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.

Current and emerging technologies for the timely screening and diagnosis of neonatal jaundice.

Neonatal jaundice is one of the most common clinical conditions affecting newborns. For most newborns, jaundice is harmless, however, a proportion of newborns develops severe neonatal jaundice requiring therapeutic interventions, accentuating the need to have reliable and accurate screening tools for timely recognition across different health settings. The gold standard method in diagnosing jaundice involves a blood test and requires specialized hospital-based laboratory instruments. Despite technological advancements in point-of-care laboratory medicine, there is limited accessibility of the specialized devices and sample stability in geographically remote areas. Lack of suitable testing options leads to delays in timely diagnosis and treatment of clinically significant jaundice in developed and developing countries alike. There has been an ever-increasing need for a low-cost, simple to use screening technology to improve timely diagnosis and management of neonatal jaundice. Consequently, several point-of-care (POC) devices have been developed to address this concern. This paper aims to review the literature, focusing on emerging technologies in the screening and diagnosing of neonatal jaundice. We report on the challenges associated with the existing screening tools, followed by an overview of emerging sensors currently in pre-clinical development and the emerging POC devices in clinical trials to advance the screening of neonatal jaundice. The benefits offered by emerging POC devices include their ease of use, low cost, and the accessibility of rapid response test results. However, further clinical trials are required to overcome the current limitations of the emerging POC's before their implementation in clinical settings. Hence, the need for a simple to use, low-cost POC jaundice detection technology for newborns remains an unsolved challenge globally.

Orange-Red-Emitting Carbon Dots for Bilirubin Detection and Its Antibacterial Activity Against Escherichia coli and Staphylococcus aureus.

In this study, orange-red-emitting carbon dots (OR-CDs) were prepared from p-phenylenediamine (p-PDA) and urea as starting precursors through the hydrothermal method. The OR-CDs exhibited bright orange-red fluorescence at 618 nm when excited at 480 nm. The obtained OR-CDs exhibited stable photophysical properties under different physiological conditions. The unique photophysical property of OR-CDs were then utilized for fluorometric determination of bilirubin. The fluorometric assay revealed that the fluorescence intensity of OR-CDs is gradually quenched upon the addition of bilirubin (1-20 µM). The mechanism of fluorescence quenching was evaluated by steady-state fluorescence analysis and time-correlated single photon counting measurements. The OR-CDs showed good selectivity and sensitivity toward bilirubin over other common interfering biomolecules. The present fluorometric assay showed a linear response toward bilirubin between 1 and 10 µM with a limit of detection of 4.80 nM. Further, a fluorescence test cotton swab-based detection probe has been successfully developed by incorporating OR-CDs for the point-of-care detection of bilirubin in biofluids. Furthermore, a light-emitting diode light that emits orange-red light was prepared by embedding the OR-CDs within the poly(vinyl alcohol) polymer matrix. Moreover, the antibacterial activity of OR-CDs was tested against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. The antibacterial efficacy of OR-CDs was demonstrated by various mechanisms, such as reactive oxygen species generation, destruction of cell structure, chemical binding to membrane, and surface wrapping. Interestingly, the survival assay against L929 fibroblast cells exhibits favorable biocompatibility and bioimaging.

A paper-based point-of-care testing device for the colourimetric estimation of bilirubin in blood sample.

A paper-based colourimetric assay for the Point-of-Care Testing (PoCT) of bilirubin has been developed based on the formation of a green-coloured copper-bilirubin complex from a blue-coloured tetraamminecopper(II) sulphate complex. The reaction was studied and optimized by UV-Visible absorption spectroscopy and translated onto a paper strip. Hydrophobic circular well patterns on Whatman chromatography paper were created by wax printing. The tetraamminecopper(II) sulphate complex was drop cast and dried on the reagent zones in the wax-patterned paper. The images of reagent zones captured using a scanner were analyzed using ImageJ software. Bilirubin spiked blood serum was tested in the concentration range of 1.2 to 950 µM. The PAD exhibited sensitivities of 0.4197 a.u/µM and 0.1040 a.u/µM for concentration ranges of bilirubin 1.2 to 96 µM and 105 to 950 µM respectively and a low detection limit of 0.799 µM. The method is highly selective to bilirubin, even in the presence of other biomarkers in serum. A plasma separation membrane incorporated PAD was fabricated for the final testing and quantification of bilirubin from whole blood.

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.

On-line SERS detection of bilirubin based on the optofluidic in-fiber integrated GO/Ag NPs for rapid diagnosis of jaundice.

