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William Henry Porter.

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tion of cupric sulphate, and add some liquor potassas; an excess of the*
potash does not interfere with the reaction. A precipitate may fall,
but it will be dissolved on shaking the tube, and the liquid will
assume a violet color. Boil the solution; no precipitate falls, but the
violet color will become deeper.

(5) Xanthoptrotein Reaction. — Add to the sample of urine some
concentrated nitric acid, and boil. Let the liquid cool, and then add
some ammonia. If the solution contains albumin, an orange color
will be produced.

15



226 ALBUMIN.

(G) Milfoil's Reaction. — Add to one drachm of urine (3.887 c.c.)ten
minims of Millon's reagent, and heat. If the sample contains albu-
min in considerable quantities, a white precipitate will fall which will

become red when heated. If only a trace of albumin be present, no
precipitate is formed, but the fluid becomes red. The red color is
produced at ordinary temperatures, but it is increased by heat.

Milfoil's Reagent is prepared by adding one drachm (3.887
grams) by weight of nitric acid (specific gravity 1.042) to one
drachm (3.887 grams) by weight of pure mercury. Dissolve the
mercury in the nitric acid at first without, and afterwards with gentle
warmth. Add to this solution twice its volume of distilled water;
let it stand for some hours, and decant the supernatant fluid from the
crystals that have formed at the bottom. Keep in a glass-stoppered
bottle.

<7) Heller's or the Nitric Acid Test.— Vouv one drachm (3.887 c.c.)
or a smaller quantity of nitric acid into a test-tube, then hold it
obliquely and drop the urine in from the bottle, or by the aid of a
medicine dropper or pipette, until an equal volume has been added
without agitation. The urine, being of less density, will naturally
float upon the top of the acid. At the line of junction of the two
liquids, all urinary samples will gi\e a brownish-color line which will
vary in intensity with the condition and amount of coloring matter
and nrea present.

This peculiar ring of color has been considered to be the result of
the action of the acid on the coloring matters of the urine. Others
hold that it is in some way influenced by the formation of the nitrate
of urea, or it may be due to this formation of urea with the property
under these circumstances of taking up the coloring matter, as is the
■case with uric acid in urine.

The only objection to this last theory is that when the crystals of
nitrate of urea are formed in abundance at this border line, as occa-
sionally happens, they can be easily taken up by a pipette, mounted
upon a slide, and examined under the microscope. They are then
iouud to be devoid of all color, and are nothing but pure white crys-
tals. After they have all been removed by the pipette, or allowed to
fall to the bottom of the tube, the color band still remains.

By others, it is held that this color band is due to the deposition of
acid urates, and it is also asserted that it is due to the formation of
hydrated uric acid. If the color band fell to the bottom with these
crystalline and solid substances, these theories might hold; but this
change appears to be the result of the action of the nitric acid upon
the coloring matter.



ALBUMIN.



227



When the color line is not very marked and no cloud forms, and
albumin is present, a decided, delicate, and perfectly vrhite band is
formed on the surface of the acid between the two solutions. If the
urine is high colored and loaded with urates, a trace of albumin may
be obscured by them, or it may lead to the belief that the whole band
is albumin, and that it may be indicative of large quantities. This diffi-
culty may be, in a measure, obviated by gently heating the solution,
when the urates will disappear and the white film of albumin become
somewhat more distinct.

Heller has advised the use of a somewhat larger dish, with slanting
edges, instead of the test-tube. The urine flowing out upon a larger
surface gives a more marked reaction.

Hoffman and Ultzmann have advised boiling the urine with an
equal bulk of officinal liquor potassa, and filtering. If still not quite
clear, add a few drops of the "magnesian fluid" (the magnesian fluid
is made by dissolving magnesium sulphate and pure ammonium chlo-
ride, each one part, in eight parts of distilled water and adding one part
of liquor ammonise), warm again, and filter. This test will detect very
■small quantities of albumin.

(8) To get rid of the two troublesome bands, Dr. A. W. Abbott, of
Minneapolis, recommends the following plan: Pour a few drops of
urine gently down the inside of a glass vessel containing acidulated
water at the boiling point. If albumin be present, a more or less
delicate film will form just at the dividing line between the fluid tested
and the clear water. This test is said to detect one-twentieth of one
per cent.

