Quantification Of Glucose Research Articles

Development of Method for Rapid Quantification of Glucose, Fructose and Sucrose in Mango Juice by Chemometric Techniques in De-noised FTIR Spectroscopic Data

Development of Method for Rapid Quantification of Glucose, Fructose and Sucrose in Mango Juice by Chemometric Techniques in De-noised FTIR Spectroscopic Data

Air\u2013liquid interface enhances oxidative phosphorylation in intestinal epithelial cell line IPEC-J2

The intestinal porcine epithelial cell line IPEC-J2, cultured under the air–liquid interface (ALI) conditions, develops remarkable morphological characteristics close to intestinal epithelial cells in vivo. Improved oxygen availability has been hypothesised to be the leading cause of this morphological differentiation. We assessed oxygen availability in ALI cultures and examined the influence of this cell culture method on glycolysis and oxidative phosphorylation in IPEC-J2 using the submerged membrane culture (SMC) and ALI cultures. Furthermore, the role of HIF-1 as mediator of oxygen availability was analysed. Measurements of oxygen tension confirmed increased oxygen availability at the medium–cell interface and demonstrated reduced oxygen extraction at the basal compartment in ALI. Microarray analysis to determine changes in the genetic profile of IPEC-J2 in ALI identified 2751 modified transcripts. Further examinations of candidate genes revealed reduced levels of glycolytic enzymes hexokinase II and GAPDH, as well as lactate transporting monocarboxylate transporter 1 in ALI, whereas expression of the glucose transporter GLUT1 remained unchanged. Cytochrome c oxidase (COX) subunit 5B protein analysis was increased in ALI, although mRNA level remained at constant level. COX activity was assessed using photometric quantification and a three-fold increase was found in ALI. Quantification of glucose and lactate concentrations in cell culture medium revealed significantly reduced glucose levels and decreased lactate production in ALI. In order to evaluate energy metabolism, we measured cellular adenosine triphosphate (ATP) aggregation in homogenised cell suspensions showing similar levels. However, application of the uncoupling agent FCCP reduced ATP levels in ALI but not in SMC. In addition, HIF showed reduced mRNA levels in ALI. Furthermore, HIF-1α protein was reduced in the nuclear compartment of ALI when compared to SCM as confirmed by confocal microscopy. These results indicate a metabolic switch in IPEC-J2 cultured under ALI conditions enhancing oxidative phosphorylation and suppressing glycolysis. ALI-induced improvement of oxygen supply reduced nuclear HIF-1α, demonstrating a major change in the transcriptional response.

Fabrication and optimisation of a fused filament 3D-printed microfluidic platform

A 3D-printed microfluidic device was designed and manufactured using a low cost ($2000) consumer grade fusion deposition modelling (FDM) 3D printer. FDM printers are not typically used, or are capable, of producing the fine detailed structures required for microfluidic fabrication. However, in this work, the optical transparency of the device was improved through manufacture optimisation to such a point that optical colorimetric assays can be performed in a 50 µl device. A colorimetric enzymatic cascade assay was optimised using glucose oxidase and horseradish peroxidase for the oxidative coupling of aminoantipyrine and chromotropic acid to produce a blue quinoneimine dye with a broad absorbance peaking at 590 nm for the quantification of glucose in solution. For comparison the assay was run in standard 96 well plates with a commercial plate reader. The results show the accurate and reproducible quantification of 0–10 mM glucose solution using a 3D-printed microfluidic optical device with performance comparable to that of a plate reader assay.

A stretchable and screen-printed electrochemical sensor for glucose determination in human perspiration

Here we present two types of all-printable, highly stretchable, and inexpensive devices based on platinum (Pt)-decorated graphite for glucose determination in physiological fluids. Said devices are: a non-enzymatic sensor and an enzymatic biosensor, the latter showing promising results. Glucose has been quantified by measuring hydrogen peroxide (H2O2) reduction by chronoamperometry at −0.35V (vs pseudo-Ag/AgCl) using glucose oxidase immobilized on Pt-decorated graphite. The sensor performs well for the quantification of glucose in phosphate buffer solution (0.25M PBS, pH 7.0), with a linear range between 0 mM and 0.9mM, high sensitivity and selectivity, and a low limit of detection (LOD). Thus, it provides an alternative non-invasive and on-body quantification of glucose levels in human perspiration. This biosensor has been successfully applied on real human perspiration samples and results also show a significant correlation between glucose concentration in perspiration and glucose concentration in blood measured by a commercial glucose meter.

