Performance evaluation of different albumin assays for the detection of analbuminemia.

Analbuminemia is characterized by the near absence of albumin in the plasma. Different methods are available for measuring albumin levels, but they do not necessarily agree with one another. It is a concern that analbuminemic samples could be falsely characterized due to the incorrect estimation of albumin. The objective of the work was to evaluate the performance of different assays in detecting analbuminemia. Albumin knockout (Alb-/-) mouse plasma was used to test the suitability of different albumin assays for their ability to properly characterize extreme albumin deficiency. Bromocresol green (BCG), bromocresol purple (BCP), enzyme-linked immunosorbent assay (ELISA), liquid chromatography-tandem mass spectrometry (LC-MS/MS), and gel electrophoresis were tested. The LC-MS/MS assay exhibited broad coverage of the amino acid sequence of albumin and indicated 8,400-fold lower (P<0.0001) albumin expression in Alb-/- than wildtype (WT), demonstrating its suitability for identifying extreme albumin deficiency. ELISA estimated albumin at 1.5±0.1 g/dL in WT and was below the detection limit in all Alb-/- samples. Gel electrophoresis yielded consistent results with LC-MS/MS and ELISA. The BCG assay overestimated albumin with apparently appreciable albumin concentrations in Alb-/- mice, yet the assay still indicated a significant difference between genotypes (Alb-/-, 1.2±0.05 g/dL, WT, 3.7±0.1 g/dL, P<0.0001). BCP drastically overestimated albumin and could not successfully identify the known analbuminemic phenotype of Alb-/- mice. By using Alb-/- plasma as a reference material and LC-MS/MS as a reference method, ELISA and gel electrophoresis appear appropriate for identifying analbuminemia, while BCG and BCP are not suitable. It is concluded that dye-binding assays should be avoided when extreme hypoalbuminemia or analbuminemia is suspected.

Whole genome sequencing analysis and Box-Behnken design for the optimization of the decolourization of mixture textile dyes by halotolerant microbial consortium.

The use of dyes in textile industries has resulted in substantially contaminated soil, water and ecosystem including fauna and flora. So, the application of eco-friendly approach for dyes removal is in great demand. The goal of this research was to develop and test a bacterial consortium for biodegrading dyes in artificial textile effluent (ATE) derived from mixture of Indigo carmine (40 mg/l); Malachite green (20 mg/l); Cotton bleu (40 mg/l); Bromocresol green (20 mg/l) and CI Reactive Red 66 (40 mg/l) dissolved in artificial seawater. The Box-Behnken design (BBD) which combine six variables with three levels each was used to determine the potential removal of dyes in ATE, by the selected microbial consortium (M31 and M69b). The experimental process indicated that decolourization of ATE reached 77.36 % under these conditions values: salinity (30 g/l), pH (9), peptone (5 g/l), inoculum size (1.5 108 CFU/ml), agitation (150 rpm) and contact time (72 h). The decolourization was confirmed by FTIR spectrum analysis of ATE before and after bacterial treatment. Bacterial strains used in this study were identified as Halomonas pacifica M31 and Shewanella algae M69b using 16 rDNA sequences. Moreover, the total genome analysis of M31 and M69b validated the implication of bacterial genes in mixture dyes removal. Therefore, the effect of the selected bacterial consortium on ATE removal was confirmed and it may be used in industrial wastewater treatment to issuing environmental safety.

Albumin determined by bromocresol green leads to erroneous results in routine evaluation of patients with chronic kidney disease.

