Analytical Approaches for the Determination of Buprenorphine, Methadone and Their Metabolites in Biological Matrices.

The abuse of buprenorphine and methadone has grown into a rising worldwide issue. After their consumption, buprenorphine, methadone and their metabolites can be found in the human organism. Due to the difficulty in the assessment of these compounds by routine drug screening, the importance of developing highly sensitive analytical approaches is undeniable. Liquid chromatography tandem mass spectrometry is the preferable technique for the determination of buprenorphine, methadone and their metabolites in biological matrices including urine, plasma, nails or oral fluids. This research aims to review a critical discussion of the latest trends for the monitoring of buprenorphine, methadone and their metabolites in various biological specimens.

Magnetic molecularly imprinted polymers for extraction of S-phenylmercapturic acid from urine samples followed by high-performance liquid chromatography.

In this study, magnetic molecularly imprinted polymers (MMIPs) were prepared and used as sorbents for extraction of S-phenylmercapturic acid (S-PMA) from urine samples, followed by high-performance liquid chromatography ultraviolet-visible (HPLC-UV/Vis) analysis. The MMIPs were synthesized by the copolymerization reaction of (phenylthio) acetic acid (template molecule), methacrylic acid (functional monomers) and ethylene glycol dimethacrylate (cross-linkers). The morphology, structure property and surface groups of the prepared MMIPs were characterized by scan electron microscopy, transmission electron microscopy, infrared spectroscopy, X-ray diffraction pattern, thermogravimetric analyses, Brunauer-Emmett-Teller and vibrating sample magnetometer. The selectivity of the MMIPs was investigated in the presence of interferents. Various parameters affecting the S-PMA extraction efficiency were investigated, including MMIPs amount, pH, sample volume, desorption solvent, as well as extraction and desorption time. The obtained optimal parameters were as follows: MMIPs amount (20 mg), pH (3.0), sample volume (5 mL), desorption solvent (methanol/acetic acid [9/1, v/v]), extraction time (30 minutes) and desorption time (2 minutes). The method was validated according to the Food and Drug Administration Guidance for Industry on Bioanalytical Method Validation. The calibration curve for the analyte was linear in the concentration range of 0.030-1.0 mg/L (r = 0.9995). The LOD and LOQ of the method were 0.0080 and 0.0267 mg/L, respectively. The enrichment factor of the MMIPs was 5. The relative standard deviations of intra- and inter-day tests were in the range of 3.8-5.1% and 3.9-6.3%, respectively. The recoveries at three different concentrations of 0.10, 0.50 and 0.80 mg/L ranged between 95.2% and 98.6%. In addition, the MMIPs could be reused for at least eight times. The proposed method was successfully applied to the determination of S-PMA in urine samples. In addition, this developed method could be used as a tool in the early screening and clinical diagnosis of benzene intoxication.

Review of LC techniques for determination of methadone and its metabolite in the biological samples.

Methadone (MTD) is a synthetic analgesic drug used for treating opioid dependence and effectively used clinically for patients with severe pain. The abuse of MTD may lead to poisoning, disorder in the central nervous system and even death. The regular monitoring of MTD in biological matrices including serum, plasma and urine samples is an effective way to control abuse of MTD. In this manner, the selection of analytical monitoring of MTD in biological matrices is of paramount importance. This study was conducted to review high-performance liquid chromatography (HPLC) techniques carried out on MTD and its main metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in the biological samples during 2015-June 2021.

Activated carbon/diatomite-based magnetic nanocomposites for magnetic solid-phase extraction of S-phenylmercapturic acid from human urine.

In this study, activated carbon/diatomite-based magnetic nancomposites (denoted as AC/DBMNs) were synthesized and applied as an adsorbent for magnetic solid-phase extraction of S-phenylmercapturic acid (S-PMA) from human urine prior to high-performance liquid chromatography. The surface morphologies and structures of AC/DBMNs were characterized by Fourier transform infrared spectroscopy, transmission and scanning electron microscopy, X-ray diffraction, Brunauer-Emmett-Teller surface area, vibrating sample magnetometer and ζ-potential measurements. The experimental parameters including sample volume, sample pH, adsorbent amount, extraction time, elution solvent and desorption time were investigated in detail. Under the optimum conditions, the method exhibited good linearity (r > 0.9993) within the concentration ranges of 0.03-1.0 mg/L. Moreover, the limits of detection and quantification were 0.01 and 0.03 mg/L, respectively. The enrichment factor was 5, and good recoveries (88.9-97.3%) with relative standard deviations in the range of 5.6-6.8% (n = 6) for inter-day and 6.3-8.1% (n = 6) for intra-day were achieved. The developed method was successfully applied to the analysis of S-PMA in urine samples. In addition, this accurate and sensitive method has great potential to be applied in the early screening and clinical diagnosis of the workers exposed to benzene.