Nanohybrids consisting of magnetic nanoparticles and gold nanoclusters as effective peroxidase mimics and their application for colorimetric detection of glucose.

Although protein-stabilized gold nanoclusters (AuNCs) have gathered recent attention as biocompatible peroxidase mimics, their practical utility has been critically limited by the low catalytic activity. Here, the authors have developed a nanohybrid material to significantly enhance the catalytic activity of AuNCs by combining them with other inorganic enzyme mimetics, Fe3O4 magnetic nanoparticles (MNPs), through electrostatic attraction. Owing to the synergistic effect by incorporating AuNCs and MNPs, the constructed nanohybrids yielded highly enhanced catalytic activity and enabled rapid catalytic oxidation of 3,3',5,5'-tetramethylbenzidine substrate to produce a blue-colored solution in proportional to the amount of H2O2. Moreover, a highly sensitive and selective glucose biosensing strategy was developed based on the coupled catalytic action between glucose oxidase and the nanohybrids. Using this method, target glucose was successfully detected in a linear concentration range from 150 to 750 μM with a detection limit as low as 100 μM. Along with excellent linearity, high precision and reproducibility were achieved by employing real human blood serums, which enables its use for the reliable quantification of glucose in practical use. Based on these results, the authors anticipate that the nanohybrids consisting of MNPs and AuNCs can serve as potent peroxidase mimics for the detection of clinically important target molecules.

Rhinacanthins-rich Extract Enhances Glucose Uptake and Inhibits Adipogenesis in 3T3-L1 Adipocytes and L6 Myotubes.

Background:Obesity is one of the imperative dynamics in the incidence and intensification of type 2 diabetes mellitus (T2DM). Rhinacanthus nasutus leaf extracts are previously reported for their antidiabetic and antiobesity potential.Objective:The present study was performed to evaluate glucose uptake stimulatory and antiadipogenic activities of a standardized rhinacanthins-rich extract (RRE) and its marker compounds namely rhinacanthin-C (RC), rhinacanthin-D (RD), and rhinacanthin-N (RN) in 3T3-L1 and L6 cells.Materials and Methods:RRE was prepared by a green extraction process, and the marker compounds (RC, RD, and RN) were isolated from the RRE using a silica gel column chromatography. Glucose uptake stimulation in both 3T3-L1 and L6 cells was performed by quantification of residual glucose in the media using glucose oxidase kit. Antiadipogenic activity in 3T3-L1 adipocytes was performed by intracellular lipids quantification using oil red O dye.Results:At the highest effective dose, RRE (20 μg/mL) exhibited satisfactory glucose uptake stimulatory effect in 3T3-L1 adipocytes that equivalent to RN (20 μg/mL) and the positive control insulin (0.58 μg/mL) but higher than RC (20 μg/mL) and RD (20 μg/mL). In addition, treatments of L6 myotubes showed that RRE (2.5 μg/mL) exhibited potent and equivalent glucose uptake stimulation (>80%) to RC (2.5 μg/mL) and the standard drugs, insulin (2.90 μg/mL) and metformin (219.5 μg/mL), but higher than RD (2.5 μg/mL) and RN (2.5 μg/mL). Furthermore, RRE (20 μg/mL) exhibited potent antiadipogenic effect in 3T3-L1 adipocytes, which equivalent to RC (20 μg/mL) but higher than RD (20 μg/mL) and RN (20 μg/mL).Conclusions:The undertaken study suggests that RRE could be used as an effective remedy in the treatment of obesity-associated T2DM.SUMMARY Rhinacanthins-rich extract and its marker compounds showed potent glucose uptake stimulatory activity in 3T3-L1 adipocytes and L6 myotubesRhinacanthins-rich extract and rhinacanthin-C showed comparable antiadipogenic effect in 3T3-L1 adipocytesRRE could be used as an effective remedy in the treatment of obesity-associated T2DM. Abbreviations used: T2DM: Type-2 diabetes mellitus; RRE: Rhinacanthins-rich extract; RC: Rhinacanthin-C; RD: Rhinacanthin-D; RN: Rhinacanthin-N; α-MEM: α-Minimum essential medium; DMEM: Dulbecco's modified Eagle's medium; HS: Horse serum; FBS: Fetal bovine serum; BSA: Bovine serum albumin; IBMX: 3-isobutyl-1-methylxanthine; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; GO: Glucose oxidase; NMR: Nuclear magnetic resonance; HPLC: High-performance liquid chromatography.

