Chemical Examination

Chemical Examination of Urine
Introduction The color reactions are interpreted by comparing the color

Routine chemical examination of urine has changed produced on the pad with a chart supplied by the
dramatically since the early days of urine testing, owing to the manufacturer.
development of the reagent strip method for chemical analysis.
The introduction of the reagent strip currently provide a simple,
rapid screening means for performing medically significant
chemical analysis of urine.
Reagent Strip
An inert plastic strip onto which reagent – impregnated test
pads are bonded.
The following are the parameters analyzed using the urinalysis
reagent strip.
pH
Protein
Glucose
Ketones
Blood
Bilirubin
Urobilinogen
Nitrite
Several colors or intensities of a color for each substance being
Leukocytes
tested appear on the chart.
Specific Gravity
A semi quantitative value of each can be reported.
Reagent strips consist of chemical – impregnated absorbent
An estimate of the milligrams per deciliter present is available
pads attached to a plastic strip.
for some of the parameters.
1
A color-producing chemical reaction takes place when the
Page
absorbent pad comes in contact with urine.
By: Ken Kawakami, RMT, MLS(ASCPi)CM

@japanesejuan
Reporting can be done by:  The strip must be placed very near the color chart but
1. In concentration (mg/dL) not making contact with it
2. Descriptive (small, moderate, large)  Specimens that have been refrigerated must be brought
3. Plus system to room temperature first before actual testing
 Negative (enzymatic reactions are temperature dependent)
 Trace  MTs who are severely color blind should not perform
 1+ reagent strip testing
 2+  Color masking by drugs and other substances my
 3+ interfere with readings (perform chemical testing instead)
 4+  Although ascorbic acid has the potential to adversely
4. Positive or Negative affect several reagent strip test results, most
manufacturers use an iodate overlay to prevent this.
Reagent Strip Technique  Specimens must be tested within 2 hours of collection
Testing Methodology
 Dip the reagent strip completely (but briefly) into a well- Handling and Storage
mixed specimen  Strips must be protected from moisture, volatile
 Remove excess urine by running the edge of the strip on chemicals, heat and light
the container, test tube or absorbent pad  Desiccants should not be removed from the bottle
 Wait for the specified time for reactions to take place  Strips should be removed prior to testing only and bottle
 Compare color reactions against manufacturer’s chart should be tightly sealed immediately
using a good light source  Bottles should not be opened in the presence of volatile
TIPS! fumes
 Improper technique can result in error.  Reagent strip bottles should be stored per manufacturer
 RBCs and WBCs sink to the bottom of the specimen instructions (usually RT)
 Do not allow the strip to remain too long with the  All reagent strips used should not be beyond expiration
specimen as this may cause removal of reagents date
 Excess urine that remains on the strip may cause  Care should be taken not to touch reagent pads when
chemical run over, hence, distortion of colors removing the strips
2
 Manufacturer’s timing should be followed for best results
Page
 A good light source is required

@japanesejuan
Quality Control
Each bottle should be checked with a positive and negative
control
 minimum once every 24 hours or when a new bottle is
opened
 When there are questionable results
 Concern about strip integrity
All quality control results should be documented
Distilled water should never be used as a negative control.
All negative controls should read negative.
All positive controls should agree ± one color block.
All QC results that do not agree should be documented and
resolved before proceeding with urine testing.
Tablet and Chemical Tests

 Each tablet or chemical test must follow manufacturer’s
directions to ensure reproducible and reliable results
 The laboratorian should know the following for each test
and compare it with reagent strip testing
Sensitivity
Specificity
Potential interferences
 Chemical and tablet tests are generally performed
1. To confirm results already obtained by reagent strip
2. As an alternative method for highly pigmented urine
3. Because some are more sensitive for the substance
(e.g. Ictotest)
4. Specificity of the test differs from that of the reagent
3
strip (e.g. SSA, Hoesch)
Page
@japanesejuan
pH
Introduction:
The renal system, the pulmonary system, and blood buffers are
major regulators of acid-base content in the body. Through
secretion of Hydrogen in the form of:
1. Ammonium ions
2. Hydrogen phosphate
3. Weak organic acids
4. Reabsorption of bicarbonate
Normal pH of urine of a healthy individual:
First morning/average person: pH 5.0-6.0
after a meal – more alkaline (alkaline tide)
Normal range: pH 4.5-8.0
Clinical Significance
Urinary pH must be considered in conjunction with other patient  An aid in determining the existence of systemic acid-
information such as: base disorders
1. Acid-base content of blood o Metabolic
2. Renal function o Respiratory
3. Infections  Management of urinary conditions
4. Dietary intake  Aids in evaluation of kidney reabsorption or secretion
5. Age of the specimen abilities
 Calculi formation
 Management of infections
 Specimen viability
4
Page
@japanesejuan
 Care must be taken to prevent run over from the protein

area as the latter is acidic and may give a false low pH
(false acidic) when testing alkaline urine
Reagent strip reaction

PRINCIPLE: Double Indicator System
 Most reagent strip pH pads measure urine pH in 0.5 or 1
unit increments from pH 5.0 to 9.0
 A double indicator system is used to differentiate the pH
range
 Methyl Red – produces a color change from red
to yellow from pH 4 – 6 NOTES:
 Bromthymol blue – turns from yellow to blue in the  pH of greater than 8.0 or less than 4.5 are physiologically
range pH 6 – 9 impossible, hence, investigation required
 Color ranges from ORANGE at pH 5.0 through yellow and if greater than 8.0
green(ph 7.0) to a final deep blue at pH 9  highly alkaline medications
 urease – producing bacteria present
following improper preservation
5
Page
 No known substance is known to interfere with pH pads

@japanesejuan
Protein
 Demonstration of urine protein does not always signify
Introduction: renal disease – additional testing is required
Of all chemical tests performed on urine, the most indicative of  Clinical proteinuria (>30mg/dL or 300mg/L)
renal disease is the protein determination. Presence of protein in o Pre – renal
urine or proteinuria is almost always associated with early renal o Renal
disease. o Post – renal
Normal urine: (Pre – renal proteinuria)