Highly luminescent N-doped carbon quantum dots as an effective multifunctional fluorescence sensing platform.

The doping of carbon quantum dots with nitrogen provides a promising direction to improve fluorescence performance and broaden their applications in sensing systems. Herein we report a one-pot solvothermal synthesis of N-doped carbon quantum dots (NCQDs) and the synthesis of a series of NCQDs with different nitrogen contents. The as-prepared NCQDs were compared with carbon quantum dots (CQDs); the introduction of nitrogen atoms largely increased the quantum yield of NCQDs and highest emission efficiency is up to 36.3 %. The fluorescence enhancement may originate from more polyaromatic structures induced by incorporated nitrogen atoms and protonation of nitrogen atoms on dots. It was found that NCQDs can act as a multifunctional fluorescence sensing platform because they can be used to detect pH values, Ag(I), and Fe(III) in aqueous solution. The fluorescence intensity of NCQDs is inversely proportional to pH values across a broad range from 5.0 to 13.5, which indicates that NCQDs can be devised as an effective pH indicator. Selective detection of Ag(I) and Fe(III) was achieved based on their distinctive fluorescence influence because Ag(I) can significantly enhance the fluorescence whereas Fe(III) can greatly quench the fluorescence. The quantitative determination of Ag(I) can be accomplished with NCQDs by using the linear relationship between fluorescence intensity of NCQDs and concentration of Ag(I). The sensitive detection of H2O2 was developed by taking advantage of the distinct quenching ability of Fe(III) and Fe(II) toward the fluorescence of NCQDs. Cellular toxicity test showed NCQDs still retain low toxicity to cells despite the introduction of a great deal of nitrogen atoms. Moreover, bioimaging experiments demonstrated that NCQDs have stronger resistance to photobleaching than CQDs and more excellent fluorescence labeling performance.

Simultaneous detection of multiple DNA targets by integrating dual-color graphene quantum dot nanoprobes and carbon nanotubes.

Simultaneous detection of multiple DNA targets was achieved based on a biocompatible graphene quantum dots (GQDs) and carbon nanotubes (CNTs) platform through spontaneous assembly between dual-color GQD-based probes and CNTs and subsequently self-recognition between DNA probes and targets.

B-doped carbon quantum dots as a sensitive fluorescence probe for hydrogen peroxide and glucose detection.

Fluorescent B-doped carbon quantum dots (BCQDs) were prepared by a facile one-pot solvothermal route. The BCQDs can be used as a novel fluorescence sensing system for hydrogen peroxide and glucose detection.

Synthesis of High-Fluorescent Diphenyl-anthracene Derivatives and Application in Detection of Nitroaromatic Explosives and Fingerprint Identification.

The development of high-intensity fluorescent materials is always the focuses and forefront projects because of their important applications in displays, sensing and detection fields. In recent years, the detection of explosives has attracted increasing attention due to security and counterterrorism issues. Herein, two diphenyl-anthracene (DPA) derivatives were designed and synthesized by introducing strong electron withdrawing fluorine atoms and cyano-groups to DPA, which exhibited strong fluorescence both in the solution and solid phase with the absolute quantum yields up to 70.4 % and 45.9 % respectively. The detection behavior of nitroaromatic explosives such as picric acid (PA), 2,4,6-trinitrotoluene (TNT) and 3-Nitropropionic acid (3-NP) also shows good sensitivity with the quenching constant as high as 6.3×104  L mol-1 . Theoretical calculation demonstrates that the fluorescence quenching behavior of the two DPA derivatives is caused by the behavior of photoinduced electron transfer (PET) and the resonance energy transfer (RET) studies explained the higher sensitivity and selectivity of both compounds towards PA than other nitro-containing explosives. Furthermore, the strong solid-state fluorescence of the DPA derivatives also shows excellent advantages in enhancing latent fingerprint recognition.