In this paper, we propose a self-assembled graphene oxide (GO)/Ag NPs SERS Raman sensor based on a novel type of optofluidic MHF as a point-of-care testing (POCT) device. This device is used to diagnose jaundice and its related diseases through on-line detection of free bilirubin content in human serum. This optofluidic Raman sensor is composed of a microstructured hollow fiber (MHF) with a microstructured channel and a suspended core, which allows the sample solution to flow in the channel while interacting with the strong evanescent field on the suspended core. Here, the suspended core was modified by a GO/Ag NPs SERS substrate. When the sample flows through the channel, and interacts with the strong evanescent field generated by the suspended core, the on-line SERS signal is generated and can be coupled back to the suspended core to be detected. In addition, both the electrostatic interaction and interference between GO/Ag NPs with the target enriched bilirubin. The results show that the detection concentration range of bilirubin aqueous, bilirubin in albumin and bilirubin in human blood are all in the range of 2 µM-100 µM, and all have a good linear response. The limit of detection reaches the order of 10-6 M. This rapid, sensitive and label-free SERS Raman sensor of free bilirubin in blood can detect excessive levels of bilirubin in the actual blood environment of the human body, providing a broad prospect for clinically accurate diagnosis of jaundice and related diseases.

Multifunctional N-Doped Carbon Dots for Bimodal Detection of Bilirubin and Vitamin B12, Living Cell Imaging, and Fluorescent Ink.

A N-doped carbon dot (NCD) has been synthesized via a simplistic one-step hydrothermal technique using l-aspartic acid and 3,6-diaminoacridine hydrochloride. The NCDs exhibit a high quantum yield (22.7%) and excellent optical stability in aqueous media. Additionally, NCDs display good solid-state yellowish-green emission and are suitable for security ink applications. The remarkable fluorescence (FL) properties of NCDs are further applied to develop a multifunctional sensor for bilirubin (BR) and vitamin B12 (VB12) via fluorescence quenching. We have systematically studied the FL quenching mechanisms of the two analytes. The primary quenching mechanism of BR is via the Förster resonant energy transfer (FRET) pathway facilitated by the H-bonding network between the hydrophilic moieties existing at the surface of BR and NCDs. In contrast, the inner filter effect (IFE) is mainly responsible for the recognition of VB12. The practicability of the nanoprobe NCDs is further tested in real-sample analysis for BR (human serum and urine samples) and VB12 (VB12 tablets, human serum, and energy drink) with a satisfactory outcome. The in vitro competency is also verified in the human cervical cancer cell line (HeLa cell) with negligible cytotoxicity and significant biocompatibility. This result facilitates the application of NCDs for bioimaging and recognition of VB12 in a living organism.

Pyrene-based prospective biomaterial: In vitro bioimaging, protein binding studies and detection of bilirubin and Fe3.

Herein, we have meticulously derived the nanosized fluorescent aggregates from pyrene Schiff base (PS) in DMSO:water (10:90) ratio. The aggregation property of PS molecule was characterized by SEM and TEM measurements, revealed the aggregated particles are in spherical shape with ~3 nm in size. Moreover, aggregates exhibit a high fluorescence quantum yield (48%) which was effectively used for the in vitro bioimaging of two different cancer cells such as A549 and MCF-7 cells in which it exhibiting excellent biocompatibility. Further, it was estimated the capability of twofold acridine orange/ethidium bromide (AO/EB) staining to identify the apoptotic associated changes in cancer cells. Additionally, the aggregates were successfully demonstrated as a luminescent probe for the perceptive biomolecule detection of bilirubin. On the other hand, the PS molecule was successfully utilized for protein binding and metal ion sensing studies. The interaction of bovine serum albumin (BSA) with PS molecule in DMSO was using fluorescence spectroscopic method and nature of interaction was also confirmed through molecular docking analysis. The PS molecule also acts as an excellent sensor for biologically important Fe3+ ion with detection limit of 336 nM. Overall, PS molecule can be a prospective material in biological field both in solution as well as aggregated forms.

Detection Methods for Bilirubin and Its Related Clinical Diseases / 中国康复理论与实践

@# Bilirubin is a kind of pigment formed from catabolism of compounds,such as hemoglobin,which had been considered to be harmful and used as an index for diagnosis of hepatobiliary diseases.Recently,the physiological functions of bilirubin has been renewed and the detection of bilirubin has been advanced.This paper reviews the methods of bilirubin examination and the relation between bilirubin content in blood and clinical diseases.