OBJECTIVES: Measurement of plasma albumin is pivotal for clinical decision-making in patients with chronic kidney disease (CKD). Routinely used methods as bromocresol green (BCG) and bromocresol purple (BCP) can suffer from aselectivity, but the impact of aselectivity on the accuracy of plasma albumin results of CKD-patients is still unknown. Therefore, we evaluated the performance of BCG-, BCP- and JCTLM-endorsed immunological methods in patients with various stages of CKD. METHODS: We evaluated the performance of commonly used albumin methods in patients with CKD stages G1 through G5, the latter divided in two groups based on whether they received hemodialysis treatment. In total, 163 patient plasma samples were measured at 14 laboratories, on six different BCG and BCP-platforms, and four different immunological platforms. The results were compared with an ERM-DA-470k-corrected nephelometric assay. The implications on outcome is evaluated by the proportion of patient results <38 g/L for the diagnosis of protein energy wasting. RESULTS: Albumin results determined with BCP- and immunological methods showed the best agreement with the target value (92.7 and 86.2 %, respectively vs. 66.7 % for BCG, namely due to overestimation). The relative agreement of each method with the target value was platform-dependent, with larger variability in agreement between platforms noted for BCG and immunological methods (3.2-4.6 and 2.6-5.3 %) as opposed to BCP (0.7-1.5 %). The stage of CKD had similar effects on the variability in agreement for the three method-groups (0.6-1.8 % vs. 0.7-1.5 % vs. 0.4-1.6 %). The differences between methods cause discrepancies in clinical decision-making, as structurally fewer patients were diagnosed with protein energy wasting upon using BCG-based albumin results. CONCLUSIONS: Our study shows that BCP is fit for the intended use to measure plasma albumin levels in CKD patients from all stages, including patients on hemodialysis. In contrast, most BCG-based platforms falsely overestimate the plasma albumin concentration.

Use of Euphorbia balsamifera Extract in Precursor Fabrication of Silver Nanoparticles for Efficient Removal of Bromocresol Green and Bromophenol Blue Toxic Dyes.

Silver nanoparticles (Ag-NPs) are attracting great attention for their use in various applications, along with methods for their green and facile production. In this study, we present a new eco-friendly approach based on the use of Euphorbia balsamifera extract (EBE) in the green synthesis of silver nanoparticles (Ag-NPs), which are then applied as a reducing and stabilizing agent for the efficient removal of water-based reactive dyes such as bromocresol green (BCG) and bromophenol blue (BPB). The as-prepared Ag-NPs are quasi-spherical in shape, with an average diameter of 20-34 nm. Diverse characterization methods, including X-ray diffractometry (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and Brunauer-Emmett-Teller (BET) analysis, were used to analyze these Ag-NPs. The results reveal that water-soluble biomolecules in the Euphorbia balsamifera extract play an important role in the formation of the Ag-NPs. The removal of toxic dyes was studied under varied operational parameters such as Ag-NP dosage, initial dye concentration, pH, stirring time, and temperature. Under the optimum investigated conditions, nearly 99.12% and 97.25% of the bromocresol green and bromophenol blue dyes, respectively, were removed. Both BCG and BPB adsorption were found to adhere to pseudo-second-order kinetics (r22 = 1 and 0.995) and fit the Langmuir isotherm models well (R12 = 0.998 and 0.994), with maximal monolayer adsorption capacities of 20.40 and 41.03 mg/g, respectively. Their adsorption processes were observed to be intrinsically endothermic. The results confirm the potential of the Euphorbia balsamifera extract as a low-cost, nontoxic, and eco-friendly natural resource for the synthesis of Ag-NPs that may be useful in the remediation of hazardous dye-contaminated water sources.

Fluorescence turn-on detection of human serum albumin based on the assembly of gold nanoclusters and bromocresol green.

