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Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines

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A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Medicinal Chemistry".

Deadline for manuscript submissions: closed (15 November 2021) | Viewed by 46152

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Special Issue Editors

Dr. Mário Barroso

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Laboratory of Forensic Chemistry and Toxicology, National Institute of Legal Medicine and Forensic Sciences, I.P. – South Branch, Rua Manuel Bento de Sousa, n.º 3, 1169-201 Lisboa, Portugal
Interests: method development and validation; hair testing; alternative specimens; drug monitoring; toxicology
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Eugenia Gallardo

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Guest Editor

1. Health Sciences Research Centre, University of Beira Interior (CICS-UBI), Covilhã, Portugal
2. Pharmaco-Toxicology Laboratory, UBIMedical, University of Beira Interior, Covilhã, Portugal
3. Centro Académico Clínico das Beiras (CACB)—Grupo de Problemas Relacionados com Toxicofilias, Covilhã, Portugal
Interests: toxicology; analytical method development; recreational drugs; natural psychoactive substances; therapeutic drug monitoring; sample preparation; alternative samples; miniaturized extraction procedures
Special Issues, Collections and Topics in MDPI journals

Dr. Luís Passarinha

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Guest Editor

1. Associate Laboratory i4HB—Institute for Health and Bioeconomy, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2819-516 Caparica, Portugal
2. UCIBIO—Applied Molecular Biosciences Unit, Chemistry Department, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal
3. CICS-UBI—Health Sciences Research Centre, University of Beira Interior, 6201-506 Covilhã, Portugal
Interests: method development and validation; electrochemical detection; proteomics; protein biomarkers
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Analytical tools to determine drugs, drugs of abuse, or biomarkers for the rapid diagnosis of intoxications or diseases are mandatory for every laboratory working in the bioanalytical field. In the last few years, the development of sensible analytical instrumentation (e.g., time of flight, high-resolution mass spectrometry, or Orbitrap mass spectrometers) has widened the scope of applications of such techniques to several areas, for instance, in the development of new therapeutic agents, industry, and others. This progress concerning instrumentation was also accompanied by important achievements related to sample preparation approaches, namely, trending in low-sample, low-cost, and low-organic solvent procedures without compromising sensitivity and allowing the analysis of alternative samples, such as exhaled breath, hair, oral fluid, or sweat.

Presenting a very broad scope, this Special Issue welcomes full papers, short communications, and review articles on, but not limited to, new developments in bioanalysis, sample preparation, and the development of new detection and quantification of disease biomarkers, toxic agents, and drug monitoring approaches with applications in different analytical fields.

This issue will be an excellent collection of papers for academia, or as a reference tool for researchers, particularly those working in the fields of medicine, biotechnology, pharmaceutical sciences, chemistry, and toxicology, including health and industry professionals as well.

We are looking forward to receiving your contributions.

Dr. Mário Barroso
Prof. Dr. Eugenia Gallardo
Prof. Dr. Luís Passarinha
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Molecules is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

bioanalysis
development of analytical methods
sample preparation techniques
biomarkers
toxics
drugs of abuse
drug monitoring

Published Papers (14 papers)

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Editorial

Jump to: Research, Review

5 pages, 215 KiB

Open AccessEditorial

Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines

by Mário Barroso, Eugenia Gallardo and Luís A. Passarinha

Molecules 2022, 27(10), 3188; https://doi.org/10.3390/molecules27103188 - 17 May 2022

Viewed by 1167

Abstract

New developments in instrumental approaches, for instance, hyphenated techniques, have allowed great advances in the bioanalytical field over the last half century, and there is no doubt that toxicology was one of the most improved areas [...] Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

Research

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17 pages, 4205 KiB

Open AccessArticle

Biosensors Based on Ion-Sensitive Field-Effect Transistors for HLA and MICA Antibody Detection in Kidney Transplantation

by Thu Zar Ma Ma Moe Min, Sonwit Phanabamrung, Woraphan Chaisriratanakul, Apirak Pankiew, Awirut Srisuwan, Kondee Chauyrod, Cholatip Pongskul, Chamras Promptmas and Chanvit Leelayuwat

