Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples

Aim. Development of an easy-to-use colorimetric sensor system for fast and accurate detection of phenol in envi- ronmental samples. Methods. Technique of molecular imprinting, method of in situ polymerization of molecularly imprinted polymer membranes. Results. The proposed sensor is based on free-s...

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Дата:	2014
Автори:	Sergeyeva, T.A., Chelyadina, D.S., Gorbach, L.A., Brovko, O.O., Piletska, E.V., Piletsky, S.A., Sergeeva, L.M., El’skaya, A.V.
Формат:	Стаття
Мова:	English
Опубліковано:	Інститут молекулярної біології і генетики НАН України 2014
Назва видання:	Вiopolymers and Cell
Теми:	Molecular and Cell Biotechnologies
Онлайн доступ:	http://dspace.nbuv.gov.ua/handle/123456789/154308
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Назва журналу:	Digital Library of Periodicals of National Academy of Sciences of Ukraine
Цитувати:	Colorimetric biomimetic sensor systemsbased on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples / T.A. Sergeyeva, D.S. Chelyadina, L.A. Gorbach, O.O. Brovko, E.V. Piletska, S.A. Piletsky, L.M. Sergeeva, A.V. El’skaya // Вiopolymers and Cell. — 2014. — Т. 30, № 3. — С. 209-215. — Бібліогр.: 21 назв. — англ.

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spelling	irk-123456789-1543082019-06-16T01:27:47Z Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples Sergeyeva, T.A. Chelyadina, D.S. Gorbach, L.A. Brovko, O.O. Piletska, E.V. Piletsky, S.A. Sergeeva, L.M. El’skaya, A.V. Molecular and Cell Biotechnologies Aim. Development of an easy-to-use colorimetric sensor system for fast and accurate detection of phenol in envi- ronmental samples. Methods. Technique of molecular imprinting, method of in situ polymerization of molecularly imprinted polymer membranes. Results. The proposed sensor is based on free-standing molecularly imprinted polymer (MIP) membranes, synthesized by in situ polymerization, and having in their structure artificial binding sites capable of selective phenol recognition. The quantitative detection of phenol, selectively adsorbed by the MIP membranes, is based on its reaction with 4-aminoantipyrine, which gives a pink-colored product. The intensity of staining of the MIP membrane is proportional to phenol concentration in the analyzed sample. Phenol can be detected within the range 50 nM–10 mM with limit of detection 50 nM, which corresponds to the concentrations that have to be detected in natural and waste waters in accordance with environmental protection standards. Stability of the MIP-membrane-based sensors was assessed during 12 months storage at room temperature. Conclusions. The sensor system provides highly-selective and sensitive detection of phenol in both mo- del and real (drinking, natural, and waste) water samples. As compared to traditional methods of phenol detection, the proposed system is characterized by simplicity of operation and can be used in non-laboratory conditions. Мета. Розробка простих у використанні колориметричних сенсорних систем для швидкого і точного визначення фенолу у зразках із довкілля. Методи. Метод молекулярного імпринтингу, метод полімеризації in situ молекулярно імпринтованих полімерних (МІП) мембран. Результати. Запропонований сенсор створено на основі МІП мембран, синтезованих методом полімеризації in situ, які мають у своїй структурі штучні рецепторні сайти зв’язування фенолу. Кількісне визначення фенолу, селективно адсорбованого МІП мембранами, грунтується на детекції забарвленого у малиновий колір продукту його реакції з 4-аміноантипірином. Інтенсивність забарвлення МІП мембран є пропорційною концентрації фенолу в аналізованому зразку. Фенол детектується у діапазоні 50 нМ–10 мМ, що відповідає концентраціям, які необхідно виявляти у природних і стічних водах. Стабільність сенсорних систем на основі МІП мембран становить12 місяців за кімнатної температури. Висновки. Сенсорні системи забезпечують високоселективний і чутливий аналіз фенолу як у модельних, так і реальних зразках (питна, природна, стічна вода). Порівняно до традиційних методів визначення фенолу пропонована система є простою у використанні та може бути застосована за польових умов. Цель. Разработка простых в использовании колориметрических сенсорных систем для быстрого и точного определения фенола в образцах из окружающей среды. Методы. Метод молекулярного импринтинга, метод полимеризации in situ молекулярно импринтированных полимерных (МИП) мембран. Результаты. Предложенный сенсор создан на основе МИП мембран, синтезированных методом