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Simultaneous spectrophotometric determination of pseudoephedrine hydrochloride and ibuprofen in a pharmaceutical preparation using ratio spectra derivative spectrophotometry and multivariate calibration techniques.

Spectrophotometric methods are described for the simultaneous determination of pseudoephedrine hydrochloride and ibuprofen in their combination. The obtained data were evaluated by using five different methods. In the first method, ratio spectra derivative spectrophotometry, analytical signals were measured at the wavelengths corresponding to either maximums and minimums for both drugs in the first derivative spectra of the ratio spectra obtained by using each other spectra as divisor in their solution in 0.1 M HCl. In the other four spectrophotometric methods using chemometric techniques, classical least-squares, inverse least-squares, principal component regression and partial least-squares (PLS), the concentration data matrix were prepared by using the synthetic mixtures containing these drugs in methanol:0.1 M HCl (3:1). The absorbance data matrix corresponding to the concentration data matrix was obtained by the measurements of absorbances in the range 240-285 nm in the intervals with deltalambda = 2.5 nm at 18 wavelengths in their zero-order spectra, then, calibration or regression was obtained by using the absorbance data matrix and concentration data matrix for the prediction of the unknown concentrations of pseudoephedrine hydrochloride and ibuprofen in their mixture. The procedures did not require any separation step. The linear range was found to be 300-1300 microg/ml for ibuprofen and 100-1300 microg/ml for pseudoephedrine hydrochloride in all five methods. The accuracy and the precision of the methods have been determined and they have been validated by analyzing synthetic mixtures. The five methods were successfully applied to tablets and the results were compared with each other.

Calibration↗

Simultaneous determination of tenoxicam and 2-aminopyridine using derivative spectrophotometry and high-performance liquid chromatography.

Derivative spectrophotometry and high-performance liquid chromatography (HPLC) were used to determine tenoxicam and one of its decomposition products (2-aminopyridine) simultaneously and in the presence of each other. The derivative procedure was based on the linear relationship between the tenoxicam concentration and the second derivative amplitudes at 390-348 nm (peak-to-trough) measurement. The 2-aminopyridine was determined through measuring the second derivative amplitude at 241 nm (zero-crossing for tenoxicam). For the HPLC procedure, a reversed-phase C8 column with a mobile phase composed of 0.02 M sodium acetate-methanol-acetonitrile (11:8:1) with 0.005 M heptane sulfonic acid sodium salt, as an ion pair, was used to separate both compounds with 2,4-dinitrochlorobenzene, as an internal standard, in reasonable time. The flow rate was 1.5 ml min-1 with a programmable ultraviolet (UV) detection at 300 and 375 nm. Both UV derivative spectrophotometric and HPLC approaches were followed for confirming the purity of tenoxicam in bulk and tablets dosage form.

Aminopyridines↗

Quantitative determination of acrivastine and pseudoephedrine hydrochloride in pharmaceutical formulation by high performance liquid chromatography and derivative spectrophotometry.

In this study, fourth derivative spectrophotometry and high performance liquid chromatography (HPLC) have been used and described for the quantitative determination of acrivastine (I) and pseudoephedrine hydrochloride (II) in their pharmaceutical capsules form (Duact). In the former method, d4A/d gamma 4 values were measured in methanol at 315 and 269 nm for (I) and (II) respectively. The relative standard deviations (RSD) for the method were found to be 1.16% for (I) and 0.94% for (II). The latter method based on reversed phase HPLC system using LiChrosorb C18 analytical column. The mobile phase used for separation of (I), (II) and internal standard (p-hydroxymethylbenzoate) were the water/acetonitrile/methanol/perchloric acid/n-octylamine (500:130:25:13:0.3 v/v) and the detection of the compounds in the capsules were at 260 nm using UV detector. The RSD for the HPLC method were determined to be 0.79 and 0.88% for (I) and (II) respectively. The proposed methods, which give thoroughly comparable data, are simple, rapid, and allow precise and accurate results and could be used for commercial formulations containing acrivastine and pseudoephedrine hydrochloride in combination.

