New colorimetric method with bromocresol purple for estimating the redox state of human serum albumin

Abstract

Oxidative damage results in protein modification and is observed in many diseases, such as heart failure and renal insufficiency. Human serum albumin is an index of oxidative change and is conventionally measured using high-performance liquid chromatography (HPLC). Although this method is more sensitive than the colorimetric method, it is time-consuming for clinical practice and the sera must be stored at –80°C before analysis. To overcome these limitations, in the present study we developed a new reagent for a more rapid and convenient quantification of oxidative stress, involving determination of the ratio of human nonmercaptalbumin to total albumin using a colorimetric method with bromocresol purple. The clinical utility of the developed reagent was confirmed by demonstrating the consistently higher oxidative stress levels in dialysis patients than in healthy control subjects, matching the results of the conventional HPLC method. This novel approach could be a valuable tool for immediate estimation of the state of oxidative stress during the course of disease and treatment, and could aid clinical treatment decisions.

Serum Albumin Redox States: More Than Oxidative Stress Biomarker

“…Muramoto et al developed a modified BCP method [ 55 ], where the difference in binding affinities was resolved through facilitating conversion of reduced ALB to oxidized ALB by adding sodium dodecyl sulfate and 5,5′-dithiobis(2-nitrobenzoic acid) to the BCP reagent. A recent study by Yoshihiro et al is notable as they applied the modified BCP method to the determination of serum ALB redox state [ 56 ]. Namely, serum samples are reacted with BCP both in the presence and absence of sodium dodecyl sulfate and 5,5′-dithiobis(2-nitrobenzoic acid) under a condition where BCP sensitively reacts with oxidized ALB.…”

Redox state as assessed using the measurement of human non-mercaptalbumin in embryo culture media is associated with successful embryo development in human <i>in vitro</i> fertilization

“…The total measurement time was 15 min per sample. (8,9) Statistical analysis. Continuous variables were expressed as the mean and SD, while categorical variables were expressed as the percentage (%).…”

“…(7) Recently, novel colorimetric method with bromocresol purple was also reported. (8) To this end, we recently reported that HNA is a useful marker for evaluating the redox state in humans, (9) and we established a reliable and less timeconsuming method for measuring HNA. (10)(11)(12) At present, many studies on oxidative stress have sought its potential roles in the pathogenesis of various diseases.…”

A simple colorimetric assay to determine the concentration and proportion of human mercaptalbumin

“…A handful of studies have explored alternative approaches using simple dye-based assays [ 3 , 23 , 24 ]. These can be adapted for high-throughput formats, such as multi-well plate readers and clinical chemistry analyzers.…”

“…These can be adapted for high-throughput formats, such as multi-well plate readers and clinical chemistry analyzers. For example, Yoshihiro and co-workers developed a simple colorimetric assay to measure the percentage of HNA using bromocresol purple (BCP) dye [ 24 ]. This assay takes advantage of the fact that the BCP dye, traditionally used to measure total serum albumin concentrations, shows greater colorimetric absorbance when it reacts with HNA than with HMA [ 25 ].…”