In vivo solid phase microextraction for bioanalysis Queiroz, Maria Eugênia Costa; Souza, Israel Donizeti de; Oliveira, Igor Gustavo de ...
TrAC, Trends in analytical chemistry (Regular ed.),
August 2022, 2022-08-00, Volume:
153
Journal Article
Peer reviewed
Solid phase microextraction (SPME) is a rapid, well-established, and solvent-free sample preparation technique. It has usually been hyphenated with GC-MS or LC-MS/MS to separate and to detect ...enriched analytes. In recent years, ambient ionization techniques have been developed fast. This has allowed MS to be used for direct and straightforward SPME analysis of complex samples, dismissing the need for chromatographic separation. SPME sampling disturbs systems to a minimum—it removes (extracts) only small fractions of analytes. Therefore, in vivo analysis is a special application area where SPME is gaining ground.
This review summarizes state-of-the-art in vivo SPME for bioanalysis (in human, rats, fishes, rhesus macaque, frog, and living cattle), including biocompatible SPME coatings, sampling approaches for in vivo SPME (skin, exhaled breath, saliva, blood, muscle, and brain sampling), analytical/detection systems, and calibration methods. This review also discusses in vivo SPME applications (pharmacokinetic studies, exposure, biomarkers, diagnosis, doping control, lipidomics, drug analysis, cosmetic dermatology, bioaccumulation, biomonitoring, environmental pollutants, and therapeutic drug monitoring) and future trends in in vivo SPME bioanalysis.
•State-of-the-art in vivo SPME for bioanalysis.•In vivo SPME applications.•Future trends in in vivo SPME bioanalysis.
Activatable (turn‐on) probes that permit the rapid, sensitive, selective, and accurate identification of cancer‐associated biomarkers can help drive advances in cancer research. Herein, a ...NAD(P)H:quinone oxidoreductase‐1 (NQO1)‐specific chemiluminescent probe 1 is reported that allows the differentiation between cancer subtypes. Probe 1 incorporates an NQO1‐specific trimethyl‐locked quinone trigger moiety covalently tethered to a phenoxy‐dioxetane moiety through a para‐aminobenzyl alcohol linker. Bio‐reduction of the quinone to the corresponding hydroquinone results in a chemiluminescent signal. As inferred from a combination of in vitro cell culture analyses and in vivo mice studies, the probe is safe, cell permeable, and capable of producing a “turn‐on” luminescence response in an NQO1‐positive A549 lung cancer model. On this basis, probe 1 can be used to identify cancerous cells and tissues characterized by elevated NQO1 levels.
An activatable chemiluminescence probe was developed for imaging endogenous NOQ1 in cells and mice without the requirement of additional components. The advantages of this probe include rapid response, good selectivity, and high signal‐to‐noise ratio.
Over the past two decades, there has been increasing focus on ascorbic acid (AA) due to its anti-oxidant and neuroprotective properties as well as its neuromodulating capability. Conventional ...analytical methods for selective in vivo monitoring of AA mainly involve complex procedures, which lower the temporal resolution and throughput of data gathering. Moreover, analytical methods for selective real-time monitoring of AA exocytosis at a single-cell level is still lacking. The lack of effective methods for AA detection in the central nervous system (CNS) has rendered difficulties in better understanding the roles of AA in brain function. AA is, in itself, electrochemically active, and thereby rationally tailoring the structure of an electrode/solution interface would offer an effective approach to selective electrochemical measurements in the CNS. Guided by this, electrochemical methods have been recently established for AA detection by combining selective electrochemical oxidation of AA at functionalized electrodes with microelectrode techniques and with in vivo microdialysis. This review mainly focuses on recent updates on in vivo detection of AA by modulating the electron transfer of AA to achieve the selectivity for its detection in the CNS, an environment with high chemical complexity. Additionally, the practical implications of the methods in selective and sensitive monitoring the dynamics of AA in different brain functions are also reviewed.