Capsules↗

Zero-crossing derivative spectrophotometry for the determination of haloperidol in presence of parabens.

First derivative spectrophotometry with a zero-crossing technique of measurement is used for the quantitative determination of haloperidol in the presence of methylparaben and propylparaben, which is added as antimicrobial preservatives in pharmaceuticals. This technique permits the quantification of haloperidol in the presence of parabens, with closely overlapping spectral bands, and without any separation step. Linear calibration graphs of first derivative values (at 255.2 nm for haloperidol) versus concentration (in the range 4.0-20.0 micrograms ml-1) were obtained with negligible intercepts. Relative standard deviation of 0.83% was obtained for intra-day precision and 1.86% for inter-day precision. The recovery of haloperidol in synthetic mixtures with parabens and pharmaceutical dosage forms is also reported.

Calibration↗

Quantitation of amphotericin B in plasma by second-derivative spectrophotometry.

A method is described for determining amphotericin B in plasma using second-derivative spectrophotometry after deproteinization. The assay was based on the absorbance at 407.5 nm. The second-derivative spectrum recorded between 350 and 450 nm allowed identification of the analyte and showed absence of drug interference. Only bilirubin interfered at high concentration (> or = 50 mumol l-1. The linear concentration ranges were 0.05 -5.0 mg l-1 (r = 0.999, slope = 2.731, intercept = 0.008). Between-day CV < or = 9.7%, within-day CV < or = 5.5%, analytical recovery close to 100% were suitable for clinical investigations. This method provides better specificity than direct absorbance, is simpler and faster than a high performance liquid chromatography assay and can be used routinely by any laboratory possessing a spectrophotometer with a derivative accessory.

Amphotericin B↗

Simultaneous determination of melatonin-pyridoxine combination in tablets by zero-crossing derivative spectrophotometry and spectrofluorimetry.

Two methods have been developed for the analysis of melatonin (M) and pyridoxine hydrochloride (PH) in combination. The first method depends on first- and second-derivative ultraviolet spectrophotometry, with the zero crossing technique of measurement. First-derivative amplitudes at 296 nm and second-derivative amplitudes at 294 and 322 nm are selected for the determination of M and PH, respectively. The second method is based on the native fluorescence of both M and PH, in methanol and 0.1 M hydrochloric acid, respectively, after a preliminary solvent extraction procedure. The relative standard deviation of both methods was less than 2.0%. The two methods have been successfully applied to the determination of both drugs in laboratory-prepared mixtures and in tablets.

Melatonin↗

Application of derivative spectrophotometry for determination of coenzyme Q10 in pharmaceuticals and plasma.

The use of derivative spectrophotometry is proposed in this work for determination of coenzyme Q10 in formulations and in human plasma. The spectrophotometric procedure is simpler and less expensive than chromatographic techniques commonly used for the analysis of coenzyme. The active compound can be determined in the range 0.25-10 ppm for standard solutions and pharmaceuticals and 0.05-1.5 ppm in plasma. The proposed method was applied for coenzyme determination in real samples. The results agree well with declared value and with these obtained by HPLC.

Antioxidants↗

Simultaneous determination of pteridines in multicomponent mixtures using derivative spectrophotometry and partial least-squares calibration.

Simple binary mixtures composed of very similar pteridines (neopterin and pterin) have been resolved by derivative spectrophotometry. Detection limits of 0.30 microgram ml-1 and 0.12 microgram ml-1 for pterin and neopterin, respectively, have been calculated. Also, different mixtures of pteridines considered as disease markers, such as pterin, neopterin, xanthopterin and isoxanthopterin, have been determined by using a partial least-squares (PLS-2) model. Calibration set containing 0-7 micrograms ml-1 for each component was used. The resolution of several mixtures and single determination was tested in artificial samples.

