<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>2215-3470</journal-id>
<journal-title><![CDATA[Uniciencia]]></journal-title>
<abbrev-journal-title><![CDATA[Uniciencia]]></abbrev-journal-title>
<issn>2215-3470</issn>
<publisher>
<publisher-name><![CDATA[Universidad Nacional, Costa Rica]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S2215-34702020000200031</article-id>
<article-id pub-id-type="doi">10.15359/ru.34-2.2</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Revisando los fundamentos del análisis de p-nitrofenol para su aplicación en la cuantificación de la actividad de lipasas. Una actualización gráfica.]]></article-title>
<article-title xml:lang="en"><![CDATA[Revisiting the fundamentals of p-nitrophenol analysis for its application in the quantification of lipases activity. A graphical update.]]></article-title>
<article-title xml:lang="pt"><![CDATA[Revisão dos fundamentos da análise do p-nitrofenol para sua aplicação na quantificação da atividade da lipase. Uma atualização gráfica.]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Syedd-León]]></surname>
<given-names><![CDATA[Randall]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Sandoval-Barrantes]]></surname>
<given-names><![CDATA[Manuel]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Trimiño-Vásquez]]></surname>
<given-names><![CDATA[Humberto]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Villegas-Peñaranda]]></surname>
<given-names><![CDATA[Luis Roberto]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Rodríguez-Rodríguez]]></surname>
<given-names><![CDATA[Gerardo]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad Estatal a Distancia Maestría en Manejo de Recursos Naturales ]]></institution>
<addr-line><![CDATA[Montes de Oca ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Universidad Naciona Escuela de Química Laboratorio de Investigación en Biorrefinería]]></institution>
<addr-line><![CDATA[Heredia ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Universidad Nacional Escuela de Química Laboratorio de Investigación en Biorrefinería]]></institution>
<addr-line><![CDATA[Heredia ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af4">
<institution><![CDATA[,Universidad Nacional Escuela de Química Laboratorio de Investigación en Biorrefinería]]></institution>
<addr-line><![CDATA[Heredia ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af5">
<institution><![CDATA[,Universidad Nacional. Heredia Laboratorio de Fitoquímica, Escuela de Química ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Costa Rica</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2020</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2020</year>
</pub-date>
<volume>34</volume>
<numero>2</numero>
<fpage>31</fpage>
<lpage>43</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_arttext&amp;pid=S2215-34702020000200031&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_abstract&amp;pid=S2215-34702020000200031&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_pdf&amp;pid=S2215-34702020000200031&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen El p-nitrofenol (pNP) es un compuesto ampliamente utilizado para la determinación analítica de esterasas (EC. 3.1.1.X), incluidas las lipasas (E.C. 3.1.1.3). La mayoría de las mediciones enzimáticas emplean sus derivados, tales como ésteres, que se descomponen por hidrólisis, liberando pNP que se cuantifica por su absorbancia a 410 nm. Aunque, este tipo de métodos se desarrolló hace algunas décadas, el análisis espectrofotométrico de pNP requiere mediciones precisas de pH y temperatura, para lograr determinaciones confiables. El objetivo de este trabajo es ofrecer una actualización gráfica de cómo el pH y la temperatura afectan la absorbancia de p-nitrofenol a diferentes longitudes de onda en medios emulsionados para ensayos con lipasas, debido a su relevancia para la determinación cuantitativa de la actividad de las lipasas bajo métodos espectrofotométricos. Para resaltar la importancia de cada variable en este análisis, se disolvió pNP en medios emulsionados (para la cuantificación de la actividad de la lipasa) a varios valores de pH de 4.00 a 11.00 y se midió su absorbancia en un rango de 270 nm a 500 nm a varias temperaturas en el rango de 25 °C a 50 °C. El comportamiento químico de pNP bajo estas condiciones se correlacionó mediante la construcción de gráficos tridimensionales. Como resultado, los gráficos 3D construidos experimentalmente, mostraron que, independientemente de la temperatura, por debajo de pH 6.00 el pNP absorbe principalmente, a 317 nm, debido a la mayor abundancia de su forma protonada que es completamente predominante a pH 3.50 e inferiores. Por otro lado, a pH 10.0 y superiores, el equilibrio se desplaza completamente a la forma aniónica pNP, que absorbe a 410 nm. Nuestros datos confirman que en valores de pH cercano a la neutralidad, el pNP muestra un efecto de dependencia a la temperatura, aumentando la absorbancia a 410 nm a temperaturas más altas. Debido a muchas determinaciones cuantitativas de las actividades enzimáticas, el pNP se libera en un medio de reacción de alrededor de pH 7.00. Es necesario recordar el papel determinante del pH y la temperatura sobre estas mediciones y cómo estas variables deben ser estrictamente consideradas.