Electrophoretic Parameters Research Articles

B-057 Evaluating the Use of Unconcentrated Urine for Protein Electrophoresis and Immunofixation: An Alternative Approach for Efficient Analysis

Abstract Background Protein electrophoresis is a widely employed technique for separating and analyzing proteins based on their charge and size. Traditionally, concentrated urine samples have been utilized to enhance protein detection sensitivity. However, this study explores the feasibility of using unconcentrated urine as an alternative sample type for protein electrophoresis (UEL) and immunofixation (UIF) to streamline the analysis process and reduce costs and turnaround time in clinical laboratories. It offers a more efficient and resource-friendly alternative. Methods Both UEL and UIF were performed on routinely ordered 80 urine specimens (random or 24 Hr.) for patient management and performed by both concentrated (X40) by Helena and unconcentrated using Sebia (HYDRAGEL 9IF gels using HYDRAGEL BJ dynamic mask procedure. The electrophoretic patterns obtained from unconcentrated urine were compared with those from concentrated urine. Various electrophoretic parameters, including resolution, sensitivity, and reproducibility, were evaluated to determine the reliability of unconcentrated urine in protein electrophoresis. In addition, retrospective analysis was performed before and after implementation on all urine specimens ordered to analyze if there were any differences in the monoclonal band distributions in the population. Results All the protein concentrations were correlated well with 95-100% agreement between the methods (Figure 1) clear separation of protein fractions. Serial dilution studies of monoclonal band analyses showed that these bands can be detected at 1.25 mg/dL using an unconcentrated method. Retrospective analysis of patient data before and after the implementation of the new method shows no differences in the identification of monoclonal proteins (Helena [69 of 872 total patients tested (8.0%)] vs. Sebia [90/800 (11.0%)]. Conclusions These results indicate that unconcentrated urine can be successfully utilized for UEL and UIF with comparable outcomes to traditional concentrated samples. Utilizing unconcentrated urine simplifies the sample preparation process and presents a promising alternative.

Diet supplementation with Saccharomyces cerevisiae influences the electrophoretic parameters in blood in young Charolaise bulls.

The objective of the research was to investigate the effect of Saccharomyces cerevisiae supplementation on some acute-phase proteins, haptoglobin and all electrophoretic parameters in young Charolaise bulls. Sixty bulls were divided into two equal groups: the control group (CG) receiving the base diet without yeast supplementation and the diet supplementation group (YG) receiving the base diet with 5g of Saccharomyces cerevisiae supplementation. The base diet was total mixed ration allocated at 11.85 kg per animal per day. Blood samples were collected from all bulls on day 0 before the start of the diet supplementation, and on days 20 and 40 after the start. Total proteins, albumin, globulin fraction (α1-, α2-, β1-, β2- and γ-globulins), albumin: globulin ratio (A: G) and haptoglobin were determined. Two-way analysis of variance showed a significant effect of the yeast feeding time on all studied parameters except α2-globulins in both groups. The YG showed a higher average concentration of total proteins, albumin and A: G and a lower average concentration of γ-globulins and haptoglobin than the CG. These results indicated the beneficial effect of the Saccharomyces cerevisiae on the inflammatory status of the young bulls, which showed an adequate response in serum levels of the acute-phase proteins tested.

Characterization of Hemoglobin Malay Phenotypes in Tertiary Hospitals

Hemoglobin (Hb) Malay is a common β hemoglobinopathy in Malaysia caused by A > G mutation in codon 19 leading to β+-thalassemia phenotype. However, screening for Hb Malay is challenging as it is undetectable by routine capillary electrophoresis (CE) or high-performance liquid chromatograpy (HPLC) methods. This study aimed to determine the Hb Malay phenotypes. The study was done on 521 cases with presumed β thalassemia from UKMMC and Hospital Selayang as well as confirmed Hb Malay cases from Hospital Sultanah Bahiyah, Kedah in over a 5-year period. Hb analysis using CE or HPLC followed by multiplex amplification refractory mutation system polymerase chain reaction and DNA sequencing were performed. Significant differences in mean values of haematological parameters among Hb Malay carriers against β thalassemia carriers were determined using one-way ANOVA and ROC analysis. A total of 482/521 cases of β globin mutations were identified. Among these, 54 Hb Malay cases were identified whereby 21 Hb Malay cases were from UKMMC and Hospital Selayang whilst 33 Hb Malay cases were from Hospital Sultanah Bahiyah, Kedah. Fifty-two were Hb Malay carriers whereas two cases were compound heterozygotes. The mean hemoglobin, mean cell volume, mean cell hemoglobin, and HbA of Hb Malay carriers were significantly higher than β° thalassemia carriers. The HbA2 range of Hb Malay carriers was wider (3.5–5.5%) with median value of 3.9%. A new HbA2 cutoff value ≤4.6% (AUC 0.717, p < 0.001) was proposed. Compound heterozygous Hb Malay/IVS1-5(G > C) showed transfusion-dependent thalassemia phenotype. Hb Malay carriers have different red cell and electrophoretic parameters than classical β° thalassemia carriers with wider HbA2 range. HbA2 of ≤4.6% should prompt a molecular confirmation for Hb Malay carrier status.

