Detection Of Bilirubin Research Articles

Turn-Off Fluorometric Determination of Bilirubin Using Facile Synthesized Nitrogen-Doped Carbon Dots as a Fluorescent Probe.

Bilirubin plays a significant role in human health management, particularly in the case of jaundice. Because of the need for the monitoring of bilirubin levels in jaundice patients, the development of a robust sensitive method becomes essential. Here, we describe the development ofa highly sensitive and selective turn-off fluorometric detectionmethod for bilirubin in blood serum samples using nitrogen-doped carbon dots (N-CDs). N-CDs was synthesized by the pyrolysis process, using citric acid and L-asparagine as the carbon and nitrogen sources, respectively. The prepared N-CDs solution showed highly intense blue emission with good stability. The HR-TEM image of N-CDs revealed spherical dot-like structures with anaverage size calculated to be 7.16nm. Further, the surface functional groups of N-CDs were analyzed by FT-IR, Raman, XRD, and XPS techniques. Fluorescence spectra showed the maximum emission intensity at 443nm (λex). The linear range of addition was performed from 1 to 150µM, and the limit of detection (LOD) was determined to be 1.97nM. The emission of N-CDs was quenched by Förster Resonance Energy Transfer (FRET) by adding bilirubin. These N-CDs showed extraordinary sensitivity and selectivity in the detection of bilirubin. Hence, this fluorescent probe has been proven successful in detecting the concentration of free bilirubin in human serum samples.

ZnS nanocrystallites as fluorescent probe for selective detection of bilirubin

Selective determination of bilirubin is much essential to identify neonatal hyperbilirubinemia. Herein, we present a simple and precise method for quantitative measuring of bilirubin levels using polyvinylpyrrolidone (PVP) capped ZnS nanocrystallites (ZSP). The crystalline structure and purity are studied from x-ray diffraction (XRD) patterns and the transmission electron microscope (TEM) images. The optical characteristics are analyzed using UV-visible absorption and photoluminescence spectra. For ZSP, the estimated value for the optical band gap is 4.06 eV, surpassing the band gap value of uncapped ZnS (3.89 eV). Upon 265 nm excitation, ZSP nanocrystallites exhibits blue emission peaked at 371 nm wavelength. The fluorescence intensity shows a consistent reduction as the bilirubin concentration rises from 3.0 μM to 80 μM. The spectral overlap between the emission spectrum of ZSP and the absorption spectrum of bilirubin, results in fluorescence attenuation due to the inner filter effect (IFE). The ZnS nanocrystallites exhibit remarkable fluorescence properties and demonstrate high sensitivity in detecting bilirubin, achieving a limit of detection (L.O.D) as low as 56 nM. The selectivity of bilirubin over certain co-existing biomolecules is also analyzed. The utility of the fluorescence probe for practical use is confirmed in spiked human serum and urine samples.

Label-Free Detection of Unbound Bilirubin and Nitrophenol Explosives in Water by a Mechanosynthesized Dual Functional Zinc Complex: Recognition of Picric Acid in Various Common Organic Media.

High levels of unconjugated bilirubin (UB) in serum lead to asymptomatic and neonatal jaundice and brain dysfunctions. Herein, we have reported the detection of UB at as low as 1 μM in an aqueous alkaline medium using a Zn(II) complex. The specificity of the complex has been validated by the HPLC in the concentration window 6-90 μM, which is rare. The sensory response of the probe at physiological pH against nitro explosives developed it as an instant-acting fluorosensor for picric acid (PA) and 2,4-dinitrophenol (2,4-DNP). Spectroscopic titration provided a binding constant of 4×105 M-1 with PA. The naked eye detection was found to be 15 μM. The solid-state photoluminescent nature of the complex enabled it for PA sensing in the solid phase. Interestingly, the probe remained fluorescent in various volatile and non-volatile organic solvents. As a result, it can also detect PA and 2,4-DNP in a wide range of common organic media. NMR studies revealed the coordination of PA, 2,4-DNP, and UB to the Zn(II) center of the probe, which is responsible for the observed quenching of the probe with the analytes.

