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.

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 [...].

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 [...].

Profilage phytochimique par chromatographie sur couche mince haute performance d'extraits de flavonoïdes totaux du Drepanoalpha® et évaluation de leur activité anti-drépanocytaire in vitro / In vitro evaluation of anti-sickling activity of crude flavonoid extracted from Drepanoalpha® and their phytochemical profiling by high performance thin layer chromatography

Contexte et objectif. Le défi le plus important dans la drépanocytose consiste à améliorer l'état de santé des patients dans les pays en développement. L'une des meilleures solutions est donc le développement de la phytomédecine basée sur la connaissance de la pharmacopée traditionnelle. L'objectif de la présente étude était d'évaluer les activités anti-drépanocytaires des flavonoïdes totaux extraits du phytomédicament Drépanoalpha® d'une part et déterminer leur profilage chimique par chromatographie sur couche mince haute performance d'autre part. Méthodes. Les flavonoïdes totaux ont été obtenus par fractionnement de l'extrait méthanolique par chromatographie flash (PURIFLASH COLUMN 30 SILICA HP - 12,0 g) et purifies à l'aide d'une cartouche (Polymeric Reversed Phase) puis caractérisés et dosés par chromatographie sur couche mince haute performance (CCMHP). L'activité anti-drépanocytaire a été mise en évidence grâce aux tests d'Emmel, de polymérisation, de rapport Fe2+/Fe3+, d'hémolyse et de la fragilité osmotique membranaire. Résultats. La poudre du Drépanoalpha® contenait une quantité de flavonoïdes totaux de 8,14 mg équivalent de quercétine/g d'extrait. Les flavonoïdes totaux extraits du Drépanoalpha® possèdent une activité antifalcémiante (avec le taux maximal de normalisation d'environ 90 % et une concentration minimale de normalisations de 11,4 µg/mL), un taux d'augmentation du rapport Fe2+/Fe3+ de 97,0 %, une activité anti-hémolytique avec une fragilité corpusculaire membranaire des érythrocytes (FCM) de 0,73 et un taux d'inhibition de la polymérisation de 77,5%. Conclusion. La pertinence des résultats de cette étude permet de confirmer les flavonoids comme phytomarqueur pour le contrôle de qualité et de standardisation de cet alicament.

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.

Antimicrobial and Antibiofilm Effect of Hydrogel with Origanum vulgare on Culture of Streptococcus mutans and Streptococcus sobrinus / Efecto Antimicrobiano y Antibiofilm de Hidrogel a Base de Origanum vulgare sobre Cultivo de Streptococcus mutans y Streptococcus sobrinus

The oral cavity is an ecosystem that provides ideal conditions for the growth of bacteria, the Streptococcus genus is important for the formation of biofilms that lead to the development of dental caries, which affects the population worldwide. The world health organization encourages the use of plants thanks to its various therapeutic actions. Origanum vulgare L. (oregano), is an aromatic plant with medicinal and culinary properties. The objective of this study was to investigate the in vitro antimicrobial and antibiofilm activity of the ethanolic extract of oregano, against the growth of Streptococcus mutans and Streptococcus sobrinus ATCC. Leaves of the plant were obtained and the ethanolic extract was made by maceration. Antimicrobial activity was evaluated using the Kirby-Bauer method and compared with 2% chlorhexidine, subsequently the extract was incorporated into a hydrogel and its effect on biofilm formation was assessed by fluorescence microscopy and the main compounds were identified. present in the extratco. The study revealed that the extract presented antimicrobial effect against both strains and at 2% it showed high antimicrobial action compared to chlorhexidine at the same concentration, with average inhibition halos of 26.3 mm and 19 mm for each microorganism analyzed, (p < 0.05). Likewise, the hydrogel prepared with 2% extract significantly eliminated the preformed Streptococcus biofilm, at 24 hours of exposure, due to the presence of a variety of chemical groups, such as sterols, triterpenes, flavonoids, flavanones, flavanonol s, lactones. sesquiterpenic, tannins and coumarins. The oregano extract presented high antimicrobial action for both species, with a greater effect towards Streptococcus mutans and an interesting antibiofilm action; These results show the importance of exploring treatment alternatives of plant origin, to be considered as interesting complementary aids in dental therapy.

