Design of a disposable μPAD for on-hand quantification of urinary creatinine
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.14/40572 |
Resumo: | In this work, a new microfluidic paper-based analytical device (μPAD) was developed for on-hand creatinine quantification in urine samples. When compared to other conventional methods, this innovative paper device is more accessible, portable, providing low-cost analysis and applicable to non-invasive biological fluids. Furthermore, this paper approach is environmentally friendly, as it uses small amounts of reagents, results in low waste production and it is disposable by incineration. Its configuration consisted in two layers of filter paper discs of 9.5 mm of diameter, in which the top layer served as a reservoir and the bottom layer contained the reagent (alkaline picrate). This hydrophilic unit consisting of two-layer of filter paper was aligned within a laminated plastic pouch to set the hydrophobic separation, with 3 mm holes for the sample insertion. When inserted, creatinine of the standard/sample reacts with alkaline picric acid (colorimetric reaction), and the colour changes from yellow to orange/red. Under optimal conditions, the developed method allowed creatinine quantification in the dynamic range of 2.20-35.0 mg/dL, with a limit of detection (LOD) of 0.66 mg/dL and a limit of quantification (LOQ) of 2.20 mg/dL. The image was scanned after 20 minutes and was processed in ImageJ software to obtain the colour intensity developed, later converted to pseudo-absorbance. The stability of the reaction product after the sample insertion was verified until 3 hours and the device could be scanned within that time. The μPAD allows to obtain linear calibration curves up to one week when stored in vacuum and placed in the fridge at approximately 4 ºC. Finally, the method was validated by the results obtained with the developed device and a comparison method, and it was demonstrated that there were no statistically significant differences between them. As so, it was possible to conclude that this novel device was successfully developed, as it proved to be sensitive and simple, as well as affordable, as it showed a cost of approximately 0.40 € (in terms of consumables). |
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Design of a disposable μPAD for on-hand quantification of urinary creatinineMicrofluidic paper-based analytical device (µPAD)Creatinine determinationUrine samplesJaffe reactionReal timeDispositivo microfluídico baseado papel (µPAD)Determinação creatininaAmostras urinaReação JaffeDeterminações tempo realDomínio/Área Científica::Engenharia e Tecnologia::Engenharia MédicaIn this work, a new microfluidic paper-based analytical device (μPAD) was developed for on-hand creatinine quantification in urine samples. When compared to other conventional methods, this innovative paper device is more accessible, portable, providing low-cost analysis and applicable to non-invasive biological fluids. Furthermore, this paper approach is environmentally friendly, as it uses small amounts of reagents, results in low waste production and it is disposable by incineration. Its configuration consisted in two layers of filter paper discs of 9.5 mm of diameter, in which the top layer served as a reservoir and the bottom layer contained the reagent (alkaline picrate). This hydrophilic unit consisting of two-layer of filter paper was aligned within a laminated plastic pouch to set the hydrophobic separation, with 3 mm holes for the sample insertion. When inserted, creatinine of the standard/sample reacts with alkaline picric acid (colorimetric reaction), and the colour changes from yellow to orange/red. Under optimal conditions, the developed method allowed creatinine quantification in the dynamic range of 2.20-35.0 mg/dL, with a limit of detection (LOD) of 0.66 mg/dL and a limit of quantification (LOQ) of 2.20 mg/dL. The image was scanned after 20 minutes and was processed in ImageJ software to obtain the colour intensity developed, later converted to pseudo-absorbance. The stability of the reaction product after the sample insertion was verified until 3 hours and the device could be scanned within that time. The μPAD allows to obtain linear calibration curves up to one week when stored in vacuum and placed in the fridge at approximately 4 ºC. Finally, the method was validated by the results obtained with the developed device and a comparison method, and it was demonstrated that there were no statistically significant differences between them. As so, it was possible to conclude that this novel device was successfully developed, as it proved to be sensitive and simple, as well as affordable, as it showed a cost of approximately 0.40 € (in terms of consumables).Neste projeto foi desenvolvido um novo dispositivo microfluídico baseado em papel (μPAD) para a quantificação de creatinina em amostras de urina. Quando comparado com outros métodos convencionais, este dispositivo inovador baseado em papel é mais acessível, portátil, e promove análises de baixo custo, podendo ser aplicado a fluídos biológicos não-invasivos. Para além disso, esta abordagem vai de encontro aos princípios de química-verde, uma vez que usa quantidades pequenas de reagentes, resulta numa produção de poucos resíduos, e pode ser descartado por inceneração. A configuração deste dispositivo consistiu em duas camadas de papel de filtro com 9,5 mm de diâmetro, onde a camada superior serviu de reservatório e a camada inferior continha o reagente (solução alcalina de ácido pícrico). Esta unidade hidrofílica composta por uma dupla camada de discos, foi alinhada dentro de uma bolsa de plástico posteriormente laminada criando separação hidrofóbica, que continha furos de 3 mm para a inserção da amostra. Quando inserida, a creatinina presente nos padrões/amostras reage com o ácido pícrico (reação colorimétrica), e a cor dos discos muda de amarelo para cor-de-laranja/vermelho. Após os estudos de otimização, o método desenvolvido permitiu uma quantificação de creatinina no intervalo dinâmico de 2.20-35.0 mg/dL, com um limite de deteção (LOD) de 0.66 mg/dL e um limite de quantificação (LOQ) de 2.20 