Less than 10mg/dL or 100 mg per 24 hours is excreted  Not an actual renal disease
 Also called overflow proteinuria
Protein excreted is mostly made up of low – molecular weight  Caused by conditions affecting plasma protein
serum proteins (filtered by the glomerulus and some proteins in  Increased filtration and exceeds capacity of the renal
the genitourinary tract) tubules to reabsorb resulting in an overflow
 Usually transient
Major protein in urine: ALBUMIN  Caused by increased levels of:
High concentration in plasma but low in urine because of the Low – molecular weight plasma proteins
filtration process (Most albumin cannot pass glomerulus and  Hemoglobin (after hemolytic episode)
those that pass are reabsorbed by the tubules)  Myoglobin (following muscle injury)
Other proteins:  Acute phase reactants (inflammation and
1. Serum and tubular microglobulins infection)
2. Tamm-Horsfall protein by tubules *Usually not discovered in routine urinalysis since
3. Prostatic protein the protein pad detects primarily albumin
4. Seminal protein
5. Vaginal secretions
6
Page
@japanesejuan
 Bence – Jones Protein (Renal Proteinuria)

An example of pre – renal proteinuria (due to  Proteinuria associated with TRUE RENAL DISEASE
increased serum protein levels)  Classified as either:
Found in patients with Multiple Myeloma (a 1. Glomerular Proteinuria
proliferative disorder of plasma cells)  Happens when the glomerular membrane is
Excretion of a unique type of protein called Bence damaged
– Jones Protein which are immunoglobulin light  Selective filtration is impaired
chains  A large amount of serum protein, RBCs and WBCs
Low molecular weight protein which exceeds the pass through and are excreted in urine
tubular reabsorption capacity and is excreted in  Conditions that damage the glomerulus
urine o Amyloid material
When BJP is suspected, a screening test is o Toxic substances
performed: o Immune complexes found in Lupus
Principle: erythematosus and Streptococcal GN
BJP has a unique solubility characteristic.  Other conditions but are reversible:
BJP coagulates when heated to 40⁰C-60⁰C and o Increased blood pressure / Hypertension
dissolves at 100⁰C. o Strenous exercise
o Dehydration
Positive result: Turbid urine at 40⁰C-60⁰C and clear at o Pregnancy (Pre-eclamptic states)
100⁰C 2. Tubular Proteinuria
 Not all patients with Multiple Myeloma excrete BJP  Happens when the tubules are no longer capable
 Diagnosis of Multiple Myeloma should be of reabsorbing normally filtered albumin
confirmed through serum electrophoresis and  Causes of tubular proteinuria:
immunoelectrophoresis o Toxic substances / Heavy metals
o Severe Viral infections
o Fanconi Syndrome
 Protein levels found in urine are slightly above
normal to 4g/day
7
 Markedly increased urinary protein is seldom seen
Page
in tubular proteinuria

@japanesejuan
NOTE:  Detection of microalbumin before needed a 24 –

Discovery of protein from random samples is not always hour urine sample
pathologic. Several cases are benign. o Results were reported in mg of albumin / 24
hours or AER (Albumin excretion) in ug/min
Other Renal Proteinuria:  Newer testing methods use Enzyme immunoassays or
1. Orthostatic (Postural) Proteinuria Immunochromographics
 Persistent benign proteinuria o Sensitivity: 0-10 mg/dL for EIA
 Functional proteinuria o Sensitivity: 1.2-8.0 mg/dL for
 Occurs frequently in young adults Immunochromographics
 Called ORTHOSTATIC or POSTURAL because urinary  Microalbumin is considered significant if found in urine
protein increases following periods in vertical position. at 30 to 300mg albumin secreted in 24 hours or AER is
 Urinary protein returns to normal when horizontal 20-200 ug/min.
position is assumed
 Caused by pressure on the renal vein (Post – Renal Proteinuria)
Testing procedure:  Protein that is added to urine ultrafiltrate as it passes
 Patient is requested to empty bladder before going to through the lower urinary tract
bed and collect a specimen immediately upon arising in Ureter
the morning Bladder
 A second specimen is collected after remaining in Urethra
vertical position for several hours Prostate
Positive result: Vagina
1st specimen is negative for protein and 2nd specimen is positive  Examples of post – renal urinary protein causes:
2. Microalbumin Bacterial infections
 A predictive indicator of renal complications Fungal infections
 Useful test for diabetic patients Inflammation (exudates)
 Diabetic nephropathy leads to reduced glomerular Menstruation
filtration and ends in renal failure (Type I and II Prostatic fluid
diabetes) Semen
8
 Microalbuminuria is also associated with increased risk
Page
of cardiovascular disease

@japanesejuan
Reagent strip reactions

Principle: PROTEIN ERROR OF INDICATORS
 Colorimetric
 Changes color in the presence of a certain protein and
not due to pH (pH held constantly at pH 3.0)
 Albumin accepts hydrogen ions from the indicator
 The test is sensitive to albumin compared to other
proteins because it has more amino groups to accept
hydrogen ions given by the indicator
 Reagents:
o Tetrabromphenol blue or
3’,3”,5’,5” – tetrachlorophenol -3,4,5,6-
tetrabromosulfonphthalein
o Acid buffer (to maintain at an acidic pH)
 At pH 3.0, indicators are yellow
 As protein concentration increases, the color progresses
through shades of green and finally blue
 Reported as
o NEGATIVE
o TRACE (usually less than 30mg/dL)
o 1+, 2+, 3+, and 4+
 Semiquantitative reporting:
o 30 mg/dL
o 100 mg/dL
o 300 mg/dL
o 2000mg/dL
9
Page
@japanesejuan
Reaction Interference/Limitation o Tolbutamide metabolites (Px history)

 Highly buffered alkaline urine (Major interference) o Cephalosporins (Px history)
o Overrides acidic environment of protein pad o Penicillins
o Produces a color change unrelated to protein o Sulfonamides
concentration  Highly alkaline urine may overcome acidity of SSA –
 Prolonged contact with urine specimen less turbidity than it should or False negative results
o May remove acid buffer o Use more concentrated SSA
o False – positive results when acid buffer is removed  SSA is sensitive to 5mg/dL to 10mg/dL protein
 Highly pigmented urine (False – positive) regardless of the type of protein present
 Contamination of quaternary ammonium compounds  When SSA is crystalline, radiographic contrast media
(False – positive) could be present and reagent strip results should be
 Detergents (False – positive) reported
 Antiseptics (False – positive)  SSA should never be used to confirm protein results
 A very high specific gravity (False – positive) from reagent strip testing because it lacks protein
 Protein other than albumin (False – negative) specificity
2. Heat and Acetic method
NOTE: Chemical confirmation of protein results: a. Place 5-10mL of urine in a clean test tube
o A very high protein result b. Boil upper 1/3 of the tube (1-2 minutes)
o Urine is very alkaline c. Add 1-2 drops of glacial acetic acid (5%)
o I. Initial turbidity after flaming is due to
2 tests that test for protein other than reagent strip: phosphates and carbonates – glacial acetic
1. Sulfosalicylic Acid Precipitation Test will clear it up
 Cold precipitation test d. Reboil the specimen
 Reacts with all types of protein e. Grade the turbidity
 Methods vary amongst laboratories
 Specimen should be supernatant of urine to remove
any extraneous proteins
10
 Examples of protein that are precipitated but are not
clinically important: (False turbidity)
Page
o Radiographic dyes (high specific gravity)