полимеризации in situ, имеющих в своей структуре синтетические рецепторные сайты связывания фенола. Количественное определение фенола, селективно адсорбированного МИП мембранами, основано на детекции окрашенного в малиновый цвет продукта его реакции с 4-аминоантипирином. Интенсивность окрашивания МИП мембран пропорциональна концентрации фенола в анализируемом образце. Фенол можно детектировать в пределах 50 нМ–10 мМ, что соответствует концентрациям, которые необходимо выявлять в природных и сточных водах. Стабильность сенсорных систем на основе МИП мембран составляет 12 месяцев при комнатной температуре. Выводы. Сенсорные системы обеспечивают высокоселективный и чувствительный анализ фенола как в модельных, так и реальных образцах (питьевая, природная и сточная вода). По сравнению с традиционными методами определения фенола предложенная система проста в использовании и может применяться в полевых условиях. 2014 Article Colorimetric biomimetic sensor systemsbased on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples / T.A. Sergeyeva, D.S. Chelyadina, L.A. Gorbach, O.O. Brovko, E.V. Piletska, S.A. Piletsky, L.M. Sergeeva, A.V. El’skaya // Вiopolymers and Cell. — 2014. — Т. 30, № 3. — С. 209-215. — Бібліогр.: 21 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.000898 http://dspace.nbuv.gov.ua/handle/123456789/154308 577.1 + 573.6 + 543.393 + 543.556 + 004.942 en Вiopolymers and Cell Інститут молекулярної біології і генетики НАН України
institution	Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection	DSpace DC
language	English
topic	Molecular and Cell Biotechnologies Molecular and Cell Biotechnologies
spellingShingle	Molecular and Cell Biotechnologies Molecular and Cell Biotechnologies Sergeyeva, T.A. Chelyadina, D.S. Gorbach, L.A. Brovko, O.O. Piletska, E.V. Piletsky, S.A. Sergeeva, L.M. El’skaya, A.V. Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples Вiopolymers and Cell
description	Aim. Development of an easy-to-use colorimetric sensor system for fast and accurate detection of phenol in envi- ronmental samples. Methods. Technique of molecular imprinting, method of in situ polymerization of molecularly imprinted polymer membranes. Results. The proposed sensor is based on free-standing molecularly imprinted polymer (MIP) membranes, synthesized by in situ polymerization, and having in their structure artificial binding sites capable of selective phenol recognition. The quantitative detection of phenol, selectively adsorbed by the MIP membranes, is based on its reaction with 4-aminoantipyrine, which gives a pink-colored product. The intensity of staining of the MIP membrane is proportional to phenol concentration in the analyzed sample. Phenol can be detected within the range 50 nM–10 mM with limit of detection 50 nM, which corresponds to the concentrations that have to be detected in natural and waste waters in accordance with environmental protection standards. Stability of the MIP-membrane-based sensors was assessed during 12 months storage at room temperature. Conclusions. The sensor system provides highly-selective and sensitive detection of phenol in both mo- del and real (drinking, natural, and waste) water samples. As compared to traditional methods of phenol detection, the proposed system is characterized by simplicity of operation and can be used in non-laboratory conditions.
format	Article
author	Sergeyeva, T.A. Chelyadina, D.S. Gorbach, L.A. Brovko, O.O. Piletska, E.V. Piletsky, S.A. Sergeeva, L.M. El’skaya, A.V.
author_facet	Sergeyeva, T.A. Chelyadina, D.S. Gorbach, L.A. Brovko, O.O. Piletska, E.V. Piletsky, S.A. Sergeeva, L.M. El’skaya, A.V.
author_sort	Sergeyeva, T.A.
title	Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples
title_short	Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples
title_full	Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples
title_fullStr	Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples
title_full_unstemmed	Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples
title_sort	colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples
publisher	Інститут молекулярної біології і генетики НАН України
publishDate	2014
topic_facet	Molecular and Cell Biotechnologies
url	http://dspace.nbuv.gov.ua/handle/123456789/154308
citation_txt	Colorimetric biomimetic sensor systemsbased on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples / T.A. Sergeyeva, D.S. Chelyadina, L.A. Gorbach, O.O. Brovko, E.V. Piletska, S.A. Piletsky, L.M. Sergeeva, A.V. El’skaya // Вiopolymers and Cell. — 2014. — Т. 30, № 3. — С. 209-215. — Бібліогр.: 21 назв. — англ.