Development of Terbium based luminescent platform as a “Turn-off” sensor for the detection of bilirubin

The sensitive and selective fluorescent probes for biomolecule detection have significant application in clinical diagnosis and bioanalysis. Hyperbilirubinemia is a disease condition in which the excessive or less production of bilirubin in blood resulting in jaundice. In this work, we have synthesised Terbium based bright green, fluorescent lanthanide complex (Tb-DPA) with emission atλem = 544 nm for the detection of Bilirubin. A one-step synthesis approach was adopted with2,6-pyridine dicarboxylic acid (DPA) as ligand and Sodium dodecyl sulphate (SDS) as surfactant. The green fluorescence of Tb-DPA was quenched with the addition of bilirubin due to the inner filter effect mechanism. The fluorescence intensity of Tb-DPA decreases as bilirubin concentration increases and a linear “turn-off” response is obtained. Even in presence of other coexisting biomolecules and ions, the Tb-DPA probe exhibited good selectivity and sensitivity for the detection of bilirubin. The limit of detection (LOD) of 2.6 mg/dL was obtained. Furthermore, the proposed fluorescent probe was applied in Human serum and urine samples with satisfactory recoveries in the range of 92–122 %. Based on the result obtained, it has probability to be used as a rapid detection method for bilirubin in biological samples at bedside.

Serum amyloid A as a biomarker for immunoglobulin resistance in Kawasaki disease

Background Intravenous immunoglobulin (IVIG) resistance is of prime importance in Kawasaki disease (KD). In this study, we examined the value and mechanism of serum amyloid A (SAA) level in predicting IVIG resistance in patients with KD. Methods SAA levels were measured in 497 consecutive patients with KD before IVIG therapy in the training set. The patients were divided into two groups (IVIG-responsive and IVIG-resistant) according to the American Heart Association (AHA) definition of IVIG resistance. Demographic, echocardiographic, and laboratory data were also retrospectively analyzed and tabulated to predict IVIG resistance. The predictive value of SAA was validated on test sets of prospective data. Cytokine microarrays were analyzed from 4 patients with resistant to IVIG, 4 patients with responsive to IVIG and 4 healthy volunteers. Results During the training set, 409 patients with KD were enrolled, of whom 43 (10.5%) were resistant to initial IVIG treatment and 47 (11.49%) had coronary artery lesions (CALs). Serum levels of SAA were higher in the IVIG resistant group compared to the IVIG responsive group, (380.00 [204.40–547.25] vs 230.85 [105.40–490.00] mg/L; p = .008). The values of total bilirubin, C-reactive protein, neutrophils, alanine aminotransferase, aspartate aminotransferase, interleukin-6(IL-6), and procalcitonin were significantly higher in the IVIG-resistant group than in the IVIG-responsive group (p < .05); however, the lymphocytes, platelets, serum sodium levels, and duration of fever before IVIG therapy were significantly lower (p < .05). There was no significant difference in SAA levels between patients with KD with and without CALs. Binary logistic regression analysis showed that SAA (p = .008), neutrophils (p < .001), total bilirubin (p = .001), platelet count (p = .004), and serum sodium level (p = .019) were independent factors influencing IVIG resistance. The optimal cutoff value of SAA for IVIG resistance prediction was 252.45 mg/L, with a corresponding clinical sensitivity of 69.8% and specificity of 54.4%. Based on receiver operating characteristic (ROC) curve analyses, the area under the curve (AUC) of combined detection with these five indicators was 0.800, clinical sensitivity was 69.8%, and specificity was 76.2%. In the prospective data, the sensitivity, specificity, and accuracy of SAA for identifying IVIG resistance KD were 77.8%,69.0%, and 70.0%, respectively. Compared with IVIG- responsive group and healthy children, the levels of IL-6 was upregulated significantly in IVIG-resistant group through cytokine microarrays. Conclusions SAA may be a potential biomarker for predicting IVIG responsiveness to KD, Combined detection of SAA levels, total bilirubin, neutrophil count, platelet count, and serum sodium levels is superior to that of any other single indicator for predicting IVIG resistance in KD. And elevated SAA may accompany with IL-6 in KD patients, its use in clinical practice may be helpful for treatment management.

