Stopped-flow fluorimetric determination of amoxycillin and clavulanic acid by partial least-squares multivariate calibration

Mixtures of amoxycillin and clavulanic acid were determined by using kinetic data in combination with partial least-squares multivariate calibration. The reaction of oxidation of these compounds with cerium(IV) in sulfuric acid medium has been monitored fluorimetrically. To follow the kinetics of the reaction, the stopped-flow mixing technique was used. Partial least-squares calibration of the kinetic data allowed the resolution of the analytes investigated in the concentration ranges between 0 and 4 mugml(-1). The method was applied satisfactorily to several pharmaceutical formulations, including Clavucid, Augmentine, Pangamox, Eupeclanic and Clamoxyl. The results obtained were validated by using an HPLC method. The percentages of recovery range from 91 to 105% for amoxycillin and from 78 to 117% for clavulanic acid, respectively.     

Simultaneous determination of acetaminophen and caffeine in tablet preparations by partial least-squares multivariate spectrophotometric calibration

The use of partial least-squares spectrophotometric calibration for the simultaneous determination of analgesic tablets in a multicomponent formulation is presented. This method is applied to the determination of acetaminophen and caffeine in tablet preparations. The results show that these components in a molar ratio of about 21:1 in tablets have been determined simultaneously with high precision.     

UV-spectrophotometric determination of beclomethasone dipropionate and phenylethyl alcohol in a nasal spray by inverse least-squares regression

Inverse least-squares (ILS) regression was used for the simultaneous UV spectrophotometric determination of the active principle (beclomethasone dipropionate) and a solvent (phenylethyl alcohol) in a pharmaceutical preparation commercially available in nasal spray form. A factorial design was used to establish the calibration equations, which enables the construction of calibration models using a minimum number of samples. The operating wavelengths were chosen by first choosing that coinciding with the absorption maximum for the analyte to be determined and then adding terms, one at a time, following the stepwise-forward procedure until no significantly improved S.E. is obtained. Single calibration equations were found for both constituents that provide satisfactory results in absorbance mode. The ILS procedure was applied to five industrial preparations with highly satisfactory results. In all, coefficients of variation values were less than 2%.     

Fast spectrophotometric determination of fluoride in ground waters by flow injection using partial least-squares calibration

The presence of sulphate constitutes a serious interference in the usual zirconium lake-based spectrophotometric method for the determination of fluoride in water. In this report, full spectral data have been recorded for the zirconium lake of 2-(parasulfophenylazo)-1,8-dihydroxy-3,6-naphthalene-disulfonate (SPADNS) in the simultaneous presence of fluoride and sulphate, as obtained with a flow injection system with a diode-array detector. The information has been processed with partial least-squares (PLS) multivariate calibration. Adequate modeling using a sixteen-sample calibration set allows fluoride to be determined in ground waters by the automated flow injection method, even in the presence of sulphate in concentrations up to 1000 mg l⁻¹. In the calibration range 0-1.50 mg l⁻¹ for fluoride, the limit of detection is 0.1 mg l⁻¹. The fluoride contents in real samples, as determined with the present method, were satisfactorily compared with those provided by ion selective potentiometry.     

Kinetic spectrophotometric determination of hydrocortisone acetate in a pharmaceutical preparation by use of partial least-squares regression

A kinetic spectrophotometric method for the determination of hydrocortisone acetate based on its condensation with isonicotinic acid hydrazide is proposed. The method is applied to the determination of hydrocortisone acetate in a commercially available pharmaceutical preparation, presented as a pomade, that also contains another corticosteroid and additional active compounds. The operating procedure involves dissolving the pomade in chloroform and the addition of the reagent solution directly to the cuvette, in this way avoiding the previous extraction of analytes from the insoluble pomade matrix required by the alternative HPLC procedure. Calibration is performed by partial least-squares regression, using absorbance or first derivative spectra values recorded each minute during the first 30 min of reaction. Use of first derivative spectra overcomes possible scattered light problems produced by excipients precipitating, and produced slightly better results than absorbance data. The relative standard deviation obtained for 11 replicates analysed on different days was approx. 1.5%. The proposed method improves both accuracy and precision of the classical initial rate method and the precision of the HPLC procedure.     

