Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.

New natural reducing agents with a lower negative impact on the environment and with a high antimicrobial potential are required for the process of obtaining silver nanoparticles through the chemical reduction method. The use of plant extracts can be a fast track in the formation of nanoparticles. I...

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Autores:
Sánchez, Manuel Alejandro
Fiscal Ladino, Jhon Alexander
Tipo de recurso:
Article of journal
Fecha de publicación:
2023
Institución:
Universidad Francisco de Paula Santander
Repositorio:
Repositorio Digital UFPS
Idioma:
eng
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oai:repositorio.ufps.edu.co:ufps/7098
Acceso en línea:
https://repositorio.ufps.edu.co/handle/ufps/7098
Palabra clave:
HPLC
quercetin
nanomaterial
green synthesis
antimicrobial agent
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openAccess
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Está bajo una licencia Creative Commons Atribución 4.0 Internacional (CC BY 4.0).
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dc.title.eng.fl_str_mv Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.
title Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.
spellingShingle Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.
HPLC
quercetin
nanomaterial
green synthesis
antimicrobial agent
title_short Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.
title_full Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.
title_fullStr Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.
title_full_unstemmed Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.
title_sort Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.
dc.creator.fl_str_mv Sánchez, Manuel Alejandro
Fiscal Ladino, Jhon Alexander
dc.contributor.author.none.fl_str_mv Sánchez, Manuel Alejandro
Fiscal Ladino, Jhon Alexander
dc.subject.proposal.eng.fl_str_mv HPLC
quercetin
nanomaterial
green synthesis
antimicrobial agent
topic HPLC
quercetin
nanomaterial
green synthesis
antimicrobial agent
description New natural reducing agents with a lower negative impact on the environment and with a high antimicrobial potential are required for the process of obtaining silver nanoparticles through the chemical reduction method. The use of plant extracts can be a fast track in the formation of nanoparticles. In this case, organic compounds such as terpenes, flavonoids, enzymes, proteins, and cofactors present in plants act as reducing agents for nanomaterials. This research evaluated the antimicrobial property of silver nanoparticles from extracts of Crescentia cujete L. The presence of quercetin (flavonoid) was determined by high-performance liquid chromatography (HPLC); the production of silver nanoparticles (AgNPs) was established by green synthesis; the size and morphology of the nanomaterials were evaluated by scanning electron microscope (SEM). The antimicrobial capacity was studied by two analysis methods: modified culture medium and surface seeding. The presence of quercetin (26.55 mg L-1) in the crude extract of Crescentia cujete L., identified by HPLC, was evidenced. Nanoparticle formation was spherical, with an average size of 250 ± 3 and 460 ± 6 nm. Microbiological cultures with treatment showed 94% microbial inhibition. It was concluded that the Crescentia cujete L., leaves shoed an acceptable concentration of quercetin to be used as a useful adjuvant to enhance the reduction of NPs synthesis. The nanoparticles produced by green synthesis proved to have a positive effect to combat pathogenic microorganisms
publishDate 2023
dc.date.issued.none.fl_str_mv 2023-03-26
dc.date.accessioned.none.fl_str_mv 2024-04-30T16:28:11Z
dc.date.available.none.fl_str_mv 2024-04-30T16:28:11Z
dc.type.spa.fl_str_mv Artículo de revista
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dc.identifier.uri.none.fl_str_mv https://repositorio.ufps.edu.co/handle/ufps/7098
dc.identifier.doi.none.fl_str_mv 10.1590/1519-6984.270215
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dc.relation.ispartof.none.fl_str_mv Brazilian Journal of Biology, 2024, vol. 84, e270215 | https://doi.org/10.1590/1519-6984.270215
dc.relation.citationedition.spa.fl_str_mv Vol.84 No. (2024)
dc.relation.citationendpage.spa.fl_str_mv 8
dc.relation.citationissue.spa.fl_str_mv (2024)
dc.relation.citationstartpage.spa.fl_str_mv 1
dc.relation.citationvolume.spa.fl_str_mv 84
dc.relation.cites.none.fl_str_mv Sánchez MA, Fiscal Ladino JA. Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L. Braz J Biol [Internet]. 2023;84:e270215. Disponible en: http://dx.doi.org/10.1590/1519-6984.270215
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dc.rights.creativecommons.spa.fl_str_mv Atribución 4.0 Internacional (CC BY 4.0)
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dc.publisher.spa.fl_str_mv Brazilian Journal of Biology
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spelling Sánchez, Manuel Alejandro5c8897da1223ede8de480ac08e6ece74600Fiscal Ladino, Jhon Alexander27e8216b5f4515c7a12b8cc0436bdf976002024-04-30T16:28:11Z2024-04-30T16:28:11Z2023-03-26https://repositorio.ufps.edu.co/handle/ufps/709810.1590/1519-6984.270215New natural reducing agents with a lower negative impact on the environment and with a high antimicrobial potential are required for the process of obtaining silver nanoparticles through the chemical reduction method. The use of plant extracts can be a fast track in the formation of nanoparticles. In this case, organic compounds such as terpenes, flavonoids, enzymes, proteins, and cofactors present in plants act as reducing agents for nanomaterials. This research evaluated the antimicrobial property of silver nanoparticles from extracts of Crescentia cujete L. The presence of quercetin (flavonoid) was determined by high-performance liquid chromatography (HPLC); the production of silver nanoparticles (AgNPs) was established by green synthesis; the size and morphology of the nanomaterials were evaluated by scanning electron microscope (SEM). The antimicrobial capacity was studied by two analysis methods: modified culture medium and surface seeding. The presence of quercetin (26.55 mg L-1) in the crude extract of Crescentia cujete L., identified by HPLC, was evidenced. Nanoparticle formation was spherical, with an average size of 250 ± 3 and 460 ± 6 nm. Microbiological cultures with treatment showed 94% microbial inhibition. It was concluded that the Crescentia cujete L., leaves shoed an acceptable concentration of quercetin to be used as a useful adjuvant to enhance the reduction of NPs synthesis. The nanoparticles produced by green synthesis proved to have a positive effect to combat pathogenic microorganisms8 Páginasapplication/pdfengBrazilian Journal of BiologyBrazilian Journal of Biology, 2024, vol. 84, e270215 | https://doi.org/10.1590/1519-6984.270215Vol.84 No. (2024)8(2024)184Sánchez MA, Fiscal Ladino JA. Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L. Braz J Biol [Internet]. 2023;84:e270215. Disponible en: http://dx.doi.org/10.1590/1519-6984.270215Está bajo una licencia Creative Commons Atribución 4.0 Internacional (CC BY 4.0).https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessAtribución 4.0 Internacional (CC BY 4.0)http://purl.org/coar/access_right/c_abf2https://www.scielo.br/j/bjb/a/bKjsqJFdjk5Gnrn9tSsBLzD/Antimicrobial evaluation of silver nanoparticles using extracts of Crescentia cujete L.Artículo de revistahttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85HPLCquercetinnanomaterialgreen synthesisantimicrobial agentADAMS, D.J., 2004. Fungal cell wall chitinases and glucanases. Microbiology vol. 150, no. 7, pp. 2029-2035. http://dx.doi. org/10.1099/mic.0.26980-0. 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 incorporada en las Obras Colectivas.