series	Вiopolymers and Cell
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fulltext	MOLECULAR AND CELL BIOTECHNOLOGIES UDC 577.1 + 573.6 + 543.393 + 543.556 + 004.942 Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples T. A. Sergeyeva1, D. S. Chelyadina1, L. A. Gorbach2, O. O. Brovko2, E. V. Piletska3, S. A. Piletsky3, L. M. Sergeeva2, A. V. El’skaya1 1Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine 03680 2Institute of Macromolecular Chemistry, NAS of Ukraine 48, Kharkivske Shosse, Kyiv, Ukraine, 02160 3University of Leicester University Road, Leicester LE1 7RH, UK t_sergeyeva@yahoo.co.uk Aim. Development of an easy-to-use colorimetric sensor system for fast and accurate detection of phenol in envi- ronmental samples. Methods. Technique of molecular imprinting, method of in situ polymerization of molecu- larly imprinted polymer membranes. Results. The proposed sensor is based on free-standing molecularly im- printed polymer (MIP) membranes, synthesized by in situ polymerization, and having in their structure artificial binding sites capable of selective phenol recognition. The quantitative detection of phenol, selectively adsorbed by the MIP membranes, is based on its reaction with 4-aminoantipyrine, which gives a pink-colored product. The in- tensity of staining of the MIP membrane is proportional to phenol concentration in the analyzed sample. Phenol can be detected within the range 50 nM–10 mM with limit of detection 50 nM, which corresponds to the concentrations that have to be detected in natural and waste waters in accordance with environmental protection standards. Stability of the MIP-membrane-based sensors was assessed during 12 months storage at room temperature. Conclusions. The sensor system provides highly-selective and sensitive detection of phenol in both model and real (drinking, natural, and waste) water samples. As compared to traditional methods of phenol detection, the proposed system is characterized by simplicity of operation and can be used in non-laboratory conditions. Keywords: phenol, molecularly imprinted polymer membranes, sensors, test-systems, colorimetry. Introduction. Contamination of environment, inclu- ding natural waters, foodstuffs and drinking water is one of the worldwide problems. Population upsurge, ur- banization, as well as intensification of agricultural and industrial development resulted in a three-fold increase in water consumption. At the same time, these factors caused a significant deterioration of water quality. Phe- nols are widespread water pollutants. These compounds are widely used as antiseptics inhibiting bacterial and fungal growth in industrial water supply systems, in pro- duction of paper, some medical preparations, phenol- formaldehyde resins, synthetic fibers, and plastics [1]. Phenols present in environment influence animals and humans health. They can be adsorbed through skin, gastrointestinal tract, respiratory system and cause burns, edemas, and intoxication. Phenols cause acute le- sions of central nervous system, liver, kidney, myocar- dium, blood, and other tissues. Moreover, phenol is an 209 ISSN 0233–7657. Biopolymers and Cell. 2014. Vol. 30. N 3. P. 209–215 doi: http://dx.doi.org/10.7124/bc.000898 � Institute of Molecular Biology and Genetics, NAS of Ukraine, 2014 210 endocrine disrupting compound, causing malfunction of endocrine system at very low concentrations [2]. The- refore, monitoring phenol content in water as well as development of easy-to-use and convenient methods for its rapid and accurate detetion is of great importance for analytical biotechnology. There are a number of tra- ditional analytical methods of phenol detection, inclu- ding HPLC [3], GC [4], these methods in combination with mass-spectrometry [5, 6], and spectrophotometric methods [7]. A number of biosensors were also propo- sed for phenol detection