Preliminary insights on the development of a continuous-flow solar system for the photocatalytic degradation of contaminants of emerging concern in water.

The ubiquity and impact of pharmaceuticals and pesticides, as well as their residues in environmental compartments, particularly in water, have raised human and environmental health concerns. This emphasizes the need of developing sustainable methods for their removal. Solar-driven photocatalytic degradation has emerged as a promising approach for the chemical decontamination of water, sparking intensive scientific research in this field. Advancements in photocatalytic materials have driven the need for solar reactors that efficiently integrate photocatalysts for real-world water treatment. This study reports preliminary results from the development and evaluation of a solar system for TiO2-based photocatalytic degradation of intermittently flowing water contaminated with doxycycline (DXC), sulfamethoxazole (SMX), dexamethasone (DXM), and carbendazim (CBZ). The system consisted of a Fresnel-type UV solar concentrator that focused on the opening and focal point of a parabolic trough concentrator, within which tubular quartz glass reactors were fixed. Concentric springs coated with TiO2, arranged one inside the other, were fixed inside the quartz reactors. The reactors are connected to a raw water tank at the inlet and a check valve at the outlet. Rotating wheels at the collector base enable solar tracking in two axes. The substances (SMX, DXC, and CBZ) were dissolved in dechlorinated tap water at a concentration of 1.0 mg/L, except DXM (0.8 mg/L). The water underwent sequential batch (~ 3 L each, without recirculation) processing with retention times of 15, 30, 60, 90, and 120 min. After 15 min, the degradation rates were as follows: DXC 87%, SMX 35.5%, DXM 32%, and CBZ 31.8%. The system processed 101 L of water daily, simultaneously removing 870, 355, 256, and 318 µg/L of DXC, SMX, DXM, and CBZ, respectively, showcasing its potential for real-world chemical water decontamination application. Further enhancements that enable continuous-flow operation and integrate highly effective adsorbents and photocatalytic materials can significantly enhance system performance.

Selective immunoglobulin aggregates removal in antivenoms by a simple chromatographic step based on a monolithic stationary phase.

Antivenom therapy is a critical intervention for treating the more than 5.000.000 envenomation accidents that occur each year around the world. These immunotherapeutic drugs are mostly produced following techniques developed more than fifty years ago with minor changes. Aggregate content has been described as one of the main causes of early adverse effects after intravenous administration of antivenoms. In this work we propose the introduction of a final polishing step to traditional antivenom manufacturing processes aimed at lowering the aggregate content in the final product. The refinement step proposed in this work is based on the selective capture of immunoglobulin aggregates by a cation exchange monolithic stationary phase. We show that this media can effectively remove aggregates in the final product under isotonic ion-strength and mildly acidic conditions following a negative chromatography strategy, thus making it a useful technique for producing higher quality products.

Qualitative Analysis of Baltic Amber Resin by Gas Chromatography Coupled with Mass Spectrometry and the Therapeutic Potential of this Fossil Resin

ABSTRACT Objective: To analyze the molecular composition of a sample of Baltic amber taken from an amber bracelet, verify the presence or absence of compounds beneficial to human health, and discuss the therapeutic potential of this fossil resin. Material and Methods: For this, a qualitative analysis was performed by gas chromatography coupled with mass spectrometry (GC-MS system), which can perform an automated separation into individual components of amber. Results: The samples comprised terpenes and terpenoids: monoterpenoids, sesquiterpenoids, diterpenoids, hydronaphthalene, succinic acid, and isopimaric acid. These have therapeutic potential for several diseases, inhibit several stages of the inflammatory process, and improve the symptoms caused by it. Conclusion: The amber of this sample tested is formed by terpenes and terpenoids, substances with anti-inflammatory and analgesic. However, an in-depth study is needed on the release and absorption of substances in human skin or their actual effectiveness.

Spectrophotometry and chromatography analyses combined with chemometrics tools to differentiate green coffee beans into special or traditional.

Green coffee is the hulled coffee bean, rich in chemical compounds indicative of quality before roasting, making the classification special or traditional. This work aimed to determine compounds in green coffee beans and find the differentiation of green coffee beans into special or traditional ones through chemometrics. For that, the levels of phenolic compounds, reducing, nonreducing, and total sugars were quantified by spectrophotometry: caffeine, trigonelline, 5-hydroxymethylfurfural (5-HMF), 3-hydroxybenzoic, 4-hydroxybenzoic, chlorogenic, caffeic, and nicotinic acids (NAs) by high-performance liquid chromatography-UV-Vis; acetaldehyde, acetone, methanol, ethanol, and isoamyl by HS-GC-FID. Principal component analysis (PCA) was used to differentiate green coffee beans through the levels obtained in spectrophotometric and chromatographic analyses. Statistically, the contents of total phenolic compounds, caffeine, nonreducing sugars, total sugars, NA, 5-HMF, acetaldehyde, ethanol, and ethanol/methanol showed significant differences. The PCA made it possible to classify green coffee beans into special and traditional, in addition to understanding the attributes that influenced the differentiation between coffees. In addition, it was possible to classify green coffee beans into special and traditional, either using all parameters evaluated or only using spectrophotometric analyses. In this way, some advantages allow classification without using a trained and experienced evaluator as their previous experience can influence the results due to their expertise in a certain type of coffee, in addition to being faster and cheaper, especially regarding spectrophotometric analyses.

