ANALYSIS OF URINE AND BODY FLUIDS

PRELIM | LECTURE

CHAPTER 2: URINALYSIS

Specimen Collection Disadvantage: interferes with acid precipitation

1. Specimen should be collected on clean, dry test for proteins.
leak-proof disposable wide mouth
containers. Boric acid
2. All specimens should be properly labeled Advantage: preserves protein and formed
with the patient’s name and identification elements well, does not interfere with routine
number, the date and time of collection, urinalysis other than pH.
and additional collection, such as patient’s Disadvantage: May precipitate crystals when
age, and location, and the requesting used in large amounts.
physician, as required by institutional Additional Information: keeps pH at 6.0,
protocol. bacteriostatic agent (not bactericidal) at 18g/L,
3. A requisition slip should be accompanied can be used for culture transport.
by the specimen delivered to the
laboratory that matched the information Formalin
written in the label of the specimen. Advantage: an excellent sediment preservative.
Disadvantage: acts as reducing agent interfering
Additional information: method of collection, with chemical test for glucose, blood, leukocyte
posible interfering medication, and the patient’s esterase, and copper reduction.
clinical information. Additional Information: Rinse specimen
container with formalin to preserve cells and
Specimen Handling casts.
1. Specimen Integrity - prior to specimen
collection, the specimen should be Toluene
delivered to the laboratory promptly Advantage: Doses do not interfere with routine
and tested within 2 hours. tests.
2. Specimen Preservation - if urine is not Disadvantage: floats on the surface of
be yet tested within 2 hours, it should be specimens and clings to pipets and testing
preserved so the bacterial growth and materials, a carcinogen.
metabolism shall minimize.
Sodium Fluoride
Types of Urine Preservatives: Advantage: prevents glycolysis.
Disadvantage: inhibits reagent strip test for
Refrigeration glucose, blood and leukocytes.
Advantage: does not interfere with chemical Additional Information: may use sodium
test. benzoate instead of fluoride for reagent strip
Disadvantage: rises sp. gr. by hydrometer, testing.
precipitates amorphous phosphates and
urates. Phenol
Additional Information: prevents bacterial Advantage: does not interfere with routine tests
growth for 24 hrs. Disadvantage: cause odor change
Additional Information: use 1 drop per ounce of
Thymol specimen.
Advantage: Preserve glucose and sediments well
ANALYSIS OF URINE AND BODY FLUIDS
PRELIM | LECTURE

Commercial Preservative Tablets Types of Urine Specimen

Advantage: are convenient when refrigeration is 1. Random Specimen
not possible, have controlled concentration to - for routine screening, the most common
minimize interference. received specimen because its ease of collection
Disadvantage: many contain one or more of the and convenience for the patient. Used as
above preservatives including sodium fluoride. screening test for obvious abnormalities but it is
Additional Information: check tablet dependent on dietary intake and physical activity
composition to determine possible effects on that can give erroneous results.
desired test.
2. First Morning Specimen
Urine C + S Transport Kit (Becton - for routine testing and pregnancy testing, the
Dickinson, Rutherford, NJ) ideal specimen for routine urinalysis because it is
Advantage: can run routine urinalysis and urine concentrated. It is also essential for preventing
culture on the same specimen false-negative pregnancy test and for diagnosing
Disadvantage: deceases pH orthostatic proteinuria.
Additional Information: preservative is boric
acid. 3. Second Morning (Fasting Specimen)
- for diabetic sreening and monitoring, it is the
Saccomano’s Fixative
second voided urine of the day prior to fasting.
Advantage: Preserve cellular elements
Additional Information: Used for cytology
4. 2 Hour-Postprandial Specimen
studies.
- for diabetic monitoring, the patient is
instructed to collect urine 2 hours after eating
Special Preservatives Used in
Clinical Microscopy routine meal and then subjected to routine
urine glucose test.
1. 10ml 40% formalin- for Addis count.
2. 10 ml conc. HCl pH 2.0- for epinephrine,
noradrenalin, cathecholamines, VMA. 5. Glucose Tolerance Specimen
3. 10 ml glacial acetic acid pH 4.5- for - accompaniment to blood samples in glucose
aldosterone. tolerance test, may include fasting blood and
4. 10g Na2CO3- for porphyrine and urine samples, ½ hour blood and urine samples,
urobilinogen. 1 hour b & u samples, 3 hour b & u samples, and
5. 25 ml glacial acetic acid pH 2.0- for probably 4 hour b & u samples. The blood and
serotonin. urine test results are interpreting the ability to
6. . 10 ml conc. HCl- for steroid, ammonia, metabolize a measured amount of glucose and
urea, total N2. are correlated with the renal threshold for
7. Chloroform- for aldosterone. glucose.
8. H2SO4- preserves calcium and other
inorganic constituents. 6. 24-hour Urine (Timed) Specimen
9. NaF or Benzoic acid- ideal for glucose - used for quantitative chemical test, on the start
analysis by preventing glycolysis. of collection the patient must empty his/her
bladder and collect the urine he produce he
whole 24 hours on clean large containers and at
least refrigirated.
ANALYSIS OF URINE AND BODY FLUIDS
PRELIM | LECTURE

