Raman mapping for determination of TiO2 in different solid food samples by multivariate curve resolution with alternating least squares.

Titanium dioxide is a food additive commonly used as a white food coloring (E171). Its wide use by the food industry associated with the nanometric size distribution of the particles of this pigment has shown high genotoxicity associated with recurrent exposure by ingestion. Therefore, the use of E171 in food products has already been banned by some industries and in the European Union. Such banishment should soon be extended to other countries around the world, making it important to establish techniques for the efficient determination of TiO2 in different food products. The association between hyperspectral images and chemometric tools can be useful in this sense, aiming to enable the use of a single method for sample preparation and analysis of different types of food. Thus, the present work aims to evaluate the use of Raman mapping associated with the resolution of multivariate curves with alternating least squares (MCR-ALS) for the determination of titanium dioxide in solid food samples with different compositions, without the need to introduce specific sample preparation. The proposed method allowed for the first-time quantification of TiO2 in different food matrices without specific sample preparation, with a simple, rapid, accurate (93% of recovery), low detection limits (0.0111% m/m) and quantification (0.0370% m/m) and adequate linearity (r = 0.9990) and precise (standard deviation around 0.020-0.030% w/w) methodology. Such results highlight the potential use of Raman mapping associated with the MCR-ALS for quantification of the nano-TiO2 in commercial samples.

Design of experiments applied to stress testing of pharmaceutical products: A case study of Albendazole.

Forced degradation tests are studies used to assess the stability of active pharmaceutical ingredients (APIs) and their formulations. These tests are performed submitting the API under extreme conditions in order to know the main degradation products in a short period of time. The results of these studies are used to assess the degradation susceptibility of APIs and to validate chromatographic analytical methods. However, most of degradation studies are performed using one-factor-at-the-time (OFAT) which does not consider the interactions between degradation variables. This work proposes the use of Design of Experiment (DoE) approach in forced degradation of albendazole (ABZ). It was used a central composite design (CCD) to evaluate the forced degradation in a multivariate way. Experiments were performed taking into account the variables pH, temperature, oxidizing agent (H2O2) and UV radiation. It was verified the influence of the variables and their interactions on the ABZ degradation. The ABZ oxidation showed to be the main degradation route for ABZ, which is strongly influenced by the temperature. The hydrolysis was relevant at alkaline medium and high temperature. LC-IT-MSn was used to identify the degradation products. It was found three known degradation products (albendazole-2-amino, albendazole sulfoxide and albendazole sulfone) and a new derivate of albendazole molecule (albendazole sulfoxide with a chlorine). This last one was isolated and characterized by UPLC-QToF-MS and NMR analyses.

Determination of B-complex vitamins in pharmaceutical formulations by surface-enhanced Raman spectroscopy.

The aim of this work was to quantify B-complex vitamins in pharmaceutical samples by surface enhanced Raman spectroscopy technique using gold colloid substrate. Synthesis of gold nanoparticles was performed according to an adapted Turkevich method. Initial essays were able to suggest the orientation of molecules on gold nanoparticles surface. Central Composite design was performed to obtain the highest SERS signal for nicotinamide and riboflavin. The evaluated parameters in the experimental design were volume of AuNPs, concentration of vitamins and sodium chloride concentration. The best condition for nicotinamide was NaCl 2.3×10-3molL-1 and 700µL of AuNPs colloid and this same condition showed to be adequate to quantify thiamine. The experimental design for riboflavin shows the best condition at NaCl 1.15×10-2molL-1 and 2.8mL of AuNPs colloid. It was possible to quantify thiamine and nicotinamide in presence of others vitamins and excipients in two solid multivitamin formulations using the standard addition procedure. The standard addition curve presented a R2 higher than 0.96 for both nicotinamide and thiamine, at orders of magnitude 10-7 and 10-8molL-1, respectively. The nicotinamide content in a cosmetic gel sample was also quantified by direct analysis presenting R2 0.98. The t-student test presented no significant difference regarding HPLC method. Despite the experimental design performed for riboflavin, it was not possible its quantification in the commercial samples.

Crystalline phase transition of ezetimibe in final product, after packing, promoted by the humidity of excipients: Monitoring and quantification by Raman spectroscopy.