POLYMORPHISM AND INHERITANCE OF GLIADIN PROTEINS IN WHEAT LANDRACES OF UZBEKISTAN

Wheat genotypes grown in Uzbekistan and other Central Asian countries cannot compete with commercial cultivars for productivity. These landraces are valuable genetic resources that could benefit in developing new cultivars enriched with nutritious microelements and other ingredients beneficial for human health. As a result of wheat research, for the first time, preserving the 31 landraces in their natural form in the remote territories of Uzbekistan ensued, including a catalog of compilation by determining their economic and morphologically valuable features. Based on cluster analysis and quantitative characteristics, local wheat genotypes attained five groupings with a compiled dendrogram. With the dendrogram, determining the morphological attributes of ancient local cultivars of wheat materialized, even though the grain color is the same, a sharp difference is noticeable, but still similar based on quantitative characteristics that ensure the grain yield. In ancient local wheat cultivars, plant height and 1000-seed weight have a weak correlation with other traits, and a strong positive association was distinct between the spike length and spikelets per spike and the number and grain weight per spike. The polymorphism of soft wheat landraces showed according to the electrophoretic spectra in the grain’s gliadin proteins. The electrophoretic parameters of gliadin proteins indicated that of the six samples of cultivar Kzyl-bugdoy listed in the catalog, only four have similar electrophoretic spectra. In cultivar Ak-bugdoy’s four specimens, three have identical spectra, while three from the cultivar Surkhak have different bands. Based on individual selection and according to electrophoretic spectra from wheat landraces, the release of the wheat cultivar Kairaktash was due to its high baking and nutritional value.

Development of Capillary Zone Electrophoresis Method for the Simultaneous Separation and Quantification of Metformin and Pioglitazone in Dosage Forms; and Comparison with HPLC Method.

A capillary zone electrophoretic (CZE) method was developed, validated, and applied for the assay of metformin (MET) and pioglitazone (PIO) in pharmaceutical formulations. The optimum running buffer composition was found to be 75 mmol/L phosphate buffer containing 30% acetonitrile (ACN) at pH 4.0. The optimum instrumental conditions were found to be injection time, 10 s; applied voltage, 25 kV; hydrodynamic injection pressure, 0.5 psi for 10 s, capillary temperature, 25 °C; and the detection wavelength, 210 nm. The quantifications were calculated based on the ratio of the peak areas of analytes to atenolol as an internal standard. The CZE method was validated in terms of accuracy (98.21-104.81%), intra- and inter-day precision of migration time and peak area (relative standard deviation ≤ 5%), linearity (correlation coefficients ≥ 0.9985), limit of detection (≤0.277 μg/mL), and limit of quantitation (≤0.315 μg/mL). The proposed method was applied for the analysis of PIO and MET both individually and in a combined dosage tablet formulation. All electrophoretic parameters were calculated and evaluated. A previously reported high-performance liquid chromatographic (HPLC) method was also applied to the same samples. A comprehensive comparison was then carried out for the analytical features of both methods CZE and HPLC. Comparable results were obtained with the advantage of reagent consumption and separation efficiency of CZE over HPLC and shorter analysis time by HPLC compared with CZE.