mg/dL. A imagem é digitalizada ao fim de 20 minutos, sendo processada no software ImageJ, de modo a obter a intensidade de cor desenvolvida, que é depois convertida a pseudo-absorbância. A estabilidade do produto de reação, depois da inserção da amostra, foi verificada até 3 horas e o dispositivo pôde ser digitalizado dentro desse período de tempo. O μPAD permite obter curvas de calibração lineares até uma semana, quando armazenado em vácuo e colocado no frigorífico, a aproximadamente 4 ºC. Por fim, o método foi validado comparando os resultados obtidos com o dispositivo desenvolvido e um método de comparação, e foi demonstrado que não havia diferenças estatisticamente significativas entre eles. Assim sendo, foi possível concluir que este novo dispositivo foi desenvolvido com sucesso, uma vez que provou ser sensível e simples, assim como económico, visto custar cerca de 0.40 € (em termos de consumíveis).Mesquita, Raquel Beatriz Ribeiro deRangel, António Osmaro Santos SilvaVeritati - Repositório Institucional da Universidade Católica PortuguesaRodrigues, Maria Manuel Pereira de Melo2022-11-242022-092024-03-14T00:00:00Z2022-11-24T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.14/40572TID:203223292enginfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-12T17:46:07Zoai:repositorio.ucp.pt:10400.14/40572Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:33:17.609555Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Design of a disposable μPAD for on-hand quantification of urinary creatinine |
title |
Design of a disposable μPAD for on-hand quantification of urinary creatinine |
spellingShingle |
Design of a disposable μPAD for on-hand quantification of urinary creatinine Rodrigues, Maria Manuel Pereira de Melo Microfluidic paper-based analytical device (µPAD) Creatinine determination Urine samples Jaffe reaction Real time Dispositivo microfluídico baseado papel (µPAD) Determinação creatinina Amostras urina Reação Jaffe Determinações tempo real Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Médica |
title_short |
Design of a disposable μPAD for on-hand quantification of urinary creatinine |
title_full |
Design of a disposable μPAD for on-hand quantification of urinary creatinine |
title_fullStr |
Design of a disposable μPAD for on-hand quantification of urinary creatinine |
title_full_unstemmed |
Design of a disposable μPAD for on-hand quantification of urinary creatinine |
title_sort |
Design of a disposable μPAD for on-hand quantification of urinary creatinine |
author |
Rodrigues, Maria Manuel Pereira de Melo |
author_facet |
Rodrigues, Maria Manuel Pereira de Melo |
author_role |
author |
dc.contributor.none.fl_str_mv |
Mesquita, Raquel Beatriz Ribeiro de Rangel, António Osmaro Santos Silva Veritati - Repositório Institucional da Universidade Católica Portuguesa |
dc.contributor.author.fl_str_mv |
Rodrigues, Maria Manuel Pereira de Melo |
dc.subject.por.fl_str_mv |
Microfluidic paper-based analytical device (µPAD) Creatinine determination Urine samples Jaffe reaction Real time Dispositivo microfluídico baseado papel (µPAD) Determinação creatinina Amostras urina Reação Jaffe Determinações tempo real Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Médica |
topic |
Microfluidic paper-based analytical device (µPAD) Creatinine determination Urine samples Jaffe reaction Real time Dispositivo microfluídico baseado papel (µPAD) Determinação creatinina Amostras urina Reação Jaffe Determinações tempo real Domínio/Área Científica::Engenharia e Tecnologia::Engenharia Médica |
description |
In this work, a new microfluidic paper-based analytical device (μPAD) was developed for on-hand creatinine quantification in urine samples. When compared to other conventional methods, this innovative paper device is more accessible, portable, providing low-cost analysis and applicable to non-invasive biological fluids. Furthermore, this paper approach is environmentally friendly, as it uses small amounts of reagents, results in low waste production and it is disposable by incineration. Its configuration consisted in two layers of filter paper discs of 9.5 mm of diameter, in which the top layer served as a reservoir and the bottom layer contained the reagent (alkaline picrate). This hydrophilic unit consisting of two-layer of filter paper was aligned within a laminated plastic pouch to set the hydrophobic separation, with 3 mm holes for the sample insertion. When inserted, creatinine of the standard/sample reacts with alkaline picric acid (colorimetric reaction), and the colour changes from yellow to orange/red. Under optimal conditions, the developed method allowed creatinine quantification in the dynamic range of 2.20-35.0 mg/dL, with a limit of detection (LOD) of 0.66 mg/dL and a limit of quantification (LOQ) of 2.20 mg/dL. The image was scanned after 20 minutes and was processed in ImageJ software to obtain the colour intensity developed, later converted to pseudo-absorbance. The stability of the reaction product after the sample insertion was verified until 3 hours and the device could be scanned within that time. The μPAD allows to obtain linear calibration curves up to one week when stored in vacuum and placed in the fridge at approximately 4 ºC. Finally, the method was validated by the results obtained with the developed device and a comparison method, and it was demonstrated that there were no statistically significant differences between them. As so, it was possible to conclude that this novel device was successfully developed, as it proved to be sensitive and simple, as well as affordable, as it showed a cost of approximately 0.40 € (in terms of consumables). |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-11-24 2022-09 2022-11-24T00:00:00Z 2024-03-14T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.14/40572 TID:203223292 |
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http://hdl.handle.net/10400.14/40572 |
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TID:203223292 |
dc.language.iso.fl_str_mv |
eng |
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eng |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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