Influencing factors and prediction of neutralizing antibodies in post-exposure rabies vaccine recipients.

To assess the factors that influencing the persistence of virus neutralizing antibody (VNA), and to establish prediction models to provide the appropriate timing for booster administration, a cohort of post-exposure rabies vaccine recipients was investigated. The VNA determined records from 2019 to 2023 and interrelated factors were analyzed, including gender, age, rabies immunoglobulin (RIG) administration, vaccine products, vaccination schedule, and vaccination intervals etc. The geometric mean of VNA titre within 1 month after primary vaccination with 2-1-1 schedule was statistically higher than that with 5-dose course (P = 0.031). The interaction between exposure and vaccination schedule was observed on primary vaccination, which showed that a decrease of 19.74 % (95 % CI: 5.99%-64.95 %, P = 0.008) of VNA titre among vaccinee with 5-dose and exposure III. Individuals with RIG administration produced lower VNA titres than those without RIG administration (P = 0.001). Vaccine products (Chengda, P = 0.015; human diploid cell, P = 0.026) and re-exposed time (P = 0.000) exhibited independent effects following booster vaccination. Based on the prediction model, the 99 % individual prediction intervals (IPI) of VNA titres were established at 3, 6, 12 and 18 months for the 12 characteristic populations respectively. The cases of VNA below 0.5 IU/ml first appeared at 6 months in group D of primary vaccinations and at 10 years in group F of boosters. We conclude that for primary vaccination 2-1-1 schedule is more efficient than 5-dose; the use of residual rabies immunoglobulin for distal intramuscular injection isn't recommended. The 99 % IPI of VNA titres could provide the appropriate timing for booster vaccination.

The structures of Arabidopsis Deg5 and Deg8 reveal new insights into HtrA proteases.

Plant Deg5 and Deg8 are two members of the HtrA proteases, a family of oligomeric serine endopeptidases that are involved in a variety of protein quality-control processes. These two HtrA proteases are located in the thylakoid lumen and participate in high-light stress responses by collaborating with other chloroplast proteins. Deg5 and Deg8 degrade photodamaged D1 protein of the photosystem II reaction centre, allowing its in situ replacement. Here, the crystal structures of Arabidopsis thaliana Deg5 (S266A) and Deg8 (S292A) are reported at 2.6 and 2.0 Å resolution, respectively. The Deg5 trimer contains two calcium ions in a central channel, suggesting a link between photodamage control and calcium ions in chloroplasts. Previous structures of HtrA proteases have indicated that their regulation usually requires C-terminal PDZ domain(s). Deg5 is unique in that it contains no PDZ domain and the trimeric structure of Deg5 (S266A) reveals a novel catalytic triad conformation. A similar triad conformation is observed in the hexameric structure of the single PDZ-domain-containing Deg8 (S292A). These findings suggest a novel activation mechanism for plant HtrA proteases and provide structural clues to their function in light-stress response.

Expression, purification, crystallization and preliminary X-ray diffraction analysis of Arabidopsis thaliana Deg8.

Arabidopsis thaliana Deg8, an ATP-independent serine endopeptidase, is involved in the repair of photosystem II (PSII), specifically the degradation of the photo-damaged PSII reaction centre D1 protein. To understand the molecular mechanism underlying the participation of Deg8 in the degradation of the photo-damaged D1 protein, the structure of Deg8 is needed. Until recently, however, no structure of Deg8 had been solved. In this study, Deg8 from A. thaliana was cloned, overexpressed and purified in Escherichia coli. Crystallization was performed at 277 K using tribasic sodium citrate as the precipitant and the crystals diffracted to 2.0 Šresolution, belonging to space group C2 with unit-cell parameters a = 129.5, b = 124.2, c = 93.3 Å, α = γ = 90, ß = 132.4°. Assuming one trimer in the asymmetric unit, the Matthews coefficient and the solvent content were calculated to be 2.35 Å(3) Da(-1) and 47.6%, respectively.

Characterization of welding fume and airborne heavy metals in electronic manufacturing workshops in Hangzhou, China: implication for occupational population exposure.