Optimizing Nanostructured Yttria-Stabilized Zirconia Electrophoretic Coatings on MCrAlY Bond-Coated Inconel 738LC Superalloy

In this research study, to apply uniform and homogenous nanostructured yttriastabilized zirconia (YSZ) coatings on the Inconel 738LC with MCrAlY bond-coat, the effect of electrophoretic parameters was evaluated. Two different suspensions of 3YSZ nanoparticles were prepared in acetylacetone and acetone separately, using an iodine dispersant. The electrophoretic deposition was conducted by applying voltages of 20 V, 50 V and 80 V for 3 min on MCrAlY coated Inconel 738LC. The coatings were sintered for 120 min at 900°, 1000° and 1100°C temperature. The results revealed that the non-uniform coating included some cracks and cavities as the applied voltage exceeded the optimal amount. The results also depicted that the YSZ coating prepared in acetylacetone medium with 0.4 g/L iodine dispersant, voltage of 50 V, coating time of 3 min, and sintering temperature of 1000°C was a uniform and crack-free structure. Furthermore, the results showed that a continuous alumina layer was formed during oxidation.

Controlled Electrophoretic Deposition Strategy of Binder-Free CoFe2O4 Nanoparticles as an Enhanced Electrocatalyst for the Oxygen Evolution Reaction.

The kinetic-sluggish oxygen evolution reaction (OER) is the main obstacle in electrocatalytic water splitting for sustainable production of hydrogen energy. Efficient water electrolysis can be ensured by lowering the overpotential of the OER by developing highly active catalysts. In this study, a controlled electrophoretic deposition strategy was used to develop a binder-free spinel oxide nanoparticle-coated Ni foam as an efficient electrocatalyst for water oxidation. Oxygen evolution was successfully promoted using the CoFe2O4 catalyst, and it was optimized by modulating the electrophoretic parameters. When optimized, CoFe2O4 nanoparticles presented more active catalytic sites, superior charge transfer, increased ion diffusion, and favorable reaction kinetics, which led to a small overpotential of 287 mV for a current density of 10 mA cm-2, with a small Tafel slope of 43 mV dec-1. Moreover, the CoFe2O4 nanoparticle electrode exhibited considerable long-term stability over 100 h without detectable activity loss. The results demonstrate promising potential for large-scale water splitting using Earth-abundant oxide materials via a simple and cheap fabrication process.

Evaluation of cyclodextrin- and cyclofructan-based chiral selectors for the enantioseparation of psychoactive substances in capillary electrophoresis.

During this study, a simple and easy-to-prepare electrophoretic method was developed for the enantioseparation of amphetamine and cathinone derivatives. Different types of β-cyclodextrin and cyclofructan-based chiral selectors (CSs), both native and derivatized, were utilized, and the most effective ones, in terms of resolution and analysis time, were identified. In addition, several electrophoretic parameters, such as background electrolyte concentration and pH, and CS concentration, were examined to optimize the separation conditions. Under the optimal electrophoretic conditions, 10 psychoactive substances were enantiomerically separated using 1mM sulfated cyclofructan-6 (SCF-6) for the amphetamine derivatives and 1mM sulfated cyclofructan-7 (SCF-7) for the cathinone derivatives dissolved in an aqueous solution of 20-mM monobasic sodium phosphate at pH2.5, a temperature of 25°C, and an applied voltage of 25kV. In addition, the method was validated by estimating the intra- and interday precision.

The Effect of Broccoli Extract in Arsenic-Induced Experimental Poisoning on the Hematological, Biochemical, and Electrophoretic Parameters of the Liver and Kidney of Rats.

Heavy metals such as arsenic contribute to environmental pollution that can lead to systemic effects in various body organs. Some medicinal plants such as broccoli have been shown to reduce the harmful effects of these heavy metals. The main aim of the present study is to evaluate the effects of broccoli extract on liver and kidney toxicity, considering hematological and biochemical changes. The experimental study was performed in 28 days on 32 male Wistar rats classified into four groups: the control group (C), a group receiving 5 mg/kg oral arsenic (AS), a group receiving 300 mg/kg broccoli (B), and a group receiving arsenic and broccoli combination (AS + B). Finally, blood samples were taken to evaluate the hematological and biochemical parameters of the liver and kidney, as well as serum proteins' concentration. Liver and kidney tissue were fixed and stained by H&E and used for histopathological diagnosis. The results demonstrated a significant decrease in white blood cells (WBC), red blood cells (RBC), and hemoglobin (Hb) in the AS group compared to other groups. However, in the B group, a significant increase in RBC and WBC was observed compared to the AS and C groups (P < 0.05). Moreover, RBC and WBC levels increased significantly in the AS + B group compared to the AS group (P = 0.046). However, in the AS group, aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, and creatinine levels increased, while total protein, albumin, and globulin decreased. This can be a result of liver and kidney damage, which was observed in the AS group. Furthermore, the increase in the concentration of albumin and globulin in the AS + B group was higher than that in the AS group. Infiltration of inflammatory cells and necrosis of the liver and kidney tissue in the pathological evaluation of the AS group were significantly higher than other groups. There was an increase in superoxide dismutases (SOD), glutathione peroxidase (GPx), and total antioxidant capacity (TAC); however, a decrease in malondialdehyde (MDA) concentration was seen in the AS + B group compared to the AS group. It seems that broccoli is highly effective at reducing liver and kidney damage and improving the hematological and biochemical factors in arsenic poisoning conditions.

