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Serum albumin assay with bromcresol purple dye.

Serum albumin was assayed with bromcresol purple using routine clinical chemical photometric analysers. Bromcresol purple and bromcresol green dyes were compared, and albumin assays using these dyes were compared to those made by radial immunodiffusion and rocket immunoelectrophoresis. Bromcresol purple seems to be insensitive to all serum proteins except albumin, and this dye is correlated linearly with radial immunodiffusion and with rocket electrophoresis.

Bromcresol Green↗

Reduction of reaction differences between human mercaptalbumin and human nonmercaptalbumin measured by the bromcresol purple method.

Although the bromcresol purple (BCP) method provides high specificity in measurements of serum albumin concentrations, we discovered a reaction difference between the values for human mercaptalbumin (HMA) and human nonmercaptalbumin (HNA) measured by the BCP. We found that the color intensity of HMA with BCP present in the reduced form in the albumin of fresh serum, was lower than for HNA. While maintaining specificity for albumin, we reduced the reaction difference between HMA and HNA, with the addition of sodium dodecylsulfate and 5,5'-dithiobis(2-nitrobenzoic acid) to the BCP reagent. The mean albumin concentration by the BCP procedure for 63 fresh sera and the 63 sera stored for 2 days at room temperature were 35.6 g/L and 38.1 g/L, respectively. Those by the modified BCP procedure were 39.8 g/L and 39.9 g/L, respectively. The difference in measured values between the fresh and stored sera which is believed to be caused by the conversion of HMA to HNA during the storage of sera was not observed in the case of the modified BCP procedure. Our modified BCP method is effective in eliminating uncertainty of the albumin concentration assigned to assay calibrators for the conventional BCP method.

Absorption↗

New automated dye-binding method for serum albumin determination with bromcresol purple.

We describe a new automated dye-binding method for serum albumin determination with bromcresol purple (BCP) that has several advantages over an existing bromcresol green (BCG) method. The continuous-flow method is sensitive, linear, and precise, with negligible sample interaction at an analytical rate of 60 samples per hour. Unlike BCG, BCP did not react with an albumin-free serum globulin preparation or pure human transferrin solutions. Reaction with serum was instantaneous; in contrast, BCG exhibits a slow nonspecific reaction with some specimens. The specificity of BCP was demonstrated by good agreement with results of "rocket" immunoelectrophoresis (EIA) where y(BCP) = 0.95X(EIA) + 1.72. The BCG method overestimated serum albumin concentration where y(BCG) = 1.01X(EIA) + 6.77. Precipitation, which affects the BCG method, was not observed with BCP. Blank corrections were negligible, salicylate did not interfere, and bilirubin affected the method only if present in very high concentration. The method offers a solution to the poor accuracy of existing BCG methods while retaining many of their desirable features.

Autoanalysis↗

Evaluation of the clinical applicability of bromcresol purple method for measuring serum albumin.

OBJECTIVE: To evaluate the clinical applicability of bromcresol purple (BCP) method in serum albumin measurement, and to investigate the possibility to render the results thus obtained in agreement with those by traditional bromcresol green (BCG) method. METHODS: The assessment of BCP method was conducted in comparison with BCG method by measuring their linearity ranges, coefficients of variation (CV) in batch and between batches, recovery rates, and resistance to interference by certain normal clinical factors. RESULTS: The linearity of BCP method was 10 to 80 g/L, CV was 0.95% in batch and 3.03% between batches, with an average recovery rate of 99.4%, showing no obvious interference by 252 micromol/L bilirubin, 4% Trilitid, or by 4 g/L hemoglobin separately. Close correlation was observed between the measurement results by the 2 methods (y=1.08x+5.98). The adult reference range was 34-49 g/L by BCP method, which was 6 g/L lower, on average, than that by traditional BCG method. CONCLUSION: BCP method is more specific than BCG method in serum albumin measurement, and the results of the former can be easily and accurately converted so as to be comparable with the results by the latter method.

Bromcresol Green↗

Underestimation of serum albumin by the bromcresol purple method and a major endogenous ligand in uremia.

In hemodialyzed patients, the serum albumin concentration determined by the bromcresol purple (BCP) method was lower than that determined by an immunological method. The degree of underestimation appeared to be well correlated to the serum concentration of 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF), a major endogenous ligand substance present in uremic serum. CMPF inhibited in vitro the binding of BCP to serum protein and human serum albumin. Our results suggest that CMPF is a major interferent in the underestimation of the serum albumin concentration by the BCP method in uremia.

