Abstract
This report describes a rapid and inexpensive colorimetric method based on biuret assay for total serum protein analysis using wax printed microzone plates and a desktop scanner. The colorimetric assay was performed in 96-microzone plates fabricated on polyester films through a wax printing protocol. The image scanning mode and the concentration of the biuret reagents were optimized to achieve the highest detectability levels. The best analytical response was obtained using the slide scanning mode as well as 4% CuSO4 and 150 mmol L-1 potassium sodium tartrate solutions. The proposed method offered linear relationship for the bovine serum albumin concentration range between 25 and 200 µmol L-1 with a limit of detection of 9.7 µmol L-1. The colorimetric approach revealed satisfactory precision (relative standard deviation values between 0.6 and 4.4%). The bioanalytical feasibility was demonstrated through the total protein analysis in serum samples collected from dogs previously diagnosed with kidney disease. The total serum protein concentrations varied from 960 ± 19 to 1661 ± 38 µmol L-1 and the achieved values revealed good accordance with those obtained through a reference method. Based on the reported results, wax printed microzone plates have demonstrated simplicity and potential to be adopted as alternative platforms for bioassays.
Highlights
Total protein analysis represents an useful parameter for routine clinical screening.[1]
We describe a rapid and inexpensive colorimetric assay based on biuret method to determine the concentration levels of total proteins in serum samples using microzone plates wax printed on polyester films
The fabrication of microzone plates on polyester films through wax printing technology has emerged as a good alternative platform for total serum protein analysis based on a biuret colorimetric assay
Summary
Total protein analysis represents an useful parameter for routine clinical screening.[1]. Printed microzones have revealed good performance for bioanalytical applications including protein analysis in different biological fluids.[27,28,29] In this current study, we describe a rapid and inexpensive colorimetric assay based on biuret method to determine the concentration levels of total proteins in serum samples using microzone plates wax printed on polyester films. The total protein assay was performed according to the procedure described by Gornall et al.[15] Briefly, 100‐μL aliquots of 4% (m/v) copper(II) sulfate pentahydrate, 3.0 mol L−1 sodium hydroxide and 0.15 mol L−1 potassium sodium tartrate solutions were mixed to prepare the chromogenic reagent, known as biuret reagent. The readout was carried out at 546 nm and the results were used to compare the data obtained on wax printed microplates
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