@japanesejuan
11
Page
@japanesejuan
Glucose
 Gestational diabetes
Hyperglycemia that occurs during pregnancy
Introduction: Disappears after delivery
The glucose test is the most frequent chemical analysis Onset is during the 6th month of pregnancy
performed on urine. Its value in the detection and monitoring of Due to action of hormones secreted by the
Diabetes mellitus is unchallenged. placenta which blocks insulin resulting in
More than half of the cases in the world are undiagnosed. resistance of insulin and hyperglycemia
Early diagnosis is the key to improved prognosis. Detection is important as glucose crosses the
placenta and insulin does not
Clinical Significance Glucose will be absorbed by the baby’s pancreas
 The kidney’s PROXIMAL CONVOLUTED TUBULE (PCT) will produce a lot of insulin converting all glucose
reabsorbs glucose almost completely into fat and stored.
 Reabsorption rate is at 160-180mg/dL (renal threshold) Baby will be at risk for obesity and type 2 diabetes
 Should glucose in the blood be too high, renal tubular Women who have gestational diabetes are also
reabsorption will be difficult and glucose will appear in prone to developing type 2 diabetes
urine
 Used for diabetes screening – fasting prior to the
collection of samples is recommended
Blood glucose levels fluctuate especially after
meals
2 hours after a meal is recommended
First morning specimens are not recommended
because they do not represent an actual
representation of the body’s ability to clear
glucose (evening meal glucose still in bladder)
 Urine glucose should be correlated with FBS
 OGTT is used to confirm diabetes
12
Page
@japanesejuan
 Examples of hyperglycemia of non diabetic origin which Reagent strip reactions

also produces glycosuria PRINCIPLE: DOUBLE SEQUENTIAL ENZYME REACTION
Pancreatitis  2 different tests used by laboratories
Pancreatic cancer Glucose oxidase
Acromegaly  Specific for glucose
Cushing’s syndrome  Used in reagent strips/impregnated in
Hyperthyroidism reagent strips
Pheochromocytoma Copper reduction
Drugs  Detects glucose and other reducing
Liver diseases substances
CNS damage  Reagent strip for glucose employ glucose oxidase by
Hormonal disorders impregnating the test area with a mixture of (refer below)
 Hormones associated with the disorders of to produce a double sequential enzyme reaction
hyperglycemia of non diabetic in origin 1. Glucose oxidase
Glucagon (increased) 2. Peroxidase
Epinephrine (increased) 3. Chromogen
Cortisol (increased) 4. Buffer
Thyroxine (increased) 1 step: Glucose oxidase rapidly catalyzes the reaction of
st
GH (increased) glucose to produce gluconic acid and hydrogen peroxide.

*all of the hormones mentioned oppose insulin function 2nd step: Peroxidase catalyzes the reaction between the
*Glycogen (fat) is converted into glucose hydrogen peroxide formed and chromogen to form an oxidized
 Glycosuria in the absence of hyperglycemia (Renal colored compound that represents the presence of glucose.
Glycosuria)
Caused by failure of tubules to reabsorb glucose
Seen in:
1. ESRD
2. Cystinosis
13
3. Fanconi Syndrome
Page
@japanesejuan
 Reagent strip manufacturers use several different  High specific gravity and low temperature decrease test
chromogens sensitivity
Potassium iodide(green to brown)  Unpreserved specimens give false – negative results
Tetramethylbenzidine (yellow to green) o Rapid glycolysis of glucose
 Urine glucose is reported in terms of:
NEGATIVE Other tests for urine glucose:
Trace 1. Copper Reduction Test
1+ One of the earliest chemical test performed on urine
2+ Test relies on the ability of glucose and other
3+ substances to reduce copper sulfate to cuprous
4+ oxide in the presence of alkali and heat
 Color charts also provide semi-quantitative Reducing sugars include:
measurements a. Glucose
Ranges from 100mg/dL to 2g/dL b. Fructose
The American Diabetes Association recommends c. Galactose
quantitative reporting d. Maltose
e. Pentose
Reaction Interference Color change progressing from a negative blue
 Glucose oxidase is specific for glucose only (CuSO4) through green, yellow, and orange/red
o other sugars are not detected (Cu2O) occurs when the reaction takes place
 Peroxide and strong oxidizing detergents give False –
positive reactions
 Strong reducing agents give False – negative reactions;
(oxidation will not proceed)
Example:
o Ascorbic acid The best example of Copper reduction is the
Ascorbic acid interference can be minimized by Benedict’s Test
14
incorporating iodate into glucose pads; iodate oxidizes
ascorbic acid
Page
@japanesejuan
The Benedict solution was developed in 1908 and IMPORTANT NOTE:

contains: Care must be taken to observe the reaction closely
o Copper sulfate as it is taking place
o Sodium carbonate High glucose OR reducing substances could cause a
o Sodium citrate buffer “PASS THROUGH” phenomenon
Procedure: PASS THROUGH PHENOMENON happens when the color
Urine + Benedict Solution + Heat = Precipitate produced passes through the orange/red stage and returns to
2. Clinitest a green-brown color, and if not observed, a high glucose level
A tablet version of the Benedict’s Test may be reported as NEGATIVE
Makes use of the ability of the reducing sugars ability An alternate method uses 2 drops instead of 5 drops
to convert cupric sulfate to cuprous oxide of urine – can minimize the occurrence of “pass
The test is based on the ability of reducing substances through”
to convert cupric sulfate to cuprous oxide  A separate color chart must be used for this
Makes use of a tablet and contains alternate method
o Anhydrous copper sulfate  The chart provides up to 5g/dL
o Sodium carbonate semiquantitation whereas the regular (5 drops
o Sodium citrate / citric acid urine) provides only up to 2g/dL
o Sodium hydroxide Sensitivity of Clinitest is 200 mg/dL
Upon addition of the tablet to water and urine, heat is Clinitest is still a nonspecific test for reducing
produced by the hydrolysis of sodium hydroxide and substances and subject to interference from several
its reaction with sodium citrate… Carbon dioxide is other reducing sugars:
released from the sodium carbonate to prevent room 1. Galactose
air interfering with the reduction reaction 2. Lactose
Tubes should be placed on a rack and should not be 3. Fructose
held by hand because the heat reaction could 4. Maltose
cause a burn 5. Pentoses
After the effervescence reaction, the tube is gently 6. Ascorbic acid
15
shaken and the color ranging from blue to 7. Drug metabolites
orange/red can be compared with the 8. Antibiotics (Cephalosporins)
Page
manufacturer’s color chart Clinitest is not a confirmatory test for urine glucose

