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PubMed2002

Speciation of Fe(II) and Fe(III) by the modified ferrozine method, FIA-spectrophotometry, and flame AAS after cloud-point extraction.

A method has been developed for the simultaneous determination of traces of Fe(III) and Fe(II) in water by on-line coupling of spectrophotometry with flame atomic absorption spectrometry (FAAS). The method involves cloud-point extraction (CPE) of both species with ammonium pyrrolidinecarbodithioate (APDC) under standard conditions, which facilitates the in situ complexation and extraction of both species. Differentiation of the oxidation states of iron is achieved by using mathematical equations to overcome the interference of Fe(III) in the spectrophotometric determination of Fe(II) when they are both present in the same solution. In this manner the time-consuming and labor-intensive steps of preoxidation of Fe(II) or reduction of Fe(III) are eliminated. By preconcentrating a 10-mL sample solution detection limits as low as 7 microg L(-1), were obtained after a single-step extraction procedure. The relative standard deviation (n=4, 30 microg L(-1)) was 2.6 % and 1.8 % for spectrophotometry and FAAS, respectively. Recoveries in the range of 96-105 % were obtained by analysis of spiked real samples. The method was further verified by analyzing a certified reference material (IMEP-9); for this the recovery was 98.5 %.

Ferrozine↗
PubMed1988

Rapid determination of kinetic constants by competitive spectrophotometry.

The use of competitive spectrophotometry to measure kinetic constants for enzyme-catalyzed reactions is described. The equation for the progress curve characterizing the kinetic behavior of an enzyme acting simultaneously on two alternative substrates is derived. By the addition of a competition term to the integrated Michaelis-Menten equation, the kinetic constants of an alternative substrate can be evaluated by measuring the competition with a substrate of known kinetic constants in a single experiment. Studies are presented involving the enzymes leucine aminopeptidase (LAP) and carboxypeptidase A (CPA). The results obtained with LAP and CPA showed that the kinetic constants determined using competitive spectrophotometry were in agreement with values cited in the literature or with values determined by single substrate enzyme kinetics.

Binding, Competitive↗
PubMed1990

Determination of tryptophan, tyrosine, and phenylalanine by second derivative spectrophotometry.

Second derivative spectrophotometry has been useful for the determination of aromatic amino acids. However, published methods produce erroneous results, because those methods measure second derivative values by the vertical distance between peak and trough which is subject to variation according to the aromatic amino acid composition of proteins. This paper presents a method of second derivative spectrophotometry which measures second derivative absorbance values by means of the vertical distance from baseline to the derivative curve at a wavelength specifically assigned to each aromatic amino acid, and makes corrections for the interference from other amino acids at the same wavelength. The Appendix describes a computational method for obtaining absolute values of second derivative absorbances directly from normal absorbance values without using the spectrophotometer's derivative mode, because most commercial instruments produce completely arbitrary second derivative values which make comparison of data obtained on two different instruments impossible.

Humans↗
PubMed1976

Differentiation between chelated and non-chelated DNA-Pt complexes by atomic absorption spectrophotometry.

As a continuation of our study of the properties of the cis-trans platinum series we have investigated the DNA-Pt complexes of [Pt(dien)Cl]Cl, cis-Pt (en)Cl2, cis-Pt (NH3)2Cl2 and K2[PtCl4] by atomic absorption spectrophotometry. The DNA-Pt complexes correspond to the saturation of the N7-(Guanine) sites. It has been found that the chelate complexes obtained with cis-Pt(en)Cl2, cis-Pt(NH3)2Cl1 and K2[PtCl4] show the same absorbance. The [Pt(dien)Cl]Cl and trans-Pt (NH3)2Cl2 which are bound to the N7 (tuanine) site only, show an absorbance greater than the chelate complexes by a factor of two. In addition, it has been possible in the case of the trans-Pt (NH3)CCl1 complex to follow the fixation of platinum to DNA by atomic absorption spectrophotometry. The result is similar to the ionic chlorine liberation procedure reported previously.

Binding Sites↗
PubMed1975

Difference-derivative absorbance spectrophotometry as a technique to measure state changes of phenylalanine residues in proteins.

