Organic polymer dot-based fluorometric determination of the activity of horseradish peroxidase and of the concentrations
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ORIGINAL PAPER
Organic polymer dot-based fluorometric determination of the activity of horseradish peroxidase and of the concentrations of glucose and the insecticidal protein toxin Cry1Ab/Ac Xin Cheng 1 & Linhao Sun 1 Jimei Ma 1
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Ruifeng Li 1 & Yan Huang 1 & Haiwei Xu 1 & Zhen Wang 1 & Zi-Long Li 1
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Hong Jiang 1
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Received: 8 May 2019 / Accepted: 14 September 2019 # Springer-Verlag GmbH Austria, part of Springer Nature 2019
Abstract Fluorescent polymer dots (PDs) with maximum excitation/emission wavelengths of 410/515 nm were prepared in water solution from 1,4-benzoquinone and ethylenediamine. The green fluorescence of these PDs is screened off by the red-colored oxidation product (PPDox, maximum absorption at 510 nm) formed by horseradish peroxidase (HRP)-catalyzed oxidation of pphenylenediamine (PPD). It causes the reduction of the fluorescence intensity of the PDs due to spectral overlap and an inner filter effect (IFE). If glucose is enzymatically oxidized under the formation of H2O2, the formed H2O2 can be quantified by the above IFE. The assay for HRP activity and glucose have detection limits of 0.2 U·L−1 and 0.1 μM, respectively. The nanoprobe was further extended to an immunosorbent assay (ELISA) for the determination of insecticidal Cry1Ab/Ac protein with a detection limit of 0.25 ng·mL−1. The ELISA was applied to rice leaf analysis. Keywords Nanoprobe . p-Phenylenediamine . Glucose oxidase . Inner filter effect . Fluorescent enzyme-linked immunosorbent assay . Rice leaves
Introduction Horseradish peroxidase (HRP), an important enzyme obtained from horseradish [1], is frequently utilized in sensing various important substances in clinical chemistry [2], environmental chemistry [2] and food industry [3]. Glucose is also closely related to HRP-catalyzed cycle in physiology [4–6]. Although plenty of methods for the detection of glucose have been developed, including colorimetry [7], fluorescent spectrometry
Xin Cheng and Linhao Sun contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-019-3831-9) contains supplementary material, which is available to authorized users. * Hong Jiang [email protected] * Jimei Ma [email protected] 1
Department of Chemistry, College of Science, Huazhong Agricultural University, Wuhan 430070, China
[8] and electrochemistry methods [9], etc. [10], only a few detection strategies for HRP was reported [11–13]. Among the analytical techniques, the fluorescent spectrometry appears to be an effective and low-cost detection tool due to its excellent fluorescence brightness, extraordinary photostability, excellent biocompatibility and good sensitivity [14, 15]. Fluorescent sensors have been commonly used for chemical and biological analysis with the detecting mechanisms including electronic energy transfer (EET), fluorescence resonance energy transfer (FRET), photoinduced electron transfer (PET), etc [14, 16, 17]. The fluorescent sensor based on these mechanisms usually involved com
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