(9) The following test has been found to work very well, and to
obviate most of the difficulties experienced with Heller's method ex-
cept as modified by Hoffman and Ultzmann:

Pour one drachm (3.887 cc.) of chemically pure nitric acid into a
clean test-tube, then carefully add one-half drachm (1.942 cc.) of dis-
tilled water. Both solutions will remain perfectly clear and separate
if carefully added, but a distinct line of separation between the two
will be easily appreciated on account of the difference in density be-
tween the two solutions. Next draw up a little of the urine to be
examined in a pipette and allow it to flow down the test-tube, which
is to be held erect and not inclined. The suspected solution passes
quickly through the water, which acts as a filter, and strikes against
the acid suddenly, when a pure white film or layer of coagulated
albumin will shoot across the surface of the acid to the opposite side.
The thickness of the precipitate will depend upon the amount of
albumin present. On account of the perfect transparency of the acid



228 ALBUMIN.

and water, if the tube is held against a dark background, the faintest
film can be detected separating the two fluids.

If the urine be high-colored or loaded with urates, it should be
diluted until almost colorless. In this way the deep color line and the
cloud produced by the urates, when acted upon by the acid, that so
often obscures the albumin in Heller's test, is almost completely over-
come, and the albuminous precipitate remains sharply outlined.

This method is much more delicate than Heller's. Its exact deli-
cacy has not been determined. After Heller's and the heat and acid
tests fail to show evidence of albumin, the sample can be diluted
many times more, and a sharp reaction will follow its application.

This method is very simple, and if carefully followed is free from
error. The reason for the precipitate being so positive is the clear-
ness of the two solutions and the precipitation of all the forms of
albumin.

(10) Roberts' Acid Brine Test. — The principle of this test is the
same as when the acetic acid and sodium sulphate are used, a different
acid and salt being amployed.

A standard test solution is made by adding one ounce (31.103 c.c.)
of dilute hydrochloric acid (IT. S. P.) to fifteen ounces (460.552 c.c.)
of a saturated solution of sodium chloride.

This test is used in the same way as Heller's nitric acid. One
drachm (3.877 c.c.) of the brine is poured into a clean test-tube, and
the urine allowed to run slowly down the tube so as to prevent undue
agitation of the two fluids and an intermingling. If albumin be pres-
ent in the sample, a white film or layer of coagulated albumin will be
found, varying in thickness with the amount present in the sample.

The advantages ascribed to this test are: (a) As a rule no color line
is formed, and if any, it is slight; (b) that it can be carried with less
danger than nitric acid; (c) it does not stain the fingers; (d) it does
not precipitate the urates; (e) it consolidates all forms of albumin,
and the sugar test applied after its use is not interfered with.

On the other side, it can be said that it is no more delicate than
the heat and less so than the nitric acid tests.

(11) Picric Acid Test. — When this acid is added to normal urine,
the only change produced is a slight yellow tinge due to the color of
the acid, but no precipitate occurs, and the solution remains perfectly
transparent. A saturated aqueous or alcoholic solution may be used
as a standard test; the former is the one commonly recommended. It
may be employed as follows. Pour one drachm (3.877 c.c.) of the
standard solution into a test-tube, then add the urine in the same



ALBUMIN. 229

manner employed in Heller's acid test; a precipitate will form be-
tween the two fluids if albumin be present.

As the picric acid has a lower specific gravity than most samples of
urine, it is better to pour the urine into the test-tube first, and then
add the acid. The same result, however, is obtained in either case.

If the urine is alkaline, the sample must first be strongly acidulated
with acetic acid. In fact, a little acid had better be added in all cases.
If the urine is turbid, it should be boiled with one-fourth its volume
of liquor potassae, and then strongly acidulated and tested as before.

This test gives the same kind of a precipitate with the salts of pot-
ash as with albumin. It also precipitates all forms of albumin as well
as serum-albumin. The application of heat will cause a disappearance
of the precipitate with some of the albumins and the potassium salts.

Taken as a whole, there is some uncertainty in relation to the id-
liability of this test.

(12) Tanret's or the Potassio-mercuric Iodide Test. — The test solu-
tion is made as follows:

Iodide of Potassium, . 49.68 grains 3.219 grams.

Bichloride of Mercury, 20.83 grains 1.349 grams.

Distilled water, a sufficient quantity to make 3.21 ounces
<99,843 o.c).

It is employed as follows:

(a) Boil the urine to be tested with liquor potassae, and filter.

(b) Acidulate the filtrate with citric or acetic acid; the former is
most strongly recommended.