La cavidad oral es un ecosistema que proporciona condiciones ideales para el crecimiento de bacterias, el género Streptococcus es importante para la formación de biopelículas que conducen al desarrollo de caries dental, que afecta a la población a nivel mundial. La organización mundial de la salud, fomenta el uso de plantas gracias a sus diversas acciones terapéuticas. Origanum vulgare L. (orégano), es una planta aromática con propiedades medicinales y culinarias. El objetivo de este estudio fue investigar la actividad antimicrobiana y antibiofilm in vitro del extracto etanólico de oregano, contra el crecimiento de Streptococcus mutans y Streptococcus sobrinus ATCC. Se obtuvierón hojas de la planta y se realizó el extracto etanólico mediante maceración. La actividad antimicrobiana se evaluó mediante el método de Kirby-Bauer y se comparó con la clorhexidina al 2 %, posteriormente se incorporó el extracto en un hydrogel y se valoró su efecto sobre la formación del biofilm mediante microscopía de fluorescencia y se identificó los principales compuestos presentes en el extratco. El estudio reveló que el extracto presentó efecto antimicrobiano contra ambas cepas y al 2 % mostró alta acción antimicrobiana en comparación con la clorhexidina a la misma concentración, con halos de inhibición promedio de 26.3 mm y de 19 mm para cada microorganismo analizado, (p < 0.05). Así mismo, el hidrogel preparado con extracto al 2 %, eliminó significativamente la biopelícula preformada de Streptococcus, a las 24 horas de exposición, debido a la presencia de una variedad de grupos químicos, como esteroles, triterpenos, flavonoides, flavanonas, flavanonoles, lactonas sesquiterpénicas, taninos y cumarinas. El extracto de orégano presentó alta acción antimicrobiana para ambas especies, con mayor efecto hacia el Streptococcus mutans y una acción antibiofilm interesante; estos resultados muestran la importancia de explorar en alternativas de tratamiento de origen vegetal, para considerarse como auxiliares complementarios interesantes en la terapia dental.

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.

Effect of reciprocating instrumentation on chlorhexidine substantivity on human dentin: chemical analysis followed by confocal laser microscopy

The present research analyzed the reciprocating instrumentation associated to chlorhexidine (CHX) substantivity as its correlation with E. faecalis viability in ex vivo root canals. Eighty extracted single-rooted human teeth were used, being 40 to high-performance liquid chromatography (HPLC) and 40 to confocal laser scanning microscopy (CLSM). In both, teeth were decoronated and the cervical third was prepared. In the CLSM analysis, the root canals were inoculated with E. faecalis for 14 days. Samples were divided into 4 groups (n=10) according to instrumentation technique: no instrumentation and irrigation with distilled water (control); manual instrumentation (K-File); rotary instrumentation (ProTaper Next); and reciprocating instrumentation (Reciproc R25). Two percent chlorhexidine was applied as irrigating substance in experimental groups. Longitudinal grooves resulted in 2 halves root and 20 proof bodies in each group. Samples were divided by chance in two groups (n=10) and the outcomes were evaluated after two days and one week. The retained chlorhexidine and live cells after instrumentation techniques in each evaluation time was measured by HPLC and CLSM, respectively. Specific analysis was applied for experimental tests (p≤0.05). Both rotary as well as reciprocating techniques significantly reduced the amount of chlorhexidine on dentin in all observation periods (p<0.05). After evaluation times, all experimental groups presented lower live cells compared to control, but without statistically difference. Intragroup comparisons in times of evaluation showed no differences in instrumentation techniques, in chlorhexidine retention and number of live cells (p>0.05). Reciprocating instrumentation does not interfere on chlorhexidine substantivity.

Studies on phenylpropanoids from Eleocharis dulcis and their hepatoprotective activities / 中国中药杂志

To study phenylpropanoids from Eleocharis dulcis and their hepatoprotective activities. The compounds were separated and purified from ethyl acetate part by conventional column chromatography and preparative liquid chromatography, and their structures were identified by various spectral techniques. The HL-7702 cells damage model of hepatocytes induced by APAP was used to screen and evaluate the hepatoprotective activities of these compounds. Sixteen compounds were isolated from ethyl acetate part of E. dulcis, and their structures were identified as 6'-(4″-hydroxy-3″-methoxy-phenylpropenyl)-1-(10-methoxy-phenylacetone)-1'-O-β-D-glucopy-ranoside(1), susaroyside A(2), clausenaglycoside B(3), clausenaglycoside C(4), clausenaglycoside D(5), emarginone A(6), emarginone B(7), thoreliin B(8), 4-O-(1',3'-dihydroxypropan-2'-yl)-dihydroconiferyl alcohol 9-O-β-D-glucopyranoside(9), 2-[4-(3-methoxy-1-propenyl)-2-methoxy-phenoxy]-propane-1,3-diol(10), 6'-O-(E-cinnamoyl)-coniferin(11), methyl 3-(2-O-β-D-glucopyranosyl-3,4,5,6-tetramethoxyphenyl) propanoate(12), clausenaglycoside A(13), 9-O-(E-cinnamoyl)-coniferin(14), 6'-O-(E-cinnamoyl)-syringin(15), 2'-O-(E-cinnamoyl)-syringin(16). Among them, compound 1 was a new compound. Compounds 2-16 were isolated from this plant for the first time. Among them, compounds 2 and 8 showed certain hepatoprotective activities.

