This document provides information on urine analysis tests, including specific gravity, pH, protein, glucose, ketone, blood, and bilirubin. It describes the principle, reagents, expected colors, sources of interference, and clinical significance of each test. The tests can be used to detect and monitor various conditions like diabetes, kidney disease, urinary tract infections, and more. Proper interpretation requires correlating the urine test results with the patient's symptoms and medical history.

1. Prepared by:
PRINCESS ALEN I. AGUILAR,RMT

2. 1. SPECIFIC GRAVITY
PRINCIPLE pKa change of polyelectrolytes
REAGENTS
M: Poly(methylvinyl ether) maleic anhydride, BTB
C: Ethyleneglycol-bis (aminoethylether), BTB
COLOR & READING TIME Blue-greenyellow (45 s)
SOURCES OF
INTERFERENCE
F(+): High concentration of protein
F(–): Highly alkaline urine
CLINICAL SIGNIFICANCE
1. Monitoring patient’s hydration.
2. Detection of loss of renal concentrating ability (in cases
of CGN, RF, ATN)
3. Diagnosis of diabetes insipidus (Hyposthenuria)
4. Determination of unsatisfactory specimens due to low
concentration

4. 2. pH
PRINCIPLE Double indicator system
REAGENTS Methyl red; bromthymol blue
COLOR & READING TIME Red-yellow-blue (60 s)
SOURCES OF INTERFERENCE Runover from adjacent pads Old specimens
CLINICAL SIGNIFICANCE
1. Respiratory or metabolic acidosis/ alkalosis 2
2. Renal tubular acidosis
3. Renal calculi formation
4. Treatment of UTI
5. Precipitation/ identification of crystals
6. Determination of unsatisfactory specimen
ADDITIONAL NOTES
Important in the identification of crystals and
determination of unsatisfatory specimens

5. pH
• Normal pH:
– Random  4.5-8.0
– 1st Morning  5.0-6.0
• pH of 9 = unpreserved urine
• Alkaline tide occurs after meals due to withdrawal
of hydrogen ions for the purpose of secretion of HCl
• Causes of acid urine: emphysema, diabetes
mellitus, starvation, diarrhea, dehydration, acid
producing bacteria, high protein diet, medications ,
Cranberry juice ((tx for UTI)
• Causes of alkaline urine: hyperventilation, vomiting,
renal tubular acidosis, urease-producing bacteria,
vegetarian diet, old specimens , after meals

6. 3. PROTEIN
PRINCIPLE
Protein (Sorensen’s) error of indicator ; indicator is
sensitive only to ALBUMIN
REAGENTS
M: Tetrabromphenol blue
C: Tetrachlorophenol tetrabromosulfonphthalein
COLOR & READING TIME Blue-green (60 s)
SOURCES OF INTERFERENCE
F(+): Highly buffered alkaline urine, pigmented
specimens, chlorhexidine, phenazopyridine, QACs
(detergents), antiseptics, loss of buffer, high SG F(–
):Proteins other than albumin, high salt concentration,
microalbuminuria
CLINICAL SIGNIFICANCE
NORMAL: (<30 mg/dL or <150 mg/day; Negative rgt
strip test)
Degrees of proteinuria:
a. Mild – < 1.0 g/day
b. Moderate – 1.0-4.0 g/day
c. Heavy – > 4.0 g/day
ADDITIONAL COMMENT
MOST INDICATIVE OF RENAL DISEASE
WHITE FOAM IN URINE

7. PROTEIN
1. ALBUMIN  Major serum protein found in the
urine
Normal values:
<10 mg/dL or 100mg/24 hrs (Strasinger)
<150mg/dL (Henry)
2. OTHER PROTEINS
a. Serum & Tubular microglobulins
b. Tamm-Horsfall protein (aka Uromodulin)
c. Protein derived from prostatic and vaginal
secretion

8. Types of proteinuria:
A. Pre-renal – caused by conditions that affect
plasma prior to its reaching the kidney
 intravascular hemolysis
 muscle injury
 severe infection and inflammation;
 Multiple Myeloma- proliferation of Igs-
producing plasma cells (BJP)
 Test  SPE, Immunofixation electrophoresis
 Urine = precipitates at 40-600C (cloudy) and
dissolves at 1000C

9. Types of proteinuria:
B. Renal
A. Glomeruular Proteinuria
1. Diabetic nephropathy- decreased GFR, may lead to renal failure
» Indicator: microalbuminuria
2. Orthostatic Proteinuria/ Cadet/ Postural Proteinuria
 Proteinuria when standing due to increased pressure to renal veins
3. Amyloidosis, Glomerulonephritis, Autoimmune Disorders, Toxic
Agents, Hypertension, Strenuous Exercise, Preeclampsia,
Dehydration,
B. Tubular Proteinuria– Reabsorption defective
 Fanconi syndrome,
 toxic agents
 severe viral infections
ORTHOSTATIC PROTEINURIA CLINICAL PROTEINURIA
First Morning NEGATIVE POSITIVE
2 hours after standing POSITIVE POSITIVE