Occupational exposure to contaminants created by electronic manufacturing process is not well characterized. The aim of this study was to carry out risk assessments of exposure to welding fume and airborne heavy metals (HMs) in electronic manufacturing workshops. Seventy-six air samples were collected from five sites in Hangzhou, China. In welding workshops, the most abundant contaminant found was welding fume, followed by Fe, Mn, Zn, Cu, Pb, Cd, and Cr. The concentration of Mn was positively correlated with Fe (r = 0.906). When compared with non-welding workshops, the Fe content in the air of welding workshops increased significantly (P < 0.05), while the Cu content decreased significantly (P < 0.05). Singapore semi-quantitative health risk assessment model and the United States Environmental Protection Agency (US EPA) inhalation risk assessment model were applied to assess the occupational exposure. In welding workshops, the levels of 8-h time weighted average (8 h-TWA) calculated for welding fume (range 0.288 ~ 6.281 mg/m3), Mn (range Nd ~ 0.829 mg/m3), and Fe (range 0.027 ~ 2.234 mg/m3) partly exceeded the permissible limits. While, in non-welding workshops, the average of 8 h-TWA for Cu (0.411 mg/m3) was higher than the limit. The risk rates (RR) assessed for Pb (2.4 vs 1.7), Mn (2.0 vs 1.4), and Fe (1.4 vs 1.0) were higher in welding workshops than that in non-welding workshops, but Cu (1.0 vs 2.2) were lower. The mean excess lifetime cancer risks (ELCR) in welding (5.59E - 06 per 1000 people) and non-welding (1.88E - 06 per 1000 people) workshops were acceptable. The mean non-cancer risk (HQ) estimated for Mn was greater than 10 in both welding (HQ = 164) and non-welding (HQ = 11.1) workshops. These results indicate that there was a risk of occupational exposure implication in the electronic manufacturing workshops. Reducing contaminant exposure through engineering controls and management strategies, such as efficient ventilation and reducing exposure hours, is thus suggested.

Association between serum copper/zinc ratio and lung cancer: A systematic review with meta-analysis.

BACKGROUND: Numerous studies have investigated the relationship between serum copper/zinc ratio and lung cancer. However, the results are inconsistent. Therefore, we evaluated the association between copper/zinc ratio and lung cancer. MATERIALS AND METHODS: Observational studies reporting serum copper/zinc ratio in lung cancer patients and controls were identified from PubMed, Web of Science, EMBASE, CNKI and Wanfang databases online before December 2021. Summary standard mean difference (SMD) and the corresponding 95 % confidence interval (95 % CI) were applied to compare the serum serum copper/zinc ratio between lung cancer patients and controls using a random-effects model. RESULTS: Thirty-nine articles including 3598 lung cancer patients, 1402 benign lung diseases, and 3314 healthy controls were included in this study. The pooled results showed that the lung cancer patients had significantly higher serum copper/zinc ratio than healthy controls [SMD (95 % CI): 1.62 (1.31, 1.93)] and patients with benign lung diseases [SMD (95 % CI): 0.60 (0.36, 0.84)]. The results were robust according to sensitivity analysis. Meanwhile, consistent results were obtained both in European populations and Asian populations. Moreover, serum copper/zinc ratio was significant higher in patients with advanced stage of lung cancer than that in patients with early stage of lung cancer. CONCLUSION: The results showed that elevated serum copper/zinc ratio might be associated with increased risk of lung cancer.

Analytical techniques for monitoring of toluene and xylene exposure biomarkers hippuric acid and methylhippuric acid in human urine samples.

Quantification of hippuric acid and methylhippuric acid in human urine matrices provides information on the toluene and xylene exposure conditions. High performance liquid chromatography coupled with UV detection is the preferable technique for hippuric acid and methylhippuric acid detection in human urine. This study was conducted to present analytical techniques developed for monitoring of hippuric acid and methylhippuric acid in human urine matrices during 2016-2021.

Review of Detection and Quantification of Rabies Virus Antibodies.

Rabies is an almost invariably fatal disease. According to the World Health Organization (WHO), rabies virus neutralizing antibody (RVNA) titers of ≥0.5 IU/mL are considered adequate for rabies protection. Therefore, detection and quantification of RABV antibodies are important. Many methods have been developed for detecting RABV antibodies. In the present study, we reviewed several methods of detecting RABV antibodies in human and animal samples and evaluated and compared their performance. Of 34 methods, 5 demonstrated unsatisfactory sensitivity or specificity. The others exhibited sensitivity and specificity of ≥75%. The correlation coefficient for five of eight methods was >0.8. The Bland-Altman mean bias of five of five methods was <±2.0. The kappa values of 25 of 28 methods were higher than 0.4, demonstrating at least moderate agreement. Analysis of the performance of these methods emphasized that any new technology should be considered carefully and objectively before being used as an appropriate and applicable alternative.