@japanesejuan
Clinitest tablets are hygroscopic and should be stored

in tightly packed packages
 A strong blue color suggests deterioration of
the tablet due to moisture and should not be
used
16
Page
@japanesejuan
Ketones
 When CHO are available, ketone synthesis is inhibited
Introduction: The term ketones and ketone bodies represent and blood ketone levels are below 3mg/dL
three intermediate products of fatty acid metabolism.  Any condition that causes increased fat metabolism
1. Acetoacetic acid (1st Ketone formed) leads to ketonuria and ketonemia. Clinical reasons for
2. Acetone increased fat metabolism:
3. Beta-hydroxybutyric acid Inability to metabolize CHO (as in DM)
Normally, measurable amounts are not detected in urine Increased loss of CHO from vomiting
because all the metabolized fat are converted into CO2 and Inadequate intake of CHO associated with
H2O. However, when the use of available CHO as the major  Starvation
source becomes compromised, body stores of fat must be  Malabsorption
metabolized to supply energy. Ketones are then detected in  Testing for urinary ketones is most valuable in the
urine. management and monitoring of insulin – dependent
(type 1) DM because of the inability to use CHO
If patient has ketonuria, it shows a deficiency in
insulin and the need to regulate dosage
Ketonuria is an often an early indicator of
insufficient dosage in type 1 Diabetes
 Increased amounts of ketone in the blood leads to
Electrolyte imbalance – large amount of H20 lost
Dehydration – large amount of H20 lost
Acidosis – due to ketoacids
Diabetic coma
 All kinds of reagent multi-strips have ketone pads
incorporated because it provides valuable information
when correlated with glucose
 Ketone renal threshold is 70mg/dL.
17
When blood ketone is more than 70mg/dL,
ketonuria happens.
Page
@japanesejuan
 Patients with ketonuria have fruity or acetonic breath  Results are reported qualitatively or semi – quantitatively
odors because acetone is also eliminated by the lungs as:
NEGATIVE
Trace (5mg/dL)
Small (1+) (15mg/dL)
Moderate (2+) (40mg/dL)
Large (3+) (80-160mg/dL)
Reaction:
Chemical Tests for Ketone bodies:

1. Legal’s Test or Rothera’s Test
Reagent strip reactions  Developed by Legal in 1883 and modified by Rothera in
The 3 ketone compounds present are not in equal amounts in 1908
urine and blood. The average distribution is as follows:  A nitroprusside – reaction test
1. Acetoacetic (20%)  15 – 20 times more sensitive to acetoacetate than
a. Acetone (2%) acetone
b. beta-hydroxybutyric acid (78%)  Does not react to beta – hydroxybutyric acid
PRINCIPLE: SODIUM NITROPRUSSIDE (NITROFERRICYANIDE)  No longer performed in clinical laboratories
REACTION  More sensitive to acetoacetate and acetone than
 Acetoacetic acid (in an alkaline medium) reacts with reagent strip ketone pads
sodium nitroprusside to produce a purple color.
 The test does not measure beta-hydroxybutryric and is
only slightly sensitive to acetone (only when glycine is
present)
Not necessary to perform individual testing for
18
acetone and beta-hydroxybutyric acid
Page
@japanesejuan
Acetest tablets Reaction interference

 A type of chemical test for ketones performed when  Patients who undergo special diagnostic procedures (i.e.
there is severe ketosis and serial dilutions are done to CT scan guided with contrast) using dyes may produce
provide a more accurate amount on ketones an interfering red color in the alkaline medium
 Performed using a tablet which consists of: Phenolsulfonphthalein
o Sodium nitroprusside Bromsulphalein
o Glycine (for acetone and color enhancement)  Highly pigmented urine interfere with color reactions
o Disodium phosphate  Medications
o Lactose (provides better color differentiation) Levodopa (large amounts)
 Specimen that can be used with Acetest Sulfhydryl group drugs (atypical color reactions)
o Serum  Mercaptoethane sulfonate sodium
o Urine (MESNA)
o Other body fluids (ex. CSF, pleural, ascitic fluid,  Captopril
etc…)  Improperly timed readings
 Acetest tablets are hygroscopic  Improperly preserved specimens
 If the specimen is not absorbed in 30 seconds, a new Volatilization of acetone (False decrease or
tablet should be used negative)
 Sensitivity of the test is 5mg/dL acetoacetate (lower limit) Breakdown of acetoactic acid by bacteria (False
 Any pink, tan, or yellow color is ignored decrease or negative)
 Deterioration of nitroprusside reagent (both pad and
tablets)
Due to heat, moisture, or light
19
Page
@japanesejuan
20
Page
@japanesejuan
Blood
 Major causes of hematuria
a. Renal calculi
Introduction: b. Glomerular diseases
Blood can enter the urinary tract anywhere from the glomeruli c. Tumors
to the urethra or can be a contaminant. d. Trauma
Blood found in urine may be in the form of: e. Pyelonephritis
1. Intact (hematuria) red blood cells f. Exposure to toxic chemicals
2. Free (hemoglobinuria) products of red blood cells g. Anticoagulant therapy
Blood present in large quantities can be detected visually.  Urinalysis is frequently requested when patients
Hematuria produces a cloudy red specimen. present with certain signs and symptoms like:
Because any amount of blood greater than 5 cells/μL urine is a. Severe back pain
considered clinically significant, visual examination cannot be b. Severe abdominal pain
relied upon to detect the presence of blood.  Hematuria of nonpathologic significance is observed
Chemical tests for hemoglobin provide the most accurate following
means for determining presence of blood in urine because a. Strenuous exercise
microscopic analysis may appear negative because some b. menstruation
patients possibly have hemolytic disorders and/or lysis of red
blood cells in which free hemoglobin is produced. 2. Hemoglobinuria
Clinical Significance  Hemoglobinuria is the result of either
 The finding of a positive reagent strip test result for blood a. Lysis of red blood cells in the urinary tract
indicates (particularly in dilute alkaline urine)
Presence of red blood cells b. Intravascular hemolysis
Hemoglobin c. Subsequent filtering of hemoglobin through the
Myoglobin glomerulus
Each of which has its own clinical significance  Lysis of red blood cells usually present hematuria and
1. Hematuria hemoglobinuria
 Closely related to disorders of renal or genitourinary  Intravascular hemolysis does not show red cells in
21
origin urine
Page
 Bleeding which is the result of TRAUMA or damage to
the organs of these systems
@japanesejuan
 Under normal conditions, haptoglobin captures  Patients taking cholesterol – lowering statin
hemoglobin and forms complexes but when the medications also present with rhabdomyolysis as a
amount of free hemoglobin exceeds haptoglobin side effect
levels, hemoglobin appears in urine. As occurs in:  Heme portions of both Hemoglobin and Myoglobin
a. Hemolytic anemias are toxic to the renal tubules; therefore, high
b. Transfusion reactions concentrations will lead to acute renal failure.
c. Severe burns
d. Brown recluse spider bites
e. Infections
f. Strenuous exercise
 When large yellow – brown granulated renal tubular
epithelial cells or urine sediments are found in urine,
these are usually due to reabsorption of filtered
hemoglobin and are called FERRITIN and
HEMOSIDERIN
3. Myoglobinuria
 Myoglobin is a heme-containing protein found in
muscle tissue
 Reacts positively with the reagent strip
 Also gives urine a red-brown color
 Presence of myoglobin is suspected in patients with
rhabdomyolysis (muscle destruction)
a. Trauma
b. Crush syndromes
c. Prolonged coma
d. Convulsions
e. Muscle – wasting diseases
22
f. Alcoholism
g. Heroin abuse
Page
h. Extensive exertion