Drug Combinations↗

Simultaneous determination of salicylic acid and acetylsalicylic acid in aspirin delayed-release tablet formulations by second-derivative UV spectrophotometry.

A rapid, simple assay procedure was developed for simultaneous analysis of aspirin (ASA) and salicylic acid (SA) in aspirin delayed-release tablet formulation by 'zero crossing' second-derivative UV spectrophotometry. The zero-order absorption spectra and second derivative spectra of ASA and SA were recorded in diluting solution acetonitrile-formic acid (99:1). The accuracy of the method was demonstrated by the determination of ASA and SA in five tablets formulations (each 20 tablets of the same batch) by the described method and by high performance liquid chromatographic method, and the results were in good agreement.

Aspirin↗

Determination of miconazole in pharmaceutical creams using internal standard and second derivative spectrophotometry.

A simple method is proposed for miconazole determination in pharmaceutical creams, based on extraction and second derivative spectrophotometry. In the presence of sodium lauryl sulfate (0.5%) and sulphuric acid (0.4 mol l(-1)), the miconazole and internal standard (IS) (methylene blue) were extracted to 100 microl of methylene chloride. The organic phase was evaporated in the nitrogen stream and the dry residue was dissolved in methanol (1.5 ml). The analytical signal was obtained as the ratio between second derivative absorbances measured at 236.9 nm (miconazole) and at 663.2 nm (IS). The use of IS in such multi-stage procedure enabled quite good analytical performance in calibration range 50.0 400 mg l(-1): linear correlation coefficient 0.9995, precision (measured as CV for ten replicates) at 50.0 mg l(-1) and at 400 mg l(-1) of miconazole was 1.5 and 0.5% respectively. Four commercial pharmaceutical creams were analyzed and the results obtained were in good agreement with the results obtained by reversed-phase high performance liquid chromatography (HPLC).

Antifungal Agents↗

Determination of active ingredients in the pharmaceutical formulations containing hydrochlorothiazide and its binary mixtures with benazepril hydrochloride, triamterene and cilazapril by ratio spectra derivative spectrophotometry and vierordt's method.

Procedures were developed for the simultaneous determination of pharmaceuticals in binary mixtures, containing hydrochlorothiazide-benazepril hydrochloride, hydrochlorothiazide triamterene and hydrochlorothiazide cilazapril by ratio spectra derivative spectrophotometry and Vierordt's method. Mean recoveries, relative standard deviations and linearity ranges in calibration graphs of the methods were compared. These procedures were successfully applied to three pharmaceutical formulations for the determination of active ingredients.

Angiotensin-Converting Enzyme Inhibitors↗

Application of first-derivative UV-spectrophotometry, TLC-densitometry and liquid chromatography for the simultaneous determination of mebeverine hydrochloride and sulpiride.

Three methods are described for the simultaneous determination of mebeverine hydrochloride (MB) and sulpiride (SU) in combined pharmaceutical tablets. The first method depends on first-derivative ultraviolet spectrophotometry, with zero-crossing measurement method. The first derivative amplitudes at 214.2 and 221.6 nm were selected for the assay of MB and SU, respectively. Calibration graphs follow Beer's law in the range of 10-30 and 2-8 microg/ml(-1), and the linearity was satisfactory (r = 0.9999), for MB and SU, respectively. The second method was based on the application of the thin layer chromatographic separation of both drugs followed by the densitometric measurements of their spot areas. After separation on silica gel GF254 plates, using ethanol: diethyl ether: triethylamine (70:30:1 v/v) as the mobile phase, the chromatographic zones corresponding to the spots of MB and SU were scanned at 262 and 240 nm, respectively. The calibration function was established in the ranges of 4-12 microg for MB and 2-8 microg for SU. The third method was an internal standard procedure based on high performance liquid chromatographic separation of the two drugs on a reversed-phase, Bondapak CN column. The detection was done at 243 nm using buclizine hydrochloride as internal standard. All chromatographic methods showed good linearity, precision and reproducibility. No spectral or chromatographic interference from the tablet excipients were found. The proposed methods were successfully applied to the assay of commercial tablets and content uniformity test. The procedures were rapid, simple and suitable for quality control application.

