Sylwester Mazurek

Quantitative determination of acetylsalicylic acid and acetaminophen in tablets by FT-Raman spectroscopyElectronic Supplementary Information available. See http://www.rsc.org/suppdata/an/b1/b108240j/

A procedure for quantitative determination of acetylsalicylic acid and acetaminophen in pharmaceuticals by PLS (partial least squares) and PCR (principal component regression) treatment of FT (Fourier transform)-Raman spectroscopic data is proposed. The proposed method was tested on powdered samples. Three chemometric models were built: the first, for samples consisting of an active substance diluted by lactose, starch and talc; the second, in which a simple inorganic salt was applied as an internal standard and additions were not taken into account; and the third, in which a model was constructed for a commercial pharmaceutical, where all constituents of the tablet were known. By utilising selected spectral ranges and by changing the chemometric conditions it is possible to carry out fast and precise analysis of the active component content in medicines on the basis of the simplified chemometric models. The proposed method was tested on five commercial tablets. The results were compared with data obtained by intensity ratio and pharmacopoeial methods. To appraise the quality of the models, the relative standard error of predictions (RSEPs) were calculated for calibration and prediction data sets. These were 0.7-2.0% and 0.8-2.3%, respectively, for the different PLS models. Application of these models to the Raman spectra of commercial tablets containing acetylsalicylic acid gave RSEP values of 1.3-2.0% and a mean accuracy of 1.2-1.7% with a standard deviation of 0.6-1.2%.

Quantitative determination of diclofenac sodium in solid dosage forms by FT-Raman spectroscopy.

The FT-Raman quantification of diclofenac sodium in tablets and capsules was performed with the help of the partial least squares (PLS), principal component regression (PCR) and counter-propagation artificial neural networks (CP-ANN) methods. For the analysed tablets, calibration models were built using unnormalised spectra and spectra normalised by the intensity of a selected band of an internal standard. Different pre-processing methods were applied for the capsules. To compare the predictive ability of the models, the relative standard errors of prediction (RSEP) were calculated. The 5 x 5 CP-ANN and PLS methods gave models of comparable quality, which were usually more efficient than the PCR ones. The RSEP error values for the tablets were in the range of 2.4-3.8% for the calibration and 2.6-3.5% for the validation data sets and for the three procedures applied. For capsules, the RSEP errors were in the range of 0.8-1.9% and 1.4-1.7% for the calibration and validation samples, respectively. Five commercial products containing 25, 50 or 75 mg of diclofenac sodium per tablet/capsule were quantified. Concentrations found from the Raman data analysis agree with the results of the reference analysis and correlate strongly with the declared values with the recovery of 99.5-101.3%, 99.7-102.0% and 99.9-101.2% for the PLS, PCR and CP-ANN methods, respectively. The proposed procedure can be a fast and convenient alternative to the standard pharmacopoeial methods of diclofenac sodium quantification in solid dosage forms.

Quantification of atorvastatin calcium in tablets by FT-Raman spectroscopy.

The FT-Raman quantification of atorvastatin calcium in tablets was performed using the partial least squares (PLS), principal component regression (PCR) and counter-propagation artificial neural networks (CP-ANN) methods. To compare the predictive abilities of the elaborated models, the relative standard errors of prediction (RSEP) were calculated. The application of PLS, PCR and 6x6 CP-ANN provided models of comparable quality. RSEP error values in the range of 1.9-2.8% for calibration and validation data sets were obtained for the three procedures applied. Four commercial products containing 10, 20 or 40 mg of atorvastatin calcium per tablet were successfully quantified. Concentrations found from the Raman data analysis correlate strongly with the declared values, with a recovery of 98.5-101.3%, and with the results of reference analysis, with the recovery of 98.9-102.1%, for the different models. The proposed procedure can be a fast, precise and convenient method of atorvastatin calcium quantification in commercial tablets.

Transmission Fourier transform infrared microspectroscopy allows simultaneous assessment of cutin and cell-wall polysaccharides of Arabidopsis petals.