The first derivatives of difference absorbance spectra of several proteins were measured to examine the applicability of this technique as a tool to investigate state changes of phenylalanine residues in proteins. It was found by this technique that phenylalanine residues in insulin and those in lysozyme are exposed to more aqueous environment by denaturation with guanidine hydrochloride. Heat denaturation of collagen caused similar changes of some of its phenylalanine residues. It was thus demonstrated that difference-derivative absorbance spectrophotometry gives the information about state changes of phenylalanine residues in native proteins, which are hardly detected by common difference spectrophotometry.

Air Sacs↗
PubMed1981

Effect of dodecyl sulfate on the spectral properties of phenylalanyl residues in serum albumin detected by second derivative spectrophotometry.

The effect of sodium dodecyl sulfate (SDS) on the spectral properties of phenylalanine residues in bovine serum albumin was studied at neutral pH by second derivative spectrophotometry. It was found that phenylalanine residues in the interior of bovine serum albumin became almost completely exposed on the surface of the protein on formation of a so-called AD12 complex. This conformational change began to be significant when 4 mol SDS bound to bovine serum albumin. At higher concentrations of SDS, when so-called ADn and AD2n complexes were formed, phenylalanine residues were transferred to the hydrophobic region again. This might be due to the involvement of phenylalanine residues in micelle-like clusters. Change in the conformation of bovine serum albumin involving tryptophan residues was also measured. These studies demonstrate the value of second derivative spectrophotometry in studies on conformational change of proteins.

Animals↗
PubMed1991

Quantitation of urobilinogen in feces, urine, bile and serum by direct spectrophotometry of zinc complex.

Previous methods to quantitate urobilinogen lack precision due to either incomplete reduction of urobilin or to losses of pigment before the use of Ehrlich's aldehyde reaction or due to pigment precipitation, as occurs in Schlesinger's fluorescent assay. The present procedure modifies the latter assay to obviate described problems as it is based on direct spectrophotometry (or spectrofluorometry) of a zinc complex of urobilin in dimethylsulfoxide. The sample is extracted with dimethylsulfoxide to increase recovery of urobilinogen from samples of various origin (feces, urine, bile, serum etc.) and to prevent the precipitation of proteins. After oxidation of urobilinogen with iodine, the concentration of the resulting urobilin is directly determined from the absorption (or fluorescent) spectrum. High sensitivity and high specificity for the procedure result from the high value of absorption coefficient and by the characteristic absorption spectrum of zinc complex of urobilin, respectively. Within-day and day-to-day coefficients of variation of stool and bile samples range from 1.6 to 9.2%. The smallest concentration of urobilinogen measurable by spectrophotometry is approximately 0.5 mumol/l, by fluorometry it is 0.25 mumol/l. The recovery varies from 82.2 to 93.8% depending on re-extraction of the sample. The method is linear in the range of 1 to 35 mumol/l and of 0.5 to 17.5 mumol/l for spectrophotometric and fluorescent determinations, respectively. The results obtained with the present method correlated well with Ehrlich's determination (r2 = 0.912), but are approximately two-fold higher. Storage of the samples at -20 degrees C or extraction with dimethylsulfoxide prior to storage are good ways for sample preservation. Twenty stool samples from healthy adults were determined.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1992

Dual wavelength spectrophotometry as a diagnostic test of the pulp chamber contents.

The purpose of this in vitro study was to determine the feasibility of using dual wavelength spectrophotometry to identify teeth with pulp chambers that are either empty, filled with fixed pulp tissue, or filled with oxygenated blood. In phase I of the experiment, a human third molar was prepared so that its pulp space could be filled with oxygenated blood and later emptied. In phase II, the lower jaw of a beagle dog was removed and placed in formalin, thereby fixing the pulps of the teeth. The pulp of the right canine was removed via an apical approach, and attachments were placed in a similar position to those on the human tooth, to allow filling and emptying of the pulp space. Cavit was placed over the exposed fixed pulp in the left canine. Ten readings, which were separated by light source and detector removal and replacement, were taken of the right canine pulp space when it was empty or filled with oxygenated blood, or the left canine pulp space when it was filled with fixed tissue. Distinct and reproducible changes were measured for pulp spaces filled with air, tissue, or oxygenated blood. In phase III, simulated pulp testing on a dog tooth model was performed. Blood was introduced into the root canal space, the chamber was rinsed with water and replaced with air, according to a predetermined code. Spectrophotometer readings were recorded. The identification of pulpal contents was correctly determined in all 20 of the predetermined conditions. The findings indicate that continuous wave spectrophotometry may become a useful pulp testing method.