Molecules 2022, 27(19), 6697; https://doi.org/10.3390/molecules27196697 - 08 Oct 2022

Cited by 1 | Viewed by 1861

Abstract

This work demonstrates the ability of the Ion-Sensitive Field-Effect Transistor (ISFET)-based immunosensor to detect antibodies against the human leukocyte antigen (HLA) and the major histocompatibility complex class-I-related chain A (MICA). The sensing membrane of the ISFET devices was modified and functionalized using an APTES-GA strategy. Surface properties, including wettability, surface thickness, and surface topology, were assessed in each module of the modification process. The optimal concentrations of HLA and MICA proteins for the immobilization were 10 and 50 μg/mL. The dose-response curve showed a detection range of 1.98–40 µg/mL for anti-HLA and 5.17–40 µg/mL for anti-MICA. The analytical precision (%CV) was found to be 10.69% and 8.92% for anti-HLA and -MICA, respectively. Moreover, the electrical signal obtained from the irrelevant antibody was considerably different from that of the specific antibodies, indicating the specific binding of the relevant antibodies without noise interference. The sensitivity and specificity in the experimental setting were established for both antibodies (anti-HLA: sensitivity = 80.00%, specificity = 86.36%; anti-MICA: sensitivity = 86.67%, specificity = 88.89%). Our data reveal the potential of applying the ISFET-based immunosensor to the detection of relevant anti-HLA and -MICA antibodies, especially in the field of kidney transplantation. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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19 pages, 3235 KiB

Open AccessArticle

Stability of Cocaine, Opiates, and Metabolites in Dried Saliva Spots

by Ema Almeida, Sofia Soares, Joana Gonçalves, Tiago Rosado, Nicolás Fernández, Jesus M. Rodilla, Luís A. Passarinha, Mário Barroso and Eugenia Gallardo

Molecules 2022, 27(3), 641; https://doi.org/10.3390/molecules27030641 - 19 Jan 2022

Cited by 7 | Viewed by 2517

Abstract

Drug abuse still represents a global problem, and it is associated with an increased risk of diseases, injuries, and deaths. Cocaine (COC) and opiates are the most abused drugs and account for a significant number of fatalities. Therefore, it is important to develop methods capable of effectively identifying and quantifying these substances. The present study aims to evaluate the long-term stability of COC, ecgonine methylester (EME), benzoylecgonine (BEG), cocaethylene (COET), norcocaine (NCOC), morphine (MOR), codeine (COD) and 6-monoacetylmorphine (6-MAM) in oral fluid samples. The analytes of interest were isolated from the matrix (50 µL) using the dried saliva spots (DSS) sampling approach and were subsequently analyzed by gas chromatography coupled with tandem mass spectrometry (GC–MS/MS). The parameters that could influence the stability of the target compounds were studied, and these were storage temperature, light, use of preservatives (and respective concentrations), and time. The effects of each parameter were evaluated using the design of experiments (DOE) approach. The stability of the target analytes was improved when the DSS were stored at room temperature, in the presence of light and using 1% sodium fluoride. The best conditions were then adopted for the DSS storage and long-term stability was assessed. COD was only stable for 1 day, EME was stable for 3 days, COC, COET, NCOC and 6-MAM were stable for 7 days, MOR for 14 days and BEG remained stable throughout the study (136 days). This is the first study that evaluates the stability of these compounds in oral fluid samples after application in DSS cards, and optimizes the conditions in order to improve their stability. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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17 pages, 1965 KiB

Open AccessArticle

Validated Simple HPLC-UV Method for Mycophenolic Acid (MPA) Monitoring in Human Plasma. Internal Standardization: Is It Necessary?

by Paweł K. Kunicki and Aleksandra Wróbel

Molecules 2021, 26(23), 7252; https://doi.org/10.3390/molecules26237252 - 29 Nov 2021