in aqueous samples [8, 9]. However, traditional instrumental methods don’t provide a possibility of fast and effective in-field ana- lysis, they are time-consuming and normally need com- plicated procedure of the sample pre-treatment, i. e. pre-concentration. Biosensors are recognized to be the most effective tools of modern analytical biotechno- logy. However, instability of selective elements based on natural receptors, antibodies and enzymes is a signi- ficant drawback, which limits their wide practical appli- cation. At the same time, biosensors and sensor sys- tems based on molecularly imprinted polymers (MIPs) mimicking active sites of biological molecules can pro- vide a promising alternative [10, 11]. For instance, MIP membranes-based sensors provide high selectivity and sensitivity of the assays as well as rapid and accurate analysis in non-laboratory conditions due to their extra- ordinary stability in extreme environments [12–14]. We have shown that MIP membranes are capable of se- lective recognition of target analytes and generation of the sensor response, which can be easily registered [15, 16]. The present research is aimed at synthesis of phe- nol-selective binding sites in the structure of free-stan- ding MIP membranes and development of colorimetric sensor systems for phenol detection in drinking and en- vironmental water samples. Materials and methods. Materials. Acrylamide (AA), 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPSA), 4-aminoantipyrine, acetonitrile, ammonium hydroxide, N,N-dimethylformamide, itaconic acid (IA), ketal (2,2-dimethoxy-2-phenylacetone), o-cresol, p-cre- sol, N,N'-methylene-bisacrylamide (MBAA), methacry- lic acid (MA), 2-nitrophenol, 3-nitrophenol, 4-nitro- phenol, triethyleneglycoldimethacrylate (TEGDMA), polyethyleneglycol Mw 20 000 (PEG 20 000), pyro- catechol, potassium ferricyanide were purchased from («Sigma-Aldrich», USA). Oligourethaneacrylate (OUA) was synthesized according to [17] and kindly provided by Dr. Matyushov (Institute of Macromolecular Chemi- stry, Kyiv, Ukraine). Synthesis of MIP membranes by in situ polymeriza- tion. MIP membranes capable of selective recognition of phenols were obtained through radical photo-ini- tiated co-polymerization of a functional monomer (AA, AMPS, IA, MA), TEGDMA and OUA. Functional mo- nomers with the highest binding to phenol were selec- ted using computational modeling [16]. The ratio TEGDMA/OUA (85/15) in the monomer composition was optimized earlier [15]. Ketal (2,2-dimethoxy-2- phenylacetone) was used as an initiator of radical pho- topolymerization. To increase accessibility of phenol- selective binding sites in MIP membranes, they were formed according to the principle of semi-interpenetra- ting polymer network formation. A mixture of dime- thylformamide (50 vol%) and PEG 20 000 (15 wt%) was used as a porogen. Molar ratios phenol/functional monomer in the initial mixture of monomers were 1:1, 1:2, 1:3, and 1:4. Typical mixture of monomers for the synthesis of phenol-selective MIP membranes contai- ned 20 mg phenol, 55.3 mg IA (for the molar ratio 1:2), 293 mg TEGDMA, 51.7 mg OUA, 50 vol % DMF, and 0.5 wt% ketal. Monomer mixture was polymerized bet- ween two glass slides fixed at a distance 60 µm. Radi- cal polymerization was initiated by UV-irradiation (� = = 365 nm) and performed for 30 min. Blank membra- nes were synthesized from the same mixture of mono- mers, except for phenol. Template molecules and non- polymerized components were extracted from the fully- formed membranes with hot ethanol in Soxhlet appa- ratus for 8 h. Polymeric porogen (PEG 20 000) was re- moved by extraction in water for 8 h (until the constant weight of the samples was reached). Calibration of the colorimetric sensor system for phe- nol detection. Samples of phenol-imprinted and blank membranes (1 � 1 cm) were used for the