Animals↗
PubMed1994

Determination of guaiphenesin in anti-tussive pharmaceutical preparations containing dextromethorphan by first- and second-derivative ultraviolet spectrophotometry.

Rapid, simple and direct assay procedures based on selective first (D1)- and second (D2)-derivative spectrophotometry, using a zero-crossing technique of measurement at 279.2 and 280.0 nm, respectively, have been developed for the specific determination of guaiphenesin in the presence of dextromethorphan, drugs with closely overlapping absorption spectra, in synthetic admixtures and in pharmaceutical dosage forms (tablets and syrups). The methods do not require extraction with organic solvents and are easier to perform than their conventional counterparts. Calibration graphs were linear (r = 0.99999 for D1 and 0.99969 for D2, respectively). Good selectivity, accuracy and precision were found. However, the performance of the analysis of guaiphenesin by the second-derivative mode deteriorated when the ratio of dextromethorphan to guaiphenesin was greater than one. Thus, the first-derivative spectrophotometry is the method of choice for the assay of tablets and syrups containing the two drugs.

Antitussive Agents↗
PubMed2003

Rapid and simple methods for quantitative analysis of some antidepressant in pharmaceutical formulations by using first derivative spectrophotometry and HPLC.

Two rapid, simple and accurate first derivative spectrophotometry and HPLC method for the determination of nefazodone hydrochloride and sertraline hydrochloride in pharmaceutical formulations are discussed. The first one is a derivative spectrophotometric procedure and the second one is based on a HPLC method with a UV detector. In the first method, first derivative spectrophotometry, nefazodone hydrochloride or sertraline hydrochloride by measurement of their first derivative signals at 241.8-256.7 nm (peak-to-peak amplitude), or 271.6-275.5 nm (peak-to-peak amplitude), respectively. Calibration graphs were established for 10.0-42.0 microg ml(-1) nefazodone hydrochloride, or 8.0-46.0 microg ml(-1) sertraline hydrochloride. In the other method, HPLC, the UV detection was carried out at 265.0 nm (nefazodone hydrochloride) and 270.0 nm (sertraline hydrochloride). The samples were chromatographed on a Supercosil RP-18 column. The mobile phases were methanol:acetonitrile:phosphate buffer at pH 5.5 (10:50:40 v/v/v) (nefazodone hydrochloride) and methanol:phosphate buffer at pH 4.5 (20:80 v/v) (sertraline hydrochloride). The results obtained from first derivative spectrophotometric method were comparable with those obtained by using HPLC. It was concluded that both the developed methods are equally accurate, sensitive, and precision could be applied directly and easily to the pharmaceutical formulations of nefazodone hydrochloride and sertraline hydrochloride, respectively.

Antidepressive Agents↗
PubMed2005

Determination of Fe(II) and Zn(II) by spectrophotometry, atomic absorption spectrometry and ions chromatography methods in Vitrum.

Metrian [3-mercapto-5-(2'-hydroxynaphtylazo-1')-1,2,4-triazole]; Metriap [3-mercapto-5-(3',4'-dihydroxyphenylazo-1')-1,2,4-triazole]; Metriarez-gamma [3-mercapto-5-(2',4'-dihydroxy-3'-caroboxyphenylazo-1')-1,2,4-triazole] and Metidarez-beta [2-mercapto-5-(2',4'-dihydroxy-5'-carboxyphenylazo-1')-1,3,4-thiadiazole]-reagents were synthesized on Faculty of Pharmacy, Medical University of Lublin and used for the determination by different analytical methods of milligram quantities of Fe(II) and Zn(II) occurring together in pharmaceutical preparations, both multivitamin preparations and those containing microelements. The determination results of classical spectrophotometry (D0), spectrophotometry of derivatives (D1 and D2), atomic absorption spectrometric (AAS) and ions chromatography (IC) were analyzed statistically and compared with declared amount. The advantages of the proposed method of Fe(II) and Zn(II) determination include it's excellent precision and reproducibility of results.