•Main electrochemical approaches for selective and real-time in vivo quantification of AA in living brains are discussed.•This review offers insight into modulating the electron transfer kinetics of AA for selective in vivo quantification of AA.•Applications of in vivo analytical methods in studying the molecular mechanism in brain functions are also highlighted.
Abstract Graphene quantum dots (GQD) generate intrinsic fluorescence, and improves aqueous stability of graphene oxide (GO) while maintaining wide chemical adaptability and high adsorption capacity. ...Despite GO's remarkable advantages in bio-imaging, bio-sensing and other biomedical applications, its biosafety issues are still unclear. Here we report a detailed and systematic study on the in vitro and in vivo toxicity of GQD. The GQD sample was prepared through a facile oxidation approach and fully characterized by means of AFM, TEM, FTIR, XPS and elemental analysis. In vitro experiments showed that GQD exhibits very low cytotoxicity owing to its ultra-small size and high oxygen content. Then, the in vivo biodistribution experiment of GQD revealed no material accumulation in main organs of mice and fast clearance of GQD through kidney. In order to mimic clinic drug administration, mice were injected with GQD and GO (as comparison) multiple times for in vivo toxicity tests. We found that GQD showed no obvious influence on mice owing to its small size, while GO appeared toxic, even caused death to mice due to GO aggregation inside mice. In brief, GQD possesses no obvious in vitro and in vivo toxicity, even under multi-dosing situation.
Solid phase microextraction (SPME) is one of the most powerful sample preparation techniques for analyte extraction and enrichment from complex matrices. SPME fibers are commonly used to extract ...analytes from collected samples. Following our recent work on development of in vivo SPME swab that integrates an SPME fiber and a medical swab (Anal Chim Acta, 2020, 1124, 71-77), the multiple SPME fibers inserted into a medical swab (multiple-SPME swab) is further developed to couple with different mass spectrometry (MS) approaches for multidimensional analysis of human saliva in this work. The new features of cotton ball and SPME fiber of multiple-SPME swab are investigated. Biomarker discovery and disease diagnosis using multiple-SPME swab are also demonstrated. The present study shows that direct coupling multiple-SPME swab with different MS-based approaches could be simple and versatile in vivo method to expand the classes of analytes extracted simultaneously from human saliva.
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•Multiple SPME fibers were inserted into a cotton ball of medical swab to fabricate a multiple-SPME swab.•The cotton ball of SPME swab played a role in excluding of large and hydrophilic analytes in sampling.•Multiple-SPME swab were employed to expand the classes of analytes extracted simultaneously from the sample matrix.•Multiple-SPME swab were coupled with different MS-based approaches for multidimensional analysis of human saliva.
Owing to its important physiological functions, especially as molecular biomarkers of diseases, RNA is an important focus of biomedicine and biochemical sensing. Signal amplification detection has ...been put forward because of the need for accurate identification of RNA at low expression levels, which is significant for the early diagnosis and therapy of malignant diseases. However, conventional amplification methods for RNA analysis depend on the use of enzymes, fixation of cells, and thermal cycling, which confine their performance to cell lysates or dead cells, thus the imaging of RNA in living cells remained until recently little explored. In recent years, the advance of isothermal amplification of nucleic acids has opened paths for meeting this need in living cells. This minireview tracks the development of in situ amplification assays for RNAs in living cells, and highlights the potential challenges facing this field, aiming to improve the development of in vivo isothermal amplification as well as usher in new frontiers in this fertile research area.
RNA imaging: In recent years, the advance of isothermal amplification technologies for nucleic acids has opened paths for the amplified imaging of RNAs in living cells. This Minireview tracks the development of in situ amplification assays of RNAs in living cells, and highlights the potential challenges facing this field.