Verificação de adulterações em cúrcuma, gengibre, noz-moscada, páprica, pimenta-do-reino e colorífico, comercializados no estado de São Paulo, Brasil. (Parte II) / Verification of adulterations in turmeric, ginger, nutmeg, paprika, black pepper and colorífico marketed in the state of São Paulo, Brazil. (Part II)

Especiarias são produtos constituídos de partes de espécies vegetais com importante valor alimentício e diversos benefícios para a saúde. O objetivo deste trabalho foi pesquisar adulterações na composição de cúrcuma (Curcuma longa Linnaeus), gengibre (Zingiber officinale Roscoe), noz-moscada (Myristica fragrans Houttuyn), páprica (Capsicum annuum Linnaeus), pimenta-do-reino (Piper nigrum Linnaeus) e colorífico (mistura de urucum, Bixa orellana Linnaeus, com fubá). Foram analisadas 180 amostras adquiridas em municípios do estado de São Paulo. A investigação dos elementos histológicos foi feita por microscopia óptica, a análise dos corantes por cromatografia em papel e a quantificação da bixina por cromatografia líquida de alta eficiência. Das amostras analisadas, 16,1% apresentaram elementos histológicos estranhos ao produto, sendo que nenhuma amostra apresentou corante orgânico artificial. A concentração de bixina nas amostras de colorífico variou entre 0,6 e 105,3 mg/100g, com média de 18,9 mg/100g e desvio padrão de 17,7 mg/100g. A avaliação microscópica revelou que a maioria das adulterações ocorre pela adição de amido de Zea mays. O colorífico não apresentou adulterações, porém foi constatada a necessidade de uma padronização da concentração de bixina. Este estudo demonstrou a necessidade da intensificação do monitoramento de adulterações em especiarias para que a comercialização de alimentos fidedignos seja garantida. (AU)

Spices are products made up of parts of plant species, with important nutritional value and many health benefits. The aim of this work was to evaluate adulterations in turmeric (Curcuma longa Linnaeus), ginger (Zingiber officinale Roscoe), nutmeg (Myristica fragrans Houttuyn), paprika (Capsicum annuum Linnaeus), black pepper (Piper nigrumLinnaeus) and colorific (mixture containing Bixa orellana with cornmeal). A total of 180 samples purchased in the municipalities of the state of São Paulo were analyzed. The investigation of the histological elements was performed by optical microscopy, the analysis of the dyes was carried out using paper chromatography and the quantification of the bixin was performance by high performance liquid chromatography. Of the 180 samples analyzed, 16.1% presented strange histological elements, classified as adulterations. Among the adulterated samples, none showed organic dye. Bixin analysis was carried out on colorific samples, ranging from 0.6 ­ 105.3 mg/100g, with an average of 18.9 mg/100g and standard deviation of 17.7 mg/100g, demonstrating the need to regulate the annatto extract concentration range added into the condiment. The evaluation demonstrated the necessity to monitor adulteration in spices, so that producers and merchants provide food with quality to the consumer. (AU)

Production, Purification and Characterization of a Novel Bacteriocin Produced by Bacillus subtilis VS Isolated from Mango (Mangifera indica L)

Abstract Bacteriocin has been identified as an excellent alternative to chemical preservatives due to its astonishing antimicrobial activity against food spoiling and food-borne pathogens. So there is a need to identify the newer and potent sources of bacteriocin producers. This study aims the isolation of potent bacteriocin producing microorganism from fresh fruits and vegetables, its production, purification, and characterization. Firstly, 43 isolates were analysed for its antimicrobial potential, out of which7 were found to inhibit the growth of various pathogens. Considering the results of antimicrobial activity; the microorganism isolated from mango was regarded as the most potent one; which was identified as Bacillus subtilis VS.70% ammonium sulphate precipitated and dialysed bacteriocin was purified using DEAE cellulose and sephadex G75 chromatography. Bacteriocin was purified by 24.64 fold with 8.65% recovery and its molecular weight was found to be 31.2kDa. The Purified bacteriocin was found to be stable at broad pH and temperature. It was found to be degraded by various proteases studied confirming its proteinaceous nature. Considering all these attributes; the purified bacteriocin isolated from Bacillus subtilis VS can be exploited by various food industries.