Estudio biodirigido de metabolitos secundarios con actividad antimicrobiana y aislamiento de D-pinitol de semillas de Leucaena esculenta y L. leucocephala / Bio-guided study of secondary metabolites with antimicrobial activity and isolation of D-pinitol from Leucaena esculenta and L. leucocephala seeds

Phytochemical bio-guided studies are used to find compounds with biological activity. Flavonoids from seeds of Leucaena species have antimicrobial activity in strains of medical interest, therefore, fresh seeds were collected from the town of Tlayacapan, Morelos, Mexico. The methanolic extracts were obtained by the maceration technique, targeted fractionation was performed using adsorption and molecular exclusion chromatographic techniques; to observe the antimicrobial activity, agar diffusion techniques were used; spectrometric and spectroscopic techniques were used for the characterization of D-pinitol, resulting from the fractionation of L. leucocephala. Antimicrobial activity was found on strains of Escherichia coli CUSI and Staphylococcus aureus ATCC 29213 of the most polar fractions, identifying the responsible compounds by HPLC: caffeic acid, gallic acid, p-coumaric acid, quercetin, catechin and apigenin, these compounds can inhibit the activation enzymatic, synthesis of nucleic acids and proteins, chelating with different ions, etc.

Los estudios biodirigidos fitoquímicos son empleados para encontrar compuestos con actividad biológica. Los flavonoides de semillas de especies de Leucaena son reportados por tener actividad antimicrobiana sobre cepas de interés médico, por tanto, se colectaron semillas frescas de la localidad de Tlayacapan, Morelos, México. Se obtuvieron los extractos metanólicos mediante la técnica de maceración, el fraccionamiento dirigido se realizó empleando técnicas cromatográficas de adsorción y exclusión molecular; la actividad antimicrobiana se determinó mediante técnicas de difusión en agar; se utilizaron técnicas espectrométricas y espectroscópicas para la caracterización del D-pinitol, resultado del fraccionamiento de L. leucocephala. Se encontró actividad antimicrobiana sobre cepas de Escherichia coli CUSI y Staphylococcus aureus ATCC 29213 de las fracciones más polares, identificando los compuestos responsables vía HPLC: ácido caféico, ácido gálico, ácido p-cumárico quercetina, catequina y apigenina, dichos compuestos pueden inhibir activación enzimática, síntesis de ácidos nucleicos y proteínas, quelarse con diferentes iones, etc.

Phytochemical screening of Cordia glabrata (MART.) A.DC. extracts and its potential antioxidant, photoprotective, antimicrobial and antiviral activities / Triagem fitoquímica de extratos de Cordia glabrata (MART.) A.DC. e sua potencial atividade antioxidante, fotoprotetora, antimicrobiana e antiviral

Species of the genus Cordia have shown biological activities, such as anti-inflammatory, analgesic, antioxidant, antiviral, and antifungal activities. The species Cordia glabrata (MART) A.DC. Has no information concerning its phytochemical profile and possible biological activities. Thus, this study aimed to evaluate this profile in ethanolic extracts of young, adult and senescent leaves, as well as their antioxidant, photoprotective, antimicrobial, and virucidal potentials. Phytochemical analysis was performed by TLC (thin-layer chromatography) and showed the presence of flavonoids, tannins, and terpenes. The evaluation by UPLC-MS/MS (Ultra performance liquid chromatography - tandem mass spectrometer) evidenced the presence of caffeic (3.89 mgL-¹), p-cumaric (6.13 mgL-¹), and ferulic (0.58 mgL-¹) acids, whilst, in GC/MS (Gas chromatography–mass spectrometry) analysis there was a greater amount of palmitic (51.17%), stearic (20.34%), linoleic (9.62%), and miristic (8.16%) fatty acids. The DPPH (2,2-Diphenyl-1-picrylhydrazyl) and ABTS+ (2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radicals were used to verify the potential antioxidant activity, observing a better activity for the leaf extract in the adult phenological stage: 54.63 ± 1.06 µgmL-¹ (DPPH) and 44.21 ± 1.69 mM (ABTS). The potential photoprotective activity of the extracts was determined by spectrophotometry and the in vitro values of SPF (Sun Protection Factor) in young and adult leaves (5.47 and 5.41, respectively) showed values close to the minimum SPF of 6.0 required by ANVISA (Brazilian Health Regulatory Agency). It was not observed an antimicrobial activity for Staphylococcus aureus with a minimum inhibitory concentration of 2000 µgmL-¹, however the anti-herpetic assay against the Herpes simplex virus type 2 (HSV-2) showed a potent virucidal activity at the tested concentrations with CV50 value <0.195 µgmL-¹ and a Selectivity Index (SI = CC50 / CV50) greater than 448. The results [...].(AU)