Cited by 2 | Viewed by 1681

Abstract

The aim of the work was to prepare a simple but reliable HPLC-UV method for the routine monitoring of mycophenolic acid (MPA). Sample preparation was based on plasma protein precipitation with acetonitrile. The isocratic separation of MPA and internal standard (IS) fenbufen was made on Supelcosil LC-CN column (150 × 4.6 mm, 5 µm) using a mobile phase: CH₃CN:H₂O:0.5M KH₂PO₄:H₃PO₄ (260:700:40:0.4, v/v). UV detection was set at 305 nm. The calibration covered the MPA concentration range: 0.1–40 µg/mL. The precision was satisfactory with RSD of 0.97–7.06% for intra-assay and of 1.92–5.15% for inter-assay. The inaccuracy was found between −5.72% and +2.96% (+15.40% at LLOQ) and between −8.82% and +5.31% (+19.00% at LLOQ) for intra- and inter-assay, respectively, fulfilling acceptance criteria. After a two-year period of successful application, the presented method has been retrospectively calibrated using the raw data disregarding the IS in the calculations. The validation and stability parameters were similar for both calculation methods. MPA concentrations were recalculated and compared in 1187 consecutive routine therapeutic drug monitoring (TDM) trough plasma samples from mycophenolate-treated patients. A high agreement (r² = 0.9931, p < 0.0001) of the results was found. A Bland–Altman test revealed a mean bias of −0.011 μg/mL (95% CI: −0.017; −0.005) comprising −0.14% (95% Cl: −0.39; +0.11), whereas the Passing–Bablok regression was y = 0.986x + 0.014. The presented method can be recommended as an attractive analytical tool for medical (hospital) laboratories equipped with solely basic HPLC apparatus. The procedure can be further simplified by disapplying an internal standard while maintaining appropriate precision and accuracy of measurements. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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17 pages, 4832 KiB

Open AccessArticle

Application of Three Ecological Assessment Tools in Examining Chromatographic Methods for the Green Analysis of a Mixture of Dopamine, Serotonin, Glutamate and GABA: A Comparative Study

by Atiah H. Almalki, Ibrahim A. Naguib, Fahad S. Alshehri, Badrah S. Alghamdi, Hashem O. Alsaab, Yusuf S. Althobaiti, Sameer Alshehri and Fatma F. Abdallah

Molecules 2021, 26(18), 5436; https://doi.org/10.3390/molecules26185436 - 07 Sep 2021

Cited by 6 | Viewed by 2332

Abstract

The assessment of greenness of analytical protocols is of great importance now to preserve the environment. Some studies have analyzed either only the neurotransmitters, dopamine, serotonin, glutamate, and gamma-aminobutyric acid (GABA), together or with other neurotransmitters and biomarkers. However, these methods have not been investigated for their greenness and were not compared with each other to find the optimum one. Therefore, this study aims to compare seven published chromatographic methods that analyzed the four neurotransmitters and their mixtures using the National Environmental Method Index, Analytical Eco-Scale Assessment (ESA), and Green Analytical Procedure Index (GAPI). As these methods cover both qualitative and quantitative aspects, they offer better transparency. Overall, GAPI showed maximum greenness throughout the analysis. Method 6 was proven to be the method of choice for analyzing the mixture, owing to its greenness, according to NEMI, ESA, and GAPI. Additionally, method 6 has a wide scope of application (13 components can be analyzed), high sensitivity (low LOQ values), and fast analysis (low retention times, especially for glutamate and GABA). Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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16 pages, 1893 KiB

Open AccessArticle

Untargeted Metabolomics in Forensic Toxicology: A New Approach for the Detection of Fentanyl Intake in Urine Samples

by Eleonora Amante, Eugenio Alladio, Rebecca Rizzo, Daniele Di Corcia, Pierre Negri, Lia Visintin, Michela Guglielmotto, Elena Tamagno, Marco Vincenti and Alberto Salomone