10. Types of proteinuria:
C. Post-renal – after
1. lower UTI;
2. injury or trauma;
3. menstrual contamination;
4. prostatic fluid;
5. spermatozoa;
6. vaginal secretions

12. TESTS
FOR
PROTEIN
a) Heat and Acetic Acid Test
– Grading:
• diffused cloud (+1);
• granular cloud (+2);
• distinct flocculi (+3);
• large flocculi (+4)
b) SSA Test/ Cold Protein Precipitation

13. T
E
S
T
S
F
O
R
P
R
O
T
E
I
N
b) SSA Test/ Cold Protein Precipitation
 Reacts equally to all forms of protein
 RGT: 3mL of 3% SSA + 3mL centrifuged urine 
Cloudiness
• False (+): mucin, uric acid, penicillin, tolbutamides,
radiocontrast media, sulfonamides, cephalosporins
• False (-): highly buffered alkaline urine
• Correlate with reagent strip results
c)Tests for microalbuminuria
 Micral test and Immunodip  strip emplying Ab-
Enzyme conjugate that binds albumin
 (-) WHITE ; (+) RED
 Significant values reported as AER
Normal AER = 0-20 ug/min
Microalbuminuria = 20-200 ug/min 0r (30-
300mg/24hrs)
Clinical albuminuria = >200 ug/min

14. 4. GLUCOSE
PRINCIPLE Glucose oxidase reaction / Double sequential enzyme rxn
REAGENTS
M: Glucose oxidase, peroxidase, KI
C: Glucose oxidase, peroxidase, TMB
COLOR & READING TIME M: greenbrown C: yellowgreen (30 s)
SOURCES OF INTERFERENCE
F(+): Oxidizing agents, detergents
F(–): Ascorbic acid, ketones, high SG, low temperatures,
improperly preserved specimens
CLINICAL SIGNIFICANCE
Types of Glucosuria:
a. Hyperglycemia-associated – diabetes mellitus,
endocrine disorders,( Cushing’s, Pheochromocytoma,
Acromegaly, Hyperthyroidism) pancreatic disorders,
CNS disorders, disturbance in metabolism, liver
disease, drugs, gestational diabetes mellitus
b. Renal-associated – renal tubular dysfunction, tubular
necrosis, Fanconi syndrome, osteomalacia, pregnancy
ADDITIONAL COMMENT Most frequently tested in urine; threshold substance

15. Benedict’s test
• (Copper Reduction)
CuSO4  (+) Cu2O
(Blue) reducing subs
 Positive result: brick red precipitate
• Copper reduction
• Procedure: 5 gtts urine + 10 gtts
H2O + clinitest tablet
• Rgts:
– CuSO4 (main reacting rgt)
– NaOH, sodium citrate (for heat
production)
– NaCO3 –elliminates interfering O2
• False(+): other reducing sugars,
ascorbic acid, drug metabolites,
cephalosporin
• False (-):
– oxidizing agents (detergents),
– pass-through phenomenon
• Occurs when >2g/dL sugar is present
• To prevent, use 2gtts urine
• Correlate with reagent strip results

16.  Non-specific test for reducing sugars
• Fructose (Levulose)
• Galactose
• Lactose (Glu-Gal)
• Pentose (Xylulose, Arabinose)
• Sucrose (Glu-Fru) – non reducing sugar,
thus negative
NOTE:

17. 5. KETONE
PRINCIPLE
Sodium nitroprusside reaction
Acetoacetic acid + Na Nitroprusside  purple (+)
(Acetone) (Glycine)
REAGENTS
M: Sodium nitroprusside
C: Sodium nitroprusside and glycine (can detect acetone)
COLOR & READING TIME Purple (40 s)
SOURCES OF INTERFERENCE
F(+): Phthalein dyes, highly pigmented red urine,
levodopa, medications containing SH group
F(–): Improperly preserved specimens
CLINICAL SIGNIFICANCE
Ketone bodies: Acetone – 2% Acetoacetic acid – 20%
β-hydroxybutyric acid – 78%
Causes of Ketonuria: diabetes mellitus, starvation,
fasting, weight reduction, strenuous exercise,
malabsorption, pancreatic disorders, inborn errors of
amino acid metabolism
CONFIRMATORY TEST ACETEST TABLET TEST

18. • CONTAINS:
– Sodium nitroprusside
– Disodium phosphate (strong alkaline buffer)
– Lactose (helps to enhance the color)
• 10 times sensitive to diacetic acid than acetone
• Can detect in urine:
– 5–10 mg/dL of diacetic acid and 20–25 mg/dL of
acetone

