to compare favorably with that of the British Pharmacopoeia. Three hydrolysis products of a basic methanolic solution of the drug, namely furoic acid, diloxanide and methylfuroate could be identified by the use of TLC, HPLC, infrared and mass spectrometry.

The degradation kinetics of methanolic solution of danazol (0.020% w/v) in aqueous buffers and sodium hydroxide was investigated using stability-indicating HPLC method. The drug degrades in alkaline medium through a base-catalysed proton abstraction rather than via an oxidative mechanism involving oxygen species. The degradation followed pseudo-first-order kinetics. The rates pH-profile exhibited specific base catalysis. The stability of the drug was found to be dependent on pH, buffer concentration, buffer species (acetate, borate, phosphate) and temperature. The ionic strength did not affect the stability of the drug. The energy of activation according to Arrhenius plot was estimated to be 22.62 kcal mol(-1) at pH 12 and temperatures between 30 and 60 degrees C. The effect of simulated gastric and intestinal fluids on the drug stability was also investigated. Two major hydrolytic degradation products were separated and identified by IR, NMR and mass spectrometry and the degradative pathway suggested.

A simple and highly sensitive voltammetric method was developed for the determination of benazepril (I) and ramipril (II). The compounds were treated with nitrous acid, and the cathodic current produced by the resulting nitroso derivatives was measured. The voltammetric behavior was studied by adopting direct current (DCt), differential pulse (DPP), and alternating current (ACt) polarography. Both compounds produced well-defined, diffusion-controlled cathodic waves over the whole pH range in Britton-Robinson buffers (BRb). At pH 3 and 5, the values of diffusion-current constants (Id), were 5.90 +/- 0.40 and 6.66 +/- 0.61 for I and II, respectively. The current concentration plots for I were rectilinear over the range of 1.5-40 and 0.1-30 microg/mL in the DCt and DPP modes, respectively; for II, the range was 2-30 and 0.1-20 microg/mL in the DCt and DPP modes, respectively. The minimum detectabilities (S/N = 2) were 0.015 microg/mL (about 3.25 x 10(-8)M) and 0.012 microg/mL (about 2.88 x 10(-8)M) for I and II, respectively, adopting the DPP mode. Results obtained for the proposed method when applied to the determination of both compounds in dosage forms were in good agreement with those obtained using reference methods. Hydrochlorthiazide, which is frequently co-formulated with these drugs, did not interfere with the assay. The method was also applied to the determination of benazepril in spiked human urine and plasma. The percentage recoveries adopting the DPP mode were 96.2 +/- 1.21 and 95.7 +/- 1.61, respectively.

A simple, rapid and sensitive HPLC method has been developed for the simultaneous determination of ramipril and hydrochlorothiazide in their dosage forms. Acetonitrile: sodium perchlorate solution (0.1 M) adjusted to pH 2.5+/-0.2 with phosphoric acid (46:54 v/v), was used as the mobile phase, at a flow rate of 1.5 ml/min. A supelcosil LC-8 column (5 microm), 15 cm x 4.6 mm i.d. was utilized as stationary phase. Detection was affected spectrophotometrically at 210 nm. Clobazam was used as an internal standard. The method was also applied for the determination of ramipril in the presence of its degradation products. Linearity ranges for ramipril and hydrochlorothiazide were 4.5-45 and 0.6-14 microg/ml, respectively. Minimum detection limits (S/N = 2) obtained were 180 and 23 ng/ml for ramipril and hydrochlorothiazide, respectively. The proposed method was further applied to the analysis of tablets containing the two drugs, the percentage recoveries +/- S.D. (n = 5) were 100.45%+/-0.63 and 99.55%+/-0.78 for ramipril and hydrochlorothiazide, respectively.

A selective high-performance liquid chromatographic procedure for the stability-indicating determination of danazol in the presence of its photolytic degradation products is demonstrated. The photolysis was carried out in glass vials and quartz cell under UV light at 254 nm. Satisfactory results were obtained for the assay and recovery testing with RSD values less than 2%. Kinetic parameters evaluated comprise the order of reaction and the rate constants of the degradation of the danazol irradiated in glass vials or quartz cell.

A simple and sensitive spectrophotometric method has been developed for the determination of benazepril HCl in pharmaceutical formulations. The method is based on the reaction of the drug with potassium permanganate in the presence of sodium hydroxide to produce a bluish-green colored species measurable at 609.4 nm. The absorbance-concentration plot is linear over the range 1-8 microg ml(-1) with minimum detectability of 0.1 microg ml(-1) (2.17 x 10(-7) M). The molar absorptivity was 4.07 x 10(4) l mol(-1) cm(-1) with correlation coefficient (n = 6) of 0.9991. The different experimental parameters affecting the development and stability of the color were studied carefully and optimized. The proposed method was applied successfully to the determination of benazepril in its dosage forms, the percentage recoveries +/- SD (n = 9) were 99.79 +/- 1.40 and 100.50 +/- 1.48 for tablets containing 10 and 20 mg, respectively. The results obtained were in good agreement with those obtained using a reference spectrophotometric method. The proposed method could be applied to the determination of benazepril in presence of the co-formulated drug, hydrochlorothiazide. A proposal of the reaction pathway was presented.