Molecules 2021, 26(16), 4990; https://doi.org/10.3390/molecules26164990 - 18 Aug 2021

Cited by 4 | Viewed by 4370

Abstract

The misuse of fentanyl, and novel synthetic opioids (NSO) in general, has become a public health emergency, especially in the United States. The detection of NSO is often challenged by the limited diagnostic time frame allowed by urine sampling and the wide range of chemically modified analogues, continuously introduced to the recreational drug market. In this study, an untargeted metabolomics approach was developed to obtain a comprehensive “fingerprint” of any anomalous and specific metabolic pattern potentially related to fentanyl exposure. In recent years, in vitro models of drug metabolism have emerged as important tools to overcome the limited access to positive urine samples and uncertainties related to the substances actually taken, the possible combined drug intake, and the ingested dose. In this study, an in vivo experiment was designed by incubating HepG2 cell lines with either fentanyl or common drugs of abuse, creating a cohort of 96 samples. These samples, together with 81 urine samples including negative controls and positive samples obtained from recent users of either fentanyl or “traditional” drugs, were subjected to untargeted analysis using both UHPLC reverse phase and HILIC chromatography combined with QTOF mass spectrometry. Data independent acquisition was performed by SWATH in order to obtain a comprehensive profile of the urinary metabolome. After extensive processing, the resulting datasets were initially subjected to unsupervised exploration by principal component analysis (PCA), yielding clear separation of the fentanyl positive samples with respect to both controls and samples positive to other drugs. The urine datasets were then systematically investigated by supervised classification models based on soft independent modeling by class analogy (SIMCA) algorithms, with the end goal of identifying fentanyl users. A final single-class SIMCA model based on an RP dataset and five PCs yielded 96% sensitivity and 74% specificity. The distinguishable metabolic patterns produced by fentanyl in comparison to other opioids opens up new perspectives in the interpretation of the biological activity of fentanyl. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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16 pages, 2160 KiB

Open AccessFeature PaperArticle

Evaluation of Cocaine Effect on Endogenous Metabolites of HepG2 Cells Using Targeted Metabolomics

by Adamantios Krokos, Olga Deda, Christina Virgiliou, Helen Gika, Nikolaos Raikos, Eleni Aggelidou, Aristeidis Kritis and Georgios Theodoridis

Molecules 2021, 26(15), 4610; https://doi.org/10.3390/molecules26154610 - 29 Jul 2021

Cited by 6 | Viewed by 2745

Abstract

Cocaine toxicity has been a subject of study because cocaine is one of the most common and potent drugs of abuse. In the current study the effect of cocaine on human liver cancer cell line (HepG2) was assessed. Cocaine toxicity (IC50) on HepG2 cells was experimentally calculated using an XTT assay at 2.428 mM. The metabolic profile of HepG2 cells was further evaluated to investigate the cytotoxic activity of cocaine at 2 mM at three different time points. Cell medium and intracellular material samples were analyzed with a validated HILIC-MS/MS method for targeted metabolomics on an ACQUITY Amide column in gradient mode with detection on a triple quadrupole mass spectrometer in multiple reaction monitoring. About 106 hydrophilic metabolites from different metabolic pathways were monitored. Multivariate analysis clearly separated the studied groups (cocaine-treated and control samples) and revealed potential biomarkers in the extracellular and intracellular samples. A predominant effect of cocaine administration on alanine, aspartate, and glutamate metabolic pathway was observed. Moreover, taurine and hypotaurine metabolism were found to be affected in cocaine-treated cells. Targeted metabolomics managed to reveal metabolic changes upon cocaine administration, however deciphering the exact cocaine cytotoxic mechanism is still challenging. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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15 pages, 10739 KiB

Open AccessArticle

A Pilot Metabolomic Study on Myocardial Injury Caused by Chronic Alcohol Consumption—Alcoholic Cardiomyopathy

by Zhipeng Cao, Tianqi Wang, Wei Xia, Baoli Zhu, Meihui Tian, Rui Zhao and Dawei Guan