As the most abundant protein in plasma, human serum albumin plays a vital role in physiological processes, such as maintaining blood osmotic pressure and carrying small-molecule ligands. Since the content of albumin in the human serum can reflect the status of liver and renal function, albumin quantitation is significant in clinical diagnosis. In this work, fluorescence turn-on detection of human serum albumin (HSA) had been performed based on the assembly of gold nanoclusters and bromocresol green. Gold nanoclusters (AuNCs) capped by reduced glutathione (GSH) were assembled with bromocresol green (BCG), and the assembly was used as a fluorescent probe for HSA. After BCG assembling, the fluorescence of gold nanoclusters was nearly quenched. In acidic solution, HSA can selectively bind to BCG on the assembly and recover the fluorescence of the solution. Based on this turn-on fluorescence, ratiometric HSA quantification was realized. Under optimal conditions, HSA detection by the probe possessed a good linear relationship in the range of 0.40-22.50 mg·mL-1, and the detection limit was 0.27 ± 0.04 mg·mL-1 (3σ, n = 3). Common coexisting components in serum and blood proteins did not interfere with the detection of HSA. This method has the advantages of easy manipulation and high sensitivity, and the fluorescent response is insensitive to reaction time.

Evaluation of the two-point calibration bromocresol green method, showing reduced deviation from the bromocresol purple method in sera from patients with hypoalbuminemia.

BACKGROUND: The bromocresol green (BCG) and bromocresol purple (BCP) methods are widely used for albumin measurements in routine testing, but the BCG method is known to react with globulin fractions and to have low specificity for albumin. We evaluated a calibration method using different concentrations of human serum albumin standards (two-point calibration BCG method) with the aim of reducing the effect of globulin fractions on the BCG method in patients with hypoalbuminemia. METHOD: In the two-point calibration BCG method, two concentrations of standard solutions and their calibration values are set based on the difference in albumin concentrations measured by the BCG method (BCG-HSA method) and the modified BCP (modified BCP-HSA method) calibrated with human serum albumin standard solution (HSA). Albumin concentrations were measured in 136 patient serum samples (healthy group: 52, hypoalbuminemic group: 84) by the two-point calibrated BCG method and compared with those obtained using the modified BCP-HSA method. RESULTS: The mean albumin concentrations obtained using the two-point calibrated BCG and modified BCP-HSA methods were 39.18 ± 3.42 g/L and 39.37 ± 3.14 g/L (healthy group) and 26.20 ± 6.23 g/L and 26.23 ± 5.67 g/L (hypoalbuminemia group), respectively. The results of the two-point calibration BCG method were in a close agreement over the entire concentration range tested compared to the modified BCP-HSA method. CONCLUSIONS: Based on these results, this calibration method reduces the influence of the globulin fraction on the BCG method. In the hypoalbuminemic group, the calibration method was shown to provide results consistent with the BCP method, which is highly specific for albumin.

Wearable Dual-Signal NH3 Sensor with High Sensitivity for Non-invasive Diagnosis of Chronic Kidney Disease.

NH3 gas in human exhaled breath contains abundant physiological information related to human health, especially chronic kidney disease (CKD). Unfortunately, up to now, most wearable NH3 sensors show inevitable defects (low sensitivity, easy to be interfered by the environment, etc.), which may lead to misdiagnosis of CKD. To solve the above dilemma, a nanoporous, heterogeneous, and dual-signal (optical and electrical) wearable NH3 sensor mask is developed successfully. More specifically, a polyacrylonitrile/bromocresol green (PAN/BCG) nanofiber film as a visual NH3 sensor and a polyacrylonitrile/polyaniline/reduced graphene oxide (PAN/PANI/rGO) nanofiber film as a resistive NH3 sensor are constructed. Due to the high specific surface area and abundant NH3 binding sites of these two nanofiber films, they exhibit good NH3 sensing performance. However, although the visual NH3 sensor (PAN/BCG nanofiber film) is simple without the need of any detecting facilities and quite stable when temperature and humidity change, it shows poor sensitivity and resolution. In comparison, the resistive NH3 sensor (PAN/PANI/rGO nanofiber film) is of high sensitivity, fast response, and good resolution, but its electrical signal is easily interfered by the external environment (such as humidity, temperature, etc.). Considering that the sensing principles between a visual NH3 sensor and resistive NH3 sensor are significantly different, a wearable dual-signal NH3 sensor containing both a visual NH3 sensor and resistive NH3 sensor is further explored. Our data prove that the two sensing signals in this dual-signal NH3 sensor mask can not only work well without interference with each other but also complement each other to improve the sensing accuracy, indicating its potential application in non-invasive diagnosis of CKD.