Accelerated photochemical degradation of norfloxacin in solutions, bulk forms, and tablets has been undertaken. The kinetic order of the drug photodegradation was determined by monitoring residual drug masses as a function of time matched with initial matched mass by adopting the HPLC-method of USP-23. A new dimeric photodegradate, formed from the active decarboxylated norfloxacin monomer, could be isolated and characterized.

High performance liquid chromatographic (HPLC) separation has been investigated for the determination of intact norfloxacin in the presence of its photodegradation products. The HPLC-separation could be achieved isocratically and by gradient elution on a Micropak -NH2 column (10 microns, 30 cm x 4 mm O) using a mobile phase containing acetonitrile, tetrabutylammonium hydroxide, o-phosphoric acid and water at a rate of 2 ml.min-1 with UV-detection (278 nm) at ambient temperature. The method was applied for the drug analysis in fresh and photodegraded norfoxacin samples, as well as for assessment of the content uniformity of tablets containing the drug. The results of the proposed liquid chromatographic method were statistically matched with those obtained by adopting an official HPLC-method (USP XXII-procedure).

Direct potentiometric titration and two HPLC conditions for the simultaneous determination of amoxycillin and dicloxacillin in their capsules have been developed. One-run titration utilizing 0.05M acet. HClO(4) enables the quantification of both antibiotics. The HPLC-separation could be undertaken on reversed phase, LiChrosorb RP-18 (10 mum), and LiChrospher 100 RP-18 (5 mum), columns by using mobile phases containing acetonitrile + 1% aq, acetic acid, in proportions of 47:53 or 39:61 (v/v), respectively, at a flow rate of 1.5 ml/min with UV-detection at 240 nm. Recoveries of the individual drugs by the application of each described method were found to be fairly satisfactory.

Fundamental harmonic alternating current polarography has been described for the determination of tolmetin sodium and its capsules, (Tolectin-200 mg). The AC procedure was applied by superimposing 10 mV, rms, AC voltage on a DC ramp in the range -1200 to -1450 mV at a frequency of 50 Hz in acetate buffer of pH 5.0 containing 0.001% gelatin as a maxima suppressor. A three electrode assembly consisting of dropping mercury electrode (dme) and two Ag/AgCl/KCl (sat.) electrodes was employed. Mercury flow rate was 3 mg sec(-1) under a corrected head pressure of 80 cm. The electrode reaction was investigated by Kalousek K1 and K2 techniques which indicated the non-reversibility of the electrode process on the AC time scale. The percent result obtained by standard addition procedure was 101.4 +/- 2.0 for tolmetin sodium capsules.

A liquid chromatographic method for the simultaneous determination of amoxycillin and potassium clavulanate in tablet and suspension preparations is presented. The method specifies reversed phase column and a buffered mobile phase (CH3OH + KH2PO4-buffer pH 6 + H2O, 15:1:84) isocratically at a rate of 1.0 ml min-1, with detection at 235 nm. The suitability of the chromatographic system developed is tested using replicate injections of the sample and standard preparations. The observed relative standard deviations (RSDs) were within 2%. Recovery experiments conducted utilizing the proposed method gives results of 101.5% +/- 1.72 (n = 6) and 101.22% +/- 1.93 (n = 6) for amoxycillin in tablets and powder for oral administration, respectively. Similarly, recovery experiments for clavulanic acid gave results of 100.33 +/- 1.90 (n = 6) and 99.61 +/- 1.32 (n = 6) in the tablets and suspension powder, respectively. Comparison of the proposed method with the USP method proved it to be satisfactory. The statistical F- and t-tests observed, indicated that there were no significant differences between the two methods regarding precision and accuracy.

High performance liquid chromatographic methods for the individual determination of khellin, phenobarbitone and dipyrone in tablets are presented. The methods specify a reverse phase column: methanol + water (68 + 32) as mobile phase at a flow rate of 0.7 ml/min with detection at 254 nm for khellin, visgnagin and dipyrone; water + ammonia + methanol (94.5 + 0.5 + 5) as a mobile phase at a flow rate of 0.7 ml/min with detection at 240 nm for phenobarbitone. At sensitivity of 0.01 AUFS, linearity ranges were found to be 0.5-4 micrograms/ml for khellin, 2.5-12.5 micrograms/ml for dipyrone and 1-7 micrograms/ml for phenobarbitone and with relative standard deviations less than 2%. The mean percentage recoveries +/- SD of khellin, dipyrone and phenobarbitone added to tablets were found to be 101.0 +/- 0.65, 100.0 +/- 0.74 and 99.9 +/- 0.74, respectively. The system can detect 2% w/w visnagin in khellin.

