Spectrophotometric multicomponent analysis of a mixture of metamizol, acetaminophen and caffeine in pharmaceutical formulations by two chemometric techniques

Inverse least squares (ILS) and factor-based (principal component analysis (PCA)) techniques were proposed for the spectrophotometric multicomponent analysis of a ternary mixture consisting of metamizol, acetaminophen and caffeine, without prior separation. In these chemometric techniques, the measurements of the absorbance values were realized in the spectral range from 225 to 285 nm in the intervals of Deltalambda=5 nm at the 13 wavelengths in the zero-order spectra of the different ternary mixtures of these active ingredients in 0.1 M HCl. The prepared calibrations of both techniques using the absorbance data and concentration matrix data sets were used to predict the concentration of the unknown concentrations of metamizol acetaminophen and caffeine in their ternary mixture. The 'MAPLE V' software was used for the numerical calculations. Mean recoveries and relative standard deviations for ILS and PCA techniques were found to be 99.8 and 1.68%, 99.9 and 1.66% for caffeine, 99.8 and 1.84%, 100.4 and 2.85% for metamizol, and 99.7 and 1.04%, 99.6 and 1.34% for acetaminophen, respectively, for the first and second techniques. The techniques were successfully applied to two pharmaceutical formulations marketed in Turkey and results were compared with a new high-performance liquid chromatography method.     

Simultaneous determination of diazepam and pyridoxine in synthetic mixtures and pharmaceutical formulations using graphical and multivariate calibration-prediction methods

Several methods are reported for the simultaneous determination of diazepam and pyridoxine: two graphical methods (zero-crossing and derivative quotient spectra); and two numerical methods (multiple linear regression and partial least-squares regression). The methods have been applied in the concentration ranges 1.4–4.0 μg ml⁻¹ diazepam and 4.0–12.0 μg ml⁻¹ pyridoxine. The accuracy and precision of the methods have been determined and they have been validated by analysing synthetic mixtures containing the two drugs in variable proportions. The methods were also applied to the determination of diazepam and pyridoxine in pharmaceutical preparations. The analytical results were quite good in all cases.     

Simultaneous determination of trimethoprim and sulphamethoxazole in veterinary formulations by chromatographic multivariate methods

A comparative chromatographic study was developed for the simultaneous quantitative resolution of trimethoprim (TMP) and sulphamethoxazole (SMX) in veterinary formulations. Multi-wavelength chromatograms were recorded by using diode array detector (DAD) system at the five-wavelength set consisting of 220, 230, 240, 250 and 260 nm. In the first step, five different calibration equations at the above wavelengths for each drug were obtained by using the relationship between concentration and peak area. These calibration graphs were used for the quantitative evaluation of TMP and SMX in samples. These single-wavelength applications were called traditional LC method. In the second step, principal component regression (PCR) and partial least-squares (PLS) calibrations were applied to the above mentioned multi-wavelength chromatograms. The amount of two investigated drugs in samples was determined by the constructed PCR and PLS calibrations. The experimental results obtained from each single-wavelength calibration graph were compared with those obtained by the chemometric approaches and chromatographic multivariate approaches give successful results more than traditional LC method.     

Spectrophotometric monitoring of nimesulide photodegradation by a combined hard-soft multivariate curve resolution-alternative least square method

Nimesulide, a Cox-2 inhibitor anti-inflammatory drug, is a light sensitive compound and its biological activity is decreased upon photodegradation. The photodegradation kinetic of nimesulide was investigated spectrophotometrically using multivariate curve resolution analysis to overcome spectral overlapping of reactant and degradation products. The absorbance spectra of the nimesulide methanolic solution, exposed to daily sunlight, were recorded at different times. Three absorbing chemical species, coexisted in the reaction system, were detected by application of factor analysis to the absorbance data matrix. The soft-modeling multivariate curve resolution-alternative least square (MCR-ALS) analysis of the evolutionary absorbance data revealed that nimesulide undergoes photodegradation through a two-step consecutive manner, where both steps obey first-order kinetic. By application of the kinetic hard model constraint to the MCR-ALS analysis, an excellent agreement was obtained between the fitted concentration profiles and those obtained by soft method. The first-order rate constants of the first and second degradation products were calculated as 0.052 (+/-0.007) and 0.009 (+/-0.001)h(-1), respectively. Finally, the pure spectra of the resolved chemical species were calculated, where that of nimesulide was the same as that obtained experimentally.     