Simultaneous determination of five different food proteins by high-performance liquid chromatography and partial least-squares multivariate calibration

A method is described for the simultaneous determination of food proteins originating from different raw food materials. The proteins are hydrolysed to amino acids which are labelled with dansyl chloride and finally separated by reversed-phase high-performance liquid chromatography. Partial least-squares multivariate calibration is used to resolve and quantify the overlapping amino-acid patterns. The method enables muscle protein, collagen, soy protein (both texturate and isolate), casein and milk protein to be quantified in both heated and raw samples from the same calibration set. The accuracies for the raw and heated samples averaged 3% and 6% relative total protein content, respectively.     

Synchronous fluorimetric determination of salicylic acid and diflunisal in human serum using partial least-squares calibration

The simultaneous determination of salicylic acid and diflunisal in human serum has been accomplished by synchronous fluorimetry, in combination with partial least-squares multivariate calibration. The total luminescence information of the analytes has been used to optimize the spectral data set for the calibration, by analysis of the three-dimensional excitation-emission matrices. The synchronous spectrum, maintaining a constant difference of Deltalambda = 128 nm between the emission and excitation wavelengths, has been selected as optimum to perform the determination. The method is based on the fluorescence of these compounds in chloroform containing 1% (v/v) acetic acid. Serum samples are treated with trichloroacetic acid to remove the proteins, and both analytes are extracted into chloroform-1% (v/v) acetic acid prior to the determination. For concentrations ranging from 60-240 mug ml(-1) of each drug, analytical recoveries range from 96% to 103% for salicylic acid and from 97% to 105% for diflunisal.     

Solid-phase spectrofluorimetric determination of acetylsalicylic acid and caffeine in pharmaceutical preparations using partial least-squares multivariate calibration

PLS-1, a variant of the partial least-squares algorithm was used for the solid-phase spectrofluorimetric determination of acetylsalicylic acid (ASA) and caffeine (CF) in pharmaceutical formulations. The method allows the simultaneous quantification of the analytes, as the closely overlapping spectral bands are efficiently solved. Sample preparation prior to analysis is not required. The calibration set consisted of 83 samples with 50-170 mg g⁻¹ ASA plus 5-20 mg g⁻¹ CF; another set of 25 samples was used for external validation. Agreement between predicted and experimental concentrations was fair (r = 0.987 and 0.974 for ASA and CF models). For both models, the prediction performance was evaluated in terms of the coefficient of variability (CV), relative predictive determination (RPD), and ratio error range (RER). The final PLS-1 models were used for the determination of ASA and CF in pharmaceutical formulations.     

A flow-through fluorimetric sensing device for determination of alpha- and beta-naphthol mixtures using a partial least-squares multivariate calibration approach

A single flow-through optosensor spectrofluorimetric system is proposed for the resolution of mixtures of alpha- and beta-naphthol at mug l(-1) levels using a partial least-squares (PLS) calibration approach. The sensor was developed in conjunction with a monochannel flow-injection analysis system with fluorimetric detection using Sephadex QAE A-25 resin as an active sorbent substrate in the flow cell and the second derivative of the native synchronous fluorescence spectra of analytes as analytical signal. In the manifold, the solutions of naphthol (at pH 10.0) were injected in a carrier stream of KCl (0.15 M)/NaOH (10(-2) M). Because of the strong spectral overlap, the mixture could not be resolved by conventional spectrofluorimetry. The non-additive behaviour of the fluorescence signals revealed an interaction in the system, which was not found by working in the solution only (without the sorbent support). This interaction, probably due to the environment of the analytes on the solid phase, made impossible their simultaneous determination. So, the use of synchronous fluorescence spectroscopy or even its derivative signal could not resolve satisfactorily the mixture. The simultaneous determination of both naphthol has been carried out by recording the signal of the second-derivative synchronous fluorescence (Deltalambda=170 nm) spectra between 200 and 450 nm and a PLS multivariate calibration treatment. The optimum number of factors was selected by using the cross-validation method. After validating the proposed method, it was applied to the determination of these compounds in natural waters with different amounts of each chemical.     