A dual enzyme-inorganic hybrid nanoflower incorporated microfluidic paper-based analytic device (μPAD) biosensor for sensitive visualized detection of glucose.

A novel microfluidic paper-based analytic device (μPAD) biosensor is developed for sensitive and visualized detection of glucose. This biosensor is easily fabricated using the wax printing technique, with a hybrid nanocomplex composed of dual enzymes glucose oxidase (GOx) and horseradish peroxidase (HRP) and Cu3(PO4)2 inorganic nanocrystals incorporated in the detection zones. The hybrid nanocomplex is found to exhibit a flower-like structure, which allows co-immobilization of these two enzymes in a biocompatible environment. These nanoflowers not only preserve the activity and enhance the stability of the enzymes, but also facilitate the transport of the substrates between the two enzymes. The biosensor is demonstrated to enable rapid and sensitive quantification of glucose in the concentration range of 0.1-10 mM with a limit of detection (LOD) of 25 μM. It is also shown to be applicable to colorimetric quantitative detection of glucose in human serum and whole blood samples, implying its potential for clinical applications.

Insulin overlapping in whole blood FTIR spectroscopy in blood glucose measurements

For the last decade, several studies on mid-IR spectroscopy for blood glucose quantification have not considered the compounds involved in the glucose regulation mechanism, in which insulin plays an important role. This work shows how insulin overlaps in the same mid-IR region in which glucose is quantified. This optical absorption interference is an important factor to be considered for this type of studies, in the scope of whole blood modeling for spectroscopy applications and the possible use of computer based metrics.

Non-enzymatic glucose sensing on copper-nickel thin film alloy

A simple and cost efficient glucose sensor was constructed using 3D printing having as active material a copper-15 at.% nickel thin film thermally co-evaporated on copper plated circuit boards. The glucose detection in alkaline solution was studied in detail by cyclic voltammetric and chronoamperometric measurements. The sensor suitability for being used in both quantitative and qualitative glucose detection was demonstrated and calibration of its response to various amounts of glucose revealed two linear regimes with different sensitivities. Glucose levels between 0 and 10mM are most efficiently quantified as indicated by an amperometric signal increase of 240μAcm−2 for each 1mM increase of glucose concentration. The potentiostatic stability of the sensor was evaluated and its complete insensitivity after 7h was solely attributed to the irreversible transformation of glucose into gluconolactone. A sensor life time of 20 cycles was demonstrated during potentiodynamic cycling when the sensor response remains constant at its maximum level. The magnitude of possible glucose quantification errors were evaluated as interferences induced by additions of ascorbic and uric acids. A worst case scenario of 96 % accuracy of glucose levels quantification was demonstrated using 25 times higher concentrations of interfering substances as compared to the glucose level.

A Robust, Enzyme-Free Glucose Sensor Based on Lysine-Assisted CuO Nanostructures.