(c) Add a few drops of this standard solution to the acidulated
urine. If only a trace of albumin be present, a slight turbidity is all
that can be detected. When the quantity of albumin is large, a de-
cided cloud results. If it is abundant, a very decided precipitate will
be produced. The cloud resembles that which occurs when the phos-
phates are thrown down by heat. If employed in the same manner
as the nitric acid test, the two solutions will be separated by a bluish
disk of albumin, varying in thickness according to the quantity con-
tained in the sample.

When only a small quantity is suspected, pour one drachm (3.877
c.c.) of the test solution into a test-tube, and add the urine drop by
drop until an equal volume has been added. A color disk will form
"between the two solutions that will be either bluish-white, bluish-
yellow, or bluish, depending upon the amount of albumin present.
This test, like most of the preceding, precipitates the peptones,
he alkaloids, the urates when abundant, and forms a cloud when



230 ALBUMIN.

mucus is present. All, however, disappear when heat is applied.
This test requires time and some little skill in order to obviate all
the possible sources of error, consequently it cannot be considered as
practical for general use as the heat and nitric acid tests.

(13) Olivers or the Sodium Tungstate Test. — This substance, like
most of the other tests, is used after the manner of Heller's.

The standard test solution is made as follows: Prepare a saturated
solution of sodium tungstate, one part in tour of water, add a satu-
rated solution of citric acid, ten parts to six of water. Add together
equal quantities of the two, and it is ready for using. The urine or
the test solution may be poured into the test-tube first, followed by
the one remaining. It is a very sensitive test, and said to be free
from all objections. Further trial, however, may prove that it pre-
cipitates other substances, the same as the foregoing.

(14) Pavey's Test Pellets. — This test consists in using little pellets
instead of solutions. They are more easily carried. They are com-
posed of sodic ferrocyanide, and are acidulated with citric acid. One
of them is crushed and placed in the bottom of a test-tube, and a
little urine added, or the urine can be first poured into the test-tube,
and the powdered pellet dropped into the suspected solution. This
test will produce the precipitation of albumin without the applica-
tion of heat.

Oleo-resins act in the same manner.

(15) TeH Papers. — These have been recommended by Dr. Oliver,
of Harrowgate, on account of the ease of transportation and their
certainty of action. They are prepared as follows: Chemically inert
filter paper is soaked, either in double iodide of mercury, sodium
tungstate, or ferrocyanide of potassium, allowed to dry and then it is
cut into slips of suitable size. Papers charged with citric acid are
prepared in a similar manner.

If the urine be turbid, it should be boiled with liquor potassae and
filtered; if clear, this need not be done. In either case the acid papers
are added until a decided acid reaction is obtained.

Having accomplished this, one of the first-named test-papers is
held in the urine. If only a trace of albumin be present a cloud will
form around the test-paper, but if more abundant, a decided cloud
will be formed, and the coagulated albumin will fall gradually to
the bottom of the tube.

These papers are said to be very delicate, and to give evidence of
albumin when heat and nitric acid fail.

It appears as if bedside testing of urine has been very much overesti-



ALUUMIN. 231

mated. For a complete chemical and microscopic analysis is abso-
lutely necessary, if anything like accuracy in diagnosis is essential.

(16) Carbolic Acid Test. — This acid has been recommended to be
used in the same manner as Heller's nitric acid test. But the sources
of error are numerous, which renders it impracticable.

The exact nicety of a number of these tests have been given by Dr.
G-. B. Fowler, of this cit} r , as follows:

Heat and acidulation, . . . .018 of one per cent.
Acidulated brine, . . . .018

Picric acid, 008

Nitric acid, .... .008

Potassium ferrocyanide and acetic acid, .008 "
Potassio-mercuric iodide, . . .004 "

In all cases the test-tube and its contents should be viewed by re-
flected light against a black background as suggested by Dr. John P.
Munn, of this city.

Instead of using an ordinary \ test-tube, take a conical test-glass 1 .
and fill it two-thirds full of the test solution to be used. Then
add the urine very carefully to the test solution in the glass, so that
it may flow out upon this broad surface and not mingle with it. In
this way a large precipitation is likely to be the result, and its pres-
ence is more easily detected.

The tests which experience has proved to be the most satisfactory
are the heat with acidulation by acetic or nitric acid, the nitric
acid alone, with an equal volume of water as recommended in the
text, the xanthoprotein reaction, and the acid brine test.