Espécies do gênero Cordia apresentam atividades biológicas, como anti-inflamatória, analgésica, antioxidante, antiviral e antifúngica. Para a espécie Cordia glabrata (MART) A.DC., ainda não existem informações sobre seu perfil fitoquímico e possíveis atividades biológicas, deste modo, o presente estudo teve como objetivo avaliar este perfil em extratos etanólicos de folhas jovens, adultas e senescentes, bem como o potencial antioxidante, fotoprotetor, antimicrobiano e virucida. A análise fitoquímica foi realizada por CCD (Cromatografia em Camada Delgada), mostrando a presença de flavonóides, taninos e terpenos. Na avaliação por CLAE EM/EM (Cromatografia Líquida de Ultra Eficiência acoplada a Espectrometria de Massas) foi evidenciado a presença dos ácidos caféico (3,89 mgL-¹), p-cumárico (6,13 mgL-¹) e ferúlico (0,58 mgL-¹), paralelamente, na CG/EM (Cromatografia Gasosa acoplada a Espectrometria de Massas) verificou-se maior quantidade dos ácidos graxos palmítico (51,17%), esteárico (20,34%), linoléico (9,62%) e mirístico (8,16%). Os radicais DPPH (2,2-Difenil-1-picrilhidrazil) e ABTS+ (2′-Azino-bis (ácido 3-etilbenzotiazolina-6-sulfônico)) foram utilizados para verificar o potencial antioxidante, observando se uma atividade superior para o extrato da folha em sua fase fenológica adulta: 54,63 ± 1,06 µgmL-¹ (DPPH) e 44,21 ± 1,69 mM (ABTS+). A potencial atividade fotoprotetora dos extratos foi determinada espectrofotometricamente e os valores in vitro de FPS (Fator de Proteção Solar) em folhas jovens e adultas (5,47 e 5,41 respectivamente) apresentaram valores próximos ao FPS mínimo de 6,0 exigido pela ANVISA (Agência Nacional de Vigilância Sanitária). Não foi observada atividade antimicrobiana para Staphylococcus aureus sendo a concentração inibitória mínima de 2000 µgmL-¹, no entanto o ensaio anti-herpético contra o vírus Herpes simplex tipo 2 (HSV-2) mostrou uma potente atividade virucida nas concentrações testadas [...].(AU)

Phytochemical screening of Cordia glabrata (MART.) A.DC. extracts and its potential antioxidant, photoprotective, antimicrobial and antiviral activities / Triagem fitoquímica de extratos de Cordia glabrata (MART.) A.DC. e sua potencial atividade antioxidante, fotoprotetora, antimicrobiana e antiviral

Species of the genus Cordia have shown biological activities, such as anti-inflammatory, analgesic, antioxidant, antiviral, and antifungal activities. The species Cordia glabrata (MART) A.DC. Has no information concerning its phytochemical profile and possible biological activities. Thus, this study aimed to evaluate this profile in ethanolic extracts of young, adult and senescent leaves, as well as their antioxidant, photoprotective, antimicrobial, and virucidal potentials. Phytochemical analysis was performed by TLC (thin-layer chromatography) and showed the presence of flavonoids, tannins, and terpenes. The evaluation by UPLC-MS/MS (Ultra performance liquid chromatography - tandem mass spectrometer) evidenced the presence of caffeic (3.89 mgL-¹), p-cumaric (6.13 mgL-¹), and ferulic (0.58 mgL-¹) acids, whilst, in GC/MS (Gas chromatography–mass spectrometry) analysis there was a greater amount of palmitic (51.17%), stearic (20.34%), linoleic (9.62%), and miristic (8.16%) fatty acids. The DPPH (2,2-Diphenyl-1-picrylhydrazyl) and ABTS+ (2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radicals were used to verify the potential antioxidant activity, observing a better activity for the leaf extract in the adult phenological stage: 54.63 ± 1.06 µgmL-¹ (DPPH) and 44.21 ± 1.69 mM (ABTS). The potential photoprotective activity of the extracts was determined by spectrophotometry and the in vitro values of SPF (Sun Protection Factor) in young and adult leaves (5.47 and 5.41, respectively) showed values close to the minimum SPF of 6.0 required by ANVISA (Brazilian Health Regulatory Agency). It was not observed an antimicrobial activity for Staphylococcus aureus with a minimum inhibitory concentration of 2000 µgmL-¹, however the anti-herpetic assay against the Herpes simplex virus type 2 (HSV-2) showed a potent virucidal activity at the tested concentrations with CV50 value <0.195 µgmL-¹ and a Selectivity Index (SI = CC50 / CV50) greater than 448. The results [...].