The production of a nanomaterial with enhanced and desirable electrocatalytic properties is of prime importance, and the commercialization of devices containing these materials is a challenging task. In this study, unique cupric oxide (CuO) nanostructures were synthesized using lysine as a soft template for the evolution of morphology via a rapid and boiled hydrothermal method. The morphology and structure of the synthesized CuO nanomaterial were characterized using scanning electron microscopy (SEM) and X-ray diffraction (XRD), respectively. The prepared CuO nanostructures showed high potential for use in the electrocatalytic oxidation of glucose in an alkaline medium. The proposed enzyme-free glucose sensor demonstrated a robust response to glucose with a wide linear range and high sensitivity, selectivity, stability, and reproducibility. To explore its practical feasibility, the glucose content of serum samples was successfully determined using the enzyme-free sensor. An analytical recovery method was used to measure the actual glucose from the serum samples, and the results were satisfactory. Moreover, the presented glucose sensor has high chemical stability and can be reused for repetitive measurements. This study introduces an enzyme-free glucose sensor as an alternative tool for clinical glucose quantification.

Glucose‐induced and fructose‐induced deboronation reaction of 4‐mercaptophenylboronic acid assembled on silver investigated by surface‐enhanced Raman scattering

We examined the deboronation reaction of 4‐mercaptophenylboronic acid (4MPBA) via fructose and glucose on silver surfaces by means of surface‐enhanced Raman scattering (SERS) at the excitation wavelengths of 488, 514, and 633 nm. The SERS spectra on silver nanoparticles clearly exhibited specific spectral signatures of thiophenol (TP) peaks, indicating a deboronation reaction of 4MPBA on the surfaces, whereas no strong TP peaks were observed on gold nanoparticles. The vibrational bands at 417, 999, 1021, and 1574 cm−1 in the Ag SERS spectra could correspond to the in‐plane aromatic ring modes in TP. X‐ray photoelectron spectroscopy also supported the surface reaction on Ag by referring the B1s peaks at ~193 eV. The ratiometric Raman measurements of the band at 1574 cm−1, with respect to that at 1587 cm−1, revealed fructose and glucose quantification in the concentration range of 1–10 mm. We did not identify such changes for mannose, sucrose, and sialic acid. The SERS peaks of 4MPBA on roughened Ag plates also exhibited TP bands to show the time‐dependent spectral change. Our findings indicate that the deboronation of 4MPBA and conjugation with fructose and glucose may be facilitated efficiently on silver surfaces for their quantification. Copyright © 2016 John Wiley & Sons, Ltd.

Nonenzymatic Glucose Sensor Based on In Situ Reduction of Ni/NiO-Graphene Nanocomposite.

Ni/NiO nanoflower modified reduced graphene oxide (rGO) nanocomposite (Ni/NiO-rGO) was introduced to screen printed electrode (SPE) for the construction of a nonenzymatic electrochemical glucose biosensor. The Ni/NiO-rGO nanocomposite was synthesized by an in situ reduction process. Graphene oxide (GO) hybrid Nafion sheets first chemical adsorbed Ni ions and assembled on the SPE. Subsequently, GO and Ni ions were reduced by hydrazine hydrate. The electrochemical properties of such a Ni/NiO-rGO modified SPE were carefully investigated. It showed a high activity for electrocatalytic oxidation of glucose in alkaline medium. The proposed nonenzymatic sensor can be utilized for quantification of glucose with a wide linear range from 29.9 μM to 6.44 mM (R = 0.9937) with a low detection limit of 1.8 μM (S/N = 3) and a high sensitivity of 1997 μA/mM∙cm−2. It also exhibited good reproducibility as well as high selectivity.

Cerebral metabolic rate of glucose quantification with the aortic image-derived input function and Patlak method: numerical and patient data evaluation.