Anticonvulsants↗

Misclassification due to body hair and seasonal variation on melanin density estimates for skin type using spectrophotometry.

Recent advances have enabled quite accurate estimations of cutaneous melanin density by spectrophotometry using reflectance of light at wavelengths 400 and 420 nm. Our purpose was to assess the effect of body hair and seasonal variation at the upper inner arm and buttock on measurements of melanin density. We estimated melanin density of 104 volunteers at 3-monthly intervals over 12 months both before and after shaving. Removing body hair at the upper inner arm had no effect, but substantially reduced melanin estimates at the buttock in men. Significant seasonal variation was only observed at the upper inner arm, with highest readings in summer-autumn. In case-control studies, misclassification due to body hair at the buttock and seasonal variation at the upper inner arm could affect the observed odds ratio substantially. However, both sources of error can be reduced by careful attention to key aspects of study design.

Adult↗

Determination of acidity constants of acid-base indicators by second-derivative spectrophotometry.

A method for calculation of acid-base dissociation constants of monoprotic weak organic acids whose acid and base species have overlapping spectra from absorptiometric and pH measurements is described. It has been shown that the second-derivative spectrophotometry can effectively be used for determining the dissociation constants, when dissociation constants obtained for methyl orange and bromothymol blue were compared with the values given in the literature.

Azo Compounds↗

A reexamination of aluminum in Alzheimer's disease: analysis by energy dispersive X-ray microprobe and flameless atomic absorption spectrophotometry.

We have attempted to verify the presence of increased aluminum (Al) levels in Alzheimer's disease (AD) brains by energy dispersive X-ray microanalysis (EDX) and flameless atomic absorption spectrophotometry (AAS). Tissue from seven AD brains, mounted on carbon polymerized coverslips, were stained with Congo-red or treated immunohistochemically to allow optical localization of AD-associated lesions during EDX. Despite a demonstrated sensitivity of 20-25 ppm, we were unable to detect Al in either plaque cores or neurons containing neurofibrillary tangles. For AAS, wet weight samples (ranging from 48-144 mg) from six of the seven AD brains and four controls were selected from regions similar to those studied under EDX, i.e., Brodmann areas 9, 11, 28, 46, 47, and the hippocampus. The tissue surrounding each sample site was sectioned and stained for thioflavin S. Both controls and AD samples revealed similar levels of Al ranging from undetectable to 1.80 ng/mg wet wt. (mean AD: 0.28 +/- 0.39 (SD), control: 0.54 +/- 0.58 (SD], independent of degree of histopathology or age of the case. We conclude that the combined strengths of these two techniques on similar tissue specimens demonstrate that abnormal Al levels are not required to produce the histologic findings of AD and that this element may not accumulate in the aging brain. It is unlikely, therefore, that Al is essential in the etiology of pathogenesis of plaques and tangles in AD. Al's role as a primary or secondarily associated event, when present, needs further delineation.

Aged↗

Optical difference spectrophotometry as a probe of rat brain nitric oxide synthase heme-substrate interaction.