7. Catheterized Specimen 12. Drug Specimen Collection

- for bacterial culture, the specimen should be - patient is subjected to Chain of Custody
collected under sterile conditions by passing a Program (COC) for the collection of urine
hollow tube (Catheter) through the urethra into samples and for the reporting and counseling.
the bladder.
Test to Identify Urine
8. Midstream Clean-Catch Specimen - the presence of considerable amount of urea
- for routine screening, bacterial culture, it and nitrogen and creatinine is highly suggestive
provides a safer, less traumatic, minimal of urine since most other body fluids contain
contaminated method for obtaining urine for only small amount of these substances.
bacterial culture and routine urinalysis.
Fistulas - abnormal connection of the intestines
9. Suprapubic Aspiration (rectum) to the bladder, the urine is often
- bladder urine for bacterial culture and cytology, contaminated with feces, and the urine is usually
the physician collect urine through external brown in color, may contain residues, plant
introduction of needle through the abdomen material, or striated muscles and many bacteria.
into the bladder.
Composition of Urine:
10. Three-Glass Collection
- for diagnosing prostatic infection, instead of A. 97% Liquid
discarding the first passing urine, it is collected B. 3% solute (both inorganic and organic)
on the first sterile container, another container
for the midstream, and then the prostate will be 1. Chiefly Organic
massaged so the prostate fluid will pass with the a. Urea- 25g/L
remaining urine in the and be collected to b. Creatinine- 1.5g/L
another sterile container. c. Uric acid- 0.6g/L
d. Ammonia- 0.6g/L
All tubes- for quantitative culture, e. Undetermined N2- 0.6g/L
▪ 1st tube and 3rd tube are examined f. Sugar- traces (Negative to Benedicts Test)
microscopically. g. Ketones- Trace
▪ 2nd tube is the control for bladder and h. Carbonates, bicarbonates, and free carbonic
renal infection. acid- traces
i. pigments, mucin-like substances, diastases-
In case of prostatic infection, the 3rd tube will traces.
have a white blood cell/high power field count
and bacterial count 10 times to that of the 2. Inorganic Constituents
first specimen. a. Chloride- 9g/L
b. Sodium- 4g/L
If 2nd tube is positive to infection, the 3rd tube c. Phosphorous- 2g/L
result will be invalid and subject it to d. Potassium- 2g/L
contamination to infected urine. e. Total Sulfur- 1.5g/L
f. Calcium- 0.2g/L
11. Pediatric Specimen g. Magnesium- 0.2g/L
- The use of wii bag. h. Iron- 0.003g/L
ANALYSIS OF URINE AND BODY FLUIDS
PRELIM | LECTURE

Urinalysis: Physical Examination e. cortical and tubular necrosis

f. calculus or tumor of the kidney
I. Volume g. thrombosis of vein and artery
Urinary Volume Depends On: h. acute renal failure
a. Cardiac and renal status of the individual. i. acute tubular necrosis due to
b. Amount of fluid intake. hemoglobinuria, myoglobinuria, shock and
c. Load of solute to be secreted primarily Na+ trauma
and Urea. j. disease that interferes with circulation to the
d. Loss of fluid in perspiration and exhaled air. kidneys like poorly compensated heart disease
Normal Values:
Newborn: birth to 3 days= 20 – 50 ml/24hrs 3. Anuria
5 – 10 days=100 – 350 ml/24hrs - total suppression of urine production due to
Children: 1 year= 300 – 600 ml/24hrs failure of kidneys to secrete urine or to
10 years= 750 – 1200 ml/24hrs obstruction of urinary passages.
Adults: 750 – 2000 ml/24hrs Found in:
a. collapse with systolic pressure below 70
Volume Variation: mmHg
b. severe acute nephritis
1. Polyuria c. kidney failure (renal agenesis, bilateral
- abnormal increase in urine output (over 2000 cortical necrosis, acute glomerulonephritis)
ml/24hrs) seen in: d. poisoning with bichloride of mercury.
a. Diabetes melitus (high sp. Gr.) and Diabetes e. obstructive uropathy- obstruction in the
insipidus (low sp. Gr) urinary system (eg. Urolithiasis, calculi in the
b. Chronic kidney disease ureters, constriction of the urinary bladder neck)
c. Mental disorders f. Prostatic hyperthropy
d. During absorption of exudates and edema
e. Hyperparathyroidism 4. Nocturia
f. In response to excessive fluid intake, - excretion of urine >500 ml by an adult with
administration of diuretics or the consumption sp.gr. Of <1.018 at night.
of diuretic drinks
g. during state of anxiety II. Odor
h. tumors with inappropriate ADH secretion - normal odor of the urine is aromatic due to the
URINOD and volatile acids. May help the
Cystitis - non-pathologic, requires frequent technologist whether the urine is fresh or an old
emptying of bladder and must not be mistaken specimen.
for polyuria
Variations:
2. Oliguria 1. Ammoniacal odor - present in Proteus
- abnormal decrease in urine excretion infections and other bacterial decomposition.
(<500ml/day) 2. Mousy odor - present in phenylketonuria.
Found in: 3. Mercaptan odor - due to ingestion of
a. dehydration due to diminished water intake asparagus, garlic.
or increase renal loss. 4. Fecaloid odor - in rectovesicular fistula.
b. renal insufficiency 5. Fruity odor - presence of acetone of diacetic
c. uremia acid in diabetic urine, starvation and
d. acute nephritis dehydration.
ANALYSIS OF URINE AND BODY FLUIDS
PRELIM | LECTURE