Quantitative maps of cerebral metabolic rate of glucose (CMRGlu) from fluorine-18 fluorodeoxyglucose-PET are useful in brain studies, but are challenging to acquire because of technical constraints, which hinder their use in clinical routine. Aortic image-derived input functions (IDIFs) combined with Sokoloff's method were proposed as a suitable solution. However, Sokoloff's method requires the use of standard kinetic constants, which may produce biased estimates. Patlak's method would be more appropriate, but concern can arise when used with an aortic IDIF from unavailability of a complete brain curve acquired starting from injection. The aim of this study was to develop a CMRGlu quantification technique that combines Patlak's method with aortic IDIFs in a clinical setting. A simple acquisition protocol for aortic IDIF measurement was developed and applied on a sample of patients with different degrees of hypometabolism (one healthy control, four patients with a neurodegenerative condition, and one coma patient). CMRGlu estimates in vivo were obtained with both the Sokoloff method and the Patlak method. Computer simulations were performed to assess the causes of bias affecting Sokoloff and Patlak estimates and interpret the results obtained in patients. Simulations showed that Sokoloff's method is less stable than Patlak's method as the extent of bias changed across different physiological states, potentially leading to misinterpretation of clinical data. In clinical patients, Sokoloff and Patlak estimates were correlated on the whole, but deviations emerged for critical physiological states. CMRGlu quantification with the Patlak method and aortic IDIF is feasible, easy to implement in clinical practice, and superior to Sokoloff's method from a personalized medicine perspective.

A single dual-emissive nanofluorophore test paper for highly sensitive colorimetry-based quantification of blood glucose

Fluorescent test papers are promising for the wide applications in the assays of diagnosis, environments and foods, but unlike classical dye-absorption-based pH test paper, they are usually limited in the qualitative yes/no type of detection by fluorescent brightness, and the colorimetry-based quantification remains a challenging task. Here, we report a single dual-emissive nanofluorophore probe to achieve the consecutive color variations from blue to red for the quantification of blood glucose on its as-prepared test papers. Red quantum dots were embedded into silica nanoparticles as a stable internal standard emission, and blue carbon dots (CDs) were further covalently linked onto the surface of silica, in which the ratiometric fluorescence intensity of blue to red is controlled at 5:1. While the oxidation of glucose induced the formation of Fe3+ ions, the blue emission of CDs was thus quenched by the electron transfer from CDs to Fe3+, displaying a serial of consecutive color variations from blue to red with the dosage of glucose. The high-quality test papers printed by the probe ink exhibited a dosage-sensitive allochromatic capability with the clear differentiations of ~5, 7, 9, 11mM glucose in human serum (normal: 3–8mM). The blood glucose determined by the test paper was almost in accordance with that measured by a standard glucometer. The method reported here opens a window to the wide applications of fluorescent test paper in biological assays.

Development of novel bienzymatic assay for quantification of glucose in human serum

Aim: The main objective of this investigation was to design and to develop a simple, sensitive, and convenient assay for quantification of glucose in human serum sample. Materials and Methods: A novel bienzymatic assay for the quantification of glucose by spectrophotometry has been developed. The method is based on the coupling of 4-aminoantipyrine and iminodibenzyl in the presence of bienzyme peroxidase/glucose oxidase (GOx) to form a blue colored product which is monitored for absorbance at 620 nm at 23°C in 100 mmol/L KH2PO4/NaOH buffer of pH 6.3. Results and Discussion: Under optimized assay conditions, glucose could be quantified in a linear range from 1.15 to 562 μmol/L and 2.31 to 370 μmol/L by kinetic and fixed time method. The limit of detection and limit of quantification were measured. Michaelis–Menten constant for glucose was found to Km = 357 μmol/L and maximum rate of the reaction (Vmax) was 0.117 × 10−6 mol/L/min. The proposed method was applied for the quantification of glucose in human serum samples, which has a good regression coefficient of 0.9992 with the enzymatic kit method. The inter-day precision was 0.53-2.9% (n = 10) and intra-day precision was 1.5-3.03% (n = 10). The accuracy range for glucose was fond to be 90-104%. The glucose recovery ranged from 91.92% to 108.5%. Conclusion: The present assay was rapid, sensitive, and convenient for quantification of glucose in human serum sample with good recovery, high accuracy, and minimum interference.