Molecules 2021, 26(8), 2177; https://doi.org/10.3390/molecules26082177 - 09 Apr 2021

Cited by 12 | Viewed by 2902

Abstract

Chronic alcohol consumption leads to myocardial injury, ventricle dilation, and cardiac dysfunction, which is defined as alcoholic cardiomyopathy (ACM). To explore the induced myocardial injury and underlying mechanism of ACM, the Liber-DeCarli liquid diet was used to establish an animal model of ACM and histopathology, echocardiography, molecular biology, and metabolomics were employed. Hematoxylin-eosin and Masson’s trichrome staining revealed disordered myocardial structure and local fibrosis in the ACM group. Echocardiography revealed thinning wall and dilation of the left ventricle and decreased cardiac function in the ACM group, with increased serum levels of brain natriuretic peptide (BNP) and expression of myocardial BNP mRNA measured through enzyme-linked immunosorbent assay and real-time quantitative polymerase chain reaction (PCR), respectively. Through metabolomic analysis of myocardium specimens, 297 differentially expressed metabolites were identified which were involved in KEGG pathways related to the biosynthesis of unsaturated fatty acids, vitamin digestion and absorption, oxidative phosphorylation, pentose phosphate, and purine and pyrimidine metabolism. The present study demonstrated chronic alcohol consumption caused disordered cardiomyocyte structure, thinning and dilation of the left ventricle, and decreased cardiac function. Metabolomic analysis of myocardium specimens and KEGG enrichment analysis further demonstrated that several differentially expressed metabolites and pathways were involved in the ACM group, which suggests potential causes of myocardial injury due to chronic alcohol exposure and provides insight for further research elucidating the underlying mechanisms of ACM. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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13 pages, 5358 KiB

Open AccessArticle

A Proteomics Study on the Mechanism of Nutmeg-Induced Hepatotoxicity

by Wei Xia, Zhipeng Cao, Xiaoyu Zhang and Lina Gao

Molecules 2021, 26(6), 1748; https://doi.org/10.3390/molecules26061748 - 20 Mar 2021

Cited by 3 | Viewed by 2969

Abstract

Nutmeg is a traditional spice and medicinal plant with a variety of pharmacological activities. However, nutmeg abuse due to its hallucinogenic characteristics and poisoning cases are frequently reported. Our previous metabolomics study proved the hepatotoxicity of nutmeg and demonstrated that high-dose nutmeg can affect the synthesis and secretion of bile acids and cause oxidative stress. In order to further investigate the hepatotoxicity of nutmeg, normal saline, 1 g/kg, 4 g/kg nutmeg were administrated to male Kunming mice by intragastrical gavage for 7 days. Histopathological investigation of liver tissue, proteomics and biochemical analysis were employed to explore the mechanism of liver damage caused by nutmeg. The results showed that a high-dose (4 g/kg) of nutmeg can cause significant increased level of CYP450s and depletion of antioxidants, resulting in obvious oxidative stress damage and lipid metabolism disorders; but this change was not observed in low-dose group (1 g/kg). In addition, the increased level of malondialdehyde and decreased level of glutathione peroxidase were found after nutmeg exposure. Therefore, the present study reasonably speculates that nutmeg exposure may lead to liver injury through oxidative stress and the degree of this damage is related to the exposure dose. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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26 pages, 1992 KiB

Open AccessFeature PaperArticle

A Systematic Study of the In Vitro Pharmacokinetics and Estimated Human In Vivo Clearance of Indole and Indazole-3-Carboxamide Synthetic Cannabinoid Receptor Agonists Detected on the Illicit Drug Market

by Andrew M. Brandon, Lysbeth H. Antonides, Jennifer Riley, Ola Epemolu, Denise A. McKeown, Kevin D. Read and Craig McKenzie

Molecules 2021, 26(5), 1396; https://doi.org/10.3390/molecules26051396 - 05 Mar 2021