Facile Preparation of Activated Carbon from Peanut Shell for Determination of Bisphenol A in Human Urine by High-Performance Liquid Chromatography.

Herein, a facile and low-cost method for the preparation of activated carbon from peanut shell was developed for the first time for the fast extraction and determination of Bisphenol A in human urine. Bisphenol A was separated by EC-C18 column (250 mm×4.6 mm, 4 µm) and was detected by VWD, with retention time for qualitative analysis and peak area for quantitation. The parameters, pH values of the urine, adsorbent dose, adsorption time and so on, were optimized to achieve the excellent extraction performance. The detection limit of Bisphenol A in human urine was 1.0 ng · mL-1 (S/N = 3), and the standard curve was linear in the range of 5.0 ng · mL-1˜200.0 ng · mL-1 (r = 0.9993). The average recovery of Bisphenol A was 78.5˜96.2% at three spiked levels in the range of 5.00 ng · mL-1˜200.00 ng·mL-1. The method was proved simple, practical and highly sensitive, which could satisfy the request for the determination of Bisphenol A in human urine.

Si-doped carbon quantum dots: a facile and general preparation strategy, bioimaging application, and multifunctional sensor.

Heteroatom doping of carbon quantum dots not only enables great improvement of fluorescence efficiency and tunability of fluorescence emission, but also provides active sites in carbon dots to broaden their application in sensor. Silicon as a biocompatible element offers a promising direction for doping of carbon quantum dots. Si-doped carbon quantum dots (SiCQDs) were synthesized through a facile and effective approach. The as-prepared Si-doped carbon quantum dots possess visible fluorescence with high quantum yield up to 19.2%, owing to fluorescence enhancement effect of introduced silicon atoms into carbon dots. The toxicity test on human Hela cells showed that SiCQDs have lower cellular toxicity than common CQDs, and bioimaging experiments clearly demonstrated their excellent biolabelling ability and outstanding performance in resistance to photobleaching. Strong fluorescence quenching effect of Fe(III) on SiCQDs can be used for its selective detection among general metal ions. Specific electron transfer between SiCQDs and hydrogen peroxide enables SiCQDs as a sensitive fluorescence sensing platform for hydrogen peroxide. The subsequent fluorescence recovery induced by removal of hydrogen peroxide from SiCQDs due to formation of the stable adducts between hydrogen peroxide and melamine was taken advantage of to construct effective sensor for melamine.

Human thioesterase superfamily member 2 (hTHEM2) is co-localized with beta-tubulin onto the microtubule.

Human thioesterase superfamily member 2 (hTHEM2) belongs to the hotdog-fold enzyme superfamily but its biological function remains unknown. Tissue specific expression in mouse showed that the encoding gene is highly expressed in the kidney, and moderately expressed in the liver, brain, large intestine, and small intestine. Small interference RNA silencing in cell line HCT116 shows that the hthem2 gene is essential for the cell sustained proliferation. Immunostaining and GFP-Tag experiments show that hTHEM2 is co-localized with microtubules.

Crystal structure of human thioesterase superfamily member 2.

Hotdog-fold has been identified in more than 1000 proteins, yet many of which in eukaryotes are less studied. No structural or functional studies of human thioesterase superfamily member 2 (hTHEM2) have been reported before. Since hTHEM2 exhibits about 20% sequence identity to Escherichia coli PaaI protein, it was proposed to be a thioesterase with a hotdog-fold. Here, we report the crystallographic structure of recombinant hTHEM2, determined by the single-wavelength anomalous dispersion method at 2.3A resolution. This structure demonstrates that hTHEM2 indeed contains a hotdog-fold and forms a back-to-back tetramer as other hotdog proteins. Based on structural and sequence conservation, the thioesterase active site in hTHEM2 is predicted. The structure and substrate specificity are most similar to those of the bacterial phenylacetyl-CoA hydrolase. Asp65, located on the central alpha-helix of subunit B, was shown by site-directed mutagenesis to be essential to catalysis.