On-line Analysis System Research Articles

Simultaneous metabolomics and lipidomics analysis based on 4in1 online analysis system reveal metabolic signatures in atherosclerotic mice

Developing efficient and comprehensive analysis methods for metabolomics and lipidomics in the biological tissues and body fluids is essential for understanding the disease mechanisms. Although various two-dimensional liquid chromatography-mass spectrometry (2D-LC-MS) methods have been proposed to expand metabolite coverage, achieving higher efficiency in integrated metabolomics and lipidomics studies remains a technical challenge. In this work, a novel 4in1 online analysis system with excellent reproducibility and mass accuracy was constructed for metabolomics and lipidomics study in various biological samples from atherosclerotic mice. This system enabled the simultaneous detection in both positive and negative ion modes with extensive polarity separation in a single analytical run. Using the 4in1 online analysis system, we identified distinct but complementary metabolic signatures associated with atherosclerosis in different biological samples. Specifically, a total of 230 and 170 differential metabolites or lipids were detected in mice plasma samples and aortic tissue samples, respectively, including glycerophospholipids, sphingolipids, fatty acyls, glycerolipids, carboxylic acids, and pyrimidine nucleosides. Additionally, atherosclerosis-related metabolic pathways involved in biosynthesis of unsaturated fatty acids, sphingolipid metabolism, cholesterol metabolism, glycerophospholipid metabolism, and choline metabolism further revealed. These findings demonstrate that the novel 4in1 online analysis system is a faithful, stable and powerful tool for comprehensive metabolomics and lipidomics studies in complex biological matrices.

A Comprehensive Online Analytical System Coupled with Standardized Data Analysis for the Electrochemical Reduction of CO2

The electrochemical reduction of CO2 (CO2RR) is a promising way to convert detrimental CO2 emissions into sustainable fuels and chemicals, and thus to achieve a circular carbon economy. 1 Depending on catalyst type and reaction conditions, different gaseous and liquid products can be obtained from the CO2RR, and their production rates vary with reaction time. It becomes thus imperative, for any catalytic performance analysis, to be able to assess the product distribution online during CO2RR. 2,3 However, while gaseous products are readily analyzed online by gas chromatography, liquid products are typically only assessed at the end of the reaction due to the lack of suitable automated liquid sampling and analysis methods. In addition, reaction parameters such as CO2 mass flow rates, temperatures and pressures are seldom recorded, causing the loss of significant information on catalysts, electrodes, and electrolyzers behavior.To overcome these issues, we assemble a comprehensive analytical system coupling online gas and liquid product analysis by gas and liquid chromatography leveraging a special automated liquid sampling valve with electrochemical protocols that assess CO2RR performance, electrolyzer cell resistance and electrode surface area. In addition, we record CO2 mass flow rates, electrolyzer temperatures, as well as gas and liquid flow pressures. 4 To rapidly and reproducibly handle the large and heterogeneous data volume obtained we implement a standardized data pipeline based on our own open-source software,5 which automatically parses the numerous different raw data files, composes a data set following FAIR practices,6 and post-processes and plots the data in a standardized way. We validate the analytical system by carrying out CO2RR at 200 mA/cm2 on Cu gas diffusion electrodes, following the changes in selectivity with reaction time for > 10 gaseous and liquid products, and recording mass flow rates, electrolyzer temperatures and pressures. The modular nature of our analytical system, combined with the standardized data pipeline, allows us to freely increase the number and type of sensors used with minimal impact on the data analysis time, as well as to multiplex our analysis to 8 parallel electrolyzer cells, paving the way for a much deeper and faster understanding of the function of CO2RR catalysts, electrodes, and electrolyzers.

Sensor model fusion reveals the dynamics of lactobacillus fermentation with real time concentrations of lactic and acetic acids in ensiling of maize and ryegrass