NO synthase (NOS) is a family of enzymes that catalyzes the NADPH-dependent formation of NO and citrulline from L-arginine and molecular oxygen. The reaction involves an initial hydroxylation of L-arginine to form the isolable intermediate NG-hydroxy-L-arginine (NOHArg). The subsequent incorporation of a second atom of oxygen during the metabolism of NOHArg is required to yield the final products NO and citrulline. NOS contains heme iron, FAD, FMN, and tetrahydrobiopterin prosthetic groups. To examine the interaction of substrates with the heme prosthetic group, substrate perturbation difference spectrophotometry was employed. By analogy with substrate binding interactions with cytochromes P450, NOS exhibits "type I" substrate perturbation difference spectra with the substrates L-arginine and NOHArg and the inhibitor NG-methyl-L-arginine (NMA). These spectral perturbations are characterized by the appearance in the difference spectrum of a peak at approximately 380 nm, a trough with an absorbance minimum at approximately 420 nm, and an isosbestic point at approximately 405 nm. The spectral binding constants, Ks, for L-arginine and NMA were determined to be approximately 2.5 microM. These values are in agreement with the reported kinetic constants for these compounds. The "apparent Ks" values for NOHArg were 0.4 microM (2.0 microM NOS) and 0.8 microM (3.5 microM NOS), respectively. Furthermore, NOS exhibits "type II" difference spectra upon titration with imidazole, characterized by the appearance of a peak at approximately 430 nm and a trough at approximately 395 nm, with a spectral binding constant of approximately 160 microM.

Amino Acid Oxidoreductases↗

Reaction of indole and analogues with amino acid complexes of Escherichia coli tryptophan indole-lyase: detection of a new reaction intermediate by rapid-scanning stopped-flow spectrophotometry.

The effects of indole and analogues on the reaction of Escherichia coli tryptophan indole-lyase (tryptophanase) with amino acid substrates and quasisubstrates have been studied by rapid-scanning and single-wavelength stopped-flow spectrophotometry. Indole binds rapidly (within the dead time of the stopped-flow instrument) to both the external aldimine and quinonoid complexes with L-alanine, and the absorbance of the quinonoid intermediate decreases in a subsequent slow relaxation. Indoline binds preferentially to the external aldimine complex with L-alanine, while benzimidazole binds selectively to the quinonoid complex of L-alanine. Indole and indoline do not significantly affect the spectrum of the quinonoid intermediates formed in the reaction of the enzyme with S-alkyl-L-cysteines, but benzimidazole causes a rapid decrease in the quinonoid peak at 512 nm and the appearance of a new peak at 345 nm. Benzimidazole also causes a rapid decrease in the quinonoid peak at 505 nm formed in the reaction with L-tryptophan and the appearance of a new absorbance peak at 345 nm. Furthermore, addition of benzimidazole to solutions of enzyme, potassium pyruvate, and ammonium chloride results in the formation of a similar absorption peak at 340 nm. This complex reacts rapidly with indole to form a quinonoid intermediate very similar to that formed from L-tryptophan. This new intermediate is formed faster than catalytic turnover (kcat = 6.8 s-1) and may be an alpha-aminoacrylate intermediate bound as a gem-diamine.

Amino Acids↗

Photoreaction cycle of phoborhodopsin studied by low-temperature spectrophotometry.

The photochemical and subsequent thermal reactions of phoborhodopsin (pR490), which mediates the negative phototaxis (phobic reaction) of Halobacterium halobium, were investigated by low-temperature spectrophotometry. At room temperature, the absorption spectrum of pR490 displayed vibrational structure with a maximum at 490 nm and a shoulder at 460 nm, which were remarkably sharpened by cooling, resulting in the appearance of two well-separated peaks. On irradiation of pR490 at -170 degrees C, a photo-steady-state mixture composed of pR490 and two photoproducts, P520 and P480, was formed. P480 had an absorption maximum at 480 nm and thermally converted to pR490 above -160 degrees C, while P520 had an absorption maximum at 515 nm and thermally converted to P350, the next intermediate, above -60 degrees C. Above -30 degrees C, P350 was converted to P530, and then reverted to pR490. P520, P350, and P530 may correspond to K, M, and O intermediates of bacteriorhodopsin, respectively, on the basis of their absorption spectra, but the intermediates corresponding to L and N intermediates were not observed. On the basis of these results, a new scheme of the photoreaction cycle of pR490 was presented.

Archaeal Proteins↗