6. Maple syrup odor - Maple Syrup Syndrome 11. Milky White - pus, phosphates, some urates,
(branched chained amino acid disorder) in fats
infants. 12. Green, Blue, Orange - drugs, medication,
7. Cabage odor - methionine malabsorption. food stuffs, Pseudomonas infection (green)
8. Rancid odor - tyrosemia. 13. Pale greenish with high sp.gr. - diabetes
9. Sweaty feet - isovaleric academia mellitus
10. Rotting fish - trimethyl amynuria 14. Brownish yellow/green with yellow foam
when shaken - bile, bilirubin oxidized to
III. Color biliverdin.
- normal urine color varies from almost colorless 15. Burgundy red - increased hematoporphyrin
or straw (low sp.gr. & large quantity) to reddish 16. Blue-green - Pseudomonas infection,
yellow or amber (high sp.gr and small volume). Amitriptyline, Methocarbamol (Robaxin) that
maybe green-brown, Clorets, Indican,
Pigments that impart color to urine: Methylene blue, phenol ingestion.
▪ Urochrome (main pigment) 17. Pink-red - Beets, Rifampin, Menstrual
▪ Uroerythrin contamination.
▪ Urobilin
▪ Porphyrin/Uresein IV. Transparency
D. mellitus- darker urine, lesser volume (conc. - freshly voided urine is clear. Diffused clouding
Urine, high sp.gr.) may occur upon standing or precipitation of
D. Insipidus - lighter color, larger volume (diluted solutes at the bottom of the container.
urine, low sp.gr.) Nubecula - a faint cloud mucus, leukocyte,
epithelial cells, bacteria, and alkaline salts that
Variations in Color of Urine: settle at the bottom of the urine upon standing.

1. Pale/Straw/Colorless - diluted urine, recent Causes of Cloudiness or Turbidity in Urine:

fluid consumption, polyuria, d. insipidus, and d.
mellitus. 1. Amorphous phosphates and Carbonates in
2. Dark yellow/brown-red - concentrated urine alkaline urine
3. Yellow-brown/Beer-brown/Amber - bilirubin, - dissolve on addition of 5 – 10%Hac, with gas
acriflavine (negative bile test with fluorescent formation.
green pigment), phenazopyridine (pyridium) 2. Amorphous urates in acidic urine
that forms orange foam, nitrofurantoin, - a white topink cloud which dissolves upon
phenindione. heating. If CHONS are present, the turbidity may
4. Orange-red/red-brown - urobilin increase formng brick-red deposits.
5. Clear red - hemoglobin 3. Oxalates - cleared by the addition of 12% HCl
6. Bright orange - pyridium (aminopyrine drugs) 4. Pus, Blood, and Epithelial Cells
7. Cloudy red - red blood cells - in alkaline urine, pus is usually mucoid, crumbly
8. Dark-red/Port Wine color - porphyrins in acidic urine. About 200 wbc/mm3 or about
9. Clerk dark brown - myoglobin as in 500 rbc/mm3 that produce turbidity.
rhabdomyolysis, and lead poisoning 5. Bacteria - produce uniform cloudiness which
10. Dark brown and black - melanin, does not settle out and cannot be filtered.
homogentisic acid (alkaptonuria), phenol Addition of Toluene or Kaolin may be added to
poisoning, Methhemoglobin, Argyrol, urine.
methyldopa or levodopa, Metronidazole (Flagyl) 6. Fat (Lipuria) - gives milky appearance, this may
be removed by extracting with ether.
ANALYSIS OF URINE AND BODY FLUIDS
PRELIM | LECTURE