Evaluation of Residual Monomer Release After Polymerization of Colored Compomer Materials

ABSTRACT Objective: To evaluate the amount of residual monomers released after polymerization by the compomers in different colors and viscosities over time. Material and Methods: The compomer samples of different colors and viscosities (flowable compomers; blue-pink and packable compomers; A2-blue-pink-gold) were prepared in molds with an inner diameter of 5 mm and a height of 2 mm. In polymerization of samples, a LED unit was used. The amount of monomers released from the samples kept in 75% ethanol/water solution was measured by a high-performance liquid chromatography (HPLC) instrument in the 10th minute, in the 1st hour, and in the 1st, 7th, and 14th days. For statistical analyses, the paired sample t-test, independent sample t-test, and one-way ANOVA with Tukey's post hoc test were used. Results: The amount of residual monomers released from all materials increased over time. At the end of the 14th day, the most released monomer from all compomer samples was BisGMA. The total amounts of released monomers from the packable compomers were Gold>A2>blue>pink. The amount of residual monomers released from flowable compomers was higher in blue than in pink. Conclusion: The color and the viscosity are the factors affecting the residual monomer release in compomers.

Actividad antimicrobiana de propóleos de abejas sin aguijón en combinación con ajo, Allium sativum (Amaryllidaceae) / Stingless bees propolis antimicrobial activity in combination with garlic, Allium sativum (Amaryllidaceae)

The excessive use of antibiotics has increased pathogenic microorganisms resistance, which derives in patient mortality. Therefore, the strategies for searching new natural and unconventional strategies has become constant and important. Objective: To determine the antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) against stingless bees propolis. Methods: We evaluated nine (20 %) propolis from stingless bees ethanolic extracts from different regions: Melipona beecheii, Melipona solani, Tetragonisca angustula and Scaptotrigona mexicana. The chemical characterization was performed by liquid chromatography (HPLC) and microbial resistance tests by the macrodilution method, to determine the effect of the combinations. Results: The compounds that were of interests and the most abundant were the following; hydroxycinnamic acids, flavanones, flavonoids and the glycosylated derivatives. Four of the nine propolis were effective against MRSA, which came from the following species; one from M. solani and three from S. mexicana. The minimum inhibitory concentration for S. mexicana was in the range of 3-8 mg mL-1 and for M. solani it was 4 mg mL-1. Isobolographic studies resulted in an additive effect (ɤ = 1) for the combination of Allium sativum with the S. mexicana propolis sambles and an antagonistic effect (ɤ > 1) for the combination of A. sativum with the propolis of M. solani. Conclusions: the combination of extracts with lower concentrations of A. sativum, may be the most effective, than those that were individually tested. More detailed studies are required to define the mechanisms of stingless bees propolis as well as their combination with other organic substances.

Introducción: El uso excesivo de antibióticos ha traído el aumento de resistencia en los microorganismos patógenos que provocan mortalidad de los pacientes. Por consiguiente, la búsqueda de nuevas estrategias naturales y no convencionales se ha vuelto constante e importante. Objetivo: Determinar la actividad antimicrobiana contra Staphylococcus aureus resistente a meticilina (SARM) de los propóleos por abejas sin aguijón. Métodos: Nueve fueron los extractos etanolicos de propóleos al 20 % de las abejas sin aguijón de distintas regiones: de Melipona beecheii, de Melipona solani, de Tetragonisca angustula y de Scaptotrigona mexicana. Se realizó la caracterización química de los compuestos por cromatografía liquida y las pruebas de resistencia microbiana por el método de macrodilución para determinar el efecto de las combinaciones. Resultados: Los compuestos de interés fueron detectados, y destacan como los más abundantes los ácidos hidroxicinámicos, flavanonas, flavonoides y sus derivados glicosilados. Cuatro de los nueve propóleos resultaron efectivos contra SARM, los cuales provinieron, uno de Melipona solani y tres de Scaptotrigona mexicana. La CMI para S. mexicana está en el rango de 3-8 mg mL-1 y para M. solani fue de 4 mg mL-1. Los estudios isobolográficos dieron como resultado un efecto aditivo (ɤ = 1) para la combinación de Allium sativum con los 3 propóleos de S. mexicana y un efecto antagónico (ɤ > 1) para la combinación de A. sativum con el propóleos de M. solani. Conclusiones: la combinación de extractos menores puede ser más efectiva que usando la CE50 de los extractos de forma individual. Se requieren estudios más detallados para definir los mecanismos de los propóleos de las abejas sin aguijón, así como su combinación con otras sustancias orgánicas.