b.	Distribuir copias o fonogramas de las Obras, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública, incluyéndolas como incorporadas en Obras Colectivas, según corresponda.

c.	Distribuir copias de las Obras Derivadas que se generen, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública.
Los derechos mencionados anteriormente pueden ser ejercidos en todos los medios y formatos, actualmente conocidos o que se inventen en el futuro. Los derechos antes mencionados incluyen el derecho a realizar dichas modificaciones en la medida que sean técnicamente necesarias para ejercer los derechos en otro medio o formatos, pero de otra manera usted no está autorizado para realizar obras derivadas. Todos los derechos no otorgados expresamente por el Licenciante quedan por este medio reservados, incluyendo pero sin limitarse a aquellos que se mencionan en las secciones 4(d) y 4(e).

4. Restricciones.
La licencia otorgada en la anterior Sección 3 está expresamente sujeta y limitada por las siguientes restricciones:

a.	Usted puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra sólo bajo las condiciones de esta Licencia, y Usted debe incluir una copia de esta licencia o del Identificador Universal de Recursos de la misma con cada copia de la Obra que distribuya, exhiba públicamente, ejecute públicamente o ponga a disposición pública. No es posible ofrecer o imponer ninguna condición sobre la Obra que altere o limite las condiciones de esta Licencia o el ejercicio de los derechos de los destinatarios otorgados en este documento. No es posible sublicenciar la Obra. Usted debe mantener intactos todos los avisos que hagan referencia a esta Licencia y a la cláusula de limitación de garantías. Usted no puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra con alguna medida tecnológica que controle el acceso o la utilización de ella de una forma que sea inconsistente con las condiciones de esta Licencia. Lo anterior se aplica a la Obra incorporada a una Obra Colectiva, pero esto no exige que la Obra Colectiva aparte de la obra misma quede sujeta a las condiciones de esta Licencia. Si Usted crea una Obra Colectiva, previo aviso de cualquier Licenciante debe, en la medida de lo posible, eliminar de la Obra Colectiva cualquier referencia a dicho Licenciante o al Autor Original, según lo solicitado por el Licenciante y conforme lo exige la cláusula 4(c).