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract p-Nitrophenol (pNP) is a widely used compound for analytical determinations of several esterases (EC. 3.1.1.X), including lipases (E.C. 3.1.1.3). Most enzymatic measurements employ pNP derivatives such as esters, which are broken down by enzymatic hydrolysis, releasing pNP that is quantified by its absorbance at 410 nm. Although this type of methods was developed a few decades ago, the spectrophotometric analysis of pNP requires analytical measurements of pH and temperature to achieve reliable determinations. The aim of this paper is to offer a graphical update of how pH and temperature affect the p-nitrophenol absorbance at different wavelengths in lipase emulsified media, due to its relevance for the quantitative determination of lipase activity using spectrophotometric methods. To highlight the importance of each variable involved in this analysis, we dissolved pNP in emulsified media (for lipase activity quantification) at several pH values from 4.00 to 11.00, and measured its absorbance in a range of 270 nm - 500 nm and at several temperatures from 25°C to 50°C. The absorption patterns of pNP under the established conditions were graphed in 3D plots. The constructed 3D plots showed that, regardless of the temperature, below pH 6.00, pNP predominantly absorbs at 317 nm, due to the greater abundance of its protonated form, which is completely predominant at pH 3.50 and below. On the other hand, at pH 10.0 and above, the major absorption occurs at about 401 nm, confirming that the equilibrium is completely shifted to the pNP anionic form. These results also indicate that close to neutral pH value pNP, it displays a temperature dependence effect, increasing absorbance to 410 nm at higher temperatures. Due to many analytical determinations of enzymatic activities, the release of pNP is carried around pH 7.00. It is necessary to consider the determinant role of both pH and temperature over these measurements, how these variables must be strictly controlled, and how the calibration curves and blanks should take the reaction media pH and temperature into account.¨]]></p></abstract>
<abstract abstract-type="short" xml:lang="pt"><p><![CDATA[Resumo O p-nitrofenol (pNP) é um composto amplamente utilizado para a determinação analítica de esterases (EC. 3.1.1.X), incluindo lipases (E.C. 3.1.1.3). A maioria das medições enzimáticas utiliza seus derivados, como ésteres, que se decompõem por hidrólise, liberando pNP quantificado por sua absorvência a 410 nm. Embora esses tipos de métodos tenham sido desenvolvidos há algumas décadas, a análise espectrofotométrica de pNP requer medições precisas de pH e temperatura para obter determinações confiáveis. O objetivo deste trabalho é oferecer uma atualização gráfica de como o pH e a temperatura afetam a absorvência do p-nitrofenol em diferentes longitudes de onda em meios emulsionados para ensaios de lipase, devido a sua relevância para a determinação quantitativa da atividade das lipases sob métodos espectrofotométricos. Para destacar a importância de cada variável nesta análise, o pNP foi dissolvido em meio emulsionado (para quantificação da atividade da lipase) em vários valores de pH de 4,00 a 11,00 e sua absorvência foi medida na faixa de 270 nm a 500 nm a várias temperaturas na faixa de 25 °C a 50 °C. O comportamento químico do pNP nessas condições foi correlacionado mediante a construção de gráficos tridimensionais. Como resultado, os gráficos 3D construídos experimentalmente mostraram que, independentemente da temperatura, abaixo de pH 6,00, o pNP absorve principalmente a 317 nm, devido à maior abundância de sua forma protonada, que é completamente predominante em pH 3,50 e inferiores. Por outro lado, o pH 10,0 e superiores, o equilíbrio muda completamente para a forma aniônica pNP, que absorve a 410 nm. Nossos dados confirmam que em valores de pH próximos à neutralidade, o pNP mostra um efeito de dependência da temperatura, aumentando a absorvência a 410 nm em temperaturas mais altas. Em virtude de muitas determinações quantitativas de atividades enzimáticas, o pNP é liberado em um meio de reação de cerca de pH 7,00. É necessário lembrar o papel determinante do pH e da temperatura nessas medições e como essas variáveis &#8203;&#8203;devem ser estritamente consideradas.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[p-Nitrophenol]]></kwd>
<kwd lng="en"><![CDATA[lipase]]></kwd>
<kwd lng="en"><![CDATA[absorbance pH-dependence]]></kwd>
<kwd lng="en"><![CDATA[temperature dependent dissociation]]></kwd>
<kwd lng="en"><![CDATA[3D pH-wavelength-absorbance spectra.]]></kwd>
<kwd lng="es"><![CDATA[p-nitrofenol, lipasa]]></kwd>
<kwd lng="es"><![CDATA[absorbancia dependiente del pH]]></kwd>
<kwd lng="es"><![CDATA[disociación dependiente de la temperatura]]></kwd>
<kwd lng="es"><![CDATA[espectro 3D de absorbancia]]></kwd>
<kwd lng="es"><![CDATA[pH]]></kwd>
<kwd lng="es"><![CDATA[longitud de onda]]></kwd>
<kwd lng="pt"><![CDATA[p-nitrofenol]]></kwd>
<kwd lng="pt"><![CDATA[lipase]]></kwd>
<kwd lng="pt"><![CDATA[absorvência dependente do pH]]></kwd>
<kwd lng="pt"><![CDATA[dissociação dependente da temperatura]]></kwd>
<kwd lng="pt"><![CDATA[espectro de absorvência 3D]]></kwd>
<kwd lng="pt"><![CDATA[pH]]></kwd>
<kwd lng="pt"><![CDATA[longitude de onda]]></kwd>
</kwd-group>
</article-meta>
</front><back>
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