Simultaneous determination of naproxen, salicylic acid and acetylsalicylic acid by spectrofluorimetry using partial least-squares (PLS) multivariate calibration

Determination of naproxen, salicylic acid and acetylsalicylic acid has been carried out in mixtures of up to three components by recording emission fluorescence spectra between 300 and 520 nm with an excitation wavelength of 290 nm. The excitation-emission spectra of these compounds are strongly overlapped, which does not permit their direct determination without previous separation by conventional methodologies. Here, a method is proposed for the determination of these chemicals by the use of a full-spectrum multivariate calibration method, partial least-squares (PLS). The experimental calibration matrix was designed with 18 samples. The concentrations were varied between 0.1 and 1.0 mug ml(-1) for naproxen, 0.5 and 5.0 mug ml(-1) for salicylic acid and from 2.0 to 12.0 mug ml(-1) for acetylsalicylic acid. The cross-validation method was used to select the number of factors. To check the accuracy of the proposed method, the optimized model, obtained using PLS-1, was applied to the determination of these compounds in pharmaceuticals and human serum samples previously spiked with different amounts of each chemical.     

Application of partial least-squares calibration to phosphorimetric data for determination of polycyclic aromatic hydrocarbons in spiked environmental samples

A partial least-squares calibration method is proposed, for the first time, for phosphorescence signals. The proposed method is based on the determination of phenanthrene, fluoranthene, and benz[a]anthracene by room temperature phosphorimetry, using microemulsion solutions. The emission and first-derivative emission spectra of the ternary mixtures were tested to perform the calibration matrix. Improved recoveries were found for the prior differentiation step in the analysis of ternary mixtures of these polycyclic aromatic hydrocarbons in road dust samples. The proposed method yielded recoveries ranging from 93.2 to 115.3%, with relative standard deviations of < 6.8%.     

Simultaneous spectrophotometric determination of Fe(III), Al(III) and Cu(II) by partial least-squares calibration method

A spectrophotometric method for simultaneous determination of Fe(III), Al(III) and Cu(II) using Alizarin Red S as a chelating agent was developed. The parameters controlling behavior of the system were investigated and optimum conditions were selected. A partial least-squares multivariate calibration method was used for the analysis of ternary mixtures of Fe(III), Al(III) and Cu(II) over the range of 450-6000, 140-4000 and 450-15000 ng ml(-1), respectively. Absorbance data were taken between 400 and 800 nm. Applying this method to simultaneous determination of these metal ions in several synthetic alloy solutions with total relative standard error of less than 5% validated the proposed method.     

Comparison of UV derivative-spectrophotometry and partial least-squares (PLS-1) calibration for determination of methotrexate and leucovorin in biological fluids

Binary mixtures of methotrexate (MTX) and leucovorin (LV) have been resolved by application of first-derivative spectrophotometry and partial least squares calibration (PLS-1). By measuring the first-derivative signals of MTX and LV at 354 and 300 nm, respectively, simultaneous determination was possible. The mean recoveries for urine samples were 91 and 96% for MTX and LV, respectively. Partial least squares (PLS-1) multivariate calibration has been applied to the determination of these compounds in serum and in urine without pretreatment of the samples. The absorption spectra of serum or urine samples spiked with methotrexate and/or leucovorin, were used to optimize the calibration matrixes by the PLS-1 method. The sensitivity and selectivity of the proposed procedures were calculated. Mean recoveries were 101 and 97% for MTX and LV, respectively, for serum samples, and 101 and 98% for MTX and LV, respectively, for urine samples.     

Simultaneous spectrophotometric determination of drugs in pharmaceutical preparations using multiple linear regression and partial least-squares regression, calibration and prediction methods

Two new methods for the simultaneous determination of acetylsalicylic acid, acetaminophen and caffeine based on total absorbance measurements and their processing by multiple linear regession and partial least-squares regression are proposed. The concentration ranges used to construct the calibration matrix were 4.0-12.0, 2.0-10.0 and 0.9-6.0 μg ml⁻¹ for acetylsalicylic acid, acetaminophen and caffeine respectively. The proposed methods were validated by using a set of synthetic sample mixtures and subsequently applied to the determination of the three active principles in three different pharmaceutical preparations.     