A Study on the Relation Between Voltage and Bilirubin Concentration in Artificial Blood Solution by Bilirubin Detector

This research project focuses on investigating the correlation between electric potential difference and bilirubin concentration within an artificial blood solution, utilizing the principles of light absorption. The bilirubin detector's design and construction incorporate a blue light source with a 460 nm wavelength and a green light source with a 560 nm wavelength, enabling interaction with bilirubin concentrations ranging from 2 to 14 mg/dl. Analysis of the project's outcomes revealed a direct relationship between the bilirubin detector's output voltage and the concentration of the bilirubin solution. Notably, an interaction with bilirubin was observed at the 460 nm wavelength, influencing light absorption. In cases of higher bilirubin concentration, light transmittance was diminished, leading to a heightened voltage difference. Conversely, lower bilirubin concentrations facilitated increased light transmission, resulting in a reduced voltage difference. This study contributes to a comprehensive understanding of bilirubin detection methodologies and their potential applications in medical diagnostics.

A sensitive “turn-off-on” fluorescence platform for bilirubin detection based on conjugated polymer nanoparticles and ferric ions

Bilirubin is an important biomarker of jaundice, and the quantitative detection of bilirubin is of great significance in the diagnosis and prevention of early jaundice. In this paper, Fe3+ quenched conjugated polymer nanoparticles were used as a "turn-off-on" fluorescence platform to detect bilirubin for the first time. The CPNsPBMC-COOH were prepared through a nanoprecipitation method based on polystyrene maleic anhydride and PBMC, a conjugated polymer synthesized by our research group. The fluorescence of CPNsPBMC-COOH can be efficiently quenched closely to 95% by Fe3+. The optimized fluorescence platform was obtained (CPNsPBMC-COOH: 10 μg/mL, Fe3+: 4.5 μM). The platform to detect bilirubin has a higher sensitivity with a 5 nM of LOD. The detection range was in the 1.32–26.68 μM, which can cover the concentration of bilirubin in normal adult serum (<25 μM). Moreover, the stoichiometries and binding constants of CPNsPBMC-COOH, bilirubin, and Fe3+ are obtained. The binding ability is better for bilirubin and Fe3+. The static quenching mode is found by the absorption spectra and the fluorescence lifetimes. Lastly, this method has obtained satisfactory results in real biological samples (mouse serum and mouse plasma). This work can be used for daily rapid clinical bilirubin detection.

Multi-dimensional plasmonic coupling system for efficient enrichment and ultrasensitive label-free SERS detection of bilirubin based on graphene oxide-Au nanostars and Au@Ag nanoparticles

Rapid and sensitive detection of free bilirubin (BR) is essential for early diagnosis of jaundice and other hepatobiliary diseases. Inspired by sandwich immunoassay strategy, a multi-dimensional plasmonic coupling SERS platform composed of graphene oxide-Au nanostars nanocomposites (GANS NCs) and Au@Ag nanoparticles (NPs) was designed for label-free detection of BR. Specifically, GANS NCs were first prepared, and their excellent SERS activity was ascribed to synergistic enhancement effect of electromagnetic enhancement and chemical enhancement. Furthermore, SERS spectroscopy was used to monitor the adsorption process of BR. Subsequently, secondary reinforcing Au@Ag NPs were directly added, ultimately resulting in a multi-dimensional plasmonic coupling effect. The SERS enhancing mechanism of coupled system was discussed through electromagnetic field simulations. Interestingly, the high-density hotspots generated by strong plasmonic coupling in GANS-Au@Ag substrate could lead to more extraordinary SERS enhancing behavior compared to GANS NCs. Sensing efficiency of the SERS platform was examined by BR with a detection limit down to 10−11 M. Besides, GANS-Au@Ag NCs performed high uniformity and reproducibility. This work not only opens up a new avenue for construction of multi-dimensional plasmonic coupling system, but also offers a new biosensing technology for label-free diagnosis of BR-related diseases, thereby expecting to be applied in clinical diagnosis.