Anaerobic lactobacillus (LAB) fermentation is key to production of silage. Tracking the dynamics of bacterial metabolism is important for the improvement of LAB fermentation and for evaluation of potential additives. This is currently limited by available ex situ analytical methods, while commercial sensors of lactic and acetic acid content remain unavailable. Here we validate an in situ pH sensor and devise a model-sensor package and a mini-fermenter system (1.5 L) for on-line analysis of LAB fermentation. By the fusion of the model based on mirror mapping principle and the time course of pH measured in situ, a general solution of the dynamic accumulation of organic acid (DAOA) and percentage yield of organic acid (PYOA, 0 ∼ 100 %) is obtained as a function of time. We link PYOA to the initial and final values of lactic and acetic concentrations analyzed ex situ to obtain specific solutions for the time courses of lactic and acetic acid production. We demonstrate the model-sensor system with fermentation of both maize and ryegrass, capturing the dynamic patterns of fermentation, including the rapid depletion of O2 in the initial aerobic phase (maize: ≈ 1.5 h, ryegrass: ≈ 9.2 h), the exponential decline of pH (R2 ≥ 0.99, RMSE ≤ 0.043) in the subsequent anaerobic phase, and metabolic feedbacks of pH on instantaneous acid production (ΔpH; inflection at pH 5), and on cumulative yields of total acidity and specific organic acids. These are all previously unavailable data streams.

Modular pharmacology: research progress and development of analysis platform

Based on the systematic deconstruction of multi-dimensional and multi-target biological networks, modular pharmacology explains the complex mechanism of diseases and the interactions of multi-target drugs. It has made progress in the fields of pathogenesis of disease, biological basis of disease and traditional Chinese medicine(TCM) syndrome, pharmacological mechanism of multi-target herbs, compatibility of formulas, and discovery of new drug of TCM compound. However, the complexity of multi-omics data and biological networks brings challenges to the modular deconstruction and analysis of the drug networks. Here, we constructed the "Computing Platform for Modular Pharmacology" online analysis system, which can implement the function of network construction, module identification, module discriminant analysis, hub-module analysis, intra-module and inter-module relationship analysis, and topological visualization of network based on quantitative expression profiles and protein-protein interaction(PPI) data. This tool provides a powerful tool for the research on complex diseases and multi-target drug mechanisms by means of modular pharmacology. The platform may have broad range of application in disease modular identification and correlation mechanism, interpretation of scientific principles of TCM, analysis of complex mechanisms of TCM and formulas, and discovery of multi-target drugs.

Stereoscopic imaging of volatile organic compounds distribution in the region and tracing emission sources of volatile organic compounds using a novel movable single-photon ionization time-of-flight mass spectrometer.

This paper presents a newly developed high-performance mobile single-photon ionization time-of-flight mass spectrometry (M-SPI-TOFMS) system for on-line analysis and stereoscopic monitoring of complex gas mixtures. The system is designed for stereoscopic imaging to map the distribution of volatile organic compounds (VOCs) and trace their emission sources in urban areas and industrial parks. It mainly consists of a SPI-TOFMS instrument, a customized commercial vehicle, a meteorological five-parameter monitor with GPS, a high-power generator, and an uninterruptible power supply. The SPI technique, using a 118 nm VUV lamp, can ionize compounds with an ionization potential below 10.78 eV. Mass spectra obtained using this technique show the profiles of various VOCs and some inorganic compounds. The VOCs composition information and mobile location data are simultaneously sent to the GIS software. In GIS software, this data is used for real-time stereoscopic imaging of VOC distribution and precise tracking of VOC movement. The system can achieve a spatial data resolution of 0.69 mm at 25 km/h due to the microsecond detection speed of the M-SPI-TOFMS instrument. The laboratory test provides a rapid overview characterization of benzene, toluene, and xylene. The M-SPI-TOFMS has limits of detection and mass resolution of 33.7 pptv and 1060, respectively. Several field applications were carried out using M-SPI-TOFMS at various locations to identify VOC sources near different factories. The M-SPI-TOFMS system has a navigation monitoring speed of 25 km/h with a time resolution of 1 s. The widespread use of this system will provide accurate data to support environmental management departments in formulating VOCs pollution control policies and improving control efficiency.