A Co(II)-based metallo-supramolecular polymer as a novel enzyme immobilization matrix for electrochemical glucose biosensing

A Co(II)-based metallo-supramolecular polymer (polyCo) was used as a novel matrix to immobilize glucose oxidase (GOx) for enzymatic glucose sensing. A facile fabrication of a modified glassy carbon electrode (GCE) was achievable by a one-step co-immobilization method. UV–vis spectroscopy and cyclic voltammetry were used to characterize the electrochemical biosensor and the sensing mechanism of the reaction on the polyCo-modified GCE. After the optimization of composition and loading of the polyCo/GOx GCE, the fabricated sensor demonstrated excellent amperometric quantification of glucose, with high sensitivity (47.1μAmM−1cm−2) in the linear range of up to 1.4mM. The selectivity, based on the enzyme matrix without any pretreatment, was also sufficiently high. The outstanding stability of the sensor supported our assumption that metallo-supramolecular polymers can be used as electrode materials for glucose biosensors.

Non-enzymatic detection of glucose using poly(azure A)-nickel modified glassy carbon electrode

A simple, sensitive and selective non-enzymatic glucose sensor was constructed in this paper. The poly(azure A)-nickel modified glassy carbon electrode was successfully fabricated by the electropolymerization of azure A and the adsorption of Ni2+. The Ni modified electrode, which was characterized by scanning electron microscope, cyclic voltammetry, electrochemical impedance spectra and X-ray photoelectron spectroscopy measurements, respectively, displayed well-defined current responses of the Ni(III)/Ni(II) couple and showed a good activity for electrocatalytic oxidation of glucose in alkaline medium. Under the optimized conditions, the developed sensor exhibited a broad linear calibration range of 5 μM–12mM for quantification of glucose and a low detection limit of 0.64μM (3σ). The excellent analytical performance including simple structure, fast response time, good anti-interference ability, satisfying stability and reliable reproducibility were also found from the proposed amperometric sensor. The results were satisfactory for the determination of glucose in human serum samples as comparison to those from a local hospital.

Su1350 Analytical Performance of a Routinely Utilized Serum/Plasma Spectrophotometric Method for Glucose Quantification in Pancreatic Cyst Fluid

Su1350 Analytical Performance of a Routinely Utilized Serum/Plasma Spectrophotometric Method for Glucose Quantification in Pancreatic Cyst Fluid

Ultrasensitive analysis of glucose in serum by capillary electrophoresis with LIF detection in combination with signal amplification strategies and on-column enzymatic assay.

A highly specific and sensitive method for glucose quantification in human serum samples based on on-column enzymatic assay is described. In this method, the head of the capillary was used as a nanoliter-microreactor, the diluted samples spiked with a novel fluorogenic reagent named 2-[6-(4'-amino) phenoxy-3H-xanthen-3-on-9-yl] benzoic acid (APF), and the mixed enzyme solutions of glucose oxidase (GOx) and horseradish peroxidase (HRP), were individually injected into the capillary. Hydrogen peroxide (H2 O2 ) generated in situ by catalytic reaction between GOx and glucose, activates APF in the presence of HRP to form a highly fluorescent product, which was electrophoretically separated from the unreacted APF and detected by the LIF detector. The proposed method allowed the determination of glucose down to 10 nM in real samples, with RSD values lower than 3.5%, which also has the potential for measurements of multicomponents in many other systems including measurement of α-glucosidase activity and screening for its inhibitors.

Simple alternative to sialic acid determination in meningococcal polysaccharides W or Y

Physicochemical methods are the primary tests used to ensure that batches of meningococcal polysaccharide (PS) antigens are manufactured consistently to those shown to be safe and effective in clinical trials. Although modern physicochemical methods of analysis providing structural information about the antigens have been developed and used, simpler assays, which can be readily validated, are still in use for polysaccharide batch release. The simple and cheap method for Neisseria meningitidis serogroup W or Y polysaccharide (MenW or MenY PS) content quantification has been developed. This colorimetric method is based on the galactose or glucose quantification in MenW or MenY PS hydrolysate, respectively. Intra- and inter-assay precision and accuracy of the novel method have been demonstrated, in comparison to the same properties of the current regulatory approved method for the same purpose — sialic acid quantification. We provided the calculation of the possible future regulatory requirement for the galactose or glucose content in MenW or MenY PS, respectively, and revealed in detail the stoichiometric calculation behind it.