Spurious capillary zone electrophoresis pattern in hypercholesterolemic dogs.

Capillary zone electrophoresis (CZE) is a relatively new serum protein electrophoresis method with higher resolution than other electrophoretic techniques. Hypercholesterolemic dogs exhibit a peculiar CZE pattern. Specifically, they have a shoulder or peak immediately next to the albumin peak. We investigated the prevalence of this spurious peak in hypercholesterolemic dogs and its correlation with the serum cholesterol concentration. Moreover, possible discrepancies between the CZE and spectrophotometric (bromocresol green [BCG] method) albumin concentrations in those animals were evaluated, as well as the accuracy in measuring albumin by a different CZE fractionation system. We retrospectively enrolled 500 hypercholesterolemic and normotriglyceridemic dogs. Each electrophoretic curve was inspected visually to identify a spurious peak (prevalence of 68.8%). We chose 120 dogs to further investigate the albumin concentration; CZE albumin was significantly higher than measured using the BCG method. A weak but significant correlation (r = 0.412; p <0.0001) was observed between the magnitude of the spurious peak and the serum cholesterol concentration. Finally, the significant difference between CZE and BCG albumin measurement disappeared (p = 0.92) when the spurious peak was considered as α1-globulins instead of albumin.

High Spatial Resolution Chemical Imaging of Implant-Associated Infections with X-ray Excited Luminescence Chemical Imaging Through Tissue.

Microbial infections associated with implantable medical devices are a major concern in fracture fixation failure. Early diagnosis of such infection will allow successful eradication with antibiotics without an extra cost for a second surgery. Herein, we describe XELCI as a technique with high X-ray resolution, implant specificity, and chemical sensitivity to noninvasively image chemical concentrations near the surface of implanted medical devices. The devices are coated with chemically reporting surfaces. This chemically responsive surface consists of two layers coated on an implantable medical device; a pH-sensitive layer (bromothymol blue or bromocresol green incorporated hydrogel) which is coated over a red-light emitting scintillator (Gd2O2S: Eu) layer for monitoring. A focused X-ray beam irradiates a spot on the implant, and the red light generated by the scintillator (with 620 nm and 700 nm peaks) is transmitted through the sensing layer which alters the spectral ratio depending on the pH. An image is generated by scanning the X-ray beam across the implant and measuring the spectral ratio of light passing through the tissue point-by-point. We used this imaging technique for monitoring implant-associated infections previously on the bone surface of the femur with a modified implantable plate sensor. Now we are studying pH changes that occur from tibial intramedullary rod infections. Two different types of intramedullary rod designs are used in pre-pilot rabbit studies, and we learned that the XELCI technique could be used to monitor any chemical changes that occur not only on the bone surface but also inside the bone. Thus, this enables noninvasive, high spatial resolution, low background local pH imaging to study implant-associated infection biochemistry.

Assessing the Impacts of Preanalytical Field Sampling Challenges on the Reliability of Serum Aflatoxin B1-Lysine Measurements by Use of LC-MS/MS.