@japanesejuan
Hemoglobin vs Myoglobin
 The laboratory is uncommonly requested to differentiate
between the presence of hemoglobin and myoglobin in
a urine specimen
Myoglobin is more toxic to the renal tubules than
hemoglobin
 Reasons for differentiation:
Diagnosis
Predicting risk for renal failure
Treatment options
 Diagnosis of myoglobinuria is usually based on:
Patient history
Elevated CK (Creatinine kinase)
Elevated LDH (Lactic dehydrogenase)
 The appearance of patient’s plasma can also aid in the
differentiation (but of limited value)
Myoglobin – clear plasma (myoglobin is rapidly
cleared by kidneys)
Hemoglobin – red plasma (haptoglobin-
hemoglobin complex imparts a red color)
 Myoglobin in the urine must be at least 25 mg/dL before
red pigmentation can be visualized
 At concentrations 25 mg/dL or more, a precipitation test
called Blondheim Test may be performed to screen for
myoglobin.
23
Page
@japanesejuan
Blondheim Test procedure: Reagent strip reactions

- Add 2.8g of 80% ammonium sulfate to 5mL urine (freshly PRINCIPLE: PSEUDOPEROXIDASE ACTIVITY OF HEMOGLOBIN
voided) Hemoglobin peroxidase catalyzing a reaction between H2O2
o Hemoglobin is precipitated out of the solution and the chromogen TETRAMETHYLBENZIDINE to produce an
o Myoglobin stays dissolved in supernatant oxidized chromogen, which is blue-green (from yellow)
- Mix
- Allow the specimen to sit for 5mins.
- Centrifuge specimen mixture
- Test the supernatant using the blood reagent pad  Blood testing areas are incorporated with
Peroxide
Positive for Myoglobin: Supernatant tests positive on reagent Tetramethylbenzidine
strip and retains a red color Buffer
 Color charts for differentiation are provided
Positive for Hemoglobin: Red precipitate and supernatant tests Hematuria
negative on blood reagent strip Hemoglobinuria/ Myoglobinuria
 Free myoglobin/hemoglobin give a uniform color ranging
Principle of Blondheim Test: from a negative yellow to green to a strongly positive
Based on the fact that the larger hemoglobin molecules are green – blue appear on the pad
precipitated by the ammonium sulfate and myoglobin remains  Intact RBCs are lysed when they come in contact with
in the supernatant. the pad and liberating hemoglobin showing an isolated
*Clinically no longer useful because of latest testing procedures reaction which is a speckled/dotted/mottled pattern
NOTE:  Reagent strips can detect concentrations as low as 5
Myoglobin is not stable in very acidic urine and, if denatured, cells/μL
may precipitate with ammonium sulfate. Must be correlated with microscopic analysis
In cases where specimen processing is delayed, neutralization  Reporting of results:
and freezing it would be proper. NEGATIVE
Immunoassay procedures are available to measure serum Trace
24
myoglobin levels. Small / 1+
Page
Moderate / 2+
Large / 3+

@japanesejuan
Reaction Interference
 False – positive reactions may be seen in:
Menstruating women
Strong oxidizing reagents in specimen containers
Vegetable peroxidase
Bacterial enzymes (E. coli peroxidase)
 False – negative reactions
High ascorbic acid (25 mg/dL)
 Directly reacts with H2O2 and removes it
Can be minimized when an iodate – mesh or an
“iodate – scavenger pad” is used
High specific gravity
 Red cells crenate and do not lyse when
they come in contact with the pad
 Decreased reactivity of pad
Formalin used as preservative
Patient taking Captopril
High concentrations of nitrite (greater than
10mg/dL)
Failure to mix specimen properly
 Red cells settle to the bottom of the
specimen – ensure proper mixing
 If hemoglobin is present, supernatant urine
and uncentrifuged specimens will still react
with the test pad
25
Page
@japanesejuan
Bilirubin
form water – soluble bilirubin diglucuronide (conjugated
bilirubin)
Introduction: - Conjugated bilirubin directly passes through the bile
Bilirubin is an intensely orange – yellow pigment that when ducts and into the intestine
present in significant amount causes a characteristic coloration - In the intestine, intestinal bacteria reduce bilirubin to
of plasma and urine. urobilinogens
The principal source of bilirubin (85%) is hemoglobin released - Urobilinogens are oxidized and excreted in the feces in
from the breakdown of senescent red blood cells in the RES. the form of UROBILIN
Other sources come from RBC precursors from bone marrow
and other heme – containing proteins such as myoglobin and
cytochromes.
Presence of bilirubin in urine can provide an early indication of
liver disease. It is often detected long before the development
of jaundice.
Production of Bilirubin
- Under normal conditions, RBC life span is 120 days
- After 120 days, RBCs are sequestered in the spleen and
liver by phagocytic cells of the RES(reticuloendothelial)
- Liberated hemoglobin is broken down into its component
parts;
1. Iron (body reuses)
2. Protein (body reuses)
3. Protoporphyrin (converted to bilirubin by RES)
- Bilirubin (water - insoluble) is released into blood
circulation
Hemoglobin degradation
- Bilirubin binds with ALBUMIN and transported to the LIVER
26
- Bilirubin then undergoes conjugation with GLUCURONIC
NOTE: Kidneys cannot clear bilirubin bound to albumin (large
Page
ACID by the action of GLUCURONYL TRANSFERASE to
and water insoluble)

