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in two test tubes. To the first tube add 3 drops of 1-1,000
solution of potassium permanganate; the other tube use
as a control. A distinct canary yellow indicates a positive

Iodine Test for Bile

To 10 c.c. of urine in a test tube add 2 c.c. of tincture
of iodine (dilute i-io in alcohol). The presence of bili-
rubin is indicated by a distinct emerald green ring at the
point of contact.

Urobilin Test

Fill a test tube with 12 c.c. of urine, add i drop of con-
centrated hydrochloric acid, shake gently, and allow to
separate by standing. Pour off the supernatant fluid and
add three times its volume of alcohol. To this alcoholic
extract add i drop of a $% solution of zinc chloride and
i drop of ammonium hydroxide. Zinc hydrate will be
precipitated and should be filtered off. A green fluores-
cence indicates the presence of urobilin.

Urobilin is an abnormal product and is increased in
carcinoma, appendicitis, Addison's disease, acute infec-
tious diseases, and pancreatic disease.

Urine 37


To 10 c.c. of urine add 2 c.c. of H 2 SO 4 . A brownish
red color indicates the presence of urophsein.


Add 10 c.c. of HC1 to 10 c.c. of urine and heat the mix-
ture. The presence of urochrome is indicated by a bright
red color.


Place a drop of urine on a slide, add a drop of nitric
acid, and partially evaporate by warming gently. Crys-
tals of urea nitrate will show if urea is present.

Weyl's Test for Creatinine

To 5 c.c. of urine add 10 drops of sodium nitro-
prusside (saturated solution). Mix and add 10 drops of
potassium hydroxide (20%). A ruby-red color results
which soon turns yellow.

To the yellow solution obtained in Weyl's test above
add an excess of acetic acid and apply heat. A green
color results and is in turn displaced by a blue color.
If creatinine is present, a precipitate of Prussian blue will

Uric Acid

To 30 c.c. of urine add 2 c.c. HC1 ; let it stand 24 hours.
Crystals of uric acid, if present, will separate out.

Murexid Test

Place 2 c.c. of urine in a porcelain dish, add I drop of
nitric acid, and evaporate to dryness. Cool and add I
drop of ammonia. A blue or violet color indicates the
presence of uric acid or urates.


Clinical Laboratory Technic

Urinary Sediments

The urinary sediment is^tjie deposit which is found in
the urine after standing. This' sediment is classified as




i. Triple Phosphate. 2. Calcium Oxalate. 3. Calcium Car-
bonate. 4. Uric Acid. 5. (a) Ammonium Urate ; (b) Sodium
Urate. 6. Leucin. 7. Cystin. 8. Tyrosin. 9. Cholesterin.

non-organized, or chemic, and organized, or anatomic

The sediment examination is useful in showing the
presence of abnormal elements excreted by the kidneys.

When the concentrated sediment has been obtained,
either by centrifugalization or sedimentation, a drop is
taken from the tube with a pipette. Place the index finger
on the upper opening of the pipette and insert it into the
tube, carrying it down into the fluid to the sediment ; then

Urine 39

release the pressure of the finger and allow a few drops
to enter the pipette, maintaining a firm pressure of the
finger on the upper end while withdrawing the pipette.
Place a drop on a slide and examine with a low power
lens and subdued light, then with high power and slightly
more light.

The non-organized sediments are usually crystalline,
although a few are amorphous. The crystalline deposits
are important only when found in freshly voided urine ;
they may be precipitated in any urine which stands and
undergoes fermentation.

The organic sediments consist of various cells, casts,
yeast fungi, spermatozoa, and bacteria.

Non-organized Sediments Occurring in Normal

URIC ACID CRYSTALS : Rhombic prisms, rosettes, and
hexagonal plates, usually yellow may be colorless. Solu-
ble when heated with NaOH.

CALCIUM OXALATE CRYSTALS: Colorless, envelope and
dumb-bell shaped. They are derived from various foods.

found in alkaline urine, sometimes in slightly acid urine.
It crystallizes in the form of granules, spicules, and

phates " ; these occur in sediment in two forms, the pris-
matic form of crystals and the amorphous, feathery type.
In faintly acid urine they have the coffin-lid appearance,
but in alkaline urine they take a variety of shapes. These
are pathological when found in fresh urine.