Espécies do gênero Cordia apresentam atividades biológicas, como anti-inflamatória, analgésica, antioxidante, antiviral e antifúngica. Para a espécie Cordia glabrata (MART) A.DC., ainda não existem informações sobre seu perfil fitoquímico e possíveis atividades biológicas, deste modo, o presente estudo teve como objetivo avaliar este perfil em extratos etanólicos de folhas jovens, adultas e senescentes, bem como o potencial antioxidante, fotoprotetor, antimicrobiano e virucida. A análise fitoquímica foi realizada por CCD (Cromatografia em Camada Delgada), mostrando a presença de flavonóides, taninos e terpenos. Na avaliação por CLAE EM/EM (Cromatografia Líquida de Ultra Eficiência acoplada a Espectrometria de Massas) foi evidenciado a presença dos ácidos caféico (3,89 mgL-¹), p-cumárico (6,13 mgL-¹) e ferúlico (0,58 mgL-¹), paralelamente, na CG/EM (Cromatografia Gasosa acoplada a Espectrometria de Massas) verificou-se maior quantidade dos ácidos graxos palmítico (51,17%), esteárico (20,34%), linoléico (9,62%) e mirístico (8,16%). Os radicais DPPH (2,2-Difenil-1-picrilhidrazil) e ABTS+ (2′-Azino-bis (ácido 3-etilbenzotiazolina-6-sulfônico)) foram utilizados para verificar o potencial antioxidante, observando se uma atividade superior para o extrato da folha em sua fase fenológica adulta: 54,63 ± 1,06 µgmL-¹ (DPPH) e 44,21 ± 1,69 mM (ABTS+). A potencial atividade fotoprotetora dos extratos foi determinada espectrofotometricamente e os valores in vitro de FPS (Fator de Proteção Solar) em folhas jovens e adultas (5,47 e 5,41 respectivamente) apresentaram valores próximos ao FPS mínimo de 6,0 exigido pela ANVISA (Agência Nacional de Vigilância Sanitária). Não foi observada atividade antimicrobiana para Staphylococcus aureus sendo a concentração inibitória mínima de 2000 µgmL-¹, no entanto o ensaio anti-herpético contra o vírus Herpes simplex tipo 2 (HSV-2) mostrou uma potente atividade virucida nas concentrações testadas [...].

Mono-PEGylated lysozyme purification with increased productivity and isomer differentiation through heparin monolith chromatography.

PEGylated protein purification with the required quality attributes has represented a bioengineering challenge and Affinity Monolith Chromatography (AMC) has never been exploited for this goal. This work reports the generation of a heparin-modified affinity monolith disk by reductive alkylation with raised ligand density for its use as chromatographic support in the separation of lysozyme PEGylation reactions (LPRs) with three different PEG sizes (1, 20 and 40 kDa). For immobilized heparin determination a modified toluidine colorimetric assay adapted to microplate format was proposed. The heparin modified-disk was able to differentiate positional isomers of 20 kDa mono-PEGylated lysozyme at neutral pH using a salt linear gradient. Identity of PEG-conjugates was verified by SDS-PAGE and positional isomers were partially characterized by peptide mapping mass spectrometry. 20 kDa mono-PEGylated lysozyme conjugate purity (99.69 ± 0.05%) was comparable with traditional chromatographic methods while productivity (0.0964 ± 0.0001 mg/mL*min) was increased up to 6.1 times compared to that obtained in heparin packed-bed affinity chromatography procedures. The proposed AMC method represents a reliable, efficient, easy-handling, fast and single-step operation for the analysis or preparative isolation of PEGylated proteins containing a heparin binding domain.

The isolation of water-soluble natural products - challenges, strategies and perspectives.

Covering period: up to 2019Water-soluble natural products constitute a relevant group of secondary metabolites notably known for presenting potent biological activities. Examples are aminoglycosides, ß-lactam antibiotics, saponins of both terrestrial and marine origin, and marine toxins. Although extensively investigated in the past, particularly during the golden age of antibiotics, hydrophilic fractions have been less scrutinized during the last few decades. This review addresses the possible reasons on why water-soluble metabolites are now under investigated and describes approaches and strategies for the isolation of these natural compounds. It presents examples of several classes of hydrosoluble natural products and how they have been isolated. Novel stationary phases and chromatography techniques are also reviewed, providing a perspective towards a renaissance in the investigation of water-soluble natural products.

Polymer monoliths for the concentration of viruses from environmental waters: A review.

Even at low concentrations in environmental waters, some viruses are highly infective, making them a threat to human health. They are the leading cause of waterborne enteric diseases. In agriculture, plant viruses in irrigation and runoff water threat the crops. The low concentrations pose a challenge to early contamination detection. Thus, concentrating the virus particles into a small volume may be mandatory to achieve reliable detection in molecular techniques. This paper reviews the organic monoliths developments and their applications to concentrate virus particles from waters (waste, surface, tap, sea, and irrigation waters). Free-radical polymerization and polyaddition reactions are the most common strategies to prepare the monoliths currently used for virus concentration. Here, the routes for preparing and functionalizing both methacrylate and epoxy-based monoliths will be shortly described, following a revision of their retention mechanisms and applications in the concentration of enteric and plant viruses in several kinds of waters.