Non-invasive detection of haemoglobin, platelets, and total bilirubin using hyperspectral cameras

Hyperspectral imaging has emerged as a promising high-resolution and real-time imaging technology with potential applications in medical diagnostics and surgical guidance. In this study, we developed a high-speed hyperspectral camera by integrating a Fabry-Perot cavity filter on each CMOS pixel. We used it to non-invasively detect three blood components (haemoglobin, platelet, and total bilirubin). Specifically, we acquired transmission images of the subject's fingers, extracted spectral signals at each wavelength, and used dynamic spectroscopy to obtain non-invasive blood absorption spectra. The prediction models were established using the PLSR method and were modelled and validated based on the standard clinical-biochemical test values. The experimental results demonstrated excellent performance. The best predictions were obtained for haemoglobin, with a high related coefficient (R) of 0.85 or more in both the calibration and prediction sets and a mean absolute percentage error (MAPE) of only 5.7%. The results for total bilirubin were also ideal, with R values exceeding 0.8 in both sets and a MAPE of 10.6%. Although the prediction results for platelets were slightly less satisfactory, the error was still less than 15%, indicating that the results were also acceptable. Overall, our study highlights the potential of hyperspectral imaging technology for the development of portable and affordable devices for blood analysis, which can be used in various settings.

Non-invasive detection of total bilirubin based on multi-wavelength PPG signal

Background and objectiveAbnormal bilirubin metabolism can result in various liver function disorders. Current clinical practice for bilirubin level detection involves invasive blood collection from patients, which is time-consuming, painful, and poses infection risks. Thus, there is a pressing need for non-invasive bilirubin detection methods. This study aims to develop a non-invasive total serum bilirubin(TSB) detection method in humans based on multi-wavelength photoplethysmography (PPG) signals. MethodsThe experimental instrument includes a light source and a spectrometer. PPG signals are collected from the subjects' fingers, and the samples are selected based on the PPG deviation degree screening method. The absorption spectrum of blood is extracted from the PPG signal using dynamic spectroscopy. Finally, locally developed software calculates the total bilirubin value. The instrument is modeled and validated according to the clinical-biochemical test values. ResultsThe results of the prediction set (correlation coefficient is 0.91, RSMEP is 2.32 umol/L, average absolute error percentage is 9.3%) show that our method has a strong correlation with the detection results of clinical-biochemical analysis instruments. The Bland-Altman test showed that the device deviated from the data detected by biochemical methods in the clinic with a mean deviation of about 0.12 umol/L and a 95% confidence interval between -2.95 umol/L and 2.7 umol/L. ConclusionsThis study's non-invasive bilirubin detection method has high accuracy, which can meet the needs of continuous non-invasive total bilirubin detection in clinical practice.

A novel ratiometric fluorescent probe for the detection of bilirubin based on the copper nanoclusters-metal organic frameworks hybrids

Bilirubin is a key biomarker in the diagnosis of jaundice. Herein, we proposed a simple and convenient quantitative method for the fluorescent detection of bilirubin. We developed a copper nanoclusters coated by zinc-based metal-organic frameworks (Cu NCs@MOF). The prepared Cu NCs@MOF exhibited bright blue fluorescence excited with ultra-violet light and well stability at various environments. More importantly, Cu NCs@MOF was used for bilirubin detection based on the emerging red fluorescence emission peak induced by bilirubin. Furthermore, Cu NCs@MOF were made into test strip for the visual detection of bilirubin by the fluorescence color variation.

Design and Characterization of Nanoflowers-based Biosensor for Estimation of Bilirubin in Jaundice.

Nanoflowers (NFs) are flower-shaped nanoparticulate systems with a higher surface-to-volume ratio and good surface adsorption. Jaundice indicates yellow discoloration of skin, sclera, and mucus membrane and is a clinical indication of bilirubin accumulation in the blood which occurs as a consequence of the incapability of the liver to excrete bilirubin in the biliary tree or conjugate bilirubin and higher production of bilirubin in the body. Several methods have been developed so far for bilirubin estimation in jaundice like the spectrophotometric method, chemiluminescence method, etc., but biosensing methods provide advantages over traditional methods concerning the surface area, adsorption, particle size, and functional characteristics. The primary objective of the present research project was to formulate and examine the adsorbent nanoflowers-based biosensor for accurate, precise, and sensitive detection of bilirubin in jaundice. The particle size of adsorbent nanoflowers was found to be in the range of 300-600nm with the surface charge (zeta potential) in the range of -1.12 to -15.42 mV. Transmission electron microscopy and scanning electron microscopy images confirmed the flower-like morphological structure of adsorbent NFs. The adsorption efficiency of NFs for bilirubin adsorption was maximum at 94.13%. Comparative studies of bilirubin estimation in the pathological sample with adsorbent NFs and diagnostic kit displayed bilirubin concentration to be 1.0 mg/dL in adsorbent nanoflowers and 1.1 mg/dL with diagnostic kit indicating effective detection of bilirubin with adsorbent NFs. The nanoflower-based biosensor acts as a smart approach to elevate adsorption efficiency on the surface of nanoflower due to a higher surface-to-volume (SV) ratio. Graphical Abstract.