The acid brine and nitric acid tests are both very ready methods
for estimating the amount in a volumetric way. If, for instance, the
albuminous precipitate occupies one-half of the fluid above the acid
or the brine, it maybe said to indicate fifty per cent; if only one-
fourth, twenty-five per cent. As a comparative test from day to day,
this is quite a practical method, but in reference to the actual per-
centage of albumin contained in a given specimen, it is, like all the
other tests recommended, absolutely inaccurate.

The only method by which anything like accuracy can be obtained
in estimating the quantity is by the gravimetric method. This con-
sists in the precipitation of the albumin upon a weighed filter, and then
deducting the increase in the weight of the filter from the total weight of
the urine from which the albumin was precipitated. The only source of
error in this method that cannot easily be avoided is the precipitation
of a small amount of coloring-matter and the earthy phosphates along-



232 ALBUMIN.

with the albumin. As most of the coloring matter and phosphates
are removed by thoroughly washing while on the filter, this error is
not sufficiently great to be considered. The method is of no practi-
cal value to the general practitioner from the fact that it requires a
large amount of delicate apparatus, and also takes considerable time
and skill to arrive at an accurate result.

There are a huge number of methods offered by different writers,
all of which appear to be too complicated and uncertain to be absolutely
practical. They will be named and the references given only. By
circunipolarization; the method of Bodeker; Vogel's optical method,
Lang, Ilaebler, and Bornhardt Menu's method, method of P. Liborius,
L. Girgensohn's method, W. Roberts' dilution method.

As the actual quantity lost is so small, the knowledge of the loss
being y,,^,-,, or T -J- - more or less, is of no practical value in relation to
the symptoms, prognosis, or treatment. The volumetric analysis in
the test-tube from day to day is certainly a very sure index of the
rise and fall in the quantity of albumin and gives all the information
required.

Clinical significance. — The first thing to decide is whether the
albuminuria is of the adventitious or intrinsic variety. This can be
accomplished by exclusion. If the albumin persists without any
further evidence of structural change in the kidneys it is reasonable
to suppose that it is of the adventitious form. The absence of serum-
albumin and the detection -of the peptones or some of the derived
albumins in abundance would confirm the diagnosis.

If, on the other hand, the serum-albumin predominates, with casts
and other evidences of a structural lesion of the kidneys, the albumin-
uria is of the intrinsic variety.

Prognosis. — This of necessity depends upon the kind of albuminuria.
In the adventitious, if its cause can be removed, the prognosis is good,
otherwise it may terminate in the intrinsic variety.

In inherent albuminuria, the prognosis also depends largely upon
the cause. In connection with the fevers, acute and chronic metallic
poisoning, and the acute renal lesions, recovery is possible and not in-
frequent, but in connection with chronic venous obstruction or in a
well-established renal lesion the prognosis is always bad.

Treatment. — This is exclusively confined to the removal of the
causes. In the adventitious variety, if due to the abstinence from
sodium chloride, it should be added to the diet; if due to imbibing too
much egg-albumen, the quantity taken should be diminished; if to a
faulty intestinal or hepatic digestion, this should be overcome.



ALBUMIN. 233

The treatment of the intrinsic variety has boon covered in treating
-of the various renal lesions in which it occurs.

The adventitious variety will often disappear under the influence of
the bile pill and the nerve tonic composed, of nux vomica and damnum,
see pages 26 and. 89. The diet should always be of the albuminous
type and limited to those substances most easily and completely di-
gested and assimilated.



CHAPTER IV.
GLUCOSE OR GRAPE SUGAR. TESTS FOR GLUCOSE.

Sugar. — Glucose is found iu the urine in the condition known as
glycosuria or diabetis mellitus. It also occurs as the result of poison-
ing by urari. carbonic oxide, with a sufficient dose of morphia
and amyl nitrite. It has also been met with in -disturbances of the
medulla or base of the brain, occasionally in health and sometimes
after surgical operations. A trace of glucose is said to be normally
present both in the blood and urine, but siuce the observations of
Prof. Satterthwaite, who found that both uric acid and kreatin caused
the same reaction with the glucose tests as sugar, it is quite possible
that this may not be true.

A large number of tests hare been suggested from time to time.

They are, first, Moore's or Heller's. — To one drachm (3.887 c.c.) of
urine add an equal quantity of caustic soda or potassa, and boil. If
glucose be present, the mixture will at first turn to a light jellow, then
to an amber color, and lastly to a dark orange-brown color.
PI This, however, is an uncertain test, as many other organic com-
pounds frequently found in the urine cause the same discoloration
under similar circumstances.