Spectrophotometric simultaneous determination of triamterene and hydrochlorothiazide in triamterene-H tablets by multivariate calibration methods

The multivariate calibration methods of partial least-square regression and principal component regression were applied for the simultaneous spectrophotometry determination of triamterene (TRM) and hydrochlorothiazide (HYD) in their mixtures. The parameters of the chemometric procedure were optimized, and the proposed methods were validated with synthetic samples and applied to analyze these drugs in pharmaceutical products with good accuracy and precision. The results were compared with those given by the British Pharmacopoeia (BP) method. The square of the correlation coefficients (R(2)) for predicted TRM and HYD with the proposed method in a test sample were 0.9994 and 0.9992, respectively. The relative standard deviation for commercial tablets in the proposed method and BP standard method were 0.405 and 2.142%, respectively.     

A case of an accidental exposure to a veterinary insecticide product formulation

A veterinary technician while opening a package was accidentally exposed to a commercial canned product formulation containing insecticides and solvents. The patient was twice briefly treated and released as an outpatient from 2 different hospitals on the first and second day after the exposure. However, on the fourth day, as some of the symptoms (headache, nausea, vomiting, diarrhea, difficult breathing) persisted, the patient was admitted to another hospital. The patient was treated for exposure to organophosphates and solvents and was released after 13 days. The patient developed diabetes insipidus, a condition which lasted for approximately 1 year. The cause of the temporary development of diabetes insipidus is not understood. There is a need to prevent and minimize such accidental exposures in future.     

Chemometric resolution of a mixture containing hydrochlorothiazide and amiloride by absorption and derivative spectrophotometry

Four chemometric techniques, classical least squares (CLS) and inverse least squares (ILS) and principal component regression (PCR) and partial least squares regression (PLSR) were applied to the absorption and derivative spectrophotometric determinations of amiloride and hydrochlorothiazide in a pharmaceutical preparation. Four chemometric calibrations for both zero-order and first derivative spectra were constructed by measuring the absorbance and their dA/dlambda values at 34 points in the wavelength range 205-395 nm for a training set containing 2-10 microg/ml amiloride and 4-28 microg/ml hydrochlorothiazide corresponding to 25 point mixture design. The building chemometric calibrations were confirmed by using the synthetic mixtures containing two drugs. The results obtained by the proposed techniques based on the use of the measurements at the absorption spectra and at the first derivative spectra were statistically compared with each other.     

Evaluation of different formulation studies on air-filled polymeric microcapsules by multivariate analysis

Air-filled polymeric microcapsules have been prepared by freeze-drying of emulsions containing the wall-forming polymer in the organic phase of oil in water emulsions. Echogenic air-filled microcapsules were prepared from emulsions containing either (-)-camphene, cyclohexane or cyclooctane as the solvent in the organic phase. Formulation studies have been performed to improve the yield and acoustic quality of the microcapsule suspensions. The yield was measured as particle concentration or efficacy, i.e. normalised attenuation at 3.5 MHz, related to the amount of polymer used.No overall conclusion could be made for all the variables when visually comparing the results from the different investigations. Multivariate analyses (PCA and PLS) were therefore necessary to be able to reveal any relevant systematic information from all the investigations. Different parameters describing the formulation, the production process and parameters describing the characterisation of the intermediates and the final product were set as independent X-variables.Three to four percent (w/v) of polymer was found to be the appropriate concentration of wall forming polymer. Including PEG 3000 resulted in improved freeze-dried product and suspension. Quenching of the emulsions by freezing in dry ice/methanol prior to freeze-drying was not necessary.Process parameters for homogenising and freeze-drying should be optimised with regard to the single systems, due to the different physico-chemical properties of the different solvents, especially melting point and vapour pressure.     

Optimization of a formulation containing viable lactic acid bacteria

In the present study, gastric juice resistant tablet formulations of lactic acid bacteria (LAB) were developed, using hydroxypropylmethylcellulose acetate succinate (HPMCAS) as well as alginates, apple pectin and Metolose as matrix forming components. To optimize the formulation-using survival rate in acid medium, and disintegration time in intestinal fluid as test parameters-tablets were modified with respect to LAB content, amount of applied excipients per tablet, and compaction forces. A decrease of viable cells of not more than one log unit after 2h of incubation in acid medium was desired, as well as a disintegration time of 1h in phosphate buffer pH 6.8. It was found that the amount of HPMCAS in the tablet correlates with gastric juice resistance. As HPMCAS also leads to a decrease of disintegration time in intestinal fluid, slight amounts of this excipient were preferred. The best protective qualities against artificial gastric juice were observed when tablets were prepared from compaction mixtures of LAB, HPMCAS and sodium alginate.     