Online monitoring system for qualitative and quantitative analysis of bioaerosols by combined ATP bioluminescence assay with loop-mediated isothermal amplification

Rapid detection of airborne pathogens is crucial in preventing respiratory infections and allergies. However, technologies aiming to real-time analysis of microorganisms in air remain limited due to the sparse and complex nature of bioaerosols. Here, we introduced an online bioaerosol monitoring system (OBMS) comprised of integrated units including a rotatable stainless-steel sintered filter-based sampler, a lysis unit for extracting adenosine triphosphate (ATP), and a single photon detector-based fluorescence unit. Through optimization of the ATP bioluminescence method and establishment of standard curves between relative luminescence units (RLUs) and ATP as well as microbial concentration, we achieved simultaneous detection of bioaerosols' concentration and activity. Testing OBMS with four bacterial and two fungal aerosols at a sampling flow rate of 10 to 50 L/min revealed an outstanding collection efficiency of 95 % at 30 L/min. A single OBMS measurement takes only 8 min (sampling: 5 min; lysis and detection: 3 min) with detection limits of 3 Pcs/ms photons (2.9 × 103 and 292 CFU/m3 for Staphylococcus aureus and Candida albicans aerosol). In both laboratory and field tests, OBMS detected higher concentrations of bioaerosol compared to the traditional Andersen impactor and liquid biosampler. When combined OBMS with loop-mediated isothermal amplification (LAMP), the bioaerosol can be qualitative and quantitative analyzed within 40 min without the cumbersome procedures of sample pretreatment and DNA extraction. These results offer a high compressive and humidity resistance membrane filtration sampler and validate the potential of OBMS for online measurement of bioaerosol concentration and composition.

COMPLICAÇÕES NEONATAIS CONCERNENTE A MORBIDADE MATERNA: DIABETES GESTACIONAL

The monitoring of this morbidity brings with it the need at public health level to emphasize regular monitoring through prenatal care, aiming at early diagnosis and treatment for Obstetric Diabetes (OD) or gestational diabetes mellitus (GDM), thus being able to reduce complications for the mother and fetus binomial. The aim of this study is to analyze these results among fetal or neonatal complications in women with GDM, bringing in the outcomes of premature birth, birth weight, neonatal morbidity and mortality, and the synergistic effect of concomitant risk factors and poor obstetric outcomes. A literature review was carried out using the following databases: Online Medical Literature Search and Analysis System (MEDLINE/PUBMED), Nursing Database - Brazilian Bibliography (BDENF), Web of Science and Cochrane Library (SCOPUS), CAPES Journals and Latin American and Caribbean Health Sciences Literature (LILACS). The search was carried out using descriptors combined with the Boolean operator “and”. From the searches, it was possible to see that the authors corroborate each other in relation to the main neonatal outcomes: Complications include fetal macrosomia, birth injuries, increased caesarean section rates, neonatal hospitalizations, neonatal hypoglycemia, neonatal respiratory distress syndrome (RDS), prematurity and fetal death. It can be concluded that for the mother the impacts, such as the risk of urinary system infections, are high, as well as bleeding in the postpartum period and tearing of the vaginal canal. For the child, hypoglycemia, prematurity and fetal macrosomia are some of the complications resulting from diabetes.

Silver nanoparticle-functionalized melamine-formaldehyde aerogel for online in-tube solid-phase microextraction of polycyclic aromatic hydrocarbons followed by HPLC-DAD analysis

Based on the π-metal interaction between silver nanoparticles (AgNPs) and aromatic compounds, AgNPs were in-situ grown to melamine-formaldehyde (MF) aerogel for improving the extraction performance to polycyclic aromatic hydrocarbons (PAHs). The AgNPs/MF aerogel was regulated through varing the concentration of reactants, and characterized by scanning electron microscopy, X-ray photoelectron spectroscopy and X-ray powder diffraction. As a new extraction coating, the AgNPs/MF aerogel was coated to stainless-steel wires for in-tube solid-phase microextraction (IT-SPME). The extraction effects of MF aerogels before and after the modification of AgNPs were compared, and the AgNPs greatly improved the extraction ability for PAHs reaching to 166.4 %. Combining IT-SPME with high performance liquid chromatographic detection, an online analytical system was constructed. Furthermore, the sampling volume and rate, concentration of organic solvent, and desorption time were optimized factor by factor. The online analytical method with low detection limits (0.003–0.010 μg L−1) and efficient enrichment factors (1998–3237) for PAHs was established, which fastly detected trace level of PAHs in drinking and environmental water samples. Compared with other methods, the method was comparable or better in the detection limit and linear range, indicating prospective application of the AgNPs/MF aerogel for sample preparation.