Caracterización de los ácidos grasos y actividad antimicrobiana del extracto en metanol de Holothuria princeps (Holothuriida: Holothuriidae) / Characterization of fatty acids and antimicrobial activity of the methanol extract of Holothuria princeps (Holothuriida: Holothuriidae)

Introduction: In recent decades, studies related to the search and characterization of bioactive molecules in marine organisms have increased exponentially, demonstrating the enormous wealth of secondary metabolites of diverse structural composition that cannot be found in organisms present in the terrestrial environment. A significant number of the new marine natural compounds discovered have contributed to solving some of the problems of humanity, mainly those related to human health. Objective: The purpose of this research is to evaluate the bactericidal and fungicidal activities of the methanolic extract of sea cucumber Holothuria princeps collect from the bay of Cispatá in the Colombian Caribbean, in addition to chemically identifying its fatty acids. Methods: A methanolic extraction was performed from the collected biological material, by the cold maceration method. The extract obtained was fractionated using chromatographic techniques and the fatty acids were obtained, which were derivatized and identified by means of gas chromatography in coupling with mass spectrometry. The antibacterial and antifungal activities of the methanolic extract of Holothuria princeps was performed through the microdilution method against reference strains and clinical isolates. Results: We found 16 fatty acids present in Holothuria princeps according to the analysis of their mass spectra. Antibacterial activity showed that Enterococcus faecalis was the most susceptible to the extract at low concentrations, while Pseudomonas aeruginosa was the highest at the higher concentrations. In antifungal treatment, the fungus with the highest inhibition was the clinical isolate of Candida albicans (blood sample). Conclusions: Taking into account previous studies in the genus Holothuria, it is considered that the environment plays a fundamental role in the presence and diversity of fatty acids. The evaluation of the antibacterial activity against reference strains of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Enterococcus faecalis demonstrated the existence of a considerable effect in the reduction of bacterial growth by the extracts applied, mainly at low concentrations (less than 1 000 ppm). On the other hand, the antifungal activity against the reference strain of Candida albicans and the clinical isolates of Candida albicans (blood sample) and Candida krusei (catheter sample), the extract showed that the best results were presented at higher concentrations (above 1 500 ppm).

Introducción: En las últimas décadas los estudios relacionados con la búsqueda y caracterización de moléculas bioactivas en organismos marinos han aumentado de una manera exponencial, lo que demuestra la enorme riqueza de metabolitos secundarios de diversa composición estructural que no pueden ser encontrados en organismos presentes en el medio terrestre. Estas nuevas moléculas halladas poseen numerosas actividades biológicas que ayudan a resolver muchos problemas que ha tenido el hombre a lo largo de su existencia, lo que las convierte en productos de gran importancia para la humanidad. Objetivo: El propósito de este estudio es la identificación de los ácidos grasos presentes en el pepino de mar Holothuria princeps recolectado en costas del Caribe colombiano, además del análisis de las actividades antibacterianas y antifúngicas de su extracto metanólico frente a cepas de referencia y aislados clínicos. Métodos: Del material biológico recolectado se realizó una extracción metanólica usando el método de maceración en frío. El extracto obtenido se fraccionó usando cromatografía en columna y se lograron obtener los ácidos grasos, los cuales fueron derivatizados e identificados por medio de cromatografía de gases en acople con espectrometría de masas. La actividad antibacteriana y antifúngica del extracto metanólico de Holothuria princeps se realizó a través del método de microdilución. Resultados: Los resultados arrojaron la identificación de 16 ácidos grasos presentes en Holothuria princeps de acuerdo con el análisis de sus espectros de masas. La actividad antibacteriana mostró que Enterococcus faecalis fue la bacteria más susceptible al efecto del extracto a bajas concentraciones, mientras que a las más altas lo fue Pseudomonas aeruginosa. A nivel general en el tratamiento antifúngico, el hongo que presentó una mayor inhibición fue el aislado clínico de Candida albicans (muestra de sangre). Conclusiones: Teniendo en cuenta estudios previos en organismos del mismo género, se puede considerar en cuanto a los ácidos grasos identificados, que el entorno juega un papel fundamental en la presencia y composición de estos compuestos. La evaluación de la actividad antibacteriana contra cepas de referencia de Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae y Enterococcus faecalis, demostró la existencia de un efecto considerable en la reducción del crecimiento bacteriano por parte de los extractos utilizados, principalmente a bajas concentraciones (menos de 1 000 ppm). En cuanto

Actividad antiproliferativa de Aldama arenaria (Baker) E. E. Schill. & Panero / Antiproliferative activity from Aldama arenaria (Baker) E. E. Schill. & Panero

El extracto de cloroformo (CE) y las fracciones obtenidas de las raíces de Aldama arenaria se evaluaron para determinar su actividad antiproliferativa in vitro contra 10 líneas celulares tumorales humanas [leucemia (K-562), mama (MCF-7), ovario que expresa un fenotipo resistente a múltiples fármacos (NCI/ADR-RES), melanoma (UACC-62), pulmón (NCI-H460), próstata (PC-3), colon (HT29), ovario (OVCAR-3), glioma (U251) y riñón (786-0)]. CE presentó actividad antiproliferativa débil a moderada (log GI50 medio 1.07), mientras que las fracciones 3 y 4, enriquecidas con diterpenos de tipo pimarane [ent-pimara-8 (14), ácido 15-dien-19-oico y ent-8(14),15-pimaradien-3ß-ol], presentaron actividad moderada a potente para la mayoría de las líneas celulares, con un log GI50 medio de 0.62 y 0.59, respectivamente. Los resultados mostraron una acción antiproliferativa in vitro prometedora de las muestras obtenidas de A. arenaria, con los mejores resultados para NCI/ADR-RES, HT29 y OVCAR-3, y valores de TGI que van desde 5.95 a 28.71 µg.mL-1, demostrando que los compuestos de esta clase pueden ser prototipos potenciales para el descubrimiento de nuevos agentes terapéuticos.