Analysis of a formulation containing lincomycin and spectinomycin by liquid chromatography with pulsed electrochemical detection

A reversed phase ion-pair liquid chromatographic method using a base deactivated column and pulsed electrochemical detection on a gold electrode is described. It allows the separation of a mixture of spectinomycin sulfate, lincomycin hydrochloride and their related substances. A step gradient was necessary to obtain a good separation together with a reasonable analysis time of 40 min. The mobile phases consisted of an aqueous solution of 3.3 or 0.55 g/l pentanesulfonic acid, 10 mM acetic acid and 20 ml/l tetrahydrofuran. Both mobile phases were adjusted to pH 4.0 with diluted sodium hydroxide. The influence of the different chromatographic parameters on the separation was investigated. Two commercial samples were analyzed using the described method. In total 12 components could be separated.     

气相色谱/质谱和多维分辨法分析仙鹤草挥发性成分

目的研究不同产地仙鹤草挥发油成分。方法利用水蒸气蒸馏法提取仙鹤草挥发油,用气相色谱/质谱(GC-MS)检测。先用子窗口因子分析法(SFA)定性定量分析湖南产仙鹤草的挥发油成分,然后用光谱相关色谱法(SCC)提取浙江产仙鹤草挥发油中的共有成分。结果在湖南和浙江产仙鹤草挥发油中分别鉴定出68和65个化学成分,其中有51个共有成分,主要组分都为雪松醇、α-蒎烯、芳樟醇、1-(2-呋喃基)-己酮、α-松油醇、乙酸龙脑酯和桉油精等,但含量有一定差别。结论该方法不仅鉴定的化合物数目增加,而且提高了定性准确度,可用于不同产地仙鹤草的分析和质量控制。     

Simultaneous resolution of a binary mixture of captopril and hydrochlorothiazide in tablets by bivariate and multivariate spectral calibrations

The multivariate spectral calibration methods, two-linear regression-calibration (bivariate calibration (BC)) and multi-linear regression-calibration (MLRC) are proposed for the simultaneous resolution of a binary mixture of hydrochlorothiazide (HCT) and captopril (CTP), which have closely overlapping spectra. The BC and MLRC calibration algorithms are described for the two-component system, HCT-CTP. Some alternative methods, classical least squares (CLS), inverse least squares (ILS), principal component regression (PCR) and principal least squares (PLS) methods, were also used to determine HCT and CTP in the mixture. Using a synthetic mixture of the two drugs, all the methods were validated and applied to tablets. The BC and MLRC methods which are very rapid, and easy to apply, yet not expensive, are powerful tools with very simple mathematical contents for the quantitative analysis. Data treatment was performed using MAPLE V, EXCEL and SPSS 10.0 Software.     

Spectrophotometric analysis of the multicomponent system: dihydralazine sulfate, hydrochlorothiazide and reserpine

With respect to their performance a computer aided spectrometric analysis of multicomponent system dihydralazine sulfate hydrochlorothiazide and reserpine was made under various conditions. To find out the advantageous measuring points the difference spectra were determined. The influence of the number of measuring points, the single and mixture calibration and the dimensiation of calibration matrix to accuracy and precision was investigated and the limits of the determination for the three component system are discussed.     

Liquid chromatographic analysis of a formulation containing polymyxin,gramicidin and neomycin

The development of a liquid chromatographic assay system for the stability study of a formulation containing polymyxin, gramicidin and neomycin is described. For the determination of each group of antibiotics, poly(styrenedivinylbenzene) is used as the stationary phase. The mobile phase for the determination of polymyxin consists of an aqueous solution containing 7 g l⁻¹ of sodium sulfate, 50 ml l⁻¹ of 1 M phosphoric acid and 160 ml l⁻¹ of acetonitrile. UV detection is performed at 215 nm. Gramicidin is chromatographed with a mixture of tetrahydrofuran-water (38:62) and detected by UV at 222 nm. An aqueous solution containing 70 g l⁻¹ of sodium sulfate, 1.4 g l⁻¹ of sodium octanesulfonate and 50 ml l⁻¹ of 0.2 M phosphate buffer pH 3.0 is used as the mobile phase for the determination of neomycin. Since neomycin has no UV-absorbing chromophore, pulsed electrochemical detection is chosen to determine neomycin.For each method, the influence of the different chromatographic parameters on the separation, the selectivity towards the other active compounds and the excipients, the repeatability and the linearity were investigated. The stability of the formulation was examined at 0, 6 and 12 months.     