Application of online teaching-based classroom behavior capture and analysis system in student management

Application of online teaching-based classroom behavior capture and analysis system in student management

Image processing-based online analysis and feedback control system for droplet dripping process

Droplets find wide application across diverse industries, where maintaining their quality is paramount. Precise control over the substance content within droplets demands non-destructive and online analysis techniques, such as Process Analytical Technology (PAT), often integrated with control strategies. In this context, the present study focuses on the example of controlling droplet quality during the dripping process of pills. Leveraging the dripping and image acquisition systems established in previous research, a novel feedback control system centered on image processing was devised for the quality control of dripping pills. The system was developed and its efficacy was assessed, yielding satisfactory outcomes. The proposed system facilitates real-time monitoring of pill weight through the analysis of droplet images during the dripping process, thereby offering real-time feedback control of pill weight. Importantly, this system holds potential for broader applications beyond the scope of this study.

IMPACTS OF INDUSTRIAL PAINTINGS ON WORKERS’ HEALTH

Objective: to describe the impacts of industrial paintings on workers’ health. Methods: an integrative literature review study was conducted. Articles recovered through secondary databases Virtual Health Library (VHL), Latin American and Caribbean Literature on Health Sciences (LILACS), Scientific Electronic Library Online (Scielo) and Online Medical Literature Search and Analysis System (MEDLINE) were analyzed from the descriptors paintings, worker’s health and occupational medicine. Studies published in Portuguese, English or Spanish were considered and dealt directly with the theme object of study. Results: the main impacts of industrial paintings on workers’ health are skin diseases, allergic reactions, eye irritation and respiratory problems, neoplasms, myelodysplastic syndromes, neuropathies, olfactory disorders, saturnism and acute and chronic effects on the central nervous system. Conclusion: industrial paintings can cause real risks to the health of workers in this service sector, which requires the performance of occupational health professionals for timely prevention and care.

Hydrogen-rich syngas production and insight into morphology-kinetics correlation for furfural residue steam gasification in a bubbling fluidized bed

Biomass steam gasification technology has attracted considerable interest for its potential of low-cost and high-efficiency in hydrogen production. However, there is still a large knowledge gap regarding morphology-kinetic correlation. To avoid the technical difficulties of thermogravimetric analysis and fixed bed studying steam gasification kinetics, this work pursues the design and building of a visualized bubbling fluidized bed with an online analysis system. According to the morphology evolution of biomass real-time observed by a high-speed camera, the shrinkage ratio during high-temperature gasification in a fluidized bed has been first deduced. Moreover, based on accurate monitoring of the gas products releasing behavior, multiple kinetic models were applied to evolve the activation energy at different carbon conversion efficiencies (CCE). The results showed that the synergistic regulation of steam concentration and temperature achieved a high H2/CO ratio (2.46) and H2 yield (869.5 L/kg). A comparative analysis of the coupling relationship between shrinkage ratio, activation energy and CCE successfully separated the volatile precipitation and char gasification processes. It was also revealed that the shrinkage rate peaked at the transition from pyrolysis to char gasification (20 % CCE). At CCE below 20 %, the reaction mechanism tended towards a nucleation and growth mechanism. As CCE reached 20 %, the reaction form shifted. Subsequently, the chemical reaction mechanism controlled the main reaction, and the average apparent activation energies of H2, CO2 and CO were 86.76, 31.27 and 39.90 kJ/mol, respectively. This work provides new insights and a theoretical basis for understanding and modeling the biomass steam gasification process in fluidized beds.

Neutron moderator structure improvement of a PGNAA online analysis system based on DT neutron generator

Neutron moderator structure improvement of a PGNAA online analysis system based on DT neutron generator

Online pharmaceutical process analysis of Chinese medicine using a miniature mass spectrometer: Extraction of active ingredients as an example

The automation of traditional Chinese medicine (TCM) pharmaceuticals has driven the development of process analysis from offline to online. Most of common online process analytical technologies are based on spectroscopy, making the identification and quantification of specific ingredients still a challenge. Herein, we developed a quality control (QC) system for monitoring TCM pharmaceuticals based on paper spray ionization miniature mass spectrometry (mini-MS). It enabled real-time online qualitative and quantitative detection of target ingredients in herbal extracts using mini-MS without chromatographic separation for the first time. Dynamic changes of alkaloids in Aconiti Lateralis Radix Praeparata (Fuzi) during decoction were used as examples, and the scientific principle of Fuzi compatibility was also investigated. Finally, the system was verified to work stably at the hourly level for pilot-scale extraction. This mini-MS based online analytical system is expected to be further developed for QC applications in a wider range of pharmaceutical processes.

