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QUANTITATIVE ESTIMATION OF PROTEID IN URINE. 793
in the spinal marrow. It gives a precipitate on heating to 40-60° C,
which on further heating to boiling dissolves again more or less completely,
depending upon the reaction and upon the amount of salt present. In
salt-free solution the precipitate is not dissolved, on heating to boiling,
at least not always. It does not separate on dialysis, but can be pre-
cipitated from the urine by double the volume of a saturated ammonium-
sulphate solution or by alcohol. It has also been obtained as crystals
(Grutterink and de Graaff, Magnus-Levy 2
). This body shows a
varying behavior in the different cases in which it has been found and its
nature has not been explained. From the investigations of the above-
mentioned and other experimenters (Moitessier, Abderhalden and
Rostoski) we can draw the conclusion that this proteid is similar to the
proteoses in several reactions, but that nevertheless it stands close to the
genuine protein bodies. It also yields primary as well as secondary
proteoses on peptic digestion (Grutterink and de Graaff), and yields
the same hydrolytic cleavage products as the other proteins (Abderhalden
and Rostoski).
Quantitative Estimation of Proteid in Urine. Of all the methods pro-
posed thus far, the coagulation method (boiling with the addition of
acetic acid) when performed with sufficient care gives the best results.
The average error need never amount to more than 0.01 per cent, and it
is generallv smaller. With this method it is best to first find how much
acetic acid must be added to a small portion of the urine, which has been
previously heated on the water-bath, to completely separate the pro-
teid so that the filtrate will not respond to Heller’s test. Then coagulate
20-50-100 cc. of the urine. Pour the urine into a beaker and heat on
the water-bath, add the required quantity of acetic acid slowly, stirring
constantly, and heat at the same time, Filter while warm, wash first
with water, then with alcohol and ether, dry and weigh, incinerate and
weigh again. In exact determinations the filtrate must not give Hel-
ler’s test.
The separate estimation of globulins and albumins is done by carefully
neutralizing the urine and precipitating with MgS04 added to saturation (Hammar-
stex), or simply by adding an equal volume of a saturated neutral solution of
ammonium sulphate (HoVmeister and Pohl 2
). The precipitate consisting of
globulin is thoroughly washed with a saturated magnesium-sulphate or half-
saturated ammonium-sulphate solution, dried continuously at 110° C, boiled
with water, extracted with alcohol and ether, then dried, weighed, incinerated,
and weighed again. The quantity of albumin is calculated as the difference
between the quantity of globulin and the total proteids.
Approximate Estimation of Proteid in Urine. Of the methods suggested for
this purpose none has been more extensively employed than Esbach’s.
’Magnus-Levy, Zeitschr. f. physiol. Chem., 30 (literature); Grutterink and de
Graaff, ibid., 34 and 36; Moitessier, Compt. rend. soc. biolog., 57; Ville and Derrien,
ibid., 62; Abderhalden and Rostoski, Zeitschr. f. physiol Chem., 46; see also Hopkins
and Savory, Journ. of Physiol., 42.
2
Hammarsten, Pfliiger’s Arch., 17; Hofmeister and Pohl, Arch. f. exp. Path. u.
Pharm., 20.
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