Chemistry, Pharmaceutical↗
PubMed2005

Ratio spectra derivative spectrophotometry for the determination of furosemide and spironolactone in a capsule formulation.

The determination of furosemide and spironolactone in a capsule formulation has been investigated using techniques such as Vierordt's method and derivative spectroscopy dA/d lambda and d2A/d lambda2 applying the zero-crossing technique following reported methods. In our hands, using standard mixtures, these methods gave unreliable results. We have therefore investigated the use of ratio spectra derivative spectrophotometry for this determination. The technique of ratio spectra derivative spectrophotometry was developed in 1990, and has recently been used for a number of analyses of co-formulated products. The method was applied to the analysis of standard mixtures of the two drugs and the combined contents of 20 capsules resulting in values (mean +/- standard deviation) of 102.1 +/- 1.9% and 101.4 +/- 4.0% of the stated content for furosemide and spironolactone, respectively. Similarly, the analysis of individual capsules resulted in values of 101.5 +/- 1.6% and 102.2 +/- 1.4% of the stated content for furosemide and spironolactone, respectively.

Calibration↗
PubMed2005

Free flap monitoring using simultaneous non-invasive laser Doppler flowmetry and tissue spectrophotometry.

BACKGROUND: Early identification of flap failure is an indispensable prerequisite for flap salvage. Although many methods of free flap monitoring are available, there is still no single reliable non-invasive technique for early recognition of flap failure and for differentiation between arterial occlusion and venous congestion. The aim of this study was to investigate the benefits of the tissue oxygen analysis system O(2)C for monitoring patients undergoing maxillofacial reconstruction with fasciocutaneous radial forearm flaps. MATERIAL AND METHODS: In a prospective clinical study the microcirculatory parameters of blood flow, flow velocity, haemoglobin concentration (AU, Arbitrary Units) and oxygen saturation (%) were assessed by clinical means, by laser Doppler flowmetry and tissue spectrophotometry in 61 patients intraoperatively. Measurements were carried out before flap harvest, in the separated radial forearm flap, immediately after anastomoses and up to 14 days after reconstruction. RESULTS: Following anastomosis, blood flow and flow velocity exceeded the level before flap elevation and reached significant differences by the third postoperative day (p<0.05). Oxygen saturation decreased significantly by the third postoperative day and haemoglobin oxygenation showed stable values after performing anastomosis. Simultaneous, non-invasive laser-Doppler flowmetry and tissue spectrophotometry detected vascular complications in all cases with no false positive or false negative results and prior to clinical assessment. CONCLUSION: For the first time this new device allows reliable prediction of venous congestion by an increase of haemoglobin-concentration, and of arterial occlusion by a decrease in blood flow parameters and oxygen saturation. It can thus differentiate the mechanisms of flap failure before clinical assessment.

Adolescent↗
PubMed2006

Comparison of FT-NIR transmission and UV-vis spectrophotometry to follow the mixing kinetics and to assay low-dose tablets containing riboflavin.

For several years, near-infrared spectroscopy (NIRS) has become an analytical technique of great interest for the pharmaceutical industry, particularly for the non-destructive analysis of dosage forms. The goal of this study is to show the capacity of this new technique to assay the active ingredient in low-dosage tablets. NIR spectroscopy is a rapid, non-destructive technique and does not need any sample preparation. As an example, a binary mixture of microcrystalline cellulose and riboflavin was used to prepare tablets of different weights by direct compression. A prediction model was built by using a partial least square regression fit method. The NIR assay was performed by transmission. The results obtained by NIR spectroscopy were compared with a conventional UV-vis spectrophotometry method. The study showed that tablets can be individually analysed by NIR with high accuracy. It was shown that the variability of this new technique is less important than that of the conventional method which is the UV-vis spectrophotometry.

Kinetics↗
PubMed1985

Oxygen supply to the liver in patients with alcoholic liver disease assessed by organ-reflectance spectrophotometry.