7. Chyle (Chyluria) - due to parasitic (filarial) pH Depends on:

infection, or to non-parasitic (thocracic duct a. Changes in GFR, and renal blood flow
obstruction, trauma/shock and tumor) in origin b. Bicarbonate reabsorption
and imparts a cream color to the urine. c. Ammonia formation
8. Colloidal Particles, Spermatozoa, Prostatic d. Titrable acid filtration
Fluid e. H+ secretion by kidney tubules
- not cleared by acidification or heating.
Clinical Importance of pH Determination in
Donne’s Test - used to differentiate phosphate Urine:
and pus by addition of caustic soda to the
sediment. a. It is a reflection of the ability of the kidney to
Results: PO4- cleared, Pus Cells- form gelatinous maintain the normal pH in the plasma and ECF.
mass. b. It portrays a defective tubular function if the
capacity to exchange H+ ion for cationic and the
V. pH formation of NH3 is decreased.
- reaction and hydrogen concentration in the
urine. The pH of the urine is approximately 6.0 NB: It should be done on a freshly voided urine
(fasting: 5.5 – 6.5, random: 4.8 – 7.8). sample. On long standing, CO2 loss and urea are
converted to NH3.
Acidic Urine Results From:
1. Starvation VI. Specific Gravity
2. High CHONS diet - it is essential for the determination of the
3. Metabolism of fats quantity of solute dissolved in the urine
4. Metabolic and respiratory acidosis compared to solute-free distilled water.
5. Drugs used in prevention of CaCO3 or - it is the ratio of the weight of a fixed volume of
Ca3(PO4)2 stone formation a solution to that of the same volume of water at
6. Sleep a specified teperature. (usually 20OC).
NV:
Alkaline Urine Results From: Newborn: random: 1.002 – 1.004
1. Vegetable diet Adults: random: 1.005 – 1.030
2. Respiratory and metabolic alkalosis 24 hrs: 1.015 – 1.018
3. Drugs used in the prevention of uric acid and
oxalate stone formation. Variations in Specific Gravity:
4. Urea-splitting bacteria
5. Intake of citrus fruits 1. Hypersthenuria
6.. Renal acidosis -increased sp.gr (values >1.020)
Found in:
▪ Normally, H+ is excreted out in the form a. Dirrhea
of NH4+, acid PO4, or associated with b. Proteinuria
weak orgnic acids. c. decreased fluid intake
▪ NH4+- excretion results to conservation d. sweating
of Na, Ca, K, and Mg which are e. dextran administration
reavsorbed together with bicarbonate f. Urinary preservatives
ions. Therefore, the net results in the g. Fever
production of acid urine. h. i. vomiting
i. intracenous albumin
ANALYSIS OF URINE AND BODY FLUIDS
PRELIM | LECTURE

j. X-ray contrast media

The Use of Hydrometer
2. Hyposthenuria - use of the refractive index of the urine in the
- decreased sp.gr (< 1.009) determination of sp.gr.
Found in:
a. alkalosis How to Correct Specific Gravity:
b. sickle-cell anemia
c. Hypothermia a. Read actual specific gravity.
d. increased fluid intake b. Use the room temp. in OC, and hydrometer
e. progressive kidney failure temp of 16OC.
f. following administration of diuretics Standard Temp= Room temp – Hydrometer
temp
3. Isosthenuria c. For every 3OC increase in standard temp., add
- urine has fixed sp.gr of 1.010 0.001.
For every 3OC decrease in standard temp.,
The use of Urinometer for the Determination of subtract 0.001.
Sp.Gr:
Test for Sp.Gr:
Urinometer - a special type of hydrometer
calibarted to measure the sp.gr of urine at a 1. Urinometer
definite temperature (16OC) 2. Refractometry - measure the refractive index
(degree of the bending of light).
Correction for Urinometer Readings: ↓Solute = ↓Light bending
3. Harmonic Oscillation Densitometry
a. Temperature difference - change in frequency of soundwaves passes
▪ For every 3OC increase in room temp., through the solution.
add 0.001. 4. Reagent Strip
▪ For every 3OC decrease in room temp., Principle: dissociation of ions in a polyelectrolyte
subtract 0.001. solution.
To convert OF to OC Indicator: Bromthymol blue
OC= (OF – 32) x 0.555 5. Falling Drop Method
- fluid expands when warm. - measures amount of time of drops to fall.

b. Proteinuria
▪ For each gram of CHONS in the urine, the
sp.gr is increased 0.003 and should be
corrected for this increase as it does not
reflect the concentrating ability of the
kidney.
▪ for every 1% CHON (albumin), subtract
0.003.

c. Glycosuria
▪ For every 1% of glucose, subtract 0.004.