b.	Usted no puede ejercer ninguno de los derechos que le han sido otorgados en la Sección 3 precedente de modo que estén principalmente destinados o directamente dirigidos a conseguir un provecho comercial o una compensación monetaria privada. El intercambio de la Obra por otras obras protegidas por derechos de autor, ya sea a través de un sistema para compartir archivos digitales (digital file-sharing) o de cualquier otra manera no será considerado como estar destinado principalmente o dirigido directamente a conseguir un provecho comercial o una compensación monetaria privada, siempre que no se realice un pago mediante una compensación monetaria en relación con el intercambio de obras protegidas por el derecho de autor.

c.	Si usted distribuye, exhibe públicamente, ejecuta públicamente o ejecuta públicamente en forma digital la Obra o cualquier Obra Derivada u Obra Colectiva, Usted debe mantener intacta toda la información de derecho de autor de la Obra y proporcionar, de forma razonable según el medio o manera que Usted esté utilizando: (i) el nombre del Autor Original si está provisto (o seudónimo, si fuere aplicable), y/o (ii) el nombre de la parte o las partes que el Autor Original y/o el Licenciante hubieren designado para la atribución (v.g., un instituto patrocinador, editorial, publicación) en la información de los derechos de autor del Licenciante, términos de servicios o de otras formas razonables; el título de la Obra si está provisto; en la medida de lo razonablemente factible y, si está provisto, el Identificador Uniforme de Recursos (Uniform Resource Identifier) que el Licenciante especifica para ser asociado con la Obra, salvo que tal URI no se refiera a la nota sobre los derechos de autor o a la información sobre el licenciamiento de la Obra; y en el caso de una Obra Derivada, atribuir el crédito identificando el uso de la Obra en la Obra Derivada (v.g., "Traducción Francesa de la Obra del Autor Original," o "Guión Cinematográfico basado en la Obra original del Autor Original"). Tal crédito puede ser implementado de cualquier forma razonable; en el caso, sin embargo, de Obras Derivadas u Obras Colectivas, tal crédito aparecerá, como mínimo, donde aparece el crédito de cualquier otro autor comparable y de una manera, al menos, tan destacada como el crédito de otro autor comparable.

d.	Para evitar toda confusión, el Licenciante aclara que, cuando la obra es una composición musical:

i.	Regalías por interpretación y ejecución bajo licencias generales. El Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública o la ejecución pública digital de la obra y de recolectar, sea individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, SAYCO), las regalías por la ejecución pública o por la ejecución pública digital de la obra (por ejemplo Webcast) licenciada bajo licencias generales, si la interpretación o ejecución de la obra está primordialmente orientada por o dirigida a la obtención de una ventaja comercial o una compensación monetaria privada.

ii.	Regalías por Fonogramas. El Licenciante se reserva el derecho exclusivo de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, los consagrados por la SAYCO), una agencia de derechos musicales o algún agente designado, las regalías por cualquier fonograma que Usted cree a partir de la obra (“versión cover”) y distribuya, en los términos del régimen de derechos de autor, si la creación o distribución de esa versión cover está primordialmente destinada o dirigida a obtener una ventaja comercial o una compensación monetaria privada.

e.	Gestión de Derechos de Autor sobre Interpretaciones y Ejecuciones Digitales (WebCasting). Para evitar toda confusión, el Licenciante aclara que, cuando la obra sea un fonograma, el Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública digital de la obra (por ejemplo, webcast) y de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, ACINPRO), las regalías por la ejecución pública digital de la obra (por ejemplo, webcast), sujeta a las disposiciones aplicables del régimen de Derecho de Autor, si esta ejecución pública digital está primordialmente dirigida a obtener una ventaja comercial o una compensación monetaria privada.

