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Copyright 1978, Institute for Clinical Science
ABSTRACT
The biuret reaction for proteins provides a sim ple and precise m ethod for
m easuring serum proteins; Beers law is obeyed to at least 10 g per dl.
Several stable biuret reagents are available. H em oglobin is the only im por
tant cause of interference w hich cannot be m inim ized by use of a sample
blank. The m echanism of the b iu ret reaction is described and attention is
drawn to the heterogeneity of the serum proteins and to the use of a cer
tified album in standard.
Introduction
106
107
M E A SU R E M E N T O F T O T A L SER U M P R O T E IN S BY B IU R E T M E T H O D
TABLE I
Biuret Values of Some Serum Proteins18
C o n c e n tr a ti o n i n
Serum (m g /d l)
Albumin
3,500 - 4,500
ai-Antitrypsin
210 500
y-Globulins
900 - 1,500
Haptoglobin
30 190
Hemopexin
80 100
a2-Macroglobulin
220 380
"Orosomucoid"
75 100
Prealbumin
28 35
Transferrin
200 320
B i u r e t V a lu e
(A lb u m in - 1 0 0 )
100
86
110
81
80
92
62
96
95
108
LU BRAN
109
M E A SU R E M E N T O F T O T A L SER U M P R O T E IN S BY B IU R E T M E T H O D
TABLE I I
Composition of Some Biuret Reagents (g/L)
R eagent
CuSO4>5H20
NaOH
1.67
1.69
117
43
Kingsley9
Mehl11
Weichselbaum2 0
Gornall6
Reinhold*1*
Doumas4
*NaKC^H^Og*4H20
15
1.5
3
3
8
30
8
24
" T a r tr a te " *
Kl
O th e r
80 ml
ethylene
glycol
45
6
9
9
5
(1) +
5
5
tlnclusion of K1 is optional.
110
LU BRA N
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
Conclusions
The biuret m ethod has stood the test of
tim e as a convenient and precise m ethod
for m easuring total serum proteins. Con
c e n tra tio n o f p ro te in s s h o u ld b e ex
pressed in term s of album in, either hum an
or bovine. The sensitivity of the m ethod is
inadequate for fluids of low protein con
centration and it may not be suitable (un
less modified) for tissue proteins. T he cor
rectly prepared reagent has a long shelf
life at room tem perature.
Beferences
W.: Colorimetric estimation of
serum albumin and globulin in urine, ascitic
fluid and blood serum . M unch, med.
Wochenschr. 64:241-245, 1917.
2. A u t e n r i e t h , W. and M in k , F.: Colorimetric
determinations: the quantitative determination
of albumin in urine. Munch, med. Wochenschr.
62:1417-1420, 1915.
3. B o d e , C., G o e b e l l , H., and S t a h l e r , E.: The
'
elimination of errors caused by turbidity in the
1.
14.
15.
16.
17.
18.
19.
A u te n rie th ,
20.
21.