AMORPHOUS PHOSPHATES of Ca and Mg are commonly

40 Clinical Laboratory Technic

found in alkaline urine. They appear as bulky, opaque,
white deposits, and this whitish, flocculent deposit is
precipitated by heat and dissolved by acetic acid. These
earthy phosphates are the only salts of phosphoric acid
that are found in the urinary sediment.

AMMONIUM URATES: This salt of uric acid is formed
during ammoniacal fermentation of the urine, and is only
abnormal if the urine is fresh ; any urine upon standing
for several hours is apt to deposit crystals of uric acid.
Acid ammonium urate occurs as yellowish red or dark
brown spherical bodies, radially striated and studded with
fine, prismatic spicules "thorn apple crystals."

AMORPHOUS URATES are of little importance. They
have a pinkish or brick-dust appearance, due to uroeryth-
rin, a coloring matter found in the urine of acute rheu-
matism. They also occur as rods and spikes ; are dissolved
by NaOH, but not by acetic acid.

Non-organized Sediments Occurring in Abnormal

CYSTIN CRYSTALS : Colorless hexagonal plates or quad-
rilateral prisms ; soluble in HC1 and ammonia, insoluble
in acetic acid, water, alcohol, and ether.

LEUCIN CRYSTALS are highly refractive, yellow spheres,
resembling fat globules ; insoluble in ether.

TYROSIN CRYSTALS appear as fine, radiating needles.
They are colorless. Tyrosin and leucin crystals are found
together in nephritis, gout, cirrhosis, and carcinoma of the

granules, needles, and rhombic plates. They have no
clinical significance.

Uric Acid Crystals. From Purdy after Peyer

Ammonium Urates, showing Spherules and Thorn-apple-
shaped Crystals. From Ogden after Peyer.

Urine 41

CRYSTALS OF URIC ACID may be suggestive of gravel
or calculus if blood and other symptoms are present.

The presence of an excessive number of calcium oxalate
crystals may signify oxaluria.

formed in fresh ammoniacal urine, are suggestive of in-
fection of the urinary passages and calculus.

CHOLESTERIN CRYSTALS are large, regular, and irregu-
lar plates. These crystals are found in the sediment in
cystitis, nephritis, pyelitis, and chyluria.

Organized Sediment

Various forms of epithelia are found in nearly all
urines. They are the formed elements coming from the
different parts of the genito-urinary tract, and are sup-
plied by the progressive desquamation of the mucous
surfaces. An increase in number indicates inflammatory
process in the part producing them. They are more or less
granular and possess one or more nuclei. The size, shape,
and condition of their protoplasm and nuclei should be
carefully observed. If the sediment is of such volume
and density as to obscure blood cells, casts, and pus, dilute
with a drop of clear urine.

Large, round, epithelial cells, with dense refractive
protoplasm, come from the neck of the bladder; other
large, round cells may come from the membranous and
prostatic urethra.

Epithelium from the Urinary Tract

Small, round, epithelial cells may come from the renal
tubules, the pelvis of the kidney, or the urethra. They
are mononuclear and slightly larger than the leucocytes.

42 Clinical Laboratory Technic

Large, polygonal, squamous cells come from the bladder,
but they may also enter the urine from the prepuce of the
male or vulva of the vagina.







I. Squamous epithelium from the bladder. 2. Cells from the
neck of the bladder. 3. Epithelium from the prostatic urethra.
4. Cells from the seminal passages. 5. Red blood cells. 6. Leu-
cocytes. 7. Scaly epithelium. 8. Compound granule cell. 9. Ure-
thral cells. 10. Pelvic cells.

Cylindrical and caudate cells may come from the neck
of the bladder or the pelvis of the kidney.

Urine 43

Leucocytes are present in small numbers in normal
urine. An increase in number denotes an inflammatory
condition of some portion of the urinary tract. They are
recognized by their polymorphic nuclei. In acid urine
they are often shrunken, and in alkaline urine they are
often degenerated and swollen.


