Márcia Cristina Breitkreitz

Cocoa butter (CB), a natural lipid, is widely used in the food and pharmaceutical industries. However, it is characterized by a polymorphism which significantly affects its physicochemical properties under varying temperatures. Because synthesis, storage and transportation may occur under varying temperatures conditions, understanding its thermal behavior is a crucial step for its characterization and final product quality. In this study, the crystalline structure of CB was analyzed using differential scanning calorimetry (DSC) and small-angle X-ray scattering (SAXS), focusing on developed procedures to isolate form II using variations in heating/cooling rates. DSC curves were deconvoluted using Gaussian curve shapes and the results revealed clear coexistence of polymorphs in the raw material. However, different heating/cooling rates showed different behavior. Recording DSC data with cooling at 10 K min ⁻¹ affected the crystallization since the crystal memory was not completely erased, while slower rates of 2–5 K min ⁻¹ allowed more time for crystallization and organization. At these heating rates, it was possible to isolate form II at 14–15 °C. To deconvolute the peaks and trace the polymorphic transitions during the cooling step, the SAXS data were evaluated using independent components analysis (ICA). In-situ SAXS provided detailed insight into the crystallization process, demonstrating that the cooling/heating rate and the isotherm time at 0 °C were critical factors. At the same time, ICA helped to isolate signals of the different forms and allowed a direct comparison of different temperature conditions, highlighting the potential of this methodology for characterizing CB and its products.

SUMMARY: Psychotria viridis Ruiz & Pav. has gained significant attention due to its role in the preparation of ayahuasca. This study aimed to improve the understanding of the specialized metabolite profile in P. viridis leaves and to evaluate how growing conditions and seasonality impact this composition. The specimens were grown either in the open field or in the shaded environment of rubber tree (Hevea brasiliensis L.) cultivation, forming a clonal population of the mother plant. Samples were collected in all four seasons of the year. After a three‐phase extraction of the samples, the aqueous and organic phases were analyzed using an ultra‐high‐performance liquid chromatography coupled with electrospray ionization and Orbitrap mass spectrometry (UHPLC‐ESI‐Orbitrap‐MS) system. The acquired data were processed using MS‐DIAL 4.9 and MetaboAnalyst 5.0 for multivariate statistics and pathway activity analysis. Chemical variations were investigated employing principal component analysis (PCA), hierarchical cluster analysis (HCA), and partial least squares discriminant analysis (PLS‐DA). The most important identified compounds for differentiation according to seasonality were flavonoids. The pathways presenting significant variation in response to seasonality were related to energy generation through biosynthesis and consumption of carbohydrates: ascorbate and aldarate metabolism, pentose and glucuronate interconversions, and citrate cycle. Meanwhile, the biosynthesis of flavonoids, flavones, and flavonols was associated with the influence of the cultivation location in full sunlight or shade in an intercrop, indicating a plant response to oxidative stress. In our comprehensive analysis, DMT concentrations did not exhibit any significant statistical variation across the studied conditions.

In the analytical quality by design (AQbD) framework, the design of experiments (DOE) plays a very important role, as it provides information about how experimental input variables influence critical method attributes. Based on the information obtained from the DOE, mathematical models are generated and used to build the method operable design region (MODR), which is a robust region of operability. Data treatment steps are usually carried out in software such as Fusion QbD, Minitab, or StaEase 360, among others. Although there are many studies in the literature that use the DOE, none of them address important aspects of data treatment for optimization and MODR generation and compare different software calculations. The purpose of this study is to contribute to a better understanding of data treatment aspects that are frequently misread or not fully understood, such as model selection, ANOVA results, and residual analysis. The discussion will be guided by the separation of curcuminoids, using ultra-high performance liquid chromatography and eight quality attributes as responses. This study highlights the importance of carefully selecting and evaluating models, as they significantly influence the generation of the MODR. Moreover, the findings emphasize that it is essential to incorporate uncertainties into the contour plots to accurately determine the MODR in compliance with the ICH Q14 guidelines and USP General Chapter .

In the AQbD framework, DOE plays a very important role as it provides information about how experimental input variables influence the critical method attributes. Based on the information obtained from the DOE, mathematical models are generated and used to build the MODR a robust region of operability. Data treatment steps are usually carried out in software such as Fusion QbD, Minitab, StaEase 360, among others. Although there are many papers in the literature using DOE, none of them address important aspect of data treatment for optimization and MODR generation and compare different software calculations. The purpose of this study is to contribute to a better understanding of data treatment aspects that are frequently misread or not fully understood, such as model selection, ANOVA results and residual analysis. The discussion will be guided using the separation of curcuminoids by ultra-high performance liquid chromatography and eight quality attribute as responses. The results highlighted the importance of the correct selection and evaluation of the models due to their influence the generation of the MODR and emphasizes the insertion of the uncertainty in the contour plots to proper obtain the MODR regarding the definitions of the USP General Chapter and ICH Q14 guidelines.