A procedure for the simultaneous analysis of cell-wall polysaccharides, amides and aliphatic polyesters by transmission Fourier transform infrared microspectroscopy (FTIR) has been established for Arabidopsis petals. The combination of FTIR imaging with spectra derivatization revealed that petals, in contrast to other organs, have a characteristic chemical zoning with high amount of aliphatic compounds and esters in the lamina and of polysaccharides in the stalk of the petal. The hinge region of petals was particular rich in amides as well as in vibrations potentially associated with hemicellulose. In addition, a number of other distribution patterns have been identified. Analyses of mutants in cutin deposition confirmed that vibrations of aliphatic compounds and esters present in the lamina were largely associated with the cuticular polyester. Calculation of spectrotypes, including the standard deviation of intensities, allowed detailed comparison of the spectral features of various mutants. The spectrotypes not only revealed differences in the amount of polyesters in cutin mutants, but also changes in other compound classes. For example, in addition to the expected strong deficiencies in polyester content, the long-chain acyl CoA synthase 2 mutant showed increased intensities of vibrations in a wavelength range that is typical for polysaccharides. Identical spectral features were observed in quasimodo2, a cell-wall mutant of Arabidopsis with a defect in pectin formation that exhibits increased cellulose synthase activity. FTIR thus proved to be a convenient method for the identification and characterization of mutants affected in the deposition of cutin in petals.

The ABCG transporter PEC1/ABCG32 is required for the formation of the developing leaf cuticle in Arabidopsis

The cuticle is an essential diffusion barrier on aerial surfaces of land plants whose structural component is the polyester cutin. The PERMEABLE CUTICLE1/ABCG32 (PEC1) transporter is involved in plant cuticle formation in Arabidopsis. The gpat6 pec1 and gpat4 gapt8 pec1 double and triple mutants are characterized. Their PEC1-specific contributions to aliphatic cutin composition and cuticle formation during plant development are revealed by gas chromatography/mass spectrometry and Fourier-transform infrared spectroscopy. The composition of cutin changes during rosette leaf expansion in Arabidopsis. C16:0 monomers are in higher abundance in expanding than in fully expanded leaves. The atypical cutin monomer C18:2 dicarboxylic acid is more prominent in fully expanded leaves. Findings point to differences in the regulation of several pathways of cutin precursor synthesis. PEC1 plays an essential role during expansion of the rosette leaf cuticle. The reduction of C16 monomers in the pec1 mutant during leaf expansion is unlikely to cause permeability of the leaf cuticle because the gpat6 mutant with even fewer C16:0 monomers forms a functional rosette leaf cuticle at all stages of development. PEC1/ABCG32 transport activity affects cutin composition and cuticle structure in a specific and non-redundant fashion.

Analysis of milk by FT-Raman spectroscopy

Fat, protein, carbohydrates and dry matter were quantified in commercial bovine milk samples, with the relative standard errors of prediction (RSEP) in the 3.4-6.1% range, using the partial least squares (PLS) method based on Raman spectra of liquid milk samples. Results of a better quality were obtained from a PLS model derived from IR spectra registered using single reflection ATR diamond accessory, which yielded RSEP values of 2.4-4.4%. The data indicated IR single reflection ATR spectroscopy and Raman spectroscopy in combination with multivariate modelling using the PLS method, allowed for the reliable, simultaneous quantitative determination of macronutrients in milk. The low signal to noise ratio of Raman spectra affects the quality of fat quantification especially for strongly defatted milk samples.

Microheterogeneity in binary mixtures of methanol with aliphatic alcohols: ATR-IR/NIR spectroscopic, chemometrics and DFT studies

This paper reports ATR-IR and NIR spectroscopic studies on microheterogeneity in binary mixtures of methanol with five short chain aliphatic alcohols: ethanol, 1-propanol, 2-propanol, tert-butanol and cyclopentanol. The inhomogeneity distribution of molecules in the mixtures was characterized by MIR/NIR excess absorption spectra, two-dimensional correlation approach, and chemometric methods. The experimental data were compared and discussed with the theoretical calculations of homo- and heteroclusters of different size and composition. The results obtained in the present work allow us to infer that all studied mixtures deviate from the ideal and the largest effect appears at equimolar mixture. At this composition about 50% of molecules are involved in the mixed clusters. In the entire range of compositions in the mixtures coexist the clusters of pure alcohols and heteroclusters with an average mole ratio of 1 : 1. Only in methanol/tert-butanol mixture was observed an additional tert-butanol-rich cluster with an average methanol : tert-butanol mole ratio between 1 : 3 and 1 : 4. The excess absorption and two-dimensional correlation spectra reveals that methanol/ethanol mixture is the closest to the ideal one, while the largest degree of non-ideality was observed for methanol/tert-butanol mixture. The extent of deviation from the ideal mixture increases with an increase in the chain length and the order of the alcohol. These trends are correlated with decreasing degree of self-association of the alcohol.