The difference between a pus cell and a leucocyte is
only quantitative. When many leucocytes are present, the
term pus is used ; when few, they are called leucocytes.

The origin of pus is important. Pus from the kidney
is usually mixed with casts and small, round cells ; if with
squamous cells, it is probably from the bladder. A cathe-
terized specimen is necessary in order to rule out the
vagina as a source.

Red blood cells are pale, non-granular, non-nucleated
discs. In concentrated urine they may present a crenated
appearance, and in alkaline urine they are often destroyed,
forming masses of brown granules. Normal blood indi-
cates a hemorrhage in the pelvis or bladder; abnormal
blood signifies that it is higher up in the tubules.


Casts are cylindrical formations which originate in the
uriniferous tubules ; they generally indicate some kidney
disorder. They have uniform marginal outlines with well-
defined borders, and show the molded effect of the kidney

HYALINE CASTS : Transparent, their shape and char-
acter may be determined by staining with iodine solution,
gentian violet, or fuchsin. Generally albumin is present
with this type of cast.


Clinical Laboratory Technic

GRANULAR CAST: The basic substance of this cast is
hyaline, and the granules consist of albumin, degenerated
epithelial cells, erythrocytes, and leucocytes. They are

Hyaline .




\ _


Mucous shreds.


CASTS Gibson

finely granular and coarsely granular, according to the
nature and size of the granules.

EPITHELIAL CASTS: Hyaline casts covered with epi-
thelial cells from the lining of the uriniferous tubules.
These are particularly abundant in acute nephritis.

BLOOD CASTS : Hyaline basis, covered with erythrocytes.
They are characteristic of acute diffuse nephritis and

Urine 45

acute congestion of the kidney, and they denote renal

FATTY CASTS : Fat globules and fatty acid crystals are
deposited upon hyaline or granular casts. They indicate
fatty degeneration of the kidney, and are found in sub-
acute and chronic inflammation of the kidney.

WAXY CASTS: Similar to the hyaline form, but are
somewhat larger and appear more solid, having a sharper
outline and a light yellow color.

CYLINDROIDS: Flat in structure, with smaller diameter
than casts, having branching ends. These "false casts"
may become coated with granules and appear granular
in structure.

Mucous SHREDS: These shreds of mucus are long,
wavy, transparent bodies, which are much thinner than
casts or cylindroids.

Tubercle Bacilli in Urine. (J. A. M. A., March, 1915)
Acidify the urine with 30% acetic acid, 2% of its
volume with a $% solution of tannic acid. Place this in
the ice chest 24 hours. Centrif ugalize the precipitate, then
redissolve with a dilute acetic acid solution ; centr if ugalize
again, and smear the sediment on a slide and stain. The
precipitate may be treated with normal sodium hydroxide
solution, then cultivated.

Other Cellular Elements

Spermatozoa, cells from neoplasms, micrococcus ureas,
streptococcus, staphylococcus, colon, typhoid, and tubercle
bacilli may find entrance and grow in the urinary tract.
To be of diagnostic value, care should be taken to pre-
vent their entering from other sources.

46 Clinical Laboratory Technic

Examination of bacteria should be upon fresh urine
obtained by catheterization.

Molds and yeasts are sometimes found in diabetic urine.

Animal parasites, booklets, and daughter cysts of echi-
nococcus, and bilharzia hematobia ova occasionally find
their way into the urine.



i. Spermatozoa. 2. Micrococcus urese. 3. Molds. 4. Bacilli.
5. Vinegar eel. 6. Yeast fungi. 7. Bilharzia hematobia eggs.
8. Echinococcus hooklets.

Burnham's Test for Formaldehyde

To 10 c.c. of urine in a test tube slightly warmed, add
3 drops of 5% alcoholic phenylhydrazine solution, one
small crystal of nitro-prusside.

Overlay with 2 c.c. of a saturated solution of NaOH.

The excretion of formaldehyde by the kidneys of
patients taking urotropin varies, as the kidneys of some
patients do not decompose the drug, and urotropin
depends for its action on formaldehyde, into which it
is decomposed in the kidney. Positive reaction 1 will

1 If the test is negative it shows that the urotropin is not being broken
up in the body, and is doing the patient no good.