Minimum value assured by a method to determine gold in alloys by using laser-induced breakdown spectroscopy and partial least-squares calibration model

A procedure for estimating the minimum value assured by an analytical method has been developed. It is applied to the determination of gold in jewellery alloys by means of a recently proposed spectroscopy technique. The laser-induced breakdown spectroscopic data of 17 gold alloys, with gold concentration ranging between 50 and 100% were used as calibration set for carrying out the partial least-squares regression (PLS). Ninety alloys, with known gold concentration, were used to evaluate the method’s accuracy. Finally, the minimum guaranteed value of the gold content was analysed, taking into account the values for gold hallmark in Spanish regulations.     

UV partial least-squares calibration and liquid chromatographic methods for direct quantitation of levofloxacin in urine

Levofloxacin was determined in human urine samples by application of a spectrophotometric multivariate calibration partial least-squares (PLS-1) method. A calibration set consisting of standards was prepared by using a multilevel multifactor experimental design. In order to ensure accurate results, the calibration matrix included a urine sample free of levofloxacin (i.e., urine blank). The components of the calibration matrix were levofloxacin and urine. The concentration of levofloxacin ranged from 0.5 to 16.5 microg/mL. Different urine concentrations were used as the second component of the calibration matrix in order to include the information inherent in the changes in the UV spectrum for urine upon dilution. In addition, a high-performance liquid chromatographic method was proposed. In this method, a Shim-pack amino column was used at ambient temperature with a mobile phase of 25 mM potassium dihydrogen phosphate (pH adjusted to 3.1 with phosphoric acid)-acetonitrile (70 + 30, v/v), and the flow rate was 1 mL/min. UV detection at 293 nm was used for quantitation. The proposed methods were applied to the determination of the dissolution rate for tablets containing levofloxacin. The urinary excretion pattern for the cumulative amount of levoflacin excreted was also calculated.     

Kinetic-spectrophotometric determination of theophylline,dyphylline, and proxyphylline by use of partial least-squares regression

A kinetic-spectrophotometric method for the determination of theophylline, dyphylline and proxyphylline, based on their azo coupling reaction with the diazonium ion of sulfanilic acid after a treatment with alkali, is proposed. The absorbance is recorded from 340 to 600 nm every second during reaction for 90 s, and calibration is performed by partial least-squares regression, using first derivative spectra values. Mixtures containing 2.5-13 micro g mL(-1) dyphylline and proxyphylline, and 2-9 micro g mL(-1) theophylline were successfully resolved with root mean squared errors of prediction (RMSEP) of 0.4, 0.3, and 0.2 for dyphylline, proxyphylline, and theophylline, respectively. The proposed method was satisfactorily applied to the determination of the three compounds in a commercially available pharmaceutical preparation and provided results similar to those obtained by HPLC.     

Simultaneous kinetic spectrophotometric determination of spironolactone and canrenone in urine using partial least-squares regression

A method is proposed for the simultaneous determination of spironolactone and canrenone in urine based on the different rates at which they react with sulphuric acid to yield a trienone. Kinetic spectrophotometric data are processed by partial least-squares (PLS) regression. The optimum sulphuric acid concentration and temperature are determined from response surfaces, using PLS methodology to relate both variables to the relative square error of prediction (RSEP, the parameter to be minimized). The relative errors made in the quantitation of each diuretic by the proposed method are less than 5% and the overall error, as RSEP, ranges from 1.06 and 1.44%.     

Potentiality of partial least-squares multivariate calibration in the spectrophotometric analysis of binary mixtures of purine bases

Abstract  

A partial least-squares (PLS) modeling was developed for the simultaneous spectrophotometric determination of adenine (AD) and guanine (GU). The determination of these analytes is pharmacologically necessary. Multivariate calibration is used because of spectral overlapping. The calibration set contained AD and GU in the concentration range of 1.4–20.3 and 1.5–25.7 μg cm⁻³, respectively. The absorption spectra were recorded from 200 to 300 nm. The predicted residual error sum-of-squares for AD and GU was 0.0500 and 0.4000 for number of principal components 3 and 2, respectively. The root mean square error of prediction for AD and GU was 0.0913 and 0.2582, respectively. The limits of detection were 0.02 and 0.03 μg cm⁻³ for AD and GU, respectively. The proposed method allows the simultaneous determination of AD and GU in spiked real matrixes of human urine, serum, and plasma.