adsorption of phenol from 50 nM–10 mM standard phenol aqueous solutions. Phenol, which was selectively adsorbed by the binding sites in MIP membranes structure, was vi- sualized after its interaction with 4-aminoantipyrine in alkaline media in the presence of potassium ferricya- nide. The adsorption procedure was followed by wa- SERGEYEVA T. A. ET AL. shing with distilled water, containing 5 % acetonitrile. The membrane samples were wetted with the mixture of 2 % aqueous 4-aminoantipyrine and 10 % ammonium hydroxide (1/3). Then the samples were treated with 2 % aqueous K3[Fe(CN)6], which resulted in immediate formation of a pink-colored staining with the intensity, proportional to phenol concentration in the analyzed so- lutions. Intensity of staining was estimated using «Scion Image J» software («Wayne Rasband, Inc.», USA). Spectrophotometric detection of phenol. 180 µl of the 50 nM–10 mM standard phenol solution or analyzed aqueous sample, 60 µl of the mixture of 2 % aqueous 4-aminoantipyrine and 10 % NH4OH (1:3) and 30 µl of 2 % aqueous K3[Fe(CN)6] were mixed in the poly- styrene microtiter plate wells. The absorbance values were measured at � = 450 nm using microplate reader DYNEX Technologies (UK). All measurements were made in triplicate. Results and discussion. Detection of phenol, which is selectively adsorbed by artificial receptor sites in the MIP membranes structure is based on its ability to form coloured complexes with 4-aminoantipyrine in alkali- ne media in the presence of potassium ferricyanide [18]. Intensity of the membrane staining is proportional to phenol concentration in the analyzed sample. To provi- de better accessibility of the receptor sites to phenol, MIP membranes were synthesized by in situ polymeri- zation according to the principle of the semi-IPN forma- tion [19]. Influence of the type of the functional mono- mer used for the membrane synthesis as well as molar ratio between the template and a functional monomer on analytical characteristics of corresponding sensor systems was investigated. General selectivity of the sen- sor systems and effectiveness of their application for phenol analysis in natural and waste waters was analysed. It is widely recognized that binding energy between the template and functional monomers directly influen- ce affinity and selectivity of artificial receptor sites in the resulting polymer. The method of computational modelling was demonstrated to be effective for the se- lection of the optimal functional monomers for both MIPs and MIP membranes synthesis [12, 16, 20]. Ac- cording to our previous results [16], IA, AMPSA, AA, and MA, providing binding energies: –34.80 kcal/mol, –30.86 kcal/mol, –24.14 kcal/mol, and –23.17 kcal/mol, respectively are the best functional monomers for the synthesis of the phenol-selective MIPs. These mono- mers were used in the present research for the MIP membranes synthesis. It was shown that the MIP membranes formed using IA as a functional monomer were the most effective for the construction of the colorimetric sensor systems. The- se membranes revealed both the highest intensity of stai- ning as compared to the MIP membranes synthesized with the other functional monomers as well as the high- est levels of selective phenol adsorption (which were es- timated as a difference in staining of MIP and corres- ponding blank membranes) (Fig. 1). Importantly, this result was in a good accordance with data of computa- tional modelling [16]. According to these data, IA was shown to give the highest binding energy with phenol as compared to the other three functional monomers. At the same time the level of non-specific binding of phe- nol by the blank membranes was quite significant in all cases (Fig. 1). First of all, it can be associated with the high levels of non-specific phenol adsorption by MIP and blank membranes caused by hydrophobic interactions. This also can be explained by the fact that the formation of selective binding sites requires multiple interactions between monomers and a template, which is difficult to achieve for monofuncitonal [21]. Since the best recognition properties were demon- strated for MIP membranes synthesized using IA, this 211 COLORIMETRIC BIOMIMETIC SENSOR BASED ON MOLECULARLY IMPRINTED POLYMER MEMBRANES 0 0.01 0.02 0.03 In te n si ty o f st ai n in g , A .U . 