19. 1. Place the tablet on a piece of clean, dry white paper.
2. Put one drop of urine, serum, plasma, or whole blood
directly on top of the tablet.
3. For urine, compare the color of the tablet with the color
chart at 30 seconds. For serum or plasma, compare the color
after 2 minutes. For whole blood, remove the clotted blood
from the tablet after 10 minutes and compare the color of
the tablet with the chart.
Note: for serum, plasma and whole blood, the loest limit of
detection s 10mg of diacetic acid per 100mL
5-10mg/dL
Diacetic acid
30-40mg/dL
Diacetic acid
80-100mg/dL
Diacetic acid

20. 1. Gerhart’s, Lindeman’s – diacetic acid
2. Frommer’s, Walhauster’s, Lange’s,
JacksonTaylor’s, Rantzmann’s, Lieben’s –
acetone
3. Legal’s, Rothera’s, Acetest, Ketostix – diacetic
acid and acetone
4. Osterberg, Hart’s test – β-hydroxybutyric acid

21. 6. BLOOD
PRINCIPLE
Pseudoperoxidase activity of heme
Hgb
H2O2 + Chromogen  oxidized chromogen + H2O
Pseudoperoxidase
REAGENTS
M:Diisopropylbenzene dihydroperoxide TMB
C: 2,5-dimethyl 2,5dihydroperoxyhexane TMB
COLOR & READING TIME
(-) yellow ; (+)Blue-green (60 s)
Uniformed green/blue color= Hgb/Mgb
Speckled/Spotted = Hematuria (intact RBCs)
SOURCES OF INTERFERENCE
F(+): Strong oxidizing agents, bacterial peroxidases, menstrual
contamination
F(–): High SG, crenated cells, formalin, captopril, nitrite, ascorbic
acid, unmixed specimen
CLINICAL SIGNIFICANCE
a. Hematuria
b. Hemoglobinuria
c. Myoglobinuria
CONFIRMATORY TEST MICROSCOPIC ANALYSIS

22. HEMATURIA HEMOGLOBINURIA MYOGLOBINURIA
Cloudy red urine Clear red urine Clear red urine/ reddish
brown/tea-colored
Seen in:
 Glomerulonephritis
 Renal calculi
Microscopic: INTACT RBCS
Seen in:
 Intravascular hemolysis
 Transfusion rxns
 Hemolytic Anemia
Seen in: Rhabdomyolysis
Heme portion of the
myoglobin is toxic to the
renal tubules
HEMOGLOBIN vs MYOGLOBIN
TEST HEMOGLOBIN MYOGLOBIN
1. Plasma Examination  Red/Pink plasma
 Decrease Haptoglobin
 CK Slightly increased
 LD 1 & 2 increased
 Pale yellow plasma
 CK Markedly increased
and Aldolase increased
 LD 4 & 5 increased
2. Blondheim’s test
(Ammonium sulfate)
Proc: Urine + 2.8 NH4Sulfate (80%
satd)  filter centri  test
supernatant for blood with rgt strip
Precipitated at the bottom
Supernatant test (-) for blood
Not precipitated
Supernatant test (+) blood

23. OTHER TESTS FOR URINE BLOOD
• Enzyme assays
• Absorption Spectrophotometry
• Immunodiffusion Technique
• Electrophoresis

24. 7. BILIRUBIN
PRINCIPLE
Diazo reaction:
B2 + Diazonium salt  Azodye
REAGENTS
M: 2,4-dichloroaniline diazonium salt
C:2,6-dichlorobenzene diazoniumtetrafluoroborate
COLOR & READING TIME Tan, pink, or violet (30 s)
SOURCES OF INTERFERENCE
F(+): Highly pigmented urine, indican, phenazopyridine, lorpromazine
(Thorazine), metabolites of Lodine , chF(–): Specimen exposure to light,
ascorbic acid >25 mg/dL, high concentration of nitrite
CLINICAL SIGNIFICANCE
1. Diagnosis of hepatitis, cirrhosis, other liver disorders, and biliary
obstruction
2. Determination is more significant when combined with serum
bilirubin and urine urobilinogen
CONFIRMATORY TEST ICTOTEST
ADDITIONAL COMMENTS
 Conjugated bilirubin (water soluble)
 Early indication of liver disease
 Amber urine with yellow foam

25. Procedure:
1. Place five drops of urine on
one square of the special test
mat supplied with Ictotest.
2. Place a tablet in the center of
the moistened area.
3. Flow two drops of water onto
the tablet so that the water
runs off of the tablet and onto
the mat.
4. Observe the color of the mat
around the tablet at the end
of 30 seconds. If a blue or
purple color develops, the test
is positive.
 able to detect as little as
0.05 mg/dL bilirubin
Contains:
 P-nitrobenzene-diazonium
p-toluenesulfonate
 SSA (provide acidity and act
with Sodium carbonate to
provide effervescence
 Sodium carbonate
 Boric acid