Influence of deposition time and applied voltage on the properties of electrophoretically deposited nickel oxide colloidal nanoparticles thin film

ABSTRACT In the present work, nickel oxide nanoparticles (NPs) have been synthesised, using the laser ablation technique in aqueous suspension. By applying the electrophoretic deposition (EPD) method with direct current (DC), the nickel oxide NPs obtained have been deposited on an aluminium substrate. During the EPD process, different films have been produced by a variety of deposition times and applied voltages. The characterisation of the samples, including nickel oxide NPs suspension and nickel oxide deposited thin films, was carried out through different methods, such as X-ray diffractometry (XRD), scanning electron microscopy (SEM), UV–Vis spectroscopy and atomic force microscopy (AFM). The crystalline structure of the samples inspected by XRD showed that nickel oxide existed in NPs suspension and all the thin films. The morphology of the deposited films and dried NPs has been obtained by SEM. By observing the SEM micrographs, it was shown that deposition time and applied voltage change the morphology of the deposited thin films. In addition, AFM results demonstrated that the electrophoretic parameters have an effect on the average particles size, thickness and roughness of the deposited films. The optical properties of the NPs and deposited films have been analysed by UV–Vis–NIR spectroscopy.

A three-dimensional electrode fabricated by electrophoretic deposition of graphene on nickel foam for structural supercapacitors

The synthesis of a 3D rGO@NF structural electrode was optimized by adjusting the electrophoretic deposition parameters and thermal reduction time.

Study of clustered DNA lesions in the liver in acute toxic injury

Objective: to test and optimize a method for detecting clustered DNA lesions with an assessment of the quantitative characteristics of hepatic changes using an experimental model of acute toxic hepatitis.Material and methods: Laboratory C57Bl/6 mice at the age of 10 weeks were used for the study. Acute toxic liver injury was induced by means of a single intraperitoneal injection of 30 % CCI4 solution in olive oil. The withdrawal of the animals from the experiment was made after 72 hours. For the detection of clustered DNA lesions, a liver fragment was resected.Results. The electrophoretic parameters which are the most optimal for obtaining primary data for the subsequent calculation of the number of clustered DNA lesions have been proposed. The number of clusters in the DNA samples from the animals of the control group is significantly lower than in the experimental group and amounts to 54.80 [37.65; 59.24] and 76.82 [60.95; 92.41] APE1 clusters per million bp., respectively.Conclusion. Successful testing and optimization of the OCDL method for the detection of clustered lesions in liver DNA have been performed. The study has shown an increase in the number of APE1 clusters and double-strand breaks in the DNA of the C57Bl laboratory mice with induced acute toxic hepatitis, which indicates significant derangement of DNA integrity and a high risk of developing progressive liver diseases in its toxic damage.

Evaluation of the Quality Characteristics of Siirt Herby Cheese: A Traditional Turkish Variety

Herby cheese produced in the Province of Siirt is one of the Turkish cheese varieties. The special parameters applied during the production stages give Siirt Herby cheese unique taste properties. The proteolytic, lipolytic, and electrophoretic parameters of Siirt Herby cheese samples were evaluated. The mean values of water-soluble nitrogen, trichloroacetic acid-soluble nitrogen, and phosphotungstic acid-soluble nitrogen of Siirt Herby cheese samples were 9.49±2.66%, 6.33±2.39%, and 1.48±0.71%, respectively. The mean values of free amino group and lipolysis levels were established as 61.97±30.84 mM glycine, and 6.00±2.32 ADV, respectively. The electrophoretic analysis showed that sheep milk was more preferable for Siirt Herby cheese production.