Although chemotherapy regimens are well-established in clinical practice, chemoresistance and adverse side effects pose significant obstacles in cancer treatment. Paclitaxel (PTX), a widely used chemotherapeutic agent, faces formulation challenges due to its poor solubility and permeability. Research suggests that the phytochemical Cannabidiol (CBD) holds potential not only in targeting cancer cells but also in alleviating pain and nausea, thereby improving the quality of life for cancer patients. However, CBD’s clinical application is also limited by its poor solubility, low bioavailability, and susceptibility to oxidation. Nanostructured lipid carriers (NLCs) represent a promising drug delivery system for hydrophobic compounds like PTX and CBD and allows their co-encapsulation. Nonetheless, achieving a stable formulation requires identifying suitable preparation methods and excipients. The aim of this study was to develop and optimize an NLC formulation for the co-encapsulation of PTX and CBD. Using factorial design, an optimized formulation was obtained with homogeneous particle sizes (200 nm), negative zeta potentials (−17 mV), particle concentration of 10 ¹³ particles/mL, spherical morphology (TEM images), and a lipid core with low crystallinity (as confirmed by XRD). To evaluate the therapeutic potential of the drug combination, cell viability assays were conducted on murine melanoma cells (B16-F10) at different exposure times (24 and 48 hours). The NLC-CBD-PTX formulation significantly reduced cell viability in a time- and concentration-dependent manner, demonstrating at least 75% greater activity at 24 hours compared to each drug individually whether free (PTX, CBD) or encapsulated (NLC-PTX, NLC-CBD). This indicates a synergistic effect of the encapsulated drugs on cytotoxicity. In conclusion, an innovative pharmaceutical formulation co-encapsulating PTX and CBD was validated, showing potential to enhance antitumor efficacy, overcome chemoresistance, reduce side effects, and broaden therapeutic applications. The resulting NLCs exhibited favorable physicochemical properties, supporting their suitability for various routes of administration.

Nanostructured lipid carriers (NLCs) are typically composed of liquid lipids, solid lipids, and surfactants, enabling the encapsulation of lipophilic drugs. Butamben is a Class II anesthetic drug, according to the Biopharmaceutical Classification System (BCS); it has a log P of 2.87 and is considered a ‘brick dust’ (poorly water-soluble and poorly lipid-soluble) drug. This characteristic poses a challenge for the development of NLCs, as they are not soluble in the liquid lipid present in the NLC core. In a previous study, we developed an NLC core consisting of a solid lipid (Crodamol^TM CP), a lipophilic liquid with medium polarity (SR^TM Lauryl lactate), and a hydrophilic excipient (SR^TM DMI) that allowed the solubilization of ‘brick dust’ types of drugs, including butamben. In this study, starting from the NLC core formulation previously developed we carried out an optimization of the surfactant system and evaluated their performance in aqueous medium. Three different surfactants (Crodasol^TM HS HP, Synperonic^TM PE/F68, and Croduret^TM 40) were studied and, for each of them, a 2³ factorial design was stablished, with total lipids, % surfactant, and sonication time (min) as the input variables and particle size (nm), polydispersity index (PDI), and zeta potential (mV) as the response variables. Stable NLCs were obtained using Crodasol^TM HS HP and Synperonic^TM PE/F68 as surfactants. Through a comparison between NLCs developed with and without SR^TM DMI, it was observed that besides helping the solubilization of butamben in the NLC core, this excipient helped in stabilizing the system and decreasing particle size. NLCs containing Crodasol^TM HS HP and Synperonic^TM PE/F68 presented particle size values in the nanometric scale, PDI values lower than 0.3, and zeta potentials above |10|mV. Concerning NLCs’ stability, SBTB-NLC with Synperonic^TM PE/F68 and butamben demonstrated stability over a 3-month period in aqueous medium. The remaining NLCs showed phase separation or precipitation during the 3-month analysis. Nevertheless, these formulations could be freeze-dried after preparation, which would avoid precipitation in an aqueous medium.

Nanostructured lipid carriers (NLC) are typically composed of liquid lipids, solid lipids, and surfactants, enabling the encapsulation of lipophilic drugs. Butamben is a Class II anesthetic drug according to the Biopharmaceutical Classification System (BCS), it has a log P 2.87 and is considered a 'brick dust' (poorly water-soluble and poorly lipid-soluble) drug. This characteristic poses a challenge for the development of NLCs, as they are not soluble in the liquid lipid present in the NLC core. In a previous study [1] we have developed a NLC core consisting of solid lipid (CrodamolTM CP), liquid lipid of medium polarity (SRTM Lauryl lactate), and a hydrophilic excipient (SRTM DMI) that allowed the solubilization of 'brick dust' type of drugs including butamben. In this study, starting from the NLC core formulation previously developed we carried out the optimization of the surfactant system and evaluated their performance in aqueous medium. Three different surfactants (CrodasolTM HS HP, SynperonicTM PE/F68, and CroduretTM 40) were studied and, for each of them, a 2³ factorial design was stablished, with total lipids, % surfactant, and sonication time (min) as the input variables and particle size (nm), polydispersity index (PDI) and zeta potential (mV) as the response variables. Stable NLCs were obtained using CrodasolTM HS HP and SynperonicTM PE/F68 as surfactants. Through a comparison between NLCs developed with and without SRTM DMI, it was observed that besides helping the solubilization of butamben in the NLC core, this excipient helped stabilizing the system and decreasing particle size NLCs containing CrodasolTM HS HP and SynperonicTM PE/F68 presented particle size values in the nanometric scale, PDI lower than 0.3 and zeta potential above |10|mV. Concerning NLCs stability, SBTB-NLC with SynperonicTM PE/F68 and butamben demonstrated stability over a 3-month period in aqueous medium. The remaining NLCs showed phase separation or precipitation during the 3-month analysis. Nevertheless, these formulations could be freeze-dried after preparation, which would avoid precipitation in aqueous medium.