Microplate analytical method for quinones by pulse photo-irradiation and chemiluminescence detection

Abstract

Quinones are widely distributed in the nature and have various bioactivities. Besides, quinones also considered as toxicological intermediates which cause a severe dangerous effects. Hereby, sensitive, simple, and rapid method is reported for quinones determination.The proposed method employed time resolved fluorescence (TRF) microplate reader based chemiluminescent (CL) detection for the first time as a novel approach for measurement.Under pulse photo-irradiation, the unique photochemical characteristic of quinones is exploited to liberate reactive oxygen species (ROS) which reacted with photosensitized CL reagent. L-012, luminol analogue, was selected for its high sensitivity. Under our investigation, para-quinones showed high CL response when compared to ortho-quinones. A linear response was obtained for studied quinone concentrations in the range of 0.05-50 µM for 1,4-naphthquinone and of 0.05-150 µM for 2-methyl-1,4-naphthoquinone and 9,10-anthraquinone with detection limit (balnk + 3SD) of 0.01 µM. The proposed method allowed the rapid determination of large number of samples in very short time (96 sample/ 125 sec).The proposed method was successfully applied for determination of menadione in spiked human serum.

Luminol-Based Chemiluminescent Signals: Clinical and Non-clinical Application and Future Uses

“…When quinones are photo-irradiated, they liberate reactive oxygen species, which further react with photosensitized CL reagents. This method was successfully applied for determination of menadione in spiked human serum samples [173]. …”

Analytical techniques for the determination of biologically active quinones in biological and environmental samples

“…Due to the specificity of antigen-antibody interaction, the ELISA could determine menadione in fetal calf serum without any pre-treatment procedure. Recently, a microplate based CL assay was developed for the rapid determination of quinone using a time resolved fluorescence microplate reader [83]. In this assay, quinone was pulse UV irradiated to form ROS that react with luminol analogue, L-012, and the generated CL was monitored during 1500 µs.…”

Quinones as novel chemiluminescent probes for the sensitive and selective determination of biothiols in biological fluids

“…On the other hand, the important roles of quinones in biology can be attributed to their versatile oxidative and electrophilic properties which promote electron transfer in living systems and Michael addition with cellular thiols, such as free cysteine, glutathione, and cysteine residues of proteins. 13 Consequently, the nucleophilic addition of thiols to quinone moieties is well established, providing the basis for the synthesis of many biologically active compounds, 14 numerous spectroscopic and pre-column chromatographic derivatisation as well as electrochemical protocols [15][16][17][18][19] which have been assessed and successfully presented the fundamental characterization technique for the analytical determination of biological thiols. However, the reasonability of these conventions has remained a matter of theory.…”

“…The application of the chemiluminescence (CL) assay for the determination of trace and ultra-trace concentrations of organic and inorganic compounds has interestingly increased owing to the simplicity and selectivity of CL measurements. 13 Several subsidiary articles have described the CL determination of thiols using batch or flow injection methods. [20][21][22] These methods lack the selectivity required for the accurate measurement of thiols in complex biological matrices.…”

“…The proposed methods exploited the unique photochemical or redox-cycling ability of quinones to liberate powerful oxidizing agents including superoxide anion, hydroxyl radical, and hydrogen peroxide which are measured by the luminol-CL assay. 13,[26][27][28] Menadione (MQ), a representative quinone, can rapidly conjugate with plasma thiols after intraperitoneal administration forming thioether adducts. 26 Although the thioether adducts might be considered a part of the quinone metabolic fate, the adducts retain the redox reactivity of the parent quinones.…”