Ezetimibe (EZT), in its anhydrous form, is a drug used for cholesterol and lipids reduction in blood plasma. The presence of EZT monohydrate in commercial tablets can change the solubility rate of the API, decreasing its activity. The objective of this work was to verify if the humidity present in the excipients could promote the phase transition from EZT anhydrous to hydrate. Initially the stability of the pure anhydrous form was monitored by Raman, at room temperature (23°C) and relative humidity (75%). The MCR-ALS method showed that almost all EZT changed to hydrated form in 30 min. Then tablets of ezetimibe in the presence of its excipients were prepared and vacuum packed using a polyethylene film. Such tablet was monitored by Raman spectroscopy for 24h in order to quantify the mixture of the crystalline forms. A multivariate calibration model using Raman spectroscopy and Partial Least Square (PLS) regression was built, with validation and cross validation errors around 0.6% (wt/wt), for both crystalline forms, and R(2) higher than 0.96. The PLS model was used to quantify the crystalline mixture of ezetimibe in the monitored tablet, after 24h more than 70% of ezetimibe changed to the hydrated form.

Determination of acetylsalicylic acid in commercial tablets by SERS using silver nanoparticle-coated filter paper.

In this work, filter paper was used as a low cost substrate for silver nanoparticles in order to perform the detection and quantification of acetylsalicylic acid by SERS in a commercial tablet. The reaction conditions were 150mM of ammonium hydroxide, 50mM of silver nitrate, 500mM of glucose, 12min of the reaction time, 45°C temperature, pretreatment with ammonium hydroxide and quantitative filter paper (1-2µm). The average size of silver nanoparticles deposited on the paper substrate was 180nm. Adsorption time of acetylsalicylic acid on the surface of the silver-coated filter paper was studied and an adsorption time of 80min was used to build the analytical curve. It was possible to obtain a calibration curve with good precision with a coefficient of determination of 0.933. The method proposed in this work was capable to quantify acetylsalicylic acid in commercial tablets, at low concentration levels, with relative error of 2.06% compared to the HPLC. The preparation of filter paper coated with silver nanoparticles using Tollen's reagent presents several advantages such as low cost of synthesis, support and reagents; minimum amount of residuals, which are easily treated, despite the SERS spectroscopy presenting fast analysis, with low sample preparation and low amount of reactants as in HPLC analysis.

Optimization of SERS scattering by Ag-NPs-coated filter paper for quantification of nicotinamide in a cosmetic formulation.

Supported silver nanoparticles on filter paper were synthesized using Tollens' reagent. Experimental designs were performed to obtain the highest SERS enhancement factor by study of the influence of the parameters: filter paper pretreatment, type of filter paper, reactants concentration, reaction time and temperature. To this end, fractional factorial and central composite designs were used in order to optimize the synthesis for quantification of nicotinamide in the presence of excipients in a commercial sample of cosmetic. The values achieved for the optimal condition were 150 mM of ammonium hydroxide, 50 mM of silver nitrate, 500 mM of glucose, 8 min for the reaction time, 45 °C temperature, pretreatment with ammonium hydroxide and quantitative filter paper (1-2 µm). Despite the variation of SERS intensity, it was possible to use an adapted method of internal standard to obtain a calibration curve with good precision. The coefficient of determination of the linear fit was 0.97. The method proposed in this work was capable of quantifying nicotinamide on a commercial cosmetic gel, at low concentration levels, with a relative error of 1.06% compared to the HPLC. SERS spectroscopy presents faster analyses than HPLC, also complex sample preparation and large amount of reactants are not necessary.

Evaluation of analytical tools and multivariate methods for quantification of co-former crystals in ibuprofen-nicotinamide co-crystals.

Co-crystals are multicomponent substances designed by the addition of two or more different molecules in a same crystallographic pattern, in which it differs from the crystallographic motif of its co-formers. The addition of highly soluble molecules, like nicotinamide, in the crystallographic pattern of ibuprofen enhances its solubility more than 7.5 times, improving the properties of this widely used drug. Several analytical solid state techniques are used to characterize the ibuprofen-nicotinamide co-crystal, being the most used: mid-infrared (ATR-FTIR), differential scanning calorimetry (DSC), X-ray diffraction (XRPD) and Raman spectroscopy. These analytical solid state techniques were evaluated to quantify a mixture of ibuprofen-nicotinamide co-crystal and its co-formers in order to develop a calibration model to evaluate the co-crystal purity after its synthesis. Raman spectroscopy showed better result than all other techniques with a combination of multivariate calibration tools, presenting lower values of calibration and prediction errors. The partial least squares regression model gave a mean error lower than 5% for all components presented in the mixture. DSC and mid-infrared spectroscopy proved to be insufficient for quantification of the ternary mixture. XRPD presented good results for quantification of the co-formers, ibuprofen and nicotinamide, but fair results for the co-crystal. This is the first report of quantification of ibuprofen-nicotinamide co-crystal, among its co-formers. The quantification is of great importance to determine the yield of the co-crystallization reactions and the purity of the product obtained.