26. Other Tests For Bilirubin
• Foam Shake Test
• Oxidation Test (Gmelin or Fouchet’s method)
 Acidic oxidation of bilirubin into a rainbow
array of colors:
– green (biliverdin)
– blue (bilicyanin)
– yellow (choletelin)

27. 8. UROBILINOGEN
PRINCIPLE Ehrlich reaction : UBG +PDAB  (+) RED
REAGENTS
M: paradiethylaminobenzaldehyde
C: 4-methoxybenzene diazonium tetrafluoroborate
COLOR & READING TIME Red (60 s)
SOURCES OF INTERFERENCE
F(+): PBG, indican, procaine, p-aminosalicylic acid,
sulfonamides, methyldopa, chlorpromazine, pigmented urine
F(–): Old specimens, formalin, high concentration of nitrite
CLINICAL SIGNIFICANCE
1. Only a small amount is normally found in urine.
2. Useful for the early detection of liver disease and for the
diagnosis of hemolytic disorders, hepatitis, cirrhosis, and
carcinoma
3. Absence in urine may indicate biliary obstruction
TEST USED TO DIFFERENTIATE
UROBILINOGEN TO
PORPHOBILINOGEN
WATSON-SCWARTZ DIFFERENTIATION TEST
ADDITIONAL COMMENTS
 (< 1.0 mg/dL or <1.0 Ehrlich unit)
 Specimen: Afternoon urine (2-4PM due to alkaline tide)

28. 2. WATSON-SCHWARTZ TEST
• Uses extraction with organic solvents Chloroform and Butanol
• Sodium acetate  enhances reaction
1. Ehrlich’s Tube Test
• Reagent: p-dimethylaminobenzaldehyde
• (+) Result: cherry red color

29. • INVERSE EHRLICH REACTION
• Rapid screening test for Porphobilinogen
(>2 mg/dL)
Procedure:
2 gtts urine + 2mL Hoesch rgt
( Ehrlich’s rgt in 6M or 6N HCl)
• Immediate development of a cherry-red color at
the top of the mixture indicates a positive result

31. 9. NITRITE
PRINCIPLE Greiss reaction
REAGENTS
M: p-arsanilic acid, tetrahydrobenzoquinolinol
C: sulfanilamide, 3hydroxy- 1,2,3,4- tetrahydro- 7,8-
benzoquinoline
COLOR & READING TIME
Uniform Pink (+) = 100,000 org/mL
not pink spots/edge (-) (60 s)
SOURCES OF INTERFERENCE
F(+): Old specimen, highly pigmented urine
F(–):Non-reductase-containing bacteria, lack of urinary nitrate,
insufficient contact time between bacteria and nitrate,
bacteria converting nitrite to nitrogen, antibiotics, ascorbic
acid, high SG
CLINICAL SIGNIFICANCE
1. Diagnosis of cystitis and pyelonephritis
2. evaluation of antibiotic therapy
3. Monitoring of patients at high risk for UTI
4. Screening of urine culture specimens
ADDITIONAL COMMENTS
 Rapid screening test for UTI/bacteriuria
 Specimen: 1st morning or 4hr urine

32. 10. LEUKOCYTE ESTERASE
PRINCIPLE Granulocytic esterase reaction
REAGENTS
M:Derivatized pyerole amino acid ester, diazonium salt
C: Indoxylcarbonic acid ester, diazonium salt
COLOR & READING TIME
Purple (120 s) (+) WBC (exc. Lymphocyte), histiocyte,
Trichomonas vaginalis = contain esterase
SOURCES OF INTERFERENCE
F(+): Strong oxidizing agents, formalin, highly pigmented urine,
nitrofurantoin
F(–): protein, glucose, oxalic acid, ascorbic acid, gentamicin,
cephalosporins, tetracyclines
CLINICAL SIGNIFICANCE
1. Detection of bacterial and nonbacterial UTI
2. Inflammation of the urinary tract
3. Screening of urine culture specimens
ADDITIONAL COMMENTS
 Strip can detect even lysed WBCs

33. List of References
Lillian Mundt & Kristy Shanahan, Graff’s Textbook
of Urinalysis and Body Fluids, 2nd Ed.
Susan Strassinger & Marjorie Di Lorenzo,
Urinalysis and Body Fluids, 5th & 6th Ed.
Erol Coderres,RMT-AUBF notes
Roderick Balce, RMT-CEU Professor AUBF Notes
Rexson R. Julian ,RMT- PPT notes
Ridley,J. et al, Essentials of Clinical Laboratory
Science, ©2011