Three methods have been developed for the determination of nomifensine maleate alone and in capsules: a spectrophotometric, an iodine charge-transfer, and a spectrofluorimetric method. All three give linear calibration graphs, over the ranges 20-100, 1-5 and 0.1-0.5 microg/ml, respectively, with coefficients of variation of 0.8, 1.3 and 1.3%, respectively.

Reversed-phase ion-pair conditions are used for the determination of D-penicillamine and penicillamine disulphide. Two chromatographic systems were employed, one for penicillamine and the other for penicillamine disulphide. The procedures permit the determination of total penicillamine (protein-bound, free and as disulphides) in whole plasma, and total penicillamine (free and as disulphides) in plasma ultrafiltrate, using an incubation step in the presence of dithiothreitol. Free penicillamine and penicillamine disulphide may be determined independently by direct injection of plasma ultrafiltrate. Both solutes may be measured at an on-column sensitivity of 10 ng, utilizing an electrochemical detector based on a glassy carbon electrode.

Piperazine and its salts are reacted with aqueous alcoholic p-benzoquinone, buffered at pH 5.4, to give a coloured product with maximum absorption at 516nm. The piperazine base has a molar absorptivity of 0.96 x 10(4)l.mole(-1).cm(-1) and Beer's law is obeyed over the range 2-10 mug ml . When applied to three commercial preparations, the proposed method gave mean recoveries within 1% of those obtained by the official gravimetric method. The relative standard deviation was less than 1%.

A rapid, sensitive and precise liquid chromatographic method is presented for the determination of aztreonam alone, in the presence of its degradation product, and in a parenteral formulation containing l-arginine. A reverse phase column and 0.2M phosphate buffer (pH 6)-methanol (95 + 5) with mobile phase at a flow rate of 2 mL/min is used. The method is sensitive for the range of 10-50 micrograms/mL with a relative standard deviation of less than 2%. The method has been applied to a parenteral formulation containing aztreonam and l-arginine. L-Arginine is also determined by a nonaqueous titrimetric method.

The first derivative curve (D1) of the absorption spectrum of vitamin A acetate in cyclohexane possesses a trough (D11) at 348 nm and a maximum (D12) at 306 nm. D11, D12, and delta D1 (= D11 - D12) are linearly related to concentration over a range of 5-25 i.u. ml-1. When a tangent is drawn between D1 at 400 nm and D1, at 306 nm, the amplitude at 348 nm (D1 corr.) is linearly related to concentration. Ratios of magnitude of D11/magnitude of D12 and D1(corr.)/magnitude of D1 are independent of concentration, and have been used to reveal the presence of interferences in the D1 curves of vitamin A in oily capsules. Vitamin A in two oily preparations has been assayed using delta D1 and D1(corr.). The potency found was within +/- 2% from that obtained by using the B.P. method. The method is rapid, precise and accurate.

A spectrophotometric procedure for the simultaneous determination of amoxycillin and clavulanic acid in some pharmaceutical preparations has been developed. As the absorption bands of amoxycillin (274 and 227 nm) and clavulanic acid (270 nm) overlap, both Vierordt's method and derivative spectrophotometry have been investigated and evaluated. The first-derivative spectrophotometric method was found to be more accurate, direct and reproducible.

A liquid chromatographic method for the individual determination of benzoic and cinnamic acids in 2 benzoin preparations is presented. The method specifies a reverse phase column and 0.01M KH2PO4-methanol (85 + 15) as mobile phase at a flow rate of 1.8 mL/min, with detection at 254 nm. The method has been applied to 2 benzoin preparations and the results were compared with those from the British Pharmacopoeia method.

Two colorimetric methods are reported for the assay of nomifensine maleate. The methods are based on coupling between the diazotised form of nomifensine maleate and (i) N-(1-naphthyl)-ethylene diamine dihydrochloride (Bratton-Marshall reagent) and (ii) p-aminosalicylic acid (PAS). The optimum conditions for the reactions were investigated. The coupled products exhibit maximum absorbance at 470 and 435 nm for the Bratton-Marshall and PAS reagents, respectively. With PAS, a linear relationship has been established between absorbance (Amax) and concentration of nomefensine maleate over the range 2-12 micrograms ml-1. Similarly, with the Bratton-Marshall reagent, a linear relationship exists in the concentration range 2-16 micrograms ml-1. The calculated mean percent recoveries for nomifensine maleate in the commercial capsules (Merital 25 mg) respectively. Similarly, for the added recoveries, the percentage obtained were 99.01 +/- 0.46 and 100.03 +/- 1.03, respectively.