Aflatoxin exposure is endemic in developing countries with warm, humid climates that promote toxigenic mold growth on crops and foodstuffs. Estimating human aflatoxin exposure is key to identifying and abating contamination sources. Serum aflatoxin B1 bound to albumin lysine (AFB1-lys) is a preferred exposure biomarker, but field sample collection, processing, transportation, and storage logistics are challenging. We validated an improved LC-MS/MS method for serum AFB1-lys and applied it to three field sampling challenges: transportation/storage (elevated temperature); collection/processing (hemolysis); and sample type substitution (heparinized plasma). Our new LC-MS/MS method had a LOD of 0.03 ng/mL, accuracy (mean spike recovery) of 112%, total imprecision (replicate pool measurements) ≤5% at ≥0.2 ng/mL, and results that were 95.1% similar (mean percentage similarity) to an established method. AFB1-lys in human serum spiked with serum from aflatoxin-dosed rats was stable for 14 days at both ambient (22.5 °C) and elevated (38 °C) temperatures. Simulated hemolysis (adding 0.25-3 mg hemoglobin) did not affect AFB1-lys accuracy at ≥0.5 ng/mL but caused 10-25% signal suppression. Heparinized plasma AFB1-lys was 99.0% similar to serum but interfered with albumin measurements (bromocresol green) causing spurious low bias. Further investigation is warranted, but our findings suggest that AFB1-lys is pre-analytically robust.

Photocatalysis of environmental organic pollutants and antioxidant activity of flavonoid conjugated gold nanoparticles.

The unique properties of nanomaterials have the potential application in different fields of biomedical application along with the management of environmental pollutants. This research work involved the isolation of hesperidin from the orange peel and the preparation of hesperidin gold nanoparticles by the chemical reduction method. The high substrate specificity and lower band gap enable the excitation of gold nanoparticles in visible light. Hence gold nanoparticles are chosen nowadays for the management and removal of organic pollutants. The efficacy of hesperidin gold nanoparticles was evaluated by the photocatalytic activity on organic dyes and pollutants like methyl orange, methylene blue, bromocresol green, and 4 - nitro phenol with sodium borohydride as reducing agent and the antioxidant study by scavenging of free radicals of DPPH, ABTS, and hydroxyl free radicals of hydrogen peroxide. The kinetics of photocatalytic degradation of organic dyes and 4 - nitro phenol was found to follow the first order with rate constants of 10 × 10-3, 37 × 10-3, 23 × 10-3 and 49 × 10-3 min-1 for methyl orange, methylene blue, bromocresol green and 4 - nitro phenol respectively. The hesperidin gold nanoparticles showed significant antioxidant activity as compared to ascorbic acid as standard. The flavonoid conjugated gold nanoparticles can be an efficient antioxidant and photocatalyst for the management of different diseases and wastewater treatment respectively.

[Assessment and influencing factors analysis of bromocresol green colorimetry for serum albumin].

To investigate the application of bromocresol green Colorimetry (BCG) method in measuring serum albumin (ALB) and to evaluate its influencing factors in different diseases. This study was a cross-sectional study that included 128 people admitted to the department of nephrology, department of general surgery, department of infectious diseases and other departments of the Third Xiangya Hospital of Central South University in July 2021. They were divided into groups according to disease types, including chronic kidney disease group (47 cases), liver disease group (40 cases), other diseases group (41 cases), serum ALB was detected by BCG method and immunoturbidimetry at the same time, and the results were expressed as ALBBCG and ALBI respectively, each group was subdivided into three subgroups according to ALBI results: relatively high-value subgroup, relatively intermediate-value subgroup and relatively low-value subgroup of albumin. ALBI and ALBBCG were compared in all groups and subgroups. Passing-Bablok regression and Bland-Altman diagram analysis were used to evaluate the application of ALBBCG in each group. Immunoturbidimetry was used as a reference method to evaluate the bias of ALBBCG, and the differences between ALBI and ALBBCG were shown as follows:ΔALB= ALBBCG-ALBI. Pearson correlation analysis and multiple linear regression analysis were used to assess the correlation between ΔALB and ALB autoconcentration (ALBI), α1-globulin, α2-globulin, ß1-globulin, ß2-globulin, γ-globulin, creatinine (Cr), urea (UN), uric acid (UA), aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBil), direct bilirubin (DBil), and C-reactive protein (CRP) levels.The results showed that ALBBCG were higher than ALBI in the relative low subgroups of total patients group, chronic kidney disease group, liver disease group and other disease groups, and the differences were statistically significant (t value was 8.025, 6.878, 2.628, 4.915, respectively, P<0.05). In the relatively high value subgroup, ALBBCG was lower than ALBI, and the differences were statistically significant in the relative high value subgroup of total patients group, liver disease group and other disease groups (t value was -4.388, -2.927, -3.979, P<0.05). Passing-Bablok regression and Bland-Altman analysis showed that the BCG method had proportional bias. In the chronic kidney disease group, the concentrations of ALBI and Cr had the greatest influence on BCG bias, and the regression model equation was ΔALB=5.437-0.146× AlbI-0.001 ×Cr, R²=0.505. In the liver disease group, the concentrations of ALBI, α1-globulin, ß1-globulin had the greatest influence on BCG bias, and the regression model equation was ΔALB=3.652-0.230×ALBI+0.398×α1-globulin+1.171×ß1-globulin, R²=0.658. In the other disease group, the concentration of ALBI and α2-globulin had the greatest influence on BCG bias, and the regression equation was ΔALB=5.558-0.225×AlbI-0.281×α2-globulin, R²=0.646. The BCG method has a proportion error, and its bias may lead to unacceptable differences. BCG method is mainly affected by the concentration of ALB itself, and may also be affected by α1-globulin, α 2-globulin, ß1-globulin, Cr.