Second, Trommer's. — Take one drachm (3.877 c.c.) of urine and add
12 minims of a 12$ solution of cupric sulphate ( 3 i. to § i. 3.877 to
31.103 c.c.) so that a very slight green color is perceptible, then add
to the solution one and one-half drachms (5.831 c. c. ) of liquor potassae.
The previously formed precipitate of the hydrated copper oxide will
now be dissolved and the fluid will assume a rich blue tint. Now
gradually heat: before precipitation occurs, the fluid becomes opaque
and presents a yellowish-red color, but after boiling for a feAV seconds
an abundant precipitate of cuprous oxide is deposited ; if sugar be
present, the precipitate is of a brick-red color.

The same reaction will take place without the application of heat
and is then called Cappezzuole's test. In this way the reaction does not
occur for half an hour or more.

If Trommer's test is employed in the way above described, it will be



GLUCOSE OR GRAPE SUGAR. 235

found one of the most accurate, if not the most certain method for
detecting the presence of the glucose. It has been stated that this
would not reveal the presence of sugar in albuminous solutions, but

this has been found to be erroneous.

The following experiments performed by Prof. Satterthwaite, and
which, never having been published, are, by his permission, given
in detail, are interesting in showing the accuracy of this test, pro-
vided the above stipulated quantities be used as recommended by
him.

Experiment 1. — With reference to copper test for sugar in albumi-
nous solution. Took a 5$ serum-albumin solution to start with. 20
Tit (1.295 c.c.) of this 5$ solution were poured into a test-tube, and
2 TTL (0.129 c.c.) of a glucose solution added, then 4 minims (0.291
c.c.) of a 12$ sulphate of copper solution ( 3 i. to § i.) and 30 minims
(1.943 c.c.) of liquor potassae added and boiled. At first the reaction
was doubtful, the fluid turning greenish, then yellow by direct light,
but at the end of five minutes the reaction was good.

With such a large percentage of albumin, the fluid is apt to gela-
tinize and become practically solid upon boiling.

Experiment 2. — This was the same as Exp. 1, except that a 2^$ solu-
tion of albumin was used instead of the 5$. The reaction in this
case was immediately sharp and satisfactory.

Experiment 3. — Trommer's test was applied to a simple aqueous
solution, which contained only one grain of glucose to the ounce. ^To
20 minims (1.295 c.c.) of the dilute solution 4 minims (0.291 c.c.) of
a 12$ copper solution, and 30 minims (1.943 c.c.) of liquor potassaa
were added and boiled, and it gave a clear brick-red color with
a slight deposit.

Experiment 4. — Took 20 minims (1.295 c.c.) of a saturated solution
of uric acid, 4 minims (0.259 c.c.) 12$ of the copper solution, and
30 minims of liquor potassae. The brick-red copper deposit was thrown
down after boiling the solution for ten minutes.

Experiment 5. — Took 20 minims (1.295 c.c.) saturated solution of
kreatin, 4 minims (0.295 c.c.) of the 12$ copper solution, and 30
minims (1.913 c.c.) of liquor potassas. The deposit of reduced cop-
per was not thrown down until the solution had been boiled for seven
minutes, and then it was reddish-green.

Experiment 6. — Took 20 minims (1.295 c.c.) of a saturated solution
of tannic acid, 4 minims of the 12$ copper solution, and 30
minims (1.943 c.c.) of liquor potassas. A reddish deposit was thrown
down, the color being a sealing-wax red.

Experiment 7. — To a 5$ solution of serum-albumin one grain (0.064



236 GLUCOSE OR GRAPE SUGAR.

c.c.) of glucose to the ounce (31.103 c.c.) was added. Took 20
minims t 1.295 c.c.) of this saccharated and highly albuminous fluid,
and added 4 minims (0.295 c.c.) of the 12^' solution of copper,
and 30 minims (1.943 c.c.) of liquor potass», and boiled. The char-
acteristic color reaction of Trommer's test occurred, but no precipitate
fell.

Experiment 8.— Took 20 minims (1.295 c.c.) of the same saccha-
rated albuminous fluid and applied Fehling's test. Reaction ob-
tained, but less perfectly than by the use of Trommer's.



Online LibraryWilliam Henry PorterA practical treatise on renal diseases and urinary analysis → online text (page 22 of 32)