1 2 3 4 b c a Fig. 1. Dependence of phenol selective adsorption on the type of a functional monomer used for membrane synthesis: 1 – acrylamide; 2 – methacrylic acid; 3 – itaconic acid; 4 – 2-acrylamido-2-methyl-1-pro- pansulfonic acid (a – selective absorption; b – MIP; c – blank). Aqueous solution of phenol (500 µM) was used for the adsorption experiments monomer was chosen for the further investigation. Theo- retically, not all molecules of a functional monomer present in a monomer mixture are included in the spe- cific binding sites. There is a balance between high con- centrations of the monomers required to shift equilibri- um in monomer mixture toward formation of mono- mer-template complex, and between impact of «free» monomers on the high level of non-specific binding in the resulting polymer. To optimize polymer specificity, a set of MIP and corresponding blank membranes was synthesized from the monomer mixtures with the diffe- rent molar ratio phenol-IA (1:1, 1:2, 1:3, and 1:4). The ability of these membranes to adsorb phenol was analy- zed by monitoring formation of the colored complexes on their surface. The optimal recognition properties we- re observed for the MIP membranes synthesized using 1:1 ratio phenol-IA (Fig. 2). Apparently, in the case of higher content of the functional monomer in the initial mixture, random distribution of the excess of functional groups on the membrane surface results in high levels of non-specific binding, which are not associated with the effect of imprinting. Typical calibration curve of the developed colori- metric sensor system is shown in Fig. 3. It was demonst- 212 SERGEYEVA T. A. ET AL. Phenol concentration, µM In te n si ty o f st ai n in g , A .U . 0 0.01 0.02 0.03 0.04 0.05 1,00E-05 1,00E-04 1,00E-03 1,00E-02 1,00E-01 1,00E+00 1,00E+01 2 1 Fig. 3. Calibration plot of the colorimetric sensor system for phenol detection in aqueous samples: 1 – MIP membrane; 2 – blank mem- brane ! 0.01 0 0.03 0.04 0.05 0.06 1 2 3 4 Ratio phenol:itaconic acid In te n si ty o f st ai n in g , A .U . 0 0.01 0.02 1 2 3 4 c a b Fig. 2. Dependence of intensity of staining of MIP and blank memb- ranes synthesized with itaconic acid as a functional monomer on the ra- tio of phenol:functional monomer in the monomer mixture: 1 – 1:1; 2 – 1:2; 3 – 1:3; 4 – 1:4 (a – MIP; b – blank; c – selective adsorption). Aqueous solution of phenol (500 µM) was used for the adsorption experiments 0 0.01 0.02 0.03 0.04 0.05 0 5 10 25 50 100 150 NaCl, ìÌ In te n si ty o f st ai n in g , A .U . 0 0.01 0.02 0,03 0.04 3 4 5 6 7 8 9 In te n si ty o f st ai n in g , A .U . a b pH Fig. 4. Dependence of intensity of staining of phenol-selective MIP membranes synthesized using itaconic acid as a functional monomer on NaCl concentration (a) and on pH (b) of the analyzed sample. Aqueous 500 µM phenol solution was used for adsorption experiments rated, that under optimized conditions, a significant dif- ference between intensity of staining of MIP and blank membranes was observed. This indicates that phenol binding to MIP membrane is mainly determined by the presence of phenol-selective artificial receptor sites, confirming imprinting effect. The detection limit for phenol was estimated as 50 nM, while the detection ran- ge of the sensor system comprised 50 nM–10 mM. Since the main working characteristics of biosensors are often significantly influenced by