Competitive adsorption of Alizarin Red S and Bromocresol Green from aqueous solutions using brookite TiO2 nanoparticles: experimental and molecular dynamics simulation.

In this work, the effective adsorption and the subsequent photodegradation activity, of TiO2 brookite nanoparticles, for the removal of anionic dyes, namely, Alizarin Red S (ARS) and Bromocresol Green (BCG) were studied. Batch adsorption experiments were conducted to investigate the effect of both dyes' concentration, contact time, and temperature. Photodegradation experiments for the adsorbed dyes were achieved using ultraviolet light illumination (6 W, λ = 365 nm). The single adsorption isotherms were fitted to the Sips model. The binary adsorption isotherms were fitted using the Extended-Sips model. The results of adsorption isotherms showed that the estimated maximum adsorption uptakes in the binary system were around 140 mg g-1 and 45.5 mg g-1 for ARS and BCG, respectively. In terms of adsorption kinetics, the uptake toward ARS was faster than BCG molecules in which the equilibrium was obtained in 7 min for ARS, while it took 180 min for BCG. Moreover, the thermodynamics results showed that the adsorption process was spontaneous for both anionic dyes. All these macroscopic competitive adsorption results indicate high selectivity toward ARS molecules in the presence of BCG molecules. Additionally, the TiO2 nanoparticles were successfully regenerated using UV irradiation. Moreover, molecular dynamics computational modeling was performed to understand the molecules' optimum coordination, TiO2 geometry, adsorption selectivity, and binary solution adsorption energies. The simulation energies distribution exhibits lower adsorption energies for ARS in the range from - 628 to - 1046 [Formula: see text] for both single and binary systems. In addition to that, the water adsorption energy was found to be between - 42 and - 209 [Formula: see text].

[Comparison of three albumin assay methods: a need for standardization within a healthcare area]. / Comparaison de trois techniques de dosage de l'albumine : un argument pour la standardisation des pratiques au sein d'un territoire de santé.