In the present study we have investigated hepatic hemodynamics in patients with alcoholic liver disease using reflectance spectrophotometry and the hydrogen clearance method. Analysis of 38 cases has shown that estimated regional hepatic-tissue hemoglobin concentration, expressed as a difference in absorbance between 569 and 650 nm (delta Er569-650), decreased significantly with progress of fibrosis or fat accumulation in the liver. This suggests that the relative compression of the vascular compartment is due to the progress of alcoholic liver disease. Estimated hepatic hemoglobin concentration also correlated positively with prothrombin time, and negatively with serum gamma-globulin level and 15-min retention rate of indocyanine green. The difference in absorbance between 569 and 650 nm obtained by reflectance spectrophotometry was positively correlated with the regional hepatic blood flow as measured by the hydrogen clearance method. Thus, it is concluded that the estimated regional hepatic-tissue hemoglobin concentration decreases with progress of fibrosis and fat accumulation in the liver, and that this decreased oxygen supply to the liver may have an important role in the progress of alcoholic liver disease.

Absorption↗
PubMed1986

Quantification of cerebral oxygenation and haemodynamics in sick newborn infants by near infrared spectrophotometry.

New apparatus was made whereby indices of cerebral oxygenation and haemodynamics in sick newborn infants could be quantified by near infrared (NIR) spectrophotometry and displayed instantaneously at the cotside. The indices included oxygenated haemoglobin, reduced haemoglobin, oxidised cytochrome aa3, and total haemoglobin concentration: cerebral blood volume, mixed cerebral venous saturation, and changes in cerebral blood flow were then derived. Striking changes were observed in response to alterations in arterial oxygen saturation and carbon dioxide tension and to tilting of the infant. Abnormal responses were detected in cerebral oedema following birth asphyxia, patent ductus arteriosus, and cystic encephalomalacia. NIR spectrophotometry provides valuable quantitative data at the cotside for the management of sick infants and for exploring the pathophysiology of damage to the brain.

Blood Volume↗
PubMed2002

Simultaneous determination of amiloride and furosemide in pharmaceutical formulations by first digital derivative spectrophotometry.

This work presents a simple and fast method for the simultaneous determination of amiloride and furosemide by digital derivative spectrophotometry. HCl 1 x 10(-2) mol/l dissolved in ethanol was used as solvent and to extract drugs from formulations. Subsequently the samples were evaluated directly by first digital derivative spectrophotometry, using a smoothing factor of 8 and scale factor of 1 x 10(-4). The simultaneous determination of furosemide and amiloride can be carried out at 241.4 and 343.6 nm, respectively. In both cases, the zero crossing approach was used. When both compounds are present together in a sample, it is possible to quantify one in the presence of the other, without mutual interference. The determination range was found to be of 6.9 x 10(-8) to 16 x 10(-5) and 6.8 x 10(-8) to 8 x 10(-5) mol/l, for amiloride and furosemide, respectively. A good level of repeatability (RSD) of 0.9 and 0.6% was observed for amiloride and furosemide, respectively. The ingredients commonly found in commercial pharmaceutical formulations do not interfere. The proposed method was applied to the determination of these drugs in pharmaceutical formulations.

Amiloride↗
PubMed2002

Determination of zafirlukast by stability indicating LC and derivative spectrophotometry.

Two analytical methods have been developed for the determination of zafirlukast, a novel anti-asthmatic drug: high performance liquid chromatography (HPLC) and derivative spectrophotometry (DS). HPLC with ultraviolet detection at 225 nm is carried out with a Symmetry Shield RP18 column and a mobile phase constituted of acetonitrile and 0.01 M potassium dihydrogen phosphate buffer, adjusted the pH to 3.5 with 0.1 M KOH. The LC method is simple, rapid, selective and stability indicating. Indole was used as internal standard for the purpose of quantification of zafirlukast in HPLC. Spectrophotometry uses the third order derivative of the UV spectrum at 251.1 nm (deltalambda value 2.1 nm) for determination. Both methods were fully validated and a comparison was made. The results confirm that the methods are highly suitable for its intended purpose.

Chromatography, High Pressure Liquid↗