Urine 47

become deep purplish black, changing quickly to dark
green, gradually getting lighter.

Systematic Scheme in Charting

The following terms are commonly used in qualitative
reactions :

MACROSCOPICAL Slightest possible trace ; faint trace ;
trace; small amount; moderate amount; large amount;
very large amount.

MICROSCOPICAL An occasional ; a few ; a moderate
number ; many ; very many.

Quantitative Estimation of Phosphates

Place 50 c.c. of urine in a flask and add 5 c.c. of sodium
acetate solution. Heat the mixture to boiling point. A
standard solution of uranium nitrate is run into the hot
mixture by means of a burette, until a precipitate ceases
to form and a drop of the mixture assumes a brownish
red color when brought in contact with a drop of potas-
sium ferrocyanide solution in a porcelain dish.

This is the end point of the precipitation.

The number of c.c. of uranium solution used is read off
from the scale on the burette.

i c.c. of the standard uranium solution is equivalent to
0.005 gram of P 2 O 5 (phosphoric acid). Therefore the
number of grams of P 2 O 5 in 50 c.c. of urine is estimated
by multiplying by 0.005.

EXAMPLE. 10 c.c. of uranium solution were used.
Then in 50 c.c. of urine there are 10 X 0.005, or 0.05;
and in 100 c.c. of urine there are 10 X 0.005 X 2, or
0.1% P 2 O,.

Normally, 3.5 grams per day.

48 Clinical Laboratory Technic

Quantitative Estimation of Chlorides. (Volhard-
Harvey Method)

Pipette 5 c.c. of urine into a porcelain dish and dilute
with 20 c.c. of distilled water.

Precipitate the chlorides with exactly 10 c.c. of standard
silver nitrate solution; add 2 c.c. of ferric ammonium
sulphate (indicator).

A solution of standard ammonium sulphocyanate is run
in from a burette until a yellowish color appears in the

Subtract the number of c.c. of sulphocyanate solution
used from 10 c.c., the quantity of silver nitrate solution
taken, and this will give the number of c.c. of the silver
nitrate solution actually used in the precipitation of the

I c.c. of silver nitrate solution is equivalent to o.oi gram
of sodium chloride, and the number of c.c. of silver nitrate
solution used multiplied by o.oi gram will give the weight
of sodium chloride in 5 c.c. of urine. Calculate from this
the weight of sodium chloride in 24-hour amount of urine.

The weight of chlorine may be estimated by multiply-
ing by the factor 0.006.

Total Acidity. (Folin's Method.)

Use a preservative in the twenty- four hour amount of
urine in order to avoid decomposition. Place 25 c.c. of
urine in an Erlenmeyer flask (250 c.c. capacity). Add 15
grams of finely powdered potassium oxalate and 2 drops
of i% phenolphthalein solution. Shake vigorously for
one minute and immediately titrate with N/io sodium
hydroxide until a faint but permanent pink color ap-
pears. Note the number of cubic centimeters of the
N/io sodium hydroxide used and calculate the acidity.

Urine 49

EXAMPLE. 6.5 c.c. of N/io sodium hydroxide were
used; then: 25 16.5 :: 1,500 :x. 25x equals 9,750.

x equals 390 (acidity of 24-hour urine expressed in
cubic centimeters of N/io sodium hydroxide).

Each cubic centimeter of N/io sodium hydroxide con-
tains 0.004 gram of sodium hydroxide, and this is equiva-
lent to 0.00^3 gram of oxalic acid. To express the total
acidity of the 24-hour amount in equivalent grams of
sodium hydroxide, multiply x by 0.004. To express the
total acidity in grams of oxalic acid, multiply x by 0.0063.

EXAMPLE. 390 times 0.004 equals 1.560 grams of
sodium hydroxide in 24-hour amount. 390 times 0.0063
equals 2.4570 grams of oxalic acid in 24-hour amount.

It is considered more difficult to increase than decrease
the acidity of the urine. The acidity of the urine may
be increased after the administration of mineral acids,
acid phosphates, or benzoates ; in cardio-renal, and other
disorders, in fasting, and in acidosis.