Manganese‐based contrast agents (MnCAs) have emerged as suitable alternatives to gadolinium‐based contrast agents (GdCAs). However, due to their kinetic lability and laborious synthetic procedures, ...only a few MnCAs have found clinical MRI application. In this work, we have employed a highly innovative single‐pot template synthetic strategy to develop a MnCA, MnLMe, and studied the most important physicochemical properties in vitro. MnLMe displays optimized r1 relaxivities at both medium (20 and 64 MHz) and high magnetic fields (300 and 400 MHz) and an enhanced r1b=21.1 mM−1 s−1 (20 MHz, 298 K, pH 7.4) upon binding to BSA (Ka=4.2×103 M−1). In vivo studies show that MnLMe is cleared intact into the bladder through renal excretion and has a prolonged blood half‐life compared to the commercial GdCA Magnevist. MnLMe shows great promise as a novel MRI contrast agent.
We present a single‐pot template synthesis strategy for a manganese‐based MRI contrast agent, MnLMe. MnLMe is highly inert toward zinc‐transmetallation and displays enhanced T1 relaxivity upon non‐covalent interaction with serum albumin. In vivo studies show higher contrast enhancement in the liver and longer blood half‐life than that of the commercially available contrast agent Magnevist. MnLMe shows great potential for use as a blood pool agent.
Acid‐base homeostasis is crucial for normal physiology, metabolism, and functions of living organisms. Thus, the development of effective techniques to monitor it either in vitro or in vivo is in ...great demand. Herein, a series of multifunctional oxazine‐containing polyheterocycles with aggregation‐induced emission characteristics are in situ generated by metal‐free one‐pot A3 polymerizations of terminal diynes, dialdehydes, and ureas. Most of the polymers are synthesized with high molecular weights (Mw up to 49 900) in nearly quantitative yields and display intense solid‐state emission and remarkable fluorescence response to protonation and deprotonation. Based on the unique photophysical properties, a fast responsive and reversible fluorescent sensor for ammonia with a detection limit of 960 ppb is achieved and applied for detecting biogenic amines and seafood spoilage. Besides, the polymeric nanoparticles show excellent lysosome‐targeting specificity in cell imaging. The polyheterocycles show ratiometric pH sensing behavior with a broad acid‐base response window from pH 1 to 9, which shed light on visualizing physiologic pH in gastrointestinal tract. Taking freshwater Cladocera Moina macrocopa as a model organism, in vivo mapping of its intestinal pH shows an increased pH gradient approximately from 4.2 to 7.8 along the foregut, midgut, and hindgut.
Visualization of biogenic amines, food spoilage detection, lysosome‐targeted cell imaging, and in vivo ratiometric mapping of intestinal pH is realized by acid‐base responsive polyheterocycles with intense solid‐state emission, which are synthesized via facile and efficient metal‐free multicomponent polymerizations. In vitro and in vivo monitoring acid‐base homeostasis is now easily accessible in a real‐time, noninvasive, and on‐site manner.
The dynamics of DNA and RNA structures in live cells are important for understanding cell behaviors, such as transcription activity, protein expression, cell apoptosis, and hereditary disease, but ...are challenging to monitor in live organisms in real time. The difficulty is largely due to the lack of photostable imaging probes that can distinguish between DNA and RNA, and more importantly, are capable of crossing multiple membrane barriers ranging from the cell/organelle to the tissue/organ level. We report the discovery of a cationic carbon quantum dot (cQD) probe that emits spectrally distinguishable fluorescence upon binding with double‐stranded DNA and single‐stranded RNA in live cells, thereby enabling real‐time monitoring of DNA and RNA localization and motion. A surprising finding is that the probe can penetrate through various types of biological barriers in vitro and in vivo. Combined with standard and super‐resolution microscopy, photostable cQDs allow time‐lapse imaging of chromatin and nucleoli during cell division and Caenorhabditis elegans (C. elegans) growth.
Connect the dots: A cationic carbon quantum dot (cQD) probe emits spectrally distinguishable fluorescence signals upon binding to DNA (green) and RNA (red) in live cells, thereby enabling real‐time imaging of DNA and RNA localization and motion. The probe can penetrate through various types of biological barriers in cells and in vivo for super‐resolution microscopy and time‐lapse imaging of chromatin and nucleoli during cell division and C. elegans growth.
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