Pathological and mycological characterization of pulmonary Aspergillus fumigatus infection producing gliotoxin in a captive African grey parrot (Psittacus erithacus)

Aspergillosis, the main causative agent of which is Aspergillus fumigatus, causes mortality in all types of birds. Gliotoxin (GT), one of the multiple virulence factors of A. fumigatus, has a variety of immunosuppressive effects. The corpse of an African grey parrot (Psittacus erithacus) was sent for necropsy and diagnostic rule-out. The lungs were enlarged, firm, and had dark-red coloration, on the parietal faces of both lungs, some semi-circular caseous necrosis areas were observed. The caudal thoracic and abdominal air sacs were thickened and contained a fibrin-heterophilic exudate. Histopathologically, a necrotic and granulomatous bronchopneumonia was observed with intralesional hyphae with characteristics compatible with Aspergillus sp. that were positive with Grocott´s staining. Fibrinous and heterophilic airsacculitis was found in the air sacs. A. fumigatus was isolated from lungs, characterized using serial microcultures, and confirmed using polymerase chain reaction. In addition, GT production was detected in vitro from the culture filtrate in which the isolate was grown; the organic extract was analysed via thin-layer chromatography. This is the first detection of GT in a case of pulmonary aspergillosis in a parrot, which could help to understand the pathogenesis of the disease in psittacines.(AU)

Impact of glyphosate formulations and adjuvants: effects on leaf interaction, metabolism, and control of sourgrass / Impacto das formulações de glifosato e dos adjuvantes: efeitos na interação foliar, no metabolismo e no controle do capim-amargoso

The addition of commercial formulations and adjuvants to a tank mix may result in differences in the wettability on the plant surface, foliar uptake and herbicide efficacy. Thus, the objectives of this study were to evaluate the influence of glyphosate formulations and tank-mixture adjuvants on the contact angle (CA), uptake, metabolism and sourgrass control and the damage to the cuticular microstructure of this species caused by herbicide solutions. For this purpose, assays were carried out in a completely randomized design, and treatments distributed in a 2x5+1 factorial scheme with five replications. Two glyphosate formulations isopropylamine salt (SL) and ammonium salt (WG) combined or not with the adjuvant methylated soybean oil (MSO), mineral oil (MO), ethoxylated alkyl ester (EAE) or polyoxyethylenealkylphenol ether (PAE); and one control (water) were evaluated. CA measurements of the droplets deposited on a sourgrass leaf surface and on the standard surface (parafilm) were obtained using a tensiometer. Herbicide uptake and shikimate accumulation were simultaneously determined by chromatography and spectrometry. The control effect was assessed by observing plant survival dry weight reductions. The glyphosate SL and WG formulations had similar effects on the variables analyzed. However, mixing the adjuvants EAE, MO or MSO with either formulation of herbicide led to greater wettability and more severe damage to the cuticular microstructure, favoring glyphosate uptake and shikimate accumulation. Despite evidence regarding the treatments containing oils/surfactants, the control effect on sourgrass was similar. The combination of glyphosate formulations with such adjuvants is potentially more effective and guarantees satisfactory sourgrass control.

Formulações comerciais e adjuvantes adicionados à mistura em tanque podem resultar em mudanças na superfície de molhamento, absorção foliar e eficácia dos herbicidas. Assim, objetivou-se com este trabalho avaliar a influência das formulações de glifosato e adjuvantes sobre o ângulo de contato (CA), absorção, metabolismo e controle do capim-amargoso, além de investigar os danos ocasionados pelas soluções herbicidas à microestrutura cuticular dessa espécie daninha. Para tanto, ensaios foram conduzidos em delineamento inteiramente casualizado, e os tratamentos distribuídos em esquema fatorial 2x5+1, com cinco repetições. Duas formulações de glifosato (SL e WG) combinadas ou não com os adjuvantes éster metílico de óleo de soja (MSO), óleo mineral (MO), alquil ester etoxilado (EAE) e polioxietilenoalquilfenoléter (PAE); e um controle (água) foram avaliados. As medidas de CA das gotas depositadas na superfície foliar de capim-amargoso e na superfície padrão (parafilme) foram obtidas usando um tensiômetro. A absorção do herbicida e o acúmulo de chiquimato foram determinados, simultaneamente, por cromatografia e espectrometria de massas. O efeito de controle foi avaliado pela observação da sobrevivência da planta e redução do peso seco. As formulações de glifosato SL e WG apresentaram efeitos similares sobre as variáveis analisadas. Contudo, os adjuvantes EAE, MO e MSO, quando em mistura ao herbicida, em ambas as formulações, destacaram-se por apresentar maior molhabilidade e danos mais severos à microestrutura cuticular, favorecendo a absorção do glifosato e acúmulo de chiquimato. Apesar das evidências sobre a bioeficácia dos tratamentos que continham óleos/surfactantes, o controle sobre capim-amargoso foi similar. A combinação das formulações de glifosato com tais adjuvantes são potencialmente mais eficazes e garantem controle satisfatório do capim-amargoso.

Quality Control of Herbal Medicines: From Traditional Techniques to State-of-the-art Approaches.