Chloroform extract (CE) and fractions obtained from Aldama arenaria roots were evaluated for their in vitro antiproliferative activity against 10 human tumor cell lines [leukemia (K-562), breast (MCF-7), ovary expressing a multidrug-resistant phenotype (NCI/ADR-RES), melanoma (UACC-62), lung (NCI-H460), prostate (PC-3), colon (HT29), ovary (OVCAR-3), glioma (U251), and kidney (786-0)]. CE presented weak to moderate antiproliferative activity (mean log GI50 1.07), whereas fractions 3 and 4, enriched with pimarane-type diterpenes [ent-pimara-8(14),15-dien-19-oic acid and ent-8(14),15-pimaradien-3ß-ol], presented moderate to potent activity for most cell lines, with mean log GI50 of 0.62 and 0.59, respectively. The results showed promising in vitro antiproliferative action of the samples obtained from A. arenaria, with the best results for NCI/ADR-RES, HT29, and OVCAR-3, and TGI values ranging from 5.95 to 28.71 µg.mL-1, demonstrating that compounds of this class may be potential prototypes for the discovery of new therapeutic agents.

Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation

Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomotor modulation, and in the insect-blood feeding by cleaving and activating protease-activated receptors (PARs). Methods T. infestans saliva was chromatographed as previously reported for purification of triapsin, a serine protease. The cleavage activity of triapsin on PAR peptides was investigated based on FRET technology. Mass spectrometry was used to analyze the sites of PAR-2 peptide cleaved by triapsin. NO measurements were performed using the DAN assay (2,3-diaminonapthalene). The vasorelaxant activity of triapsin was measured in vessels with or without functional endothelium pre-contracted with phenylephrine (3 µM). Intravital microscopy was used to assess the effect of triapsin on mouse skin microcirculation. Results Triapsin was able to induce hydrolysis of PAR peptides and showed a higher preference for cleavage of the PAR-2 peptide. Analysis by mass spectrometry confirmed a single cleavage site, which corresponds to the activation site of the PAR-2 receptor. Triapsin induced dose-dependent NO release in cultured human umbilical vein endothelial cells (HUVECs), reaching a maximum effect at 17.58 nM. Triapsin purified by gel-filtration chromatography (10-16 to 10-9 M) was applied cumulatively to mouse mesenteric artery rings and showed a potent endothelium-dependent vasodilator effect (EC30 = 10-12 M). Nitric oxide seems to be partially responsible for this vasodilator effect because L-NAME (L-NG-nitroarginine methyl ester 300 µM), a nitric oxide synthetase inhibitor, did not abrogate the vasodilation activated by triapsin. Anti-PAR-2 antibody completely inhibited vasodilation observed in the presence of triapsin activity. Triapsin activity also induced an increase in the mouse ear venular diameter. Conclusion Data from this study suggest a plausible association between triapsin activity mediated PAR-2 activation and vasodilation caused by T. infestans saliva.(AU)

Gestión de riesgos relativo al cambio de campaña productiva durante la etapa de desarrollo tecnológico en una instalación multiproducto / Risk-management related to the productive campaign change during technological development stage in a multi-product facility

Los nuevos paradigmas de la industria farmacéutica en el siglo XXI incorporan el uso de las técnicas de gestión de la calidad, necesarias en el cumplimiento de las buenas prácticas de fabricación en el sector biotecnológico. En este caso de estudio, se aplicó la gestión de riesgo en el cambio de campaña entre los ingredientes farmacéuticos activos de futuros candidatos inmunoterapeúticos contra el cáncer, en la etapa de desarrollo tecnológico en una instalación multiproducto certificada. Las causas potenciales de mayor influencia en las fallas son: la calificación del personal de la Dirección de Desarrollo Tecnológico en los procedimientos patrones de operación de la planta, la mezcla entre los componentes y materiales no dedicados utilizados en el proceso de purificación cromatográfica, la documentación en elaboración o aprobación y el establecimiento de las técnicas analíticas en función de la etapa del proyecto. Como resultado se proponen acciones que minimizan los riesgos de la contaminación cruzada y hacen viable un adecuado cambio de campaña entre la fabricación de los inmunoterapéuticos, durante el desarrollo tecnológico en una instalación multiproducto de la industria biotecnológica(AU)

