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Abstract
Conflicting historical data for the specific refractive index increment dn/dc of human plasma/serum protein have warranted an experimental re-calibration for citrated source plasma for fractionation. Measurements by <em>Dumas</em> combustion and digital refractrometry applied the pharmacopeial protein-nitrogen ratio of 6.25. The non-protein matrix was separated by centrifugal ultrafiltration. Refractometric data agreed excellently with <em>Dumas</em>-based protein concentrations.
Highlights
We have described the refractometric calibration of albumin and of purified immunoglobulin G based on nitrogen-based protein concentration values [1]
The present work demonstrated the practical benefits of digital refractometry
The dn/dc value obtained for pooled plasma (0.0001942 mg/mL) is considerably higher than the single literature value as given at the beginning of commercial plasma fractionation [2], but practically the same as the value stated for plasma first in 1928 (0.000195 mL/mg) [10], as the experimental result determined by the permeate subtraction technique in 1923 for human serum [7], and as the value of dialyzed serum given in 1952 (0.000193 mL/mg) [12]
Summary
We have described the refractometric calibration of albumin and of purified immunoglobulin G based on nitrogen-based protein concentration values [1]. In the first 50 years of refractometry, defibrinated blood serum had been a much more common sample material than blood plasma. Analytical attention towards the latter has arisen only with the introduction of pooled plasma for transfusion [16], and the establishment of the coldethanol plasma fractionation [17]. Several theses at European hematological departments had focused on the application of refractometry on plasma and serum samples, these apparently did not find wider distribution and attention (Supplemental Material)
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