Herbal medicines are important options for the treatment of several illnesses. Although their therapeutic applicability has been demonstrated throughout history, several concerns about their safety and efficacy are raised regularly. Quality control of articles of botanical origin, including plant materials, plant extracts, and herbal medicines, remains a challenge. Traditionally, qualitative (e.g., identification and chromatographic profile) and quantitative (e.g., content analyses) markers are applied for this purpose. The compound-oriented approach may stand alone in some cases (e.g., atropine in Atropa belladonna). However, for most plant materials, plant extracts, and herbal medicines, it is not possible to assure quality based only on the content or presence/absence of one (sometimes randomly selected) compound. In this sense, pattern-oriented approaches have been extensively studied, introducing the use of multivariate data analysis on chromatographic/spectroscopic fingerprints. The use of genetic methods for plant material/plant extract authentication has also been proposed. In this study, traditional approaches are reviewed, although the focus is on the applicability of fingerprints for quality control, highlighting the most used approaches, as well as demonstrating their usefulness. The literature review shows that a pattern-oriented approach may be successfully applied to the quality assessment of articles of botanical origin, while also providing directions for a compound-oriented approach and a rational marker selection. These observations indicate that it may be worth considering to include fingerprints and their data analysis in the regulatory framework for herbal medicines concerning quality control since this is the foundation of the holistic view that these complex products demand.

Calidad composicional y concentración de ácidos grasos omega-3 (alfa-linolénico) y omega-6 (linoleico) presentes en leche bovina de tres regiones naturales del Ecuador / Compositional quality and concentration of omega-3 (alpha-linolenic) and omega-6 (linoleic) fatty acids present in bovine milk from three natural regions of Ecuador

RESUMEN La leche de vaca es un componente importante en la dieta humana y uno de sus aportes nutricionales es la fracción lipídica formada por diversos ácidos grasos, entre ellos, el ácido linoleico (AL) de familia omega-6 y el ácido alfa-linolénico (AAL) de familia omega-3, ambos constituyentes estructurales de membranas de tejidos celulares y reguladores metabólicos. Por su importancia, el objetivo de esta investigación fue determinar la concentración de ácidos grasos omega-3 (alfa-linolénico) y omega-6 (linoleico) mediante cromatografía de gases acoplada a espectrometría de masas (GC-MS), en relación con la influencia de los factores región (Costa, Sierra y Amazonia) y época (lluviosa y seca) sobre la concentración de dichos ácidos. Se trabajó con 30 centros de acopio y se recolectó según el protocolo LCL-INS-01. El análisis composicional se realizó bajo el método ISO 9622-IDF 141/2013 /LCL-PE-01 y el análisis del perfil lipídico mediante GC-MS. Los resultados obtenidos mostraron concentraciones (%) promedio de 2,72 y 0,06 para AL y AAL, respectivamente, en el perfil lipídico. En cuanto al factor región, Costa presentó 2,07%, Sierra 3,03% y Amazonía 3,06%, por lo que se evidenció alta diferencia significativa (p ≤ 0,01) para el AL, mientras que el AAL no mostró variación. En el factor época, el AL presentó 2,63% en época seca y 3,03% en época lluviosa, y el AAL 0,14 y 0,06%, respectivamente. Los resultados permitieron concluir que el factor región influye en la concentración del AL, pero no en el AAL, y el factor época no es influyente en la concentración del AL ni en la del AAL.

ABSTRACT Cow's milk is an important component in human diet and one of its nutritional contributions is the lipid fraction formed by various fatty acids, including linoleic acid (AL) of the omega-6 family and alpha-linolenic acid (AAL) of the omega-3 family, both structural constituents of cell tissue, membranes, and metabolic regulators. Due to its importance, the purpose of this research was to determine the concentration of omega-3 (alpha-linolenic) and omega-6 (linoleic) fatty acids present in bovine milk, by gas chromatography coupled to mass spectrophotometry (GC-MS), establishing a relation between the influence of the region (Costa, Sierra and Amazonia) and the season (rainy and dry), and the concentration of these acids. 30 collection centers were analyzed and collected, according to the LCL-INS-01 protocol. The compositional analysis was carried out under the method ISO 9622-IDF 141/2013 /LCL-PE-01, and the analysis of the lipid profile was made by GC-MS. The results obtained show average concentrations (%) of 2,72 and 0,06 for AL and AAL respectively on the lipid profile. Regarding the region factor, Costa presented 2,07%, Sierra 3,03% and Amazonia 3,06%, showing significant difference (p ≤ 0,01) in AL, while the AAL showed no variation. With reference to the season factor, AL showed 2,63% in the dry season and 3,03% in the rainy season, and AAL 0,14 and 0,06% respectively. The results allowed to conclude that the region factor influences AL concentration but does not influence in AAL, and the season factor is not influential neither on AL nor on AAL concentration.

Identification of type B trichothecenes and zearalenone in Chilean cereals by planar chromatography coupled to mass spectroscopy.