Albumin is a main biomarker of denutrition severity. Since 2019, the French National Authority of Health (HAS) recommends an immunologic technique as a reference for measuring albuminemia. In a context of partial recovery by the laboratory of GHBS (using colorimetric method with bromcresol purple) of the biological activity of the dialysis center, until then carried out by the Ouest biology - Biolor laboratory (using colorimetric method with bromcresol green) and in order to standardize practices in the healthcare sector, we compared 3 albumin assay techniques: immunoturbidimetric (Diagam®), BCG (Siemens®), BCP (Siemens®) on Siemens Atellica PLCs. The albumin level of 183 patient samples was measured. We show an overestimation of the dosage by the BCG (+ 5.46 g/L compared to IT) and an interference related to the inflammatory state of the patient during the dosage with this technique, as well as an underestimation of the dosage by the BCP (- 0.91 g/L relative to the IT), increased in patients with renal injury. This biological difference is added to a discrepancy in clinical conclusions. A change in dosing technique from BCG to BCP may impact the biological follow-up of patients and the PMSI score (mean difference of - 6.67 g/L between BCG and BCP while the classification of the severity of denutrition uses thresholds set in steps of 5 g/L). This variability of results should be taken into account by the clinicians and constitutes an argument in favor of the standardization of laboratories around a reference technique.

Removal of Bromocresol Green from aqueous solution by electro-Fenton and electro-Fenton-like processes with different catalysts: laboratory and kinetic model investigation.

This study presents the removal of triarylmethane dye Bromocresol Green from aqueous solution by the electro-Fenton process. As catalysts five different cations were used: Fe2+, Ce3+, Ni2+, Mn2+, and Co2+. They play crucial roles in the whole process because they react with H2O2 producing hydroxyl radicals that are capable of breaking down dye molecules. Based on this, a comparison of catalytic activity of these cations in the electro-Fenton process is made for Bromocresol Green degradation. A simple and universal kinetic model is also applied to study the catalytic activity of investigated catalysts. Due to its multidimensionality it is fitted to experimental data using a genetic algorithm. The procedure of fitting using a genetic algorithm is thoroughly described and demonstrated. The activity of utilized catalysts is compared based on both experimental and model data revealing that for Bromocresol Green removal all alternative catalysts (Ni2+, Co2+, Ce3+, Mn2+) are better than the typical one (Fe2+, 51.83% degradation). The best catalyst is Co2+ with 78.35% degradation efficiency. Moreover, the adopted kinetic model proved its universality and outlined different interactions between catalysts and dye molecules.

Ligand binding to natural and modified human serum albumin.

This paper reports evaluation of ligand binding constants for unmodified or biotinylated HSA (HSAB) for two well-known HSA binding ligands, naproxen and bromocresol green. Results demonstrate differential scanning calorimetry (DSC) is a reliable quantitative method for straight-forward and rapid evaluation of ligand binding constants for HSA and modified derivatives. DSC measured the thermodynamic stability of free and ligand-bound HSA and HSAB at pH = 6.0, 7.4 and 8.0. DSC analysis provided a quantitative gauge of responses of HSA and HSAB thermodynamic stability to ligand binding. The influence of different levels of biotinylation of HSAB on ligand binding, and how ligand binding varied as a function of pH for these molecules was also examined. In the three pH environments, biotinylation increased stability of HSAB alone compared to free HSA at pH 7.4. Stabilities of free protein and ligand-bound complexes varied with pH in the order, pH = 6.0>7.4>8.0. Our analytical approach provided very accurate estimates for known binding constants of these ligands for HSA. Results revealed, for both ligands, extent of biotinylation of HSAB affected binding, reducing binding constants from three to 100-fold. DSC analysis was able to delineate inter-relationships between molecular structure and thermodynamic stability of HSA and HSAB bound by ligands; and their variations with pH.

Bias between two methods of albumin measurement in the white rhinoceros, Ceratotherium simum.

BACKGROUND: The bromocresol green (BCG) method has been reported to overestimate serum albumin concentration in several species due to non-specific binding to globulins. As the white rhinoceros has high concentrations of serum globulins, significant differences in albumin measured by the BCG method, and the field method of agarose gel serum protein electrophoresis (SPE) are expected. OBJECTIVES: We aimed to compare the BCG and SPE methods for albumin determination in the serum of white rhinoceroses. METHODS: SPE and BCG albumin were measured in 82 white rhinoceros serum samples. Results were compared using Bland-Altman difference plots and Passing-Bablok regression analysis. RESULTS: BCG albumin showed a significant mean constant positive bias of 7 g/L, or 36%, which was more than the total allowable error of 15% and was clinically significant. Methods were not comparable within the inherent imprecision of each method. CONCLUSIONS: The BCG method overestimated albumin concentrations in this species compared with agarose gel SPE, and method-specific reference intervals should be used.