Quantification of lactose and lactulose in hydrolysed-lactose UHT milk using capillary zone electrophoresis

A new method for quantification of lactose and lactulose in hydrolysed-lactose UHT milk, using capillary zone electrophoresis with indirect detection in the ultraviolet region (CZE-UV), is proposed. A 33 Box–Behnken design for instrumental electrophoretic parameters optimisation was used, with the best conditions shown to be an injection pressure of 15 mbar 2 s, applied voltage of −20 kV, and cartridge temperature of 20 °C. Extraction procedures were investigated and protein precipitation using trichloroacetic acid solution was shown to give the best results. Limits of quantification for lactose and lactulose were 0.024 g 100 mL−1 and 0.0094 g 100 mL−1, respectively. Recovery levels ranging from 86% to 102% were obtained. The method was successful applied to several hydrolysed-lactose and regular UHT milk samples from six commercial leading brands. Results showed that all hydrolysed-lactose samples are in accordance with the legal recommendations to be classified as “lactose-free” products.

New bioanalytical microemulsion Electrokinetic chromatography method for the simultaneous determination of Trifluridine with its metabolites and Tipiracil in rat plasma: Application to pharmacokinetic studies

A Microemulsion Electrokinetic Chromatography method coupled with diode array detector (MEEKC-DAD) was developed for the first time and found to be efficient, sensitive, and selective for the simultaneous analysis of Trifluridine (FTD), and its metabolites 5-(trifluoromethyl) uracil (FTY) and 5-carboxy-2′-deoxyuridine (5CDU), and Tipiracil (TIP) in rat plasma. Sample pre-treatment involved a simple protein precipitation from plasma using acetonitrile. The separation was achieved using a fused silica capillary (65 cm total length, 55 cm effective length and 50 µm i.d.) and a microemulsion solution consisted of 1.66% sodium dodecyl sulfate (SDS), 0.91% heptane, 6.61% 1-butanol, and 90.72% borate buffer (20 mM, pH 9.5). Electrophoretic separation was carried out at 20 °C and 20 kV. The samples were injected for 40 s at 20 mbar and detected simultaneously at 205 nm. The electrophoretic parameters indicated that the developed MEEKC-DAD method permitted complete resolution of the analytes within 13 min. The developed method was fully validated according to the FDA guidelines for bioanalytical method validation. The method was linear in the range 200–4000 ng/ml for FTD, FTY, 5CDU, and 100–1000 ng/ml for TIP. The intra/inter-day accuracy and precisions were ≤4% for all drugs. Extraction recovery and stability were also assessed and were within acceptable range. After being validated, the method was applied for the determination of the studied drugs in plasma samples collected from rats injected intraperitoneally with a combination of FTD and TIP. The results obtained were used to study the pharmacokinetics of FTD with its metabolite and TIP in rat plasma.

Development and validation of a capillary electrophoresis method applied to the analysis of l-citrulline in an oral formulation for pediatric use.

A simple and highly sensitive CE-UV method was applied in the determination of l-ctrulline, which was developed from an oral formulation for pediatric use. The novel method was based on the analysis of l-citrulline for direct ultraviolet detection at 198nm. The BGE consisted of 10 mM sodium tetraborate and 50 mM SDS at pH 9, and the electrophoretic parameters were optimized. The method was validated in terms of specificity, linearity, LOD, LOQ, precision, accuracy, and robustness. The LOD and LOQ obtained were 1.36 and 4.54 μg/mL, respectively. In addition, the method offers higher sensitivity and specificity compared with the results obtained from HPLC method using UV-detectors, in which l-citrulline needs to be derivatizated. Furthermore, low cost and simplicity of the system allowed the rapid and simple quantitation of l-citrulline in the oral formulation for quality control and stability indicated method.

Application of Capillary Zone Electrophoresis Coupled with a Diode Array Detector (CZE-DAD) for Simultaneous Analysis of Ibuprofen and Phenylephrine.