Test for Ammonia

Pipette 25 c.c. of fresh urine into an Erlenmeyer flask
of 250 c.c. capacity. Add 5 c.c. of a saturated solution
of potassium oxalate that has been neutralized to phe-
nolphthalein, then add 3 drops of a i% alcoholic solution
of phenolphthalein. Run in from a graduated burette
N/io sodium hydroxide until a faint pink color appears.
Then add 5 c.c. of 40% commercial formalin that has
been neutralized to phenolphthalein and again titrate to
the same color.

50 Clinical Laboratory Technic

Each cubic centimeter of N/io sodium hydroxide used
in this last titration equals i c.c. of N/no ammonia, or
0.0017 gram of ammonia.

Multiply 0.0017 by the number of cubic centimeters
used in the last titration. This gives the number of
grams of ammonia in 25 c.c. of urine.

Calculate the quantity of ammonia in the twenty- four
hour amount of urine.

The amount of ammonia in the urine varies with the
amount of protein ingested. One or two grams in
twenty- four hours is considered a normal amount ; more
than this amount indicates an acidosis.

Renal Functional Test. (Rowntree and Gerahty,
Journal A. M. A., 1911)

METHOD. The bladder should be emptied, then give
the patient 500 c.c. of water.

Exactly i c.c. of the phenolsulphonephthalem solution
from an ampoule, which contains more than i c.c., is in-
jected into the lumbar muscles. In 10 minutes obtain a
specimen and add a few drops of alkali ; then at the end
of an hour from the time the specimen, rendered alkaline,
first- shows coloration, the entire contents of the bladder
should be carefully collected and the amount of drug
excreted accurately estimated. When used as a differen-
tial test, the secretions of the two kidneys should be
separately collected by ureter catheterization for i hour
from the time the urine from either side first shows

To open the ampoule, file the neck between the small
bulb and the body of the ampoule.

Fill the wedge-shaped cell of the colorimeter with a

Urine 51

standard solution made by diluting exactly I c.c. of
phenolsulphonephthalein solution from an ampoule with
200 c.c. of water, adding 10 c.c. of a 5% solution of
sodium hydroxide, and then sufficient water to make
i liter.

At the end of the hour from the time the specimen,
rendered alkaline, first shows coloration, collect the urine
for 2 hours, each hour's specimen being kept in separate
bottles, labelled first hour and second hour.

Dilute the specimen of urine with 200 c.c. of water and
render alkaline with 10 c.c. of $% solution of sodium
hydroxide, then further dilute to make i liter. Fill the
rectangular cup to the mark. The cup is then placed in
the apparatus and the latter manipulated until the colors,
as seen through the prism, are identical, when the per-
centage of excretion will be indicated on the scale. If
the coloration is slight, showing small excretion of the
phthalein, then the dilution should be carried only to
250 or 500 c.c. and the readings on the scale divided by
4 or 2.

EXAMPLE. First hour

Second hour 27%

Total 65% (normal)

REFERENCES: Physiological Chemistry, Hammarsten.
Text-Book of Physiology, Howell. Clinical Examination
of Urine, Ogden. Physiological Chemistry, Hawk. Grad-
wohl's Blood and Urine Chemistry.

Clinical Laboratory Technic


10.00 A.M.

11.30 A.M.

2.00 P.M.

4.30 P.M.

8 to 10 P.M.

Acute Nephritic Diet

Milk, 6 oz.

Cream (top cream), 3 oz.
Zwieback (one slice, 3x2x1)
i cubic inch butter

Milk, 6 oz.
Cream, 2 oz.
Sugar, i oz.

Milk, 6 oz.
Cream, 2 oz.
Zwieback (ii slices)
Butter (i cubic inch)

Milk, 6 oz.
Cream, 2 oz.
Sugar, i dr.

Milk, 6 oz.
Cream, 2 oz.
Rice (3 tb.)
Sugar, 4 dr.
Zwieback (i slice)


Once at night, if awake

Milk, 6 oz.
Cream, 2 oz.

Milk, 6 oz.
Cream, 2 oz.