Poly (bromocresol green) flakes-decorated pencil graphite electrode for selective electrochemical sensing applications and pharmacokinetic studies.

A square wave voltammetric method for selective determination of meropenem (MRP) and ertapenem (ERP) was developed using pencil graphite electrode modified with poly (bromocresol green) (PGE/PBCG). The modified electrode film was characterized by scanning electron microscopy and electro-chemical impedance spectroscopy. Under the optimized conditions, the prepared electrode has good linearity over concentration range 1.0-60.0 and 0.3.0-75.0 µM for MRP and ERP, respectively. The developed method was validated according to ICH guidelines. In addition, the diffusion co-efficients of MRP and ERP were estimated to be 1.24 × 10-6 and 9.09 × 10-6 cm2 s-1, respectively using chronoamperometric technique. The developed method was highly sensitive and selective for the determination of MRP or ERP in the presence of their corresponding open beta-lactam ring degradation products. Consequently, it was successfully utilized for in-vitro and in-vivo applications in spiked and real plasma samples of healthy rabbits for their pharmacokinetic studies. Furthermore, the method was applied for the assay of the available dosage forms of both drugs.

The bias between different albumin assays may affect clinical decision-making.

Differences between laboratory assays can have important clinical implications. For creatinine assays this became apparent soon after the introduction of the Modification of Diet in Renal Disease formula and resulted in international efforts towards standardization. Albumin in blood is measured by different assays, either chromogenic using Bromocresol green (BCG) or Bromocresol purple (BCP), or by an immunoassay. Since differences between these assays have received limited attention we evaluated bias and imprecision of BCG and BCP assays in comparison to the immunoassay using blood samples from patients with membranous nephropathy and nephrotic syndrome. For the BCG assay, the mean bias was high (6.2 g/l, with a standard deviation of 2.4 g/l) compared to a bias of 0.3 g/l (standard deviation 1.5 g/l) for the BCP assay. Importantly, we questioned clinical relevance by evaluating the accuracy of the decision toward the use of prophylactic anticoagulant therapy. Notably, nephrologists may reach inappropriate treatment decisions using the BGC assay in up to 59% of patients. Thus, our study should stimulate efforts towards standardization of the albumin assays.

Non-invasive textile based colorimetric sensor for the simultaneous detection of sweat pH and lactate.

A non-invasive textile-based colorimetric sensor for the simultaneous detection of sweat pH and lactate was created by depositing of three different layers onto a cotton fabric: 1.) chitosan, 2.) sodium carboxymethyl cellulose, and 3.) indicator dye or lactate assay. This sensor was characterized using field emission scanning electron microscopy and fourier transform infrared spectroscopy. Then, this sensor was used to measure pH and lactate concentration using the same sweat sample. The sensing element for sweat pH was composed of a mixture of methyl orange and bromocresol green while a lactate enzymatic assay was chosen for the lactate sensor. The pH indicator gradually shifted from red to blue as the pH increased, whereas the purple color intensity increased with the concentration of lactate in the sweat. By comparing these colors with a standard calibration, this platform can be used to estimate the sweat pH (1-14) and the lactate level (0-25 mM). Fading of the colors of this sensor was prevented by using cetyltrimethylammonium bromide (CTAB). The flexibility of this textile based sensor allows it to be incorporated into sport apparels and accessories hence potentially enabling real-time and continuous monitoring of sweat pH and lactate. This non-invasive sensing platform might open a new avenue for personal health monitoring and medical diagnosis as well as for determining of the physiological conditions of endurance athletes.