Cited by 14 | Viewed by 4868

Abstract

In vitro pharmacokinetic studies were conducted on enantiomer pairs of twelve valinate or tert-leucinate indole and indazole-3-carboxamide synthetic cannabinoid receptor agonists (SCRAs) detected on the illicit drug market to investigate their physicochemical parameters and structure-metabolism relationships (SMRs). Experimentally derived Log D_7.4 ranged from 2.81 (AB-FUBINACA) to 4.95 (MDMB-4en-PINACA) and all SCRAs tested were highly protein bound, ranging from 88.9 ± 0.49% ((R)-4F-MDMB-BINACA) to 99.5 ± 0.08% ((S)-MDMB-FUBINACA). Most tested SCRAs were cleared rapidly in vitro in pooled human liver microsomes (pHLM) and pooled cryopreserved human hepatocytes (pHHeps). Intrinsic clearance (CL_int) ranged from 13.7 ± 4.06 ((R)-AB-FUBINACA) to 2944 ± 95.9 mL min⁻¹ kg⁻¹ ((S)-AMB-FUBINACA) in pHLM, and from 110 ± 34.5 ((S)-AB-FUBINACA) to 3216 ± 607 mL min⁻¹ kg⁻¹ ((S)-AMB-FUBINACA) in pHHeps. Predicted Human in vivo hepatic clearance (CL_H) ranged from 0.34 ± 0.09 ((S)-AB-FUBINACA) to 17.79 ± 0.20 mL min⁻¹ kg⁻¹ ((S)-5F-AMB-PINACA) in pHLM and 1.39 ± 0.27 ((S)-MDMB-FUBINACA) to 18.25 ± 0.12 mL min⁻¹ kg⁻¹ ((S)-5F-AMB-PINACA) in pHHeps. Valinate and tert-leucinate indole and indazole-3-carboxamide SCRAs are often rapidly metabolised in vitro but are highly protein bound in vivo and therefore predicted in vivo CL_H is much slower than CL_int. This is likely to give rise to longer detection windows of these substances and their metabolites in urine, possibly as a result of accumulation of parent drug in lipid-rich tissues, with redistribution into the circulatory system and subsequent metabolism. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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9 pages, 1284 KiB

Open AccessArticle

Determination of Trimethylamine N-oxide and Betaine in Serum and Food by Targeted Metabonomics

by Mingshuai He, Heshui Yu, Peng Lei, Shengjie Huang, Juanning Ren, Wenjing Fan, Lifeng Han, Haiyang Yu, Yuefei Wang, Ming Ren and Miaomiao Jiang

Molecules 2021, 26(5), 1334; https://doi.org/10.3390/molecules26051334 - 02 Mar 2021

Cited by 8 | Viewed by 2940

Abstract

Trimethylamine N-oxide (TMAO), as a gut-derived metabolite, has been found to be associated with enhanced risk for atherosclerosis and cardiovascular disease. We presented a method for targeted profiling of TMAO and betaine in serum and food samples based on a combination of one-step sample pretreatment and proton nuclear magnetic resonance spectroscopy. The key step included a processing of sample preparation using a selective solid-phase extraction column for retention of basic metabolites. Proton signals at δ 3.29 and δ 3.28 were employed to quantify TMAO and betaine, respectively. The developed method was examined with acceptable linear relationship, precision, stability, repeatability, and accuracy. It was successfully applied to detect serum levels of TMAO and betaine in TMAO-fed mice and high-fructose-fed rats and also used to determine the contents of TMAO and betaine in several kinds of food, such as fish, pork, milk, and egg yolk. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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Review

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15 pages, 11247 KiB

Open AccessReview

Enzymatic Methods for Salivary Biomarkers Detection: Overview and Current Challenges

by Alonso Ornelas-González, Margarita Ortiz-Martínez, Mirna González-González and Marco Rito-Palomares

Molecules 2021, 26(22), 7026; https://doi.org/10.3390/molecules26227026 - 20 Nov 2021