Background: A validated method based on capillary zone electrophoresis coupled with a diode array detector (CZE-DAD) was investigated for analyzing binary mixture of ibuprofen (IBU) and phenylephrine (PHE) in their bulk and combined dosage form. Objective: This binary mixture is a challenging one as IBU is acidic and PHE is alkaline, which may affect their simultaneous analysis using CZE. The literature lacks any CZE report for IBU and PHE simultaneous analysis. Methods: Fused silica capillary (85 cm × 75 μm id) was used, and the electrolyte was a 50 mM borate buffer adjusted to pH 11 with 0.5 M NaOH. Results: The concentration ranges were 5-200 and 5-100 μg/mL for IBU and PHE, respectively, using CZE. High efficiency was achieved (N > 92990). Reasonable migration time (tm) was attained (tm< 8.5 min). Conclusions: Although the results obtained by the proposed CZE method and reported HPLC method were statistically comparable, the proposed method showed lower linearity ranges, higher efficiency, and a more reasonable run time. Highlights: CZE-DAD was used for the analysis of IBU and PHE in bulk and tablets, as no report was found for their determination using CZE. Binary mixture is challenging due to differences in chemical and physical properties. A detailed discussion of electrophoretic parameters optimization is included. Confirmation of peak purity was attained using DAD.

Electrophoretic Behavior in Nonaqueous Capillary Zone Electrophoresis Using Tetrahydrofuran-Based Ternary Nonaqueous Solutions

Nonaqueous capillary electrophoresis has the potential to separate water-insoluble analytes. Previously, we reported the separation of water-insoluble and neutral synthetic polymers in nonaqueous capillary zone electrophoresis using tetrahydrofuran (THF)-based ternary nonaqueous solutions. In this paper, the effect of the composition of a THF-based ternary solution (THF, acetonitrile, and methanol) containing cetyltrimethylammonium chloride (CTAC) on electrophoretic behavior is investigated using small organic compounds as prove analytes. In addition to general electrophoretic parameters such as electrophoretic mobility, analytical stability was also evaluated. The electric current, electrophoretic mobility of the benzyltriethylammonium cation (ion pair formation with chloride anion), and electrophoretic mobility of coronene (solvophobic interaction with the CTA cation) were dominated mainly by the THF content, and lower THF content resulted in higher values. In contrast, electroosmotic mobility was enhanced by an increase in THF content and was affected mainly by methanol content. Interestingly, in addition to stable electropherograms, we observed instable (fluctuation of baseline or zero current) electropherograms, which were obtained irregularly. Although a notable relation between the composition and the instable behavior was not observed, it is suggested that this “discontinuous instability” is related with the composition distribution (heterogeneous composition) of the ternary solvents in a capillary. Increasing the temperature effectively suppressed the composition distribution and has the potential to decrease the risk of instable behavior.

Radiation-induced Changes in the Electrophoretic Profile of Serum Albumin

ABSTRACT Albumin protein profiles were investigated in electrophoresis system in relation to the whole body exposition to the radiation. Two groups of rats Wistar were set up as the control (CG) and the irradiated one (IG). The IG was exposed to Co-60 at a dose of 5 Gy. After a 72-hour exposition, 300 μL of blood was collected in the inferior vena cava, renal, jugular, hepatic, and pulmonary veins and the serum separated. The albumin protein was identified by vertical electrophoresis in acrylamide Commassi blue or silver stained. The calibration procedure was applied to albumin samples with well-known concentrations. The mathematical correlation was developed involving electrophoretic parameters of band intensities and sizes from gel representation, providing values of protein concentrations in comparison with standard bands with known concentrations. There were significant differences in the physiological concentrations in the jugular and pulmonary sites in relation to renal and cava regional sites. Significant differences induced by radiation in serum albumin concentration were also found in hepatic and jugular sites. Alteration of albumin concentration was found as a nearly effect from whole body irradiation. This phenomenon points out to alterations in cell metabolism in the liver justified by a possible indication of proteomics damage from radiation.

Development and validation of a novel sensitive UV-direct capillary electrophoresis method for quantification of alendronate in release studies from biomaterials.

A simple, highly sensitive, and robust CE method applied to the determination of alendronate (ALN) was developed from matrices for tissue engineering, characterized by being highly complex systems. The novel method was based on the ALN derivatization with o-phthalaldehyde and 2-mercaptoethanol for direct ultraviolet detection at 254nm. The BGE consisted of 20mM sodium borate buffer at pH 10, and the electrophoretic parameters were optimized.The method was validated in terms of specificity, linearity, LOD, LOQ, precision, accuracy, and robustness. The LOD and LOQ obtained were 0.8 and 2.7μg/mL, respectively. In addition, the method offers higher sensitivity and specificity compared to other CE and HPLC methods using UV-detectors, as well as low cost and simplicity that allowed the rapid and simple quantitation of ALN from bone regeneration matrices.