The new paradigms of the pharmaceutical industry in the 21st century introduce the use of modern quality management techniques to comply with good manufacturing practices in the biotechnological area. In this paper, it was applied the risk management for the campaign change among the process for obtaining the active pharmaceutical ingredients to future immunotherapeutic candidates at the technological development stage in a certified multi-product facility. Particularly, the training for the development personnel in the standard operating procedures of the facility, the mixture between the components and non-dedicated materials used in the chromatographic purification process, the documentation in the preparation or approval, and the establishment of analytical techniques depending on the stage of the project are the potential causes of greater influence. As a result, actions are proposed to minimize risks and carry out an adequate campaign change feasible between the manufacture of immunotherapeutics during the technological development stage in a biotechnological multi-product facility(AU)

Aplicación del Análisis de Componentes Principales en el proceso de purificación de un biofármaco / Application of Principal Component Analysis in the purification process of a biopharmaceutical

En este trabajo se presenta la aplicación del Análisis de Componentes Principales, mediante el programa THE UNSCRAMBLER versión 8.0, a los datos registrados en un período de 2 años en la etapa de purificación de una planta de producción de Eritropoyetina Humana Recombinante que está basada en varios pasos cromatográficos, de forma similar a los procesos de purificación de proteínas recombinantes que se utilizan como vacunas preventivas o terapéuticas. Se logró reducir dimensionalidad al obtenerse dos componentes principales que explican el 81 por ciento de la varianza de 18 variables originales relacionadas con cuatro pasos cromatográficos. Como resultado se llegó a definir cuáles son las variables que mayor aporte tienen a la variabilidad del proceso en la etapa de purificación, permitiendo extraer información útil para lograr un mayor entendimiento del proceso y enriquecer las estrategias de control en la planta. Dichos resultados corroboraron experiencias prácticas de especialistas de la planta y permitieron dar recomendaciones a considerar en el plan de verificación continuada del proceso como proponer cinco variables como controles de proceso y tener en cuenta que el rendimiento del segundo paso cromatográfico es el más influyente de los rendimientos considerados en la variabilidad(AU)

This paper presents the application of the Principal Component Analysis, using the program THE UNSCRAMBLER version 8.0, to the data recorded during two years in the purification stage of a Recombinant Human Erythropoietin plant that is based on several chromatographic steps, similar to the purification process of recombinant proteins that are used as preventive or therapeutic vaccines. Dimensionality was reduced by obtaining two main components that explain 81 percent of the variance of 18 original variables related to four chromatographic steps. As a result, it was possible to define which variables have the greatest contribution to the variability of the process in the purification stage, allowing to extract useful information to achieve a greater understanding of the process and enrich the control strategies in the plant. These results corroborated practical experiences of plant specialists and allowed for recommendations to be considered in the continuous verification plan of the process, such as proposing three variables as process controls and taking into account that the performance of the second step is the most influential of the performances considered in the variability(AU)

Diferencias de especie en las concentraciones plasmáticas y tisulares de florfenicol entre ovinos y conejos / Species differences in plasma and tissue concentrations of florfenicol between sheep and rabbits

RESUMEN Objetivo. Comparar las concentraciones plasmáticas y tisulares de florfenicol (FFC) y su metabolito florfenicol amina (FFC-a) entre ovinos y conejos, posterior a la administración intramuscular de 20 mg/kg de FFC. Materiales y métodos. Cinco ovinos Suffolk Down y seis conejos Neozelandés fueron utilizados en el estudio. Se colectaron muestras de sangre, previo a la administración de FFC, y a las 0.25, 0.5, 1, 1.5, 2, 3 y 4 horas posteriores al tratamiento. A las 4 horas posteriores al tratamiento, a los animales se les aplicó la eutanasia. Las concentraciones plasmáticas y tisulares de FFC y FFC-a fueron determinadas mediante HPLC. Resultados. Las concentraciones plasmáticas máximas, tasa de absorción, vida media de absorción, tasa de distribución y área bajo la curva de FFC, fueron significativamente mayores en conejos respecto a los ovinos. Asimismo, para FFC-a, las concentraciones plasmáticas máximas y área bajo la curva de concentraciones plasmáticas en el tiempo fueron significativamente mayores en conejos respecto a los ovinos. La proporción de metabolito fue mayor en conejos (12.7±3.07%) en comparación con ovinos (3.99±0.87%) (p<0.05), al igual que las concentraciones tisulares de FFC y FFC-a. Conclusiones. Se observaron diferencias significativas en la farmacocinética y concentraciones tisulares de FFC y FFC-a entre estas dos especies. La mayor concentración de FFC-a en conejos indica un mayor nivel de metabolismo de FFC, respecto a los ovinos. Esto es importante de considerar al momento de establecer dosificaciones y frecuencia de administración de FFC en conejos.