the composition of the analyzed sample, influence of ionic strength of the samples on capability of the biomimetic sensors to ef- fective phenol binding was investigated. It has been shown that the increase in NaCl concentration in the analyzed sample up to 50 mM did not significantly af- fect the adsorption capability of the MIP membranes (Fig. 4 a). However, the further increase in salt concentration up to 150 mM caused a significant decrease in sensor response values. The influence of pH of the analyzed sample on va- lue of the sensor response was also studied. Since pH of natural waters varies from acidic (pH = 3) to alkaline (pH 9), influence of the sample pH on sensor responses was investigated in the pH range from 3 to 9. It was shown that the most effective phenol binding was achie- ved at pH 6– 8, which corresponds to pH of river and la- ke water (Fig. 4 b). General selectivity of the colorimetric sensor sys- tems based on MIP membranes was investigated using phenol structural analogues – 2-nitrophenol, 3-nitro- phenol, 4-nitrophenol, p-cresol, resorcinol, and pyroca- techol. In all cases the developed sensor system posses- sed enhanced selectivity towards phenol (Fig. 5). The developed colorimetric sensor systems were tested for phenol detection in both model and real envi- ronmental samples (drinking, natural and waste wa- ters). It was demonstrated that the composition of the analyzed samples had insignificant influence on the ac- curacy of phenol detection using MIP membranes. The results of phenol detection using sensor method were in a good accordance with the results obtained using tra- ditional spectrophotometric method (Fig. 6). 213 COLORIMETRIC BIOMIMETIC SENSOR BASED ON MOLECULARLY IMPRINTED POLYMER MEMBRANES 0 0.01 0.02 0.03 In te n si ty o f st ai n in g , A .U . 1 2 3 4 5 6 7 Fig. 5. Cross-reactivity of the colorimetric sensor system based on MIP membranes. Aqueous solutions (500 µM) of phenol and its analogues were used for the adsorption experiments: 1 – phenol; 2 – 2; 3 – 3-nit- rophenol; 4 – 4-nitrophenol; 5 – p-cresol; 6 – resorcinol; 7 – pyro- catechol 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 Samples P h e n o l co n ce n tr a ti o n , µ M 1 2 3 4 5 6 7 8 9 10 11 12 a b Fig. 6. Results of phenol detection obtained by the colorimetric sensor system based on MIP membranes (a) and traditional spectrophoto- metric method (b) in samples of tap, natural, and waste waters: 1 – borehole «Troyanda», Baryshivka, Kyiv region; 2 – tap water, Kyiv; 3 – source «Dubky», Kyiv; 4 – river water, river Syrets; 5 – milk plant «Ichnya», waste waters; 6 – pond «Ichnya»; 7 – Kyiv water channel, incoming water; 8 – Kyiv water channel, outcome water; 9 – Ukrainian Research Institute «UkrNIIPlastmash», waste waters; 10 – river Vita, v. Pyrogiv, Kyiv region; 11 – filtrate of the city dump (v. Pyrogiv, Kyiv region); 12 – river Stugna, Vasylkiv, Kyiv region The stability of the MIP-membrane-based sensors stored at room temperature for 12 months was assessed, showing negligible changes in their performance du- ring this period. As compared to the traditional instru- mental analytical methods the developed system is high- ly-sensitive, easy-to-use, and can provide express-ana- lysis of phenol content in water in real analytical appli- cations. As compared to the existing biosensor methods of phenol detection, the proposed sensor system provi- des similar sensitivity and significantly higher storage stability. Conclusions. Free-standing MIP membranes ca- pable of highly-selective phenol binding were synthe- sized by in situ polymerization and their composition optimized. The developed membranes were used in easy-to-use and inexpensive colorimetric sensor system for phenol detection in environmental samples. Their performance was characterized by low detection limit (50 nM), and wide detection range (50 