@japanesejuan
Clinical Significance  No urobilinogen enters the intestine –

 Disturbances in any aspect of bilirubin formation, hepatic negative urine urobilinogen
uptake, metabolism, storage or excretion can cause  Stool is acholic (pale white or tan)
bilirubin to appear in urine (bilirubinuria)  bilirubinuria and bilirubinemia is detected long before
 Healthy individual bilirubin in urine (0.02 mg/dL) Jaundice (blood bilirubin levels 2-3mg/dL) is seen
Increased bilirubin in urine indicates distruption or  Differentation of jaundice cases
an increase in hemoglobin catabolism Results be correlated with urobilinogen
 Three principal mechanisms of altered bilirubin
metabolism occur
Prehepatic
 Due to an increase breakdown of RBCs or
overproduction of heme to bilirubin
 Unconjugated bilirubin cannot pass
through kidney glomerulus
 Liver function is normal
 Urine bilirubin is negative; Urine Urobilinogen
high due to reabsorption from intestine
Hepatic
 Due to hepatocellular disorders or disease
 Conjugated bilirubin leaks and readily
passes glomerulus – positive urine bilirubin
 Urine urobilinogen depends on extent of
liver damage – normal or increased
Post hepatic obstruction
 Blockage of bile duct or biliary system
 Liver function is normal
 Overflow of conjugated bilirubin and
27
reverts backs into blood circulation and
cleared by kidneys – positive bilirubin
Page
@japanesejuan
Reagent Strip Reactions Chemical tests for bilirubin

 Routine testing for bilirubin using reagent strip uses the  Ictotest tablets are used for questionable results
diazo reaction or azo coupling reactions  Ictotest consist testing mats and tablets containing
 Bilirubin combines with a diazonium salt 1. P-nitrobenzene-diazonium-p-toluenesulfonate
2,4-dichloroaniline diazonium salt OR 2. SSA
2,6-dichlorobenzene-diazonium-tetrafluoroborate 3. Sodium carbonate
 Uses an acid medium to produce an azo dye 4. Boric acid
 Color ranges from light tan to beige or pink to violet  Less subject to interference
 Qualitative test results are reported as  Sensitive to 0.05 mg/dL or 0.10 mg/dL of bilirubinuria
NEGATIVE  sometimes requested to detect early stages of
1+ / Small liver disease
2+ / Moderate  Reagent strip has a lower sensitivity of 0.40 mg/dL
3+ / Large
 Most difficult to read amongst others
Pigments of other substances
Color overlapping
 Lower limit of detection is 0.5 mg/dL of conjugated
bilirubin
A 25 fold increase of urine bilirubin must be
present in order for the test to be positive
Reaction:
 The mat has special absorbent properties which allow
28
bilirubin to stick to the surface as urine is absorbed
Page
@japanesejuan
 If interfering substances are suspected, adding water  Free unconjugated bilirubin is less reactive
directly to the mat after urine has been added fixes the on reagent strips
problem. High concentrations of ascorbic acid (greater
POSITIVE REACTION: A blue to purple color appears on the mat than 25 mg/dL) and nitrite medications
when bilirubin is present  Combines with diazonium salt and prevents
NEGATIVE REACTION: Any other color reaction with bilirubin
*Interferences of Ictotest are same with reagent pad for bilirubin
since they share the same principle
Other Bilirubin Methods
1. Shake test
a. Performed when urine is beer – brown or dark
yellow – brown in color
b. Characteristic YELLOW foam appears when urine
is agitated or shaken
Reaction Interference
 False positive
Other urine pigments such as from
phenazopyridine compounds – thick pigment
chlorpromazine metabolites can react to
diazonium salts
Indican
Metabolites of Lodine (medication)
 False negative
Improperly preserved specimens (most frequent)
Photo-oxidized specimens
 When specimens are exposed to light and
bilirubin is converted to biliverdin (does not
29
react with diazo tests)
Hydrolysis of bilirubin diglucuronide (conjugated
Page
bilirubin)

@japanesejuan
Urobilinogen
 Increased urine urobilinogen (greater than 1 mg/dL) is
seen in:
Introduction: Liver disease
Urobilinogen is one of the products of bacterial reduction of  Liver impairment decreases the ability of
conjugated bilirubin. The other is Stercobilinogen, which cannot the liver to process urobilinogen
be reabsorbed and further reduced to UROBILIN, which is recirculated from the intestines
responsible for the characteristic color of stool.  Excess urobilinogen shows up in urine
Some of the urobilinogen is reabsorbed from the intestine into Hemolytic disorders
the blood, recirculates to the liver, and is excreted back into the  Jaundice due to excess unconjugated
intestine through the bile duct. bilirubin and leads to high conjugated
Urobilinogen appears in the urine because as it circulates in bilirubin entering the intestines
blood en route to the liver, it passes through the kidneys and is  Cycle goes on in which urobilinogen is
filtered by the glomerulus. Therefore, a small amount of reabsorbed
urobilinogen – less than 1mg/dL or Ehrlich unit – is normally  Liver is overworked and by time is unable to
found in the urine. process urobilinogen at a normal rate
 More urobilinogen will circulate and be
Clinical Significance presented to the kidneys for excretion
 Measurement of urine urobilinogen can be valuable in  Absence of urobilinogen in urine and feces is clinically
the detection of early liver disease significant
1% of nonhospitalized patients and 9% of Cannot be detected using reagent strip
hospitalized patients exhibit high results – owing to Represents bile duct obstruction
constipation Absence of urobilin also indicates bile duct
obstruction
30
Page
@japanesejuan
Reagent Strip Reactions and Interference Reaction Interference

 2 different urobilinogen reactions exist and are used by  Ehrlich reaction is subject to a variety of interferences
different manufacturers called Ehrlich – reactive compounds
Ehrlich’s aldehyde reaction False – positive reactions due to:
 Urobilinogen reacts with p- 1. Porphobilinogen (clinically significant)
dimethylaminobenzaldehyde (Ehrlich’s 2. Indicant
reagent) 3. P-aminosalicylic acid
 Colors produced are from light to dark pink 4. Sulfonamides
 Results are reported as Ehrlich units (EU) 5. Methyldopa
which are equal to mg/dL 6. Procaine
 Normal value: 0.2-1.0 EU 7. Chlorpromazine compounds
*Reagent strip cannot differentiate or screen for
presence of PBG
Sensitivity increases with temperature (RT)
 Highly pigmented urine cause atypical readings for both
Azo – coupling (diazo) reaction
methods
 Urobilinogen reacts with 4-
Urobilinogen is high after meals because of bile
methoxybenzene-diazonium-
salt excretion
tetrafluoroborate
 False – negative readings for both methods
 Colors produced range from white to pink
Occur in improperly preserved specimens
 More specific for urobilinogen compared to
 Urobilinogen is photo-oxidized to Urobilin
Ehrlich
Formalin used as preservative
 Results are in mg/dL
 High concentrations of nitrite interfere with azo –
coupling reactions
 REAGENT STRIPS CANNOT DETERMINE THE ABSENCE OF

