Double strand DNA-based determination of menadione using a Fe3O4 nanoparticle decorated reduced graphene oxide modified carbon paste electrode.

In this work an electrochemical label free DNA biosensor (ds-DNA) for the determination of menadione (MD) was developed. The biosensor was constructed using a modified nanocomposite consisting of Fe3O4 nanoparticles decorated reduced graphene oxide (Gr) on a carbon paste electrode (CPE). Scanning electron microscope (SEM), energy dispersive X-ray (EDAX) and Fourier transform infrared (FT-IR) spectroscopy confirmed the structure of the synthesized nanocomposites (electrode composition). The Gr-Fe3O4 nanocomposites formed a sensitive layer with large surface area. Electrochemical studies revealed that modification of the electrode surface with ds-DNA and Gr- Fe3O4 nanocomposite significantly increases the oxidation peak currents and reduces the peak potentials of MD. Under the optimum conditions, calibration curve was linear in the range of 0.3-100.0 nM with a detection limit of 0.13 nM. The relative standard deviation for 50.0 nM was 3.90% (n = 5). The proposed biosensor was successfully applied to the determination of MD.

Quantification of menadione from plasma and urine by a novel cysteamine-derivatization based UPLC-MS/MS method.

Menadione, as the crucial component of vitamin Ks, possessed significant nutritional and clinical values. However, there was still lack of favourable quantification strategies for it to date. For improvement, a novel cysteamine derivatization based UPLC-MS/MS method was presented in this work. The derivatizating reaction was proved non-toxic, easy-handling and high-efficient, which realized the MS detection of menadione under positive mode. Benefitting from the excellent sensitivity of the derivatizating product as well as the introduction of the stable isotope dilution technique, the quantification could be achieved in the range of 0.05-50.0ng/mL for plasma and urine matrixes with satisfied accuracy and precision. After analysis of the samples from healthy volunteers after oral administration of menadione sodium bisulfite tablets, the urinary free menadione was quantified for the very first time. We believe the progress in this work could largely promote the exploration of the metabolic mechanism of vitamin K in vivo.

A novel liquid chromatography-tandem mass spectrometry method for determination of menadione in human plasma after derivatization with 3-mercaptopropionic acid.

Menadione (VK3), an essential fat-soluble naphthoquinone, takes very important physiological and pathological roles, but its detection and quantification is challenging. Herein, a new method was developed for quantification of VK3 in human plasma by liquid chromatography-tandem mass spectrometry (LC-MS/MS) after derivatization with 3-mercaptopropionic acid via Michael addition reaction. The derivative had been identified by the mass spectra and the derivatization conditions were optimized by considering different parameters. The method was demonstrated with high sensitivity and a low limit of quantification of 0.03 ng mL(-1) for VK3, which is about 33-fold better than that for the direct analysis of the underivatized compound. The method also had good precision and reproducibility. It was applied in the determination of basal VK3 in human plasma and a clinical pharmacokinetic study of menadiol sodium diphosphate. Furthermore, the method for the quantification of VK3 using LC-MS/MS was reported in this paper for the first time, and it will provide an important strategy for the further research on VK3 and menadione analogs.

Development and validation of the first assay method coupling liquid chromatography with chemiluminescence for the simultaneous determination of menadione and its thioether conjugates in rat plasma.

Menadione (2-methyl-1,4-naphthoquinone, MQ), a component of multivitamin drugs with antihemorrhagic, antineoplastic, and antimalarial activity, is frequently used to investigate quinone-induced cytotoxicity. The formation of MQ conjugates with glutathione (GSH) by Michael addition and subsequent biotransformation to yield N-acetyl-l-cysteine conjugates is believed to be an important detoxification process. However, the resulting conjugates, 2-methyl-3-(glutathione-S-yl)-1,4-naphthoquinone (MQ-GS) and 2-methyl-3-(N-acetyl-l-cysteine-S-yl)-1,4-naphthoquinone (MQ-NAC), retain the ability to redox cycle and to arylate cellular nucleophiles. Although the nephrotoxicity and hepatotoxicity of MQ-thiol conjugates have been reported in vitro, methods for their determination in vivo have yet to be published. Herein, a highly sensitive, simple, and selective HPLC-chemiluminescence (HPLC-CL) coupled method is reported, allowing for the first time the simultaneous determination of MQ, MQ-GS, and MQ-NAC in rat plasma after MQ administration. Our method exploits the unique redox characteristics of MQ, MQ-GS, and MQ-NAC to react with dithiothreitol (DTT) to liberate reactive oxygen species (ROS) which are detected by a CL assay using luminol as a CL probe. To verify the proposed mechanism, MQ-GS and MQ-NAC were synthetically prepared. Specimen preparation involved solid-phase extraction on an Oasis HLB cartridge followed by isocratic elution on an ODS column. No interference from endogenous substances was detected. Linearity was observed in the range of 5-120 nM for MQ-GS and MQ-NAC and 10-240 nM for MQ, with detection limits (S/N of 3) of 1.4, 0.8, and 128 fmol for MQ-GS, MQ-NAC, and MQ, respectively. The application of our method reported here is the first to extensively study the stability and reversibility of thiol-quinones.

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

Quinones are widely distributed in nature and have various bioactivities. Besides, quinones are also considered as toxicological intermediates which cause severe dangerous effects. Hereby, a 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 (menadione) and 9,10-anthraquinone with detection limit (blank + 3SD) of 0.01 µM. The proposed method allowed the rapid determination of large number of samples in very short time (96 sample/125 s). The proposed method was successfully applied for determination of menadione in spiked human serum.