Neodymium doping MoS2 nanostructures with remarkable surface-enhanced Raman scattering activity

In recent years, the study of surface-enhanced Raman scattering (SERS) substrate on semiconductors has greatly broadened the application of SERS technology in different fields. Developing new practical semiconductor SERS substrates materials is still important work. Here, different concentrations of Nd-doped MoS2 were synthesized by a simple hydrothermal method. Compared with pure MoS2, doping Nd successfully enhanced the SERS signal. The Enhancement factor (EF) of the 2 at% Nd-doped MoS2 is up to ∼1.29 × 106 due to the energy level transition and charge transfer effect. High sensitivity SERS detection of bilirubin, dipterex, and bisphenol A (BPA) was realized, and the lowest detection concentration was 10−8 M. Moreover, 2 at% Nd doped MoS2 as SERS substrate can realize high sensitivity SERS detection of bilirubin, dipterex and bisphenol A (BPA), and the limit of detection (LOD) is 10−8 M, 10−9 M, and 10−9 M, respectively. Furthermore, 2 at% Nd-doped MoS2 SERS substrate has high uniformity and excellent reproducibility, which is beneficial for practical applications. This work not only provides an efficient technique to get sensitive, stable, and low-cost SERS substrates but also advantageous for many application scenarios.

Non-Enzymatically Colorimetric Bilirubin Sensing Based on the Catalytic Structure Disruption of Gold Nanocages.

As an essential indicator of liver function, bilirubin is of great significance for clinical diagnosis. A non-enzymatic sensor has been established for sensitive bilirubin detection based on the bilirubin oxidation catalyzed by unlabeled gold nanocages (GNCs). GNCs with dual-localized surface plasmon resonance (LSPR) peaks were prepared by a one-pot method. One peak around 500 nm was ascribed to gold nanoparticles (AuNPs), and the other located in the near-infrared region was the typical peak of GNCs. The catalytic oxidation of bilirubin by GNCs was accompanied by the disruption of cage structure, releasing free AuNPs from the nanocage. This transformation changed the dual peak intensities in opposite trend, and made it possible to realize the colorimetric sensing of bilirubin in a ratiometric mode. The absorbance ratios showed good linearity to bilirubin concentrations in the range of 0.20~3.60 μmol/L with a detection limit of 39.35 nM (3σ, n = 3). The sensor exhibited excellent selectivity for bilirubin over other coexisting substances. Bilirubin in real human serum samples was detected with recoveries ranging from 94.5 to 102.6%. The method for bilirubin assay is simple, sensitive and without complex biolabeling.

Efficient Charge-Transfer Studies for Selective Detection of Bilirubin Biomolecules Using CsPbBr3 as the Fluorescent Probe.

Bright luminescence hybrid halide perovskite nanocrystals (PNCs) as a novel fluorophore class have not been broadly explored in biological sensing. Herein, we synthesized highly fluorescent CsPbBr3 PNCs through the LARP method using oleic acid and oleyl amine as capping ligands. Morphological and optical properties of as-synthesized PNCs were studied using transmission electron microscopy, X-ray diffraction, UV-vis, and emission spectroscopic analysis. Oleic acid- and oleyl amine-capped PNCs are employed for sensitive and selective detection of bilirubin (BR). A panel of characterizations (time-correlated single-photon count spectroscopy and photoluminescence (PL)) was carried out to investigate the detailed sensing study of PNCs-BR composite for quenching the PL emission of CsPbBr3 with BR. It has been noticed that the synthesized nanoparticles are highly capable of detecting BR and thus act as a biological material sensor.