5. Representaciones, Garantías y Limitaciones de Responsabilidad.
A MENOS QUE LAS PARTES LO ACORDARAN DE OTRA FORMA POR ESCRITO, EL LICENCIANTE OFRECE LA OBRA (EN EL ESTADO EN EL QUE SE ENCUENTRA) “TAL CUAL”, SIN BRINDAR GARANTÍAS DE CLASE ALGUNA RESPECTO DE LA OBRA, YA SEA EXPRESA, IMPLÍCITA, LEGAL O CUALQUIERA OTRA, INCLUYENDO, SIN LIMITARSE A ELLAS, GARANTÍAS DE TITULARIDAD, COMERCIABILIDAD, ADAPTABILIDAD O ADECUACIÓN A PROPÓSITO DETERMINADO, AUSENCIA DE INFRACCIÓN, DE AUSENCIA DE DEFECTOS LATENTES O DE OTRO TIPO, O LA PRESENCIA O AUSENCIA DE ERRORES, SEAN O NO DESCUBRIBLES (PUEDAN O NO SER ESTOS DESCUBIERTOS). ALGUNAS JURISDICCIONES NO PERMITEN LA EXCLUSIÓN DE GARANTÍAS IMPLÍCITAS, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.

6. Limitación de responsabilidad.
A MENOS QUE LO EXIJA EXPRESAMENTE LA LEY APLICABLE, EL LICENCIANTE NO SERÁ RESPONSABLE ANTE USTED POR DAÑO ALGUNO, SEA POR RESPONSABILIDAD EXTRACONTRACTUAL, PRECONTRACTUAL O CONTRACTUAL, OBJETIVA O SUBJETIVA, SE TRATE DE DAÑOS MORALES O PATRIMONIALES, DIRECTOS O INDIRECTOS, PREVISTOS O IMPREVISTOS PRODUCIDOS POR EL USO DE ESTA LICENCIA O DE LA OBRA, AUN CUANDO EL LICENCIANTE HAYA SIDO ADVERTIDO DE LA POSIBILIDAD DE DICHOS DAÑOS. ALGUNAS LEYES NO PERMITEN LA EXCLUSIÓN DE CIERTA RESPONSABILIDAD, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.

7. Término.

a.	Esta Licencia y los derechos otorgados en virtud de ella terminarán automáticamente si Usted infringe alguna condición establecida en ella. Sin embargo, los individuos o entidades que han recibido Obras Derivadas o Colectivas de Usted de conformidad con esta Licencia, no verán terminadas sus licencias, siempre que estos individuos o entidades sigan cumpliendo íntegramente las condiciones de estas licencias. Las Secciones 1, 2, 5, 6, 7, y 8 subsistirán a cualquier terminación de esta Licencia.

b.	Sujeta a las condiciones y términos anteriores, la licencia otorgada aquí es perpetua (durante el período de vigencia de los derechos de autor de la obra). No obstante lo anterior, el Licenciante se reserva el derecho a publicar y/o estrenar la Obra bajo condiciones de licencia diferentes o a dejar de distribuirla en los términos de esta Licencia en cualquier momento; en el entendido, sin embargo, que esa elección no servirá para revocar esta licencia o que deba ser otorgada , bajo los términos de esta licencia), y esta licencia continuará en pleno vigor y efecto a menos que sea terminada como se expresa atrás. La Licencia revocada continuará siendo plenamente vigente y efectiva si no se le da término en las condiciones indicadas anteriormente.

8. Varios.

a.	Cada vez que Usted distribuya o ponga a disposición pública la Obra o una Obra Colectiva, el Licenciante ofrecerá al destinatario una licencia en los mismos términos y condiciones que la licencia otorgada a Usted bajo esta Licencia.

b.	Si alguna disposición de esta Licencia resulta invalidada o no exigible, según la legislación vigente, esto no afectará ni la validez ni la aplicabilidad del resto de condiciones de esta Licencia y, sin acción adicional por parte de los sujetos de este acuerdo, aquélla se entenderá reformada lo mínimo necesario para hacer que dicha disposición sea válida y exigible.

c.	Ningún término o disposición de esta Licencia se estimará renunciada y ninguna violación de ella será consentida a menos que esa renuncia o consentimiento sea otorgado por escrito y firmado por la parte que renuncie o consienta.

d.	Esta Licencia refleja el acuerdo pleno entre las partes respecto a la Obra aquí licenciada. No hay arreglos, acuerdos o declaraciones respecto a la Obra que no estén especificados en este documento. El Licenciante no se verá limitado por ninguna disposición adicional que pueda surgir en alguna comunicación emanada de Usted. Esta Licencia no puede ser modificada sin el consentimiento mutuo por escrito del Licenciante y Usted.
0000-0002-2126-395627e8216b5f4515c7a12b8cc0436bdf97600