Connecting the Molecular Structure of Cutin to Ultrastructure and Physical Properties of the Cuticle in Petals of Arabidopsis

GPAT6 and DCR play different roles in structuring the cell wall-cuticle continuum. The plant cuticle is laid down at the cell wall surface of epidermal cells in a wide variety of structures, but the functional significance of this architectural diversity is not yet understood. Here, the structure-function relationship of the petal cuticle of Arabidopsis (Arabidopsis thaliana) was investigated. Applying Fourier transform infrared microspectroscopy, the cutin mutants long-chain acyl-coenzyme A synthetase2 (lacs2), permeable cuticle1 (pec1), cyp77a6, glycerol-3-phosphate acyltransferase6 (gpat6), and defective in cuticular ridges (dcr) were grouped in three separate classes based on quantitative differences in the ν(C=O) and ν(C-H) band vibrations. These were associated mainly with the quantity of 10,16-dihydroxy hexadecanoic acid, a monomer of the cuticle polyester, cutin. These spectral features were linked to three different types of cuticle organization: a normal cuticle with nanoridges (lacs2 and pec1 mutants); a broad translucent cuticle (cyp77a6 and dcr mutants); and an electron-opaque multilayered cuticle (gpat6 mutant). The latter two types did not have typical nanoridges. Transmission electron microscopy revealed considerable variations in cuticle thickness in the dcr mutant. Different double mutant combinations showed that a low amount of C16 monomers in cutin leads to the appearance of an electron-translucent layer adjacent to the cuticle proper, which is independent of DCR action. We concluded that DCR is not only essential for incorporating 10,16-dihydroxy C16:0 into cutin but also plays a crucial role in the organization of the cuticle, independent of cutin composition. Further characterization of the mutant petals suggested that nanoridge formation and conical cell shape may contribute to the reduction of physical adhesion forces between petals and other floral organs during floral development.

Quantification of active ingredients in suppositories by FT-Raman spectroscopy.

An efficient method for the quantitative determination of acetaminophen (AAP) and diclofenac sodium (DS) in commercial suppositories based on partial least squares (PLS) treatment of FT-Raman spectra is described. The relative standard errors of prediction (RSEP) were calculated for calibration and validation data sets to evaluate the quality of the constructed models. In the case of DS determination, RSEP error values of 1.9 % and 2.3 % for the calibration and validation data sets, respectively, were found. For AAP these errors amounted to 1.6-2.3 % and 1.8-2.8 %, respectively, for the different calibration models. Four commercial preparations containing 5, 12.5, 16.7 and 33.3 % (w/w) AAP and one containing 5 % (w/w) DS were successfully quantified using the developed models. Concentrations derived from the developed models correlated strongly with the declared values and yielded recoveries of 99.4-100.2 % and 99.6 % for AAP and DS, respectively. The proposed procedure can be used as a fast, economic and reliable method for quantification of the active pharmaceutical ingredients in suppositories.

Microheterogeneity in binary mixtures of aliphatic alcohols and alkanes: ATR-IR/NIR spectroscopic and chemometric studies

Aliphatic alcohols form homogeneous mixtures with alkanes of similar size and structure, however at a molecular level one can expect the presence of both the homo and heteroclusters leading to the local heterogeneity. Recently, we observed this phenomenon in the binary mixtures of methanol with aliphatic alcohols [RSC Adv., 2016, 6, 37195]. This paper provides new and comprehensive information on the structure of alcohol/alkane mixtures at a molecular level. Besides, we studied the relationship between the chain structure and the deviation from the ideal mixture. A particular attention was paid for the difference between the linear and cyclic alcohols and alkanes. For studies we selected two alcohols: 1-hexanol, cyclohexanol, and two alkanes: n-hexane, cyclohexane. By combining these two pairs of compounds, we obtained four different alcohol/alkane mixtures. The inhomogeneity distribution of molecules in these mixtures and deviation from the ideality was characterized by ATR-IR/NIR excess absorption spectra and chemometric methods. Obtained results allow us to conclude that the separation at a molecular level and the extent of deviation from the ideality depends on the degree of association of the alcohol and the similarity of alkyl parts. As 1-hexanol is more associated than cyclohexanol, 1-hexanol/n-hexane mixture is the closest to the ideal mixture. In contrast, cyclohexanol/n-hexane is the most non-ideal mixture since in this case the smaller degree of self-association of cyclohexanol is coupled with different structure of alcohol and alkane chains.