Courtesy of The Boston City Hospital.

Low Salt Diet ("Salt Free")

One liter 32 oz. of milk in 24 hours.
Salt Free Bread ) ad Ub
Salt Free Butter )

Sugar, rice, potatoes, stewed fruit, fresh green vegetables, choco-
late, coffee, tea, cereals, salad without salt.
Small portions of meat.
One or two eggs daily.

Add no salt to food



Diet in Nephritis

Breakfast. (Any one food from
each group may be given)

1. Cereals :

.Cooked 4tb..

Cornflakes 8 tb.

Shredded wheat I biscuit

2. Meat :

Lamb chop I

Pork chop i

Mutton chop i

Fish 2 tb.

Oysters 8

3- Eggs I

4. Bread I slice

Muffins I

Rolls i

Supper. (Any one food from
each group may be given)

1. Cereals :

Cornflakes 8 tb.

Cooked 4 tb.

2. Vegetables :

Green corn 3 tb.

Peas 2 tb.

Prunes 8 tb.

3. Oysters 8

4- Eggs i

5. Bread i slice

6. Coffee or tea, with

milk and sugar.

Dinner. (Any one food from
groups i, 2, 4, 5, 6; any

two from group 3)

1. Soup 32 oz.

2. Meats:

Steak (2 x i x i in.)
Chops (lean) i

Fish 2 tb.

3. Vegetables. (Any two of

these may be given)
Potato 2 tb.

Mashed 4 tb.

Parsnips 4 tb.

Carrots 4 tb.

Squash 4 tb.

Turnips 4 tb.

Onions 4 tb.
Butter beans 2tb.

Lima beans 2 tb.

4. Macaroni 4tb.
Vermicelli 4 tb.

5. Puddings :

Rice 4 tb.

Tapioca 2 tb.

Bread 2 tb.

Cornstarch 2 tb.

Ice cream 2 tb.

6. Bread i slice

Rolls i

Muffins i

The following may be given
ad lib.: Butter, olive oil, olives,
tomatoes, lettuce, celery, sweet
fruits of any kind, sugar.

Courtesy of The Boston City Hospital.


Clinical Laboratory Technic

Strict Diabetic Diet


Beef, mutton, ham, bacon,
poultry, shrimp, bologna,
sausage, lamb, pork chops,
steak, tongue, pigs' feet,
brains, bone marrow, smoked
or pickled meats, scraped or
corned beef.


All kinds. No dressing con-
taining flour. Crabs, lobsters,
sardines, etc.


Clear (not containing a fari-
naceous substance). Beef



Prepared any way, with large
amounts of butter.

Butter, olive oil: large amounts.

French dressing (olive oil, vin-
egar, etc.).

Coffee, tea, without sugar.
Akoll and Alpha biscuits.

Vegetables, etc. ,with 5% or less.
Lettuce, spinach, string beans,
celery, asparagus, cucumbers,
Brussels sprouts, unspiced
pickles, olives, grape fruit,
cauliflower, tomatoes, rhu-
barb, clams, scallops.

Foods, 6% or less.

Cabbage, radishes, pumpkin,
oysters, liver.

Foods, 10% or less.

Onions, squash, turnip, car-
rots, beets, lemons, oranges,
cranberries, peaches.

100 G. Carbohydrate Diet
Strict diet o.

Vegetables (5%, 6%, 10%

groups) 10.

Cream, ? pt. 6.

Oatmeal, I gill (dry) 24.
Bread, i oz. 30.

Potato, i medium, baked

or mashed; 2 tb. 15.
Orange or grape fruit,

Courtesy of The Boston City Hospital.

Urine 55

Name Age Sex Date.




Specific Gravity.






Microscopical Examination.

24 amount.
Diacetic acid.
/3-oxybutyric acid.
Sugar quantitative.
Albumin quantitative.
Total solids.
Total nitrogen.
Uric acid.

Microscopical Examination.



The Feces are the residue which remains after com-
plete digestion and absorption in the intestines, and the
residue is different qualitatively and quantitatively accord-
ing to the variety and quantity of food. The normal stool
varies in quantity from 250 to 500 grams.

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