nM–10 mM). The sensor system demonstrated high selectivity tow- ards phenol and revealed relatively low binding of its structural analogues. The sensor system was shown to be effective for phenol detection in environmental sam- ples (natural and waste waters), with the results of the detection in a good accordance with those obtained by the traditional spectrophotometric method. Acknowledgement. Financial support from Natio- nal Academy of Sciences of Ukraine (Programme «Sen- sors for medical-ecological and industrial purposes: metrological attestation and applications») is gratefully acknowledged. Êîëîðèìåòðè÷í³ ñåíñîðí³ ñèñòåìè íà îñíîâ³ ïîë³ìåð³â- á³îì³ìåòèê³â äëÿ âèñîêîñåëåêòèâíîãî âèçíà÷åííÿ ôåíîëó ó äîâê³ëë³ T. À. Ñåðãåºâà, Ä. Ñ. ×åëÿä³íà, Ë. À. Ãîðáà÷, Î. Î. Áðîâêî, Î. Â. Ï³ëåöüêà, Ñ. À. Ï³ëåöüêèé, Ë. Ì. Ñåðãåºâà, À. Â. ªëüñüêà Ðåçþìå Ìåòà. Ðîçðîáêà ïðîñòèõ ó âèêîðèñòàíí³ êîëîðèìåòðè÷íèõ ñåí- ñîðíèõ ñèñòåì äëÿ øâèäêîãî ³ òî÷íîãî âèçíà÷åííÿ ôåíîëó ó çðàç- êàõ ³ç äîâê³ëëÿ. Ìåòîäè. Ìåòîä ìîëåêóëÿðíîãî ³ìïðèíòèíãó, ìå- òîä ïîë³ìåðèçàö³¿ in situ ìîëåêóëÿðíî ³ìïðèíòîâàíèõ ïîë³ìåðíèõ (Ì²Ï) ìåìáðàí. Ðåçóëüòàòè. Çàïðîïîíîâàíèé ñåíñîð ñòâîðåíî íà îñíîâ³ Ì²Ï ìåìáðàí, ñèíòåçîâàíèõ ìåòîäîì ïîë³ìåðèçàö³¿ in situ, ÿê³ ìàþòü ó ñâî¿é ñòðóêòóð³ øòó÷í³ ðåöåïòîðí³ ñàéòè çâ’ÿ- çóâàííÿ ôåíîëó. Ê³ëüê³ñíå âèçíà÷åííÿ ôåíîëó, ñåëåêòèâíî àäñîð- áîâàíîãî Ì²Ï ìåìáðàíàìè, ãðóíòóºòüñÿ íà äåòåêö³¿ çàáàðâëå- íîãî ó ìàëèíîâèé êîë³ð ïðîäóêòó éîãî ðåàêö³¿ ç 4-àì³íîàíòèï³ðè- íîì. ²íòåíñèâí³ñòü çàáàðâëåííÿ Ì²Ï ìåìáðàí º ïðîïîðö³éíîþ êîíöåíòðàö³¿ ôåíîëó â àíàë³çîâàíîìó çðàçêó. Ôåíîë äåòåêòóºòü- ñÿ ó ä³àïàçîí³ 50 íÌ–10 ìÌ, ùî â³äïîâ³äàº êîíöåíòðàö³ÿì, ÿê³ íå- îáõ³äíî âèÿâëÿòè ó ïðèðîäíèõ ³ ñò³÷íèõ âîäàõ. Ñòàá³ëüí³ñòü ñåí- ñîðíèõ ñèñòåì íà îñíîâ³ Ì²Ï ìåìáðàí ñòàíîâèòü12 ì³ñÿö³â çà ê³ìíàòíî¿ òåìïåðàòóðè. Âèñíîâêè. Ñåíñîðí³ ñèñòåìè çàáåçïå÷ó- þòü âèñîêîñåëåêòèâíèé ³ ÷óòëèâèé àíàë³ç ôåíîëó ÿê ó ìîäåëüíèõ, òàê ³ ðåàëüíèõ çðàçêàõ (ïèòíà, ïðèðîäíà, ñò³÷íà âîäà). Ïîð³âíÿíî äî òðàäèö³éíèõ ìåòîä³â âèçíà÷åííÿ ôåíîëó ïðîïîíîâàíà ñèñòå- ìà º ïðîñòîþ ó âèêîðèñòàíí³ òà ìîæå áóòè çàñòîñîâàíà çà ïî- ëüîâèõ óìîâ. Êëþ÷îâ³ ñëîâà: ôåíîë, ìîëåêóëÿðíî ³ìïðèíòîâàí³ ïîë³ìåðí³ ìåìáðàíè, ñåíñîðè, òåñò-ñèñòåìè, êîëîðèìåòð³ÿ. Êîëîðèìåòðè÷åñêèå ñåíñîðíûå ñèñòåìû íà îñíîâå ïîëèìåðîâ- áèîìèìåòèêîâ äëÿ âûñîêîñåëåêòèâíîãî îïðåäåëåíèÿ ôåíîëà â îêðóæàþùåé ñðåäå T. À. Ñåðãååâà, Ä. Ñ. ×åëÿäèíà, Ë. À. Ãîðáà÷, À. À. Áðîâêî, Å. Â. Ïèëåöêàÿ, Ñ. À. Ïèëåöêèé, Ë. Ì. Ñåðãååâà, À. Â. Åëüñêàÿ Ðåçþìå Öåëü. Ðàçðàáîòêà ïðîñòûõ â èñïîëüçîâàíèè êîëîðèìåòðè÷åñêèõ ñåíñîðíûõ ñèñòåì äëÿ áûñòðîãî è òî÷íîãî îïðåäåëåíèÿ ôåíîëà â îáðàçöàõ èç îêðóæàþùåé ñðåäû. Ìåòîäû. Ìåòîä ìîëåêóëÿðíîãî èìïðèíòèíãà, ìåòîä ïîëèìåðèçàöèè in situ ìîëåêóëÿðíî èìïðèí- òèðîâàííûõ ïîëèìåðíûõ (ÌÈÏ) ìåìáðàí. Ðåçóëüòàòû. Ïðåäëî- æåííûé ñåíñîð ñîçäàí íà îñíîâå ÌÈÏ ìåìáðàí, ñèíòåçèðîâàí- íûõ ìåòîäîì ïîëèìåðèçàöèè in situ, èìåþùèõ â ñâîåé ñòðóêòóðå ñèíòåòè÷åñêèå ðåöåïòîðíûå ñàéòû ñâÿçûâàíèÿ ôåíîëà. Êîëè÷å- ñòâåííîå îïðåäåëåíèå ôåíîëà, ñåëåêòèâíî àäñîðáèðîâàííîãî ÌÈÏ ìåìáðàíàìè, îñíîâàíî íà äåòåêöèè îêðàøåííîãî â ìàëèíîâûé öâåò ïðîäóêòà åãî ðåàêöèè ñ 4-àìèíîàíòèïèðèíîì. Èíòåíñèâ- íîñòü îêðàøèâàíèÿ ÌÈÏ ìåìáðàí ïðîïîðöèîíàëüíà êîíöåíòðà- öèè ôåíîëà â àíàëèçèðóåìîì îáðàçöå. Ôåíîë ìîæíî äåòåêòèðî- âàòü â ïðåäåëàõ 50 íÌ–10 ìÌ, ÷òî ñîîòâåòñòâóåò êîíöåíòðà- öèÿì, êîòîðûå íåîáõîäèìî âûÿâëÿòü â ïðèðîäíûõ è ñòî÷íûõ âî- äàõ. Ñòàáèëüíîñòü ñåíñîðíûõ ñèñòåì íà îñíîâå ÌÈÏ ìåìáðàí ñîñòàâëÿåò 12 ìåñÿöåâ ïðè êîìíàòíîé òåìïåðàòóðå. Âûâîäû. Ñåíñîðíûå ñèñòåìû îáåñïå÷èâàþò âûñîêîñåëåêòèâíûé è ÷óâñò- âèòåëüíûé àíàëèç ôåíîëà êàê â ìîäåëüíûõ, òàê è ðåàëüíûõ îáðàç- öàõ (ïèòüåâàÿ, ïðèðîäíàÿ è ñòî÷íàÿ âîäà). 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Colorimetric biomimetic sensor systems based on molecularly imprinted polymer membranes for highly-selective detection of phenol in environmental samples

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