A Comparitive Study of Detection of Serum Total and Direct Bilirubin by Jendrassik &amp; Grof and Automated Method

Background: Bilirubin is one of the major bile pigments, clinically and biologically important among the bile pigments present in the mammals. Material and Methods: The cross-sectional study was conducted in department of biochemistry in the collaboration with department medicine in Civil Hospital Tarn Taran. Out of 90 patients, 54 were males and 36 were females. Result: The comparison of Serum Total and Direct bilirubin by Jendrassik &amp; Grof and Automated method was non-Significant with p value = 0.7788and 0.5394 respectively. The comparison of Serum Total and Direct bilirubin by Malloy Evelyn and Automated method is Highly Significant with p value =0.0002 and 0.0003 respectively. The mean difference of malloy with jendrassik &amp; grof and automated was 2.394 and 2.162 respectively and the mean difference of jendrassik &amp; grof with automated methodwas -0.231. Conclusion: It was concluded that a good correlation is found between bilirubin detection by manual and automated method. Non-significant mean difference between Jendrassik &amp; Grof and Automated makesit better than Malloy Evelyn.

Noninvasive direct bilirubin detection by spectral analysis of color images using a Mini-LED light source

Urine test paper is a standard, noninvasive detection method for direct bilirubin, but this method can only achieve qualitative analysis and cannot achieve quantitative analysis. This study used Mini-LEDs as the light source, and direct bilirubin was oxidized to biliverdin by an enzymatic method with ferric chloride (FeCl3) for labeling. Images were captured with a smartphone and evaluated for red (R), green (G), and blue (B) colors to analyze the linear relationship between the spectral change of the test paper image and the direct bilirubin concentration. This method achieved noninvasive detection of bilirubin. The experimental results demonstrated that Mini-LEDs can be used as the light source to analyze the grayscale value of the image RGB. For the direct bilirubin concentration range of 0.1–2 mg/dL, the green channel had the highest coefficient of determination coefficient (R2) of 0.9313 and a limit of detection of 0.56 mg/dL. With this method, direct bilirubin concentrations higher than 1.86 mg/dL can be quantitatively analyzed with the advantage of rapid and noninvasive detection.

Functionalized MoS2: circular economy SERS substrate for label-free detection of bilirubin in clinical diagnosis.

A one-pot hydrothermal synthesis of Fe-doped MoS2 nanoflowers (Fe-MoS2 NFs) has been developed as a surface-enhanced Raman spectroscopy (SERS) substrate. The Fe-MoS2 NFs display high reproducibility, stability, and recyclability, which is beneficial for the development of the sustainable ecological environment. The SERS substrate provides a high enhancement factor of 105, which can be ascribed to the inducing defects by doping Fe that can improve the charge transfer between probe molecules and MoS2. TheFe-MoS2 NFshave beenused to detect bilirubin in serum. The Fe-MoS2 NF SERS substrate exhibits a linear detection range from 10-3 to 10-9M with a low limit of detection (LOD) of 10-8M. Thesubstrate displays an excellent selectivity to bilirubin in the presence of other potentiallyinterfering molecules (dextrose and phosphate). These results providea novel concept to synthesize ultra-sensitive SERS substrates and open up a wide range of possibilities for new applications of MoS2 in clinical diagnosis.

Green synthesis of yellow-green emissive silicon nanoparticles and their application for the sensitive fluorescence detection of bilirubin.

Bilirubin, a tetrapyrrole compound metabolized by heme, is an important biomarker for diagnosis and prognosis of patients with liver diseases. Highly sensitive detection of bilirubin is essential for disease prevention and treatment. In recent years, silicon nanoparticles (SiNPs) have received intense attention due to their excellent optical properties and environmental friendliness. In this paper, water-soluble yellow-green fluorescent SiNPs were synthesized by a mild water bath method using 2-aminophenylboronic acid hydrochloride as the reducing agent and 3-[2-(2-aminoethylamino)ethylamino]propyl-trimethoxysilane (AEEA) as the silicon source. The preparation process does not require high temperature, high pressure and complex modifications. The SiNPs possessed excellent photostability and good water dispersibility. It was found that the fluorescence of SiNPs at 536 nm could be significantly quenched by bilirubin. By using SiNPs as a fluorescent probe, a novel fluorescence method for sensitive detection of bilirubin was established with a wide linear range of 0.05-75 μM and a limit of detection (LOD) of 16.67 nM. The detection mechanism was mainly due to the internal filtration effect (IFE). More significantly, the established method could successfully determine the contents of bilirubin in biological samples with good recoveries.

A graphene oxide/polyaniline nanocomposite biosensor: synthesis, characterization, and electrochemical detection of bilirubin

The level of free bilirubin is a considerable index for the characterization of jaundice-related diseases.