Design and Experiment of Online Detection System for Water Content of Fresh Tea Leaves after Harvesting Based on Near Infra-Red Spectroscopy.

Fresh tea leaves continuously lose water after harvesting, and the level of water content directly affects the configuration of tea processing parameters. To address this problem, this study established an online detection system for the water content of fresh tea leaves after harvesting based on near-infrared spectroscopy. The online acquisition and analysis system of the temperature and humidity sensor signal data was developed based on LabVIEW and Python software platforms. Near-infrared spectral data, environmental temperature, and humidity were collected from fresh leaves after harvesting. Spectral data were combined with PLS (partial least squares) to develop a prediction model for the water content of fresh tea leaves. Simultaneously, data communication between LabVIEW and PLC was established, laying the foundation for establishing a feedback mechanism to send the prediction results to the main platform of the lower computer. This provides a more objective and accurate basis for the detection of fresh leaves before processing and regulation during processing, thereby effectively promoting the standardisation and intelligent development of tea-processing equipment.

Controlled release devices as adjuvants in nonsurgical periodontal therapy: A systematic review.

Controlled release devices (CRD) have been widely studied regarding their application in periodontal therapy. Therefore, the present study aims to evaluate the use and effectiveness of controlled-release devices in nonsurgical periodontal therapy through a systematic review. The research was carried out in six different databases, namely: Online Medical Literature Search and Analysis System (Medline-PubMed), Web of Science, Science Direct, Scopus, Scielo, and Google Scholar. The descriptors "Delayed-Action Preparation," "Therapeutics," and "Periodontitis," were used with their EntryTerms and connected through the Boolean operators AND and OR. A total of 2847 studies were found, and after applying the Preferred Reporting Items for Systematic Reviews and Meta-Analyses checklist flowchart, 34 were selected using the eligibility criteria. After that, the qualitative results were tabulated and the risk of bias in each of them was evaluated. It can be observed that the use of CRD presents itself as a successful alternative for adjuvant treatment to periodontal therapy, a fact due to its availability and local concentration in the crevicular fluid. However, further clinical research is still needed to develop devices that are effective, with an easy and quick application, as well as available at a good cost-benefit ratio.

Effect of long-term infiltration on porosity-permeability evolution in carbonate rocks: An online NMR coupling penetration test

Investigating the porosity–permeability evolution in carbonate rocks during long-term infiltration is a challenging hydrodynamic problem in rock engineering (hydroelectric engineering, CO2 geological storage, and oil/gas development). In general, we consider the evolution of porosity and permeability to be positively correlated. However, the complex pore structures in carbonate rocks include multiscale pore-fracture structures, such as intergranular pores, dissolution pores, and even fractures. Although the permeability levels of carbonate rocks are similar, the evolution of the pore structure is highly variable. Here, we use the “rock seepage coupled nuclear magnetic resonance (NMR) online analysis and imaging system” platform to conduct a long-term penetration test in carbonate rocks under the coupling of stress and reactive fluid. The variation in the multiscale pore volume was quantitatively characterized by the inversion of the T2 spectrum. Subsequently, in combination with permeability measurement, the permeability-porosity evolution during long-term infiltration can be roughly divided into three stages. The experimental results show that the evolution of permeability exhibits noticeable stress sensitivity at the initial stage of the test (0–156 h). Since the compression of the pore volume is the dominant mechanism, the permeability decreases with porosity at this stage. A negative correlation between the evolution of porosity and permeability was observed during the subsequent stage of the test (156–400 h). This phenomenon may be attributed to mineral dissolution at this stage, causing the cementation to be exfoliated from the pore surface and to re-precipitate at the pore throat, locally blocking the seepage channels. As the test continued (400–720 h), mineral dissolution transitioned from micropores to mesopores and macropores, with a synergistic increase in porosity and permeability. These results revealed the competing mechanism of stress compression and mineral dissolution during long-term infiltration, providing an accurate prediction of the seepage properties in carbonate rocks.