High-performance thin-layer chromatography (HPTLC) and HPTLC coupled with mass spectrometry (MS) methods were described for the simultaneous determination of zearalenone (ZEA); type B trichothecenes (TCT-B); nivalenol (NIV) and deoxynivalenol (DON) along with its acetylated derivatives: 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON). The extract samples were cleaned-up with Bond Elut Mycotoxin® solid-phase extraction cartridges. Then, separation was performed on HPTLC silica gel 60 F254 plates using toluene, ethyl acetate and formic acid (1:8:1 v/v/v) as mobile phase. Derivatisation was then performed with 10% aluminium trichloride in 50% methanol. Mycotoxin standards and spiked cereals grains were identified by UV spots at 366 nm, with retention factors (RF) of 0.20 (NIV), 0.39 (DON), 0.45 (15-ADON), 0.50 (3-ADON) and 0.60 (ZEA). Some parameters of validation were determined. Calibration data (n = 5) fitted a linear regression model with determination coefficients, R2 > 0.99. The recovery was determined in triplicate at two levels, ranging from 84.3 ± 2.2% to 114.2 ± 11.7%. Detection limits ranged from 80 to 120 µg kg-1 and quantification limits ranged from 120.0 to 200 µg kg-1. The analysis by HPTLC/electrospray (ESI)-MS in negative mode confirmed the presence of TCT-B and ZEA standards in Chilean cereals with mass signals at m/z 355, 371, 337, and 317 for DON, NIV, 3-ADON and 15-ADON, and ZEA, respectively.

Antifungal Activity of Chemical Constituents from Piper pesaresanum C. DC. and Derivatives against Phytopathogen Fungi of Cocoa.

In this study, the antifungal potential of chemical constituents from Piper pesaresanum and some synthesized derivatives was determined against three phytopathogenic fungi associated with the cocoa crop. The methodology included the phytochemical study on the aerial part of P. pesaresanum, the synthesis of some derivatives and the evaluation of the antifungal activity against the fungi Moniliophthora roreri, Fusarium solani and Phytophthora sp. The chemical study allowed the isolation of three benzoic acid derivatives (1-3), one dihydrochalcone (4) and a mixture of sterols (5-7). Seven derivatives (8-14) were synthesized from the main constituents, of which compounds 9, 10, 12 and 14 are reported for the first time. Benzoic acid derivatives showed strong antifungal activity against M. roreri, of which 11 (3.0 ± 0.8 µM) was the most active compound with an IC50 lower compared with positive control Mancozeb® (4.9 ± 0.4 µM). Dihydrochalcones and acid derivatives were active against F. solani and Phytophthora sp., of which 3 (32.5 ± 3.3 µM) and 4 (26.7 ± 5.3 µM) were the most active compounds, respectively. The preliminary structure-activity relationship allowed us to establish that prenylated chains and the carboxyl group are important in the antifungal activity of benzoic acid derivatives. Likewise, a positive influence of the carbonyl group on the antifungal activity for dihydrochalcones was deduced.

Polyphenolic Compounds Extracted and Purified from Buddleja Globosa Hope (Buddlejaceae) Leaves Using Natural Deep Eutectic Solvents and Centrifugal Partition Chromatography.

Chemical profiling of Buddleja globosa was performed by high-performance liquid chromatography coupled to electrospray ionization (HPLC-DAD-ESI-IT/MS) and quadrupole time-of-flight high-resolution mass spectrometry (HPLC-ESI-QTOF/MS). The identification of 17 main phenolic compounds in B. globosa leaf extracts was achieved. Along with caffeoyl glucoside isomers, caffeoylshikimic acid and several verbascoside derivatives (ß-hydroxyverbascoside and ß-hydroxyisoverbascoside) were identified. Among flavonoid compounds, the presence of 6-hydroxyluteolin-7-O-glucoside, quercetin-3-O-glucoside, luteolin 7-O-glucoside, apigenin 7-O-glucoside was confirmed. Campneoside I, forsythoside B, lipedoside A and forsythoside A were identified along with verbascoside, isoverbascoside, eukovoside and martynoside. The isolation of two bioactive phenolic compounds verbascoside and forsythoside B from Buddleja globosa (Buddlejaceae) was successfully achieved by centrifugal partition chromatography (CPC). Both compounds were obtained in one-step using optimized CPC methodology with the two-phase solvent system comprising ethyl acetate-n-butanol-ethanol-water (0.25:0.75:0.1:1, v/v). Additionally, eight Natural Deep Eutectic Solvents (NADESs) were tested for the extraction of polyphenols and compared with 80% methanol. The contents of verbascoside and luteolin 7-O-glucoside after extraction with 80% methanol were 26.165 and 3.206 mg/g, respectively. Among the NADESs tested in this study, proline- citric acid (1:1) and choline chloride-1, 2- propanediol (1:2) were the most promising solvents. With these NADES, extraction yields for verbascoside and luteolin 7-O-glucoside were 51.045 and 4.387 mg/g, respectively. Taken together, the results of this study confirm that CPC enabled the fast isolation of bioactive polyphenols from B. globosa. NADESs displayed higher extraction efficiency of phenolic and therefore could be used as an ecofriendly alternative to classic organic solvents.

Chromatography-Independent Fractionation and Newly Identified Molecular Features of the Adzuki Bean (Vigna angularis Willd.) ß-vignin Protein.

Adzuki seed ß-vignin, a vicilin-like globulin, has proven to exert various health-promoting biological activities, notably in cardiovascular health. A simple scalable enrichment procedure of this protein for further nutritional and functional studies is crucial. In this study, a simplified chromatography-independent protein fractionation procedure has been optimized and described. The electrophoretic analysis showed a high degree of homogeneity of ß-vignin isolate. Furthermore, the molecular features of the purified protein were investigated. The adzuki bean ß-vignin was found to have a native size of 146 kDa, and the molecular weight determined was consistent with a trimeric structure. These were identified in two main polypeptide chains (masses of 56-54 kDa) that are glycosylated polypeptides with metal binding capacity, and one minor polypeptide chain with a mass 37 kDa, wherein these features are absent. The in vitro analysis showed a high degree of digestibility of the protein (92%) and potential anti-inflammatory capacity. The results lay the basis not only for further investigation of the health-promoting properties of the adzuki bean ß-vignin protein, but also for a possible application as nutraceutical molecule.

