Background: Bromocresol green method (BCG) is routinely used in veterinary laboratories to measure albumin. A good agreement between BCG and electrophoresis (SPE) in healthy domestic animals is ...reported. However, in human medicine, at low albumin and high globulin concentrations, BCG also binds to α- and β-globulins fractions. Surprisingly, there is no recent literature evaluating the BCG method in diseased domestic animals. Objective: The purpose of the present study was therefore to compare the serum albumin values obtained by SPE and BCG in canine and equine patients. Methods: Albumin measured by BCG and SPE was compared in the serum from 30 canine and 30 equine patients. Results: Good correlation was found (Spearman´s rho rs = 0.91 in dogs and 0.84 in horses). However, Altman and Bland analysis of results showed a positive bias of BCG with SPE (3.41 g/l in horses and 0.41 g/l in dogs) that increased when only hypoalbuminemic patients were considered (4.17 g/l in horses and 1.31 g/l in dogs). Conclusion: Results confirm that, despite good correlation, BCG method tends to overestimate albumin, especially in hypoalbuminemic patients. This is particularly evident in horses.
Columbine serum total protein (TP) and albumin concentrations were determined using the biuret method and the bromocresol green dye binding (BCG) method or serum protein electrophoresis on cellulose ...acetate membranes (SPE). Results obtained using human and pigeon standards were compared.
When pigeon albumin was used as a standard. TP values were consistently higher compared with values obtained using human protein as a standard. However, there was a high correlation between the results obtained with the two standards.
The correlation between the BCG method and SPE for serum albumin determination was poor, irrespective of the standard used. The method cannot be recommended for pigeon blood.
For avian clinical practice it is advised to establish TP concentration using the biuret method and a human standard and to calculate albumin concentration from the results of TP and SPE.
A sensitive and selective method based on the formation of ternary complexes of berberine, quinidine and a divalent anion of dibasic acid dyes (A2-) such as Bromophenol Blue, Bromochlorophenol Blue ...and Bromocresol Green is described. Berberine was extracted in large amounts into 1, 2-dichloroethane with A2-; only in the presence of quinidine in a neutral aqueous media. Berberine could react in 1, 2-dichloroethane with A2- to give a blue product. In addition, the extractability of Berberine in a neutral aqueous media could be enhanced by the formation of bulky ternary complexes (berberine+-A2--quinidine+); the selectivity was improved because most of the amines and alkaloids did not react with A2- in the above-mentioned media. The blue complexes can be used for a sensitive and selective spectrophotometric determination of berberine in crude drugs.
Plasma albumin was determined in human, rat and dog samples by bromocresol green and bromocresol purple dye-binding methods. The bromocresol purple method produced significantly lower values in dog ...and rat plasma samples, although the use of homologous standards reduced the differences between the two methods. These observations reflect differing affinities of the two dyes for albumins of different species.
A combination of known biochemical techniques (binding of bromocresol green to human albumin, nondenaturing electrophoresis and electroelution) have been utilized in a novel purification of human ...albumin. This paper reports (a) purification of intact, nondenatured human albumin, (b) simultaneous purification of albumin and hemoglobin from a human plasma and red blood cell lysate mixture, (c) development of a purification method for the two most commonly employed protein biomarkers in human environmental epidemiology and (d) demonstration of the general technique of protein purification of colored ligand-protein complexes by nondenaturing electrophoresis. The noncovalent binding between a protein (albumin) and a colored ligand (bromocresol green) is the biochemical characteristic exploited in this novel purification scheme. This general purification method may be useful for other colored ligand or fluorescent ligand binding proteins. For small-scale electrophoresis, the amount of protein isolated, percentage yield and protein purity (estimated by SDS-PAGE) were 270 micrograms, 80% yield and > 99% purity for albumin and 217 micrograms, 54% yield and > 99% purity for hemoglobin, respectively. For large-scale electrophoresis the comparable data was 38.3 mg, 57% yield and 98% purity for albumin and 17.2 mg, 57% yield and 99% purity for hemoglobin, respectively.
To determine micro-amounts of proteins with low molecular diffusivity in biological materials, an FIA type capillary stream sensor (CSS) was developed by the dye association method using Bromocresol ...Green with high sensitivity and little interference. The CSS device was based on Korenaga-Stewart's S4-FIA system which was different from the traditional FIA systems because of the use of very slow flow rates for reagent and carrier streams. A Furue Science, Model J-P-VW, double-syringe pump was used, and the optimal conditions were examined with bovine serum albumin (BSA) as a standard at a slow flow rate region. As the result, less than 0.05 ml/min of flow rate was acceptable in case of using small-bore and short reaction tube (0.5 mm bore and 200 mm length). The method had analysis rate of 78 samples/h with relatively long residence time, but straight calibration graph was obtained for 8 μl of injection volume accompanied with high reliability and reproducibility. The detection limit and RSD were 0.3 μg and 0.2% for BSA, respectively. This research gave a useful CSS device technology at a viewpoint of analytical instrumentation hydrodynamics for small consumption of valuable reagent solutions and biological material samples.
To determine micro-amounts of proteins with low molecular diffusivity in biological materials, an FIA type capillary stream sensor (CSS) was developed by the dye association method using Bromocresol ...Green with high sensitivity and little interference. The CSS device was based on Korenaga-Stewart's S4-FIA system which was different from the traditional FIA systems because of the use of very slow flow rates for reagent and carrier streams. A Furue Science, Model J-P-VW, double-syringe pump was used, and the optimal conditions were examined with bovine serum albumin (BSA) as a standard at a slow flow rate region. As the result, less than 0.05 ml/min of flow rate was acceptable in case of using small-bore and short reaction tube (0.5 mm bore and 200 mm length). The method had analysis rate of 78 samples/h with relatively long residence time, but straight calibration graph was obtained for 8 μl of injection volume accompanied with high reliability and reproducibility. The detection limit and RSD were 0.3 μg and 0.2% for BSA, respectively. This research gave a useful CSS device technology at a viewpoint of analytical instrumentation hydrodynamics for small consumption of valuable reagent solutions and biological material samples.
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