UROBILINOGEN
31
Important indicator of biliary obstruction
Page
@japanesejuan
Chemical Tests for Urobilinogen and other Ehrlich – reactive 3. Watson – Schwartz Differentiation Test
substances  The classic test for differentiating PBG, urobilinogen and
Introduction: Ehrlich – reactive compounds
Urobilinogen or other Ehrlich – reactive tests were not done Procedure:
before because they were time consuming and nonspecific. Tube 1 Tube 2
When necessary, the following 3 were performed. 2mL urine 2mL urine
1. Ehrlich Tube Test 2mL chloroform 2mL butanol
 Normally, addition of Ehrlich reagent to urine produces a 4mL sodium acetate 4mL sodium acetate
cherry red color and adding sodium acetate enhances  Tube 1
color reaction (when urobilinogen is present) Chloroform will extract UROBILINOGEN producing a
 Using the Ehrlich Tube method, one part Ehrlich reagent is colorless URINE TOP layer and a red CHLOROFORM
added to 10 parts urine. Tube is mixed and examined for red BOTTOM
color. PBG nor other ERC are soluble in chloroform
 This test is subject to false – positive results when  Tube 2
porphobilinogen and Ehrlich – reactive compounds were Butanol will extract both UROBILINOGEN and ERCs
present. producing a red BUTANOL TOP layer and a colorless
bottom urine layer if PBG is present
2. Hoesch Screening Test for PBG PBG is not soluble in butanol
 Rapid screening for urinary PBG  Before reporting the test as positive for both substances, an
 2 drops of urine added to 2mL Hoesch reagent (Ehrlich’s additional chloroform extraction should be performed on
reagent dissolved in 6M HCl) the red urine (upper) layer in Tube 1
 6M HCl inhibits urobilinogen To make sure it’s not due to an excess of urobilinogen
POSITIVE TEST: Top solution shows red color
 The test detects approximately 2mg/dL of PBG
 False positive tests
Methyldopa (high concentrations)
Indican (high concentrations)
32
Highly pigmented urine
Page
@japanesejuan
33
Page
@japanesejuan
NITRITE
 The nitrite test is a valuable test for detecting initial
bladder infection (cystitis) because most patients are
Introduction and Clinical Significance: asymptomatic and that would not lead the physician to
 The reagent pad for nitrite provides a rapid screening order a urine culture.
test for the presence of urinary tract infections (UTI)
 UTI can involve the bladder (cystitis), the renal pelvis and PYELONEPHRITIS, a complication of cystitis, is the inflammatory
tubules (pyelonephritis), or both process of the kidney and adjacent renal pelvis.
 2 possible routes for UTI: Pyelonephritis can lead to:
1. Movement of bacteria up the urethra into the bladder 1. Renal tissue damage
(ascending infection) 2. Impairment of renal function
2. Movement of bacteria from the bloodstream into the 3. Hypertension
kidneys and urinary tract 4. Septicemia
 Most common infecting microorganisms The nitrite test can also be used to evaluate:
1. Escherichia coli 1. Success of antibiotic therapy
2. Proteus species 2. Screen people who have recurring infections
3. Enterobacter species 3. Diabetic patients
4. Klebsiella species 4. Pregnant women (high risk for UTI)
 UTI is 8 times more common in females than in males Many laboratories use the nitrite test in conjunction with the
 UTI can begin as the result of urinary obstruction leukocyte esterase test to determine necessity of performing a
1. Tumor urine culture.
2. Bladder dysfunction
3. Urine stasis
 The test is designed to detect cases in which the need for
a culture may not be apparent but not to replace it as
the primary test for diagnosing and monitoring bacterial
infections.
 Most UTI cases start in the bladder as a result of external
34
contamination and move upward to the tubules, renal
Page
pelvis and kidney. (ascending infection)

@japanesejuan
Reagent Strip Reactions Reaction Interference

PRINCIPLE: GREISS REACTION  Several major factors influence reliability of nitrite test
 Chemical basis of the nitrite test is the ability of certain  Tests with negative results in the presence of vaguely
bacteria to reduce nitrate (from diet) into nitrite (not suspicious clinical symptoms should always be thoroughly
normally found in urine) investigated
 Bacteria must produce nitrate reductase to convert  The following are major considerations when interpreting
nitrate the nitrite test.
 Nitrite is detected using Greiss reaction 1. Bacteria that lack the enzyme reductase
Nitrite, at an acidic pH, reacts with an aromatic a. Cannot reduce nitrate to nitrite
amine (para-arsanilic acid or sulfanilamide) to b. Enzyme found in some gram – negative bacteria
form a diazonium compound that then reacts with (Enterobacteriaceae)
tetrahydrobenzoquinolin compounds (azo c. Most bacteria that cause UTI are gram – negative
coupling reaction )to produce a pink – colored d. Gram – positive and yeast also cause infection but
azo dye do not reduce nitrate to nitrite
2. Bacteria must remain long enough in urine
a. Nitrite test should be performed on first morning
urine
b. Urine that remained in the bladder for at least 4
hours is another alternative for first morning urine
 To prevent false – positive reactions by externally 3. Enough nitrate diet
contaminated specimens a. Green vegetables are a good source of nitrate
Test sensitivity is standardized to 100,000 b. This is seldom a problem
organisms/mL c. False – negative if nitrate is not enough
 The test does not measure degree of bacteriuria 4. Further reduction
 Any shade of pink is considered to represent a clinically a. Nitrate to nitrite to nitrogen
significant amount of bacteria b. Occurs when large amounts of bacteria present
Positive Test: Pink reagent pad
35
Negative Test: White reagent pad
Page
@japanesejuan
5. Miscellaneous
a. When patient is already under antibiotic therapy
i. Antibiotics inhibit bacteria action of
reduction
b. Large quantities of ascorbic acid
i. Interferes with diazo reaction
c. High specific gravity decreases test sensitivity
d. All give false – negative results
36
Page
@japanesejuan
Leukocyte Esterase (LE)