Adult↗

Bromcresol purple dye-binding methods underestimate albumin that is carrying covalently bound bilirubin.

We used automated and manual versions of two commonly used dye-binding methods for albumin to quantify albumin that had been modified by covalent addition of bilirubin. Dye-binding methods based on reaction with bromcresol purple underestimated the modified albumin by as much as 29%. Dye-binding methods based on reaction with bromcresol green were unaffected by the presence of covalently attached bilirubin. Neither method was affected by the presence of noncovalently bound bilirubin.

Bilirubin↗

A portable fiber-optic pesticide biosensor based on immobilized cholinesterase and sol-gel entrapped bromcresol purple for in-field use.

A fiber-optic biosensor for the detection and determination of the pesticides carbaryl and dichlorvos was developed. The sensing bioactive material was a three-layer sandwich. The enzyme cholinesterase was immobilized on the outer layer, consisting of hydrophilic modified polyvinylidenefluoride membrane. The membrane was in contact with an intermediate sol-gel layer that incorporated bromcresol purple, deposited on an inner glass disk. The sensor operated in a static mode at room temperature and the rate of the inhibited reaction served as an analytical signal. Calibration curves were obtained for carbaryl and dichlorvos, with useful concentration ranges 0.11-8.0 mg l(-1) for carbaryl and 5.0-30 microg l(-1) for dichlorvos. The respective detection limits were 108 microg l(-1) and 5.2 microg l(-1). The method reproducibility was in the order of +/-3-5%. The method was successfully applied to the detection and determination of these pesticides in real water samples, without sample preparation steps. Recovery experiments were made and the accuracy of the method was 94.9%. No enzyme regeneration steps were applied and the sensor lifetime was 3 weeks (30% activity reduction). The bioactive mini sandwich can easily be replaced by simply unscrewing the terminal holding ring of the probe and placing a new sandwich, before the sensor is ready for use.

Biosensing Techniques↗

Bromcresol purple method for serum albumin gives falsely low values in patients with renal insufficiency.

In patients with renal insufficiency, the BCP method underestimated the serum albumin. In non-renal patients, the BCP method gave albumin results in good agreement with the immunological method. The underestimation did not appear to be related to the degree of renal impairment. The interferent appears to act by preventing the binding of BCP to albumin. The BCG method is much less susceptible to this interference.

Bromcresol Purple↗

Conversion between bromcresol green- and bromcresol purple-measured albumin in renal disease.

BACKGROUND: Albumin measured by a bromcresol purple dye-binding assay (Alb(BCP)) agrees more closely with the gold standard of immunonephelometry than does bromcresol green (Alb(BCG)) measurement. Both tests are in current clinical use. A method for converting between the two would be useful. METHODS: We measured albumin by bromcresol green and bromcresol purple in 535 patients, 155 of whom had renal disease. We randomly divided data from the patients with renal disease into two equal-sized sets, and used one set to derive, and the remaining set to validate, a regression equation relating the two values. RESULTS: The relationship Alb(BCG)=5.5+Alb(BCP) performed very well in both the renal patient validation set and in the data from 380 unselected in-patients and out-patients. Intraclass correlations for agreement between measured Alb(BCG) and predicted Alb(BCG) was 0.98 in both analyses. CONCLUSIONS: The ability to convert between these measurements will be of use in clinical situations where the absolute value of the serum albumin is important, when data from laboratories using different methodologies must be combined, and in the application of the Modification of Diet in Renal Disease formula to estimate glomerular filtration rate in patients whose albumin has been measured by bromcresol purple.

Bromcresol Green↗

Effects of serum-isolated vs synthetic bilirubin-albumin complexes on dye-binding methods for estimating serum albumin.

Bush and Reed reported (Clin Chem 1987;33:821-3) that the reaction of albumin with bromcresol purple but not with bromcresol green underestimated the concentration of albumin in synthetically obtained bilirubin-albumin (Bd) by 29%. Their unproven assumption was that chemically synthesized Bd behaved in a manner indistinguishable from the natural Bd in icteric serum. Here we verify that Bd, whether synthetically obtained or isolated from serum, causes an underestimation of albumin in the bromcresol purple but not in the bromcresol green method. The molar ratio of Bd from either source to underestimated albumin approximates 1.0, suggesting that one molecule of Bd would react equivalently to a molecule of albumin in the bromcresol purple method. This underestimation might falsely suggest hypoalbuminemia in patients with increased serum Bd.

Adult↗