Cited by 9 | Viewed by 3458

Abstract

Early detection is a key factor in patient fate. Currently, multiple biomolecules have been recognized as biomarkers. Nevertheless, their identification is only the starting line on the way to their implementation in disease diagnosis. Although blood is the biofluid par excellence for the quantification of biomarkers, its extraction is uncomfortable and painful for many patients. In this sense, there is a gap in which saliva emerges as a non-invasive and valuable source of information, as it contains many of the biomarkers found in blood. Recent technological advances have made it possible to detect and quantify biomarkers in saliva samples. However, there are opportunity areas in terms of cost and complexity, which could be solved using simpler methodologies such as those based on enzymes. Many reviews have focused on presenting the state-of-the-art in identifying biomarkers in saliva samples. However, just a few of them provide critical analysis of technical elements for biomarker quantification in enzymatic methods for large-scale clinical applications. Thus, this review proposes enzymatic assays as a cost-effective alternative to overcome the limitations of current methods for the quantification of biomarkers in saliva, highlighting the technical and operational considerations necessary for sampling, method development, optimization, and validation. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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20 pages, 559 KiB

Open AccessReview

Circular RNAs in Sudden Cardiac Death Related Diseases: Novel Biomarker for Clinical and Forensic Diagnosis

by Meihui Tian, Zhipeng Cao and Hao Pang

Molecules 2021, 26(4), 1155; https://doi.org/10.3390/molecules26041155 - 21 Feb 2021

Cited by 10 | Viewed by 3121

Abstract

The prevention and diagnosis of sudden cardiac death (SCD) are among the most important keystones and challenges in clinical and forensic practice. However, the diagnostic value of the current biomarkers remains unresolved issues. Therefore, novel diagnostic biomarkers are urgently required to identify patients with early-stage cardiovascular diseases (CVD), and to assist in the postmortem diagnosis of SCD cases without typical cardiac damage. An increasing number of studies show that circular RNAs (circRNAs) have stable expressions in myocardial tissue, and their time- and tissue-specific expression levels might reflect the pathophysiological status of the heart, which makes them potential CVD biomarkers. In this article, we briefly introduced the biogenesis and functional characteristics of circRNAs. Moreover, we described the roles of circRNAs in multiple SCD-related diseases, including coronary artery disease (CAD), myocardial ischemia or infarction, arrhythmia, cardiomyopathy, and myocarditis, and discussed the application prospects and challenges of circRNAs as a novel biomarker in the clinical and forensic diagnosis of SCD. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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21 pages, 2278 KiB

Open AccessReview

Monoclonal Antibodies Application in Lateral Flow Immunochromatographic Assays for Drugs of Abuse Detection

by Zidane Qriouet, Yahia Cherrah, Hassan Sefrioui and Zineb Qmichou

Molecules 2021, 26(4), 1058; https://doi.org/10.3390/molecules26041058 - 18 Feb 2021

Cited by 17 | Viewed by 6882

Abstract

Lateral flow assays (lateral flow immunoassays and nucleic acid lateral flow assays) have experienced a great boom in a wide variety of early diagnostic and screening applications. As opposed to conventional examinations (High Performance Liquid Chromatography, Polymerase Chain Reaction, Gas chromatography-Mass Spectrometry, etc.), they obtain the results of a sample’s analysis within a short period. In resource-limited areas, these tests must be simple, reliable, and inexpensive. In this review, we outline the production process of antibodies against drugs of abuse (such as heroin, amphetamine, benzodiazepines, cannabis, etc.), used in lateral flow immunoassays as revelation or detection molecules, with a focus on the components, the principles, the formats, and the mechanisms of reaction of these assays. Further, we report the monoclonal antibody advantages over the polyclonal ones used against drugs of abuse. The perspective on aptamer use for lateral flow assay development was also discussed as a possible alternative to antibodies in view of improving the limit of detection, sensitivity, and specificity of lateral flow assays. Full article

(This article belongs to the Special Issue Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines)

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