Analytical determinations of luteolin.

Plants, through the photosynthesis process, produce the substances necessary for all the life cycles of nature, which are called "primary metabolites." Moreover, there are some plants that synthesize, in addition to these, other substances with more specific functions, which are known as "secondary metabolites." It is inside this group that flavonoids are located, whose main function is to protect organisms from damage caused by different oxidizing agents. Luteolin (3,4,5,7-tetrahydroxy-flavone) belongs to the sub-class of flavonoids known as flavones and is one of 10,000 flavonoids currently known, being one of the most bio-active flavonoids. Its various beneficial properties for health, together with the increasing reduction in the use of synthetic antioxidants, make the study of luteolin a very active field. Within this, the quantification of this molecule has become a subject of very special interest given that it is transversal to all fields. In this review article, we aim to give the reader a broad and deep vision of this topic, focusing on the events reported in the last 5 years and covering all possible techniques related to analytical determinations. We will discuss in terms of advantages and disadvantages between techniques, selectivity, sensitivity, costs, time consumption, and reagents as well as in the complexity of operations.

Caracterización Química y Actividad Antibacteriana in vitro de un Extracto Etanólico de Propóleo Peruano Frente a Streptococcus mutans / Chemical characterization and in vitro antibacterial activity of a Peruvian propolis ethanolic extract against Streptococcus mutans

El objetivo del estudio fue caracterizar químicamente un extracto etanólico de propóleo peruano y evaluar su actividad antibacteriana frente a Streptococcus mutans. (S. mutans). Se obtuvo el extracto etanólico de propóleo (EEP) por maceración en alcohol al 70 % durante 15 días, el EEP fue sometido a cromatografía en capa fina para identificar sus componentes químicos. El EEP fue diluido con agua destilada para obtener concentraciones de 75 %, 50 % y 25 %. La actividad antibacteriana se realizó mediante la prueba de difusión en disco sobre medio Brain Heart infusion agar(BHA) inoculado con S. mutans ATCC® 25175™, se empleó clorhexidina (CHX) al 0,12 % como control positivo. Las placas fueron incubadas por 48 horas a 37ºC en condiciones de microaerofilia. Posteriormente se realizó la medición de los halos de inhibición con un compás Vernier. Los resultados mostraro n que el EEP presenta como principales componentes terpenos, diterpenos y terpenoidales. Todas las concentraciones del EEP presentaron actividad antibacteriana frente al S. mutans (25 %= 17,582 ± 2,578 mm; 50 % = 16,906 ± 1,892 mm; 75 % = 16,881 ± 2,013 mm; 100 % = 17,201 ± 1,305 mm); sin embargo, fueron menores al compararlos con CHX al 0,12 % (24,543 ± 2,486 mm) (p<0,05). Según la escala de Duraffourd, S. mutans fue sensible (+) y muy sensible (++) para todas las concentraciones del EEP, mientras que para CHX al 0,12 % fue sumamente sensible (+++) (p<0,05). Como conclusión, las diversas concentraciones de EEP peruano presentan actividad antibacteriana significativa considerada como sensible y muy sensible frente a S. mutans.

The objective of the study was to chemically characterize an ethanolic extract of Peruvian propolis and evaluate its antibacterial activity against Streptococcus mutans. (S. mutans). The ethanolic extract of propolis (EEP) was obtained by maceration in 70 % alcohol for 15 days, the EEP was subjected to thin layer chromatography to identify its chemical components. The EEP was diluted with distilled water to obtain concentrations of 75%, 50 % and 25 %. The antibacterial activity was performed by the disk diffusion test on Brain Heart infusion agar (BHA) medium inoculated with S. mutans ATCC® 25175 ™, 0.12 % chlorhexidine (CHX) was used as a positive control. Plates were incubated for 48 hours at 37°C under microaerophilic conditions. Subsequently, the inhibition halos were measured with a Vernier compass. The results showed that the EEP presents as main components terpenes, di-terpenes and terpenoidals. All concentrations of the EEP showed antibacterial activity against S. mutans (25 % = 17,582 ± 2,578 mm; 50 % = 16,906 ± 1,892 mm; 75 % = 16,881 ± 2,013 mm; 100 % = 17,201 ± 1,305 mm); however, they were lower when compared with 0.12 % CHX (24,543 ± 2,486 mm) (p <0.05). According to the Duraffourd´ scale, S. mutans was sensitive (+) and very sensitive (++) for all EEP concentrations, while for 0.12 % CHX it was highly sensitive (+++) (p < 0.05). In conclusion, all concentrations of Peruvian EEP have significant antibacterial activity considered as sensitive and very sensitive against S. mutans.