Effect of oxygen on thermal behaviors and kinetic characteristics of biomass during slow and flash pyrolysis processes

Pyrolysis is usually considered as the initial stage of solid fuel combustion and gasification, and the pyrolysis characteristics in oxidizing atmosphere greatly affect the ignition behavior, flame stability and product distribution. However, the effect of oxygen on thermal behaviors and kinetic characteristics of biomass pyrolysis was poorly understood and has been studied only based on thermogravimetric analysis (TGA). TGA has heat transfer limitations, resulting in a significant difference between the calculated kinetic results and actual ones. In this work, a bubbling fluidized bed with an on-line analysis system was developed to investigate the real-time gas releasing behaviors of biomass flash pyrolysis in inert and oxidizing atmospheres. The slow pyrolysis characteristics were investigated in TGA for comparison. The effects of oxygen on the kinetics of flash and slow pyrolysis were investigated by combining the distributed activation energy model (DAEM) isothermal model and non-isothermal models including Flynn-Wall-Ozawa (FWO), Kissinger-Akahira-Sunose (KAS) and Starink. Under slow heating conditions, the presence of oxygen led to a higher maximum pyrolysis rate in a smaller temperature interval, and the maximum pyrolysis rate shifted to the lower temperature. In oxidizing atmosphere, the pore volume and specific surface of char were about 4–5 times higher than that of the char prepared in inert atmosphere. The values of apparent activation energy (Ea) at the initial stage of pyrolysis reaction in oxidizing atmosphere were much higher than those in inert atmosphere. As the conversion rate reached 0.4–0.5, the presence of oxygen led to char gasification and combustion, as well as a considerable change in Ea values. In addition, the Ea values of biomass flash pyrolysis obtained by the fluidized bed were about 4–5 times smaller than those of slow pyrolysis obtained by TGA.

Negative Magnetophoresis Focusing Microchips Online-Coupled with ICP-MS for High-Throughput Single-Cell Analysis.

High-throughput single-cell analysis is critical to elucidate the cell heterogeneity. Recently, droplet microchips using oil/gas phases to generate single-cell encapsulated droplets have been combined with inductively coupled plasma-mass spectrometry (ICP-MS) for determination of trace elements in single cells with a throughput of dozens of cells per min. To improve the sample throughput and avoid the oil phase introduced into ICP-MS, herein, a negative magnetophoresis focusing microchip was established and online-coupled to ICP-MS for single-cell analysis. MCF-7 cells in the paramagnetic salt solution were introduced into the designed focusing microchannel, in which they were focused into a single stream under both the magnetic repulsion force and inertial lift force, and then were introduced into ICP-MS for online single-cell analysis. The important parameters including the chip design, the concentration of the paramagnetic salt solution, flow rate, cell density, and dwell time were optimized. Under the optimal conditions, a high sample throughput of 1390 cells min-1 was obtained. The established online analytical system was applied to study the uptake behaviors of MCF-7 cells for Zn2+ and ZnO nanoparticles (NPs) at a single-cell level. The single-cell analysis results indicate that MCF-7 cells displayed more remarkable heterogeneity when they were treated with ZnO NPs, and the uptake content of ZnO NPs by MCF-7 cells was less than that of Zn2+. Compared with other droplet microdevice-ICP-MS analysis systems, the developed system has the advantages of simple design and fabrication, no organic phase, a high throughput, and a low sample consumption (only 5 μL).

Effect of grape seed extract on stability of restorations with composites resin: A systematic review and meta-analysis

Grape Seed Extract (GSE), rich in proanthocyanidins (PAC), has been extensively researched to optimize the stability of the hybrid layer in dentin substrate. Therefore, this study aims to evaluate the use of grape seed extract in the stability of restorations with resinous composites in dentin, by a review and meta-analysis. The search was carried out in 4 different databases: Online Medical Literature Search and Analysis System (Medline – PubMed), Web of Science, Science Direct, and Scopus. Proanthocyanidins, Cross-Linking Reagents, and Dentin descriptors were used with their respective entry terms connected by the boolean operators AND and OR. We found 1074 articles, which, after applying the PRISMA flowchart checklist, resulted in 9 articles selected according to the eligibility criteria. We tabulated the qualitative results and assessed the risk of bias in each of them. Of the selected articles, 2 presented low risk, 5 medium risk and 2 high risk of bias. We compiled and tabulated the numerical data of bond strength to perform the meta-analysis (p < 0.05) of the articles that presented the GSE as a pretreatment and incorporated into the conditioning agent. We noted that, as a pre-treatment, the GSE improves the bonding stability, but incorporated with phosphoric acid it decreases the stability when compared to the control group.