ABSTRACT Objective. The aim of this study was to compare tissue and plasma concentrations of florfenicol (FFC) and its metabolite florfenicol amine (FFC-a) between sheep and rabbits, after intramuscular administration of 20 mg FFC/kg. Materials and methods. Five Suffolk Down sheep and six New Zealand rabbits were used in this study. Blood samples were collected before FFC administration and at 0.25, 0.5, 1, 1.5, 2, 3 and 4 hours after treatment. At 4 hours after treatment, euthanasia was applied to animals. Plasma and tissue concentrations of FFC and FFC-a were determined by HPLC. Results. For FFC, maximum plasma concentrations, absorption rate, absorption half-life, distribution rate, and area under the plasma concentration-time curve were all found to be significantly higher in rabbits than in sheep. Similarly, for FFC-a, significantly higher maximum plasma concentrations and area under the concentration-time curve were observed in rabbits as compared to sheep. The metabolite ratio was higher in rabbits (12.7±3.07%) compared to sheep (3.99±0.87%) (p<0.05), as were the tissue concentrations of FFC and FFC-a. Conclusions. Significant differences in the pharmacokinetics and tissue concentrations of FFC, and its metabolite FFC-a, were observed between these two animal species. The higher concentrations of FFC-a in rabbits indicate a greater level of FFC metabolism as compared to sheep. This should be considered when establishing dosage and frequency of FFC administration for rabbits.

Quantification of biogenic amines in fabrication steps of Gorgonzola-type cheese / Quantificação de aminas biogênicas nas etapas de fabricação do queijo tipo Gorgonzola

During the Gorgonzola-type cheese preparation there are proteolysis and lipolysis which may be influenced by the type of starter culture chosen. Six manufacturing steps were selected to identify which of them is most suitable for biogenic amines (BA) formation (1- milk, 2- lactic acid bacterial culture and fungus addition, 3- curd, 4- dry salting, 5- maturation at 30 days and maturation at 60 days); perform research on enterobacteria; accomplish the research of BA-producing bacteria (BAPB); detect and quantify the most abundant BA (putrescine, cadaverine, tyramine, histamine, spermidine and spermine) in the six steps of Gorgonzola cheese production and in bacterial isolates using high performance liquid chromatography and UV-Vis SPD/10AV detector and define if the presence of enterobacteria and BAPB would be correlated with BA production in this cheese. The bacterial culture used increased its log population by 7 log cycles and reached its highest level in batch 2 during cheese maturation. There was a decrease in the enterobacterial population in 2 log cycles after 60 days of maturation in batch 1. Tyramine was the BA with the highest concentration 306.32 mg.Kg-1 quantified in step 6 (60 days maturation) in batch 1. Criterion is requiered in bacterial starter culture selection because it is a quality determinant factor in relation to BA production and more rigor in raw material

Durante a elaboração do queijo tipo Gorgonzola ocorre proteólise a partir das bactérias e dos fungos adicionados ao leite que podem levar a formação de aminas biogênicas (AB) neste tipo de queijo. Portanto, no presente estudo foi feito o acompanhamento com coleta de amostras em seis etapas na fabricação deste queijo paraidentificar em qual delas haveria maior formação de aminas biogênicas (AB). As amostras coletadas em três diferentes lotes foram o leite cru (1), leite pasteurizado adicionado de cultura de bactérias ácido-láticas (2), massa coalhada (3), queijo após a etapa de salga seca (4), queijo após 30 dias de maturação (5) e queijo após 60 dias de maturação (6). Também foram realizadas a pesquisa de enterobactérias e bactérias ácido-láticas com característica capacidade de descarboxilação de aminoácidos e produção de aminas biogênicas (BPAB); detecção e quantificação da AB mais abundante (putrescina, cadaverina, tiramina, histamina, espermidina e espermina) nas seis etapas de fabricação do queijo tipo Gorgonzola e nos isolados bacterianos utilizando cromatografia líquida de alta eficiência e detector UV-Vis SPD/10AV e a verificação se a presença de enterobactérias e BPAB estariam correlacionadas com a produção de AB nesse queijo. A cultura bacteriana utilizada cresceu aumentando em sete ciclos logarítmicos sua população e alcançou seu maior nível no lote 2 na etapa de maturação do queijo. Houve diminuição da população enterobactérias em 2 ciclos logarítmicos após 60 dias de maturação no lote 1. A tiramina foi a AB com concentração mais elevada 306,32 mg.Kg-1 quantificada na etapa 6 (60 dias de maturação) no lote 1. É necessário dar mais atenção em duas etapas na elaboração dos queijos: mais critério na seleção da cultura bacteriana iniciadora por ser um fator determinante na qualidade em relação à produção de AB e mais rigor na seleção da matéria-prima.