A study of the photo-degradation kinetics of nifedipine by multivariate curve resolution analysis

A multivariate curve resolution method based on the combination of Kubista approach and iterative target transformation method of Gemperline has been proposed. This method is a soft model and need no information about the spectrum of the product and mechanism of the reaction. The method was used to study the degradation kinetics of nifedipin, 1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl)-3,5-pyridine dicarboxilic acid dimethyl ester, upon exposure to the light of a 40 W tungsten lamp. The spectra of the nifedipine, collected at different lighting times, were subjected to the factor analysis and two chemical components were detected in the reaction system. Pure spectra of the components involved and their concentration profiles were obtained. The results revealed that the photodecomposition kinetics of nifedipine is zero-order at the beginning of the reaction. However, when the reaction preceded more than 50%, the kinetics of reaction changed to a first-order manner. The rate constants for the zero-order and first order regions were estimated as regions (4.96+/-0.13) x 10(-9) M(-1) s(-1) and (6.22+/-0.10) x 10(-5) s(-1), respectively.     

Drying of a plasmid containing formulation: chitosan as a protecting agent

Background

Along with research on development of more efficient gene delivery systems, it is necessary to search on stabilization processes to extend their active life span. Chitosan is a nontoxic, biocompatible and available gene delivery carrier. The aim of this study was to assess the ability of this polymer to preserve transfection efficiency during spray-drying and a modified freeze-drying process in the presence of commonly used excipients.

Methods

Molecular weight of chitosan was reduced by a chemical reaction and achieved low molecular weight chitosan (LMWC) was complexed with pDNA. Obtained nanocomplex suspensions were diluted by solutions of lactose and leucine, and these formulations were spray dried or freeze dried using a modified technique. Size, polydispersity index, zeta potential, intensity of supercoiled DNA band on gel electrophoresis, and transfection efficiency of reconstituted nanocomplexes were compared with freshly prepared ones.

Results and conclusion

Size distribution profiles of both freeze dried, and 13 out of 16 spray-dried nanocomplexes remained identical to freshly prepared ones. LMWC protected up to 100% of supercoiled structure of pDNA in both processes, although DNA degradation was higher in spray-drying of the nanocomplexes prepared with low N/P ratios. Both techniques preserved transfection efficiency similarly even in lower N/P ratios, where supercoiled DNA content of spray dried formulations was lower than freeze-dried ones. Leucine did not show a significant effect on properties of the processed nanocomplexes. It can be concluded that LMWC can protect DNA structure and transfection efficiency in both processes even in the presence of leucine.     

Spectrophotometric determination of vitamin B1 in a pharmaceutical formulation using triphenylmethane acid dyes

A highly sensitive colour reaction has been developed, based on the fact that vitamin B(1) reacted with a triphenylmethane acid dye such as thymol blue, bromothymol blue, bromophenol blue, bromocresol green, phenol red or cresol red to form an ion-association complex in a weak-base aqueous solution in the presence of some solubilization agents e.g. polyvinyl alcohol, emulgent OP, Triton X-100 or Tween-20. The wavelengths of maximum absorbance of the six ion-association complexes were between 420 and 450 nm, and fading reaction appeared at the longer wavelength and the maximum fading wavelengths were between 550 and 620 nm. The reactions had highly sensitivities and their apparent molar absorptivities of the colour reactions were (0.82-1.65) x 10(5) l mol(-1) cm(-1) and those of fading reactions were (1.26-3.92) x 10(5) l mol(-1) cm(-1) depending on the different dye systems. The composition ratio of the ion-association complex was vitamin B(1):dye = 1:1 as established by Job's and equilibrium shift methods. The method had good selectivity and could be applied to direct spectrophotometric determination of vitamin B(1) in aqueous phase without using organic solvent extraction. Therefore, the method was simple and rapid. The colour reaction mechanism was discussed with the quantum chemistry method.