 Lymphocytes are not detected using LE because they
are agranular
Introduction:  Esterases are also present in Trichomonas
Prior to the development of LE, urinary leukocytes were  RBCs, bacteria, and renal tissue cells do not contain
detected only by microscopy using urine sediments. esterases
Microscopy can be subject to variation depending on the  A positive LE test result is most frequently accompanied
method used to prepare the sediment and the technical by the presence of bacteria
personnel examining the sediment. Bacteria may or may not produce a positive nitrite
Chemical testing for leukocytes in urine standardized the means reaction
for detection.  Infections caused by the following cause leukocyturia
The test is not designed to measure the concentration of without bacteriuria
leukocytes – quantitation should be done by microscopy Trichomonas
An advantage in using LE is that it detects the presence of Chlamydia
leukocytes that have been lysed, particularly in dilute, Yeast infection (Moniliasis)
hypotonic, alkaline urine, in which would not appear in Inflammation of renal tissue (interstitial nephritis)
microscopy.  Screening urine specimens using LE and nitrite determine
the necessity of performing urine cultures
Clinical Significance This can be a cost – effective measure
 Normal value: 0 – 2 /hpf up to 0 – 5 / hpf or 0-8/hpf  LE test has more reliability in the practice of medical
 Women tend to have higher numbers than men as a diagnosis than nitrite
result of vaginal contamination
 Increased urinary leukocyte is an indicator of UTI
 The test detects the presence of ESTERASE in azurophilic
granules of granulocytic white blood cells (in cytoplasm)
Neutrophils
Eosinophils
Basophils
37
 Monocytes and macrophages have granules as well,
Page
although they are not entirely granulocytic

@japanesejuan
Reagent strip reaction Reaction Interference

PRINCIPLE: LEUKOCYTE ESTERASE  False – positive reactions
 The action of LE to catalyze the hydrolysis of an acid Formalin in the collection container
ester embedded on the reagent pad to produce an Strong oxidizing reagents
aromatic compound and acid Phenazophyridine
 The aromatic compound then combines with a Beets
diazonium salt present on the pad to produce a purple  Atypical color reactions
dye Highly pigmented urine
Nitrofurantoin
 False – negative reactions
High concentration of protein (greater than 500
mg/dL)
 LE reaction requires the longest time of all the reagent High glucose (greater than 3g/dL)
strip reactions Oxalic acid
2 minutes or 120 seconds Ascorbic acid (combines with diazonium salt)
 Reactions are reported as High specific gravity
NEGATIVE  Leukocytes crenate and prevent esterase
TRACE release
SMALL / 1+ Strong oxidizing agents
MODERATE / 2+  Interfere with reaction pH
LARGE / 3+  Antibiotics (decreased sensitivity)
 Trace results may not be significant Gentamicin
Repeat test with a fresh specimen Cephalosporins
 Advantages of the LE screening test  Cephalexin
Detect the presence of intact and lysed WBCs  Cephalothin
Serve as a screening for WBC that is independent Tetracycline
of procedural variations for sediment preparation
38
 The LE Test detects 10 to 25 WBCs/μL
Page
@japanesejuan
39
Page
@japanesejuan
Specific Gravity
Reagent strip reaction
PRINCIPLE:
Introduction: CHANGE IN pKa (dissociation constant) OF A POLYELECTROLYTE
Specific gravity is a physical property of urine and an expression IN AN ALKALINE MEDIUM
of solute concentration.
The ultrafiltrate that enters the Bowman’s space of the glomeruli  The polyelectrolyte ionizes, releasing hydrogen ions in
has the same SG as protein – free plasma (1.010) – isosthenuria proportion to the number of ions in the solution
As the ultrafiltrate passes through the nephrons, solutes and  The higher the concentration of urine, the more
water are selectively absorbed and secreted thus increasing or hydrogen ions are released, thereby lowering the pH
decreasing SG.  Incorporation of the indicator bromthymol blue on the
Normal SG is from 1.002 to 1.035. Values greater or lesser than reagent pad measures the change in pH.
these require further investigation.  As the specific gravity increases, the indicator changes
SG that is 1.000 or 1.040 is physiologically impossible. from blue (1.000 [alkaline]), through shades of green to
The addition of SG to the test strip has eliminated a time – yellow (1.030 [acid])
consuming step in routine urinalysis and has provided a  Readings can be made in 0.005 intervals by careful
convenient method for routine screening. comparison with the color chart.
Osmometry and Refractometry should never be replaced for
fluid monitoring as these are more accurate compared to
reagent strip testing for SG.
40
Page
@japanesejuan
Reaction Interference Reasons for a 1.000 SG:

 Measures only ionic solutes 1. Specimen might not be urine
Thereby, eliminating interference by large organic a. Check urea and creatinine. Urine always has urea
molecules such as: and creatinine while ordinary water does not.
 Urea 2. Check QC of reagent strip
 Glucose a. Strip might be outdated or deteriorated
 Radiographic contrast media Reasons for a 1.040 SG
 Plasma expanders 1. Mannitol IV
 Analysts should consider what type of method is used
when testing for SG The use of refractometry or osmometry usually resolves problems
 Elevated concentrations of protein slightly increase the with very high or low SG (counterchecking)
readings as a result of protein anions
 Specimens with a pH of 6.5 or higher have decreased
readings
Interference with the bromthymol blue indicator
Bromthymol blue reacts best with alkaline pH and
low specific gravity
Manufacturers recommend adding 0.005 to SG
when pH is 6.5 or higher
 Correction is performed when using
automated strip readers
41
Page
@japanesejuan
Overview
System Description Semi-automated urine chemistry analyzer
Tests Measured Leukocyte, Nitrite, Protein, Blood, Glucose, Ketone, Bilirubin,
Urobilinogen, pH, Specific Gravity, Creatinine,* and Protein-to-Creatinine
Ratio*
Automatic Measurement Urine color
Test Format Dry chemistry reagent strips
Test Measurement Color change measured by reflectance photometry
Dual readings at reactive and reference wavelengths
Automatically adjusts for urine color
Sample Clarity Results entered via keyboard or bar code reader
Automatic Checks - Identification and reporting for validated Siemens strip
(Auto-Checks) types
- Humidity exposure tested on every strip with leukocyte
pad
- Sample interferences, availability dependent upon strip
type**
Sources:
(CLINITEK Advantus® Analyzer) 1. Urinalysis and Body Fluids by Susan King Strasinger and
Marjorie Schaub Di Lorenzo 5th edition
Respond to demands for higher productivity and high quality
with the CLINITEK Advantus® Analyzer. Streamline your workflow 2. Urine and Body Fluid Analysis by Nancy A. Brunzel 3rd
with flexible operation. edition
Immediate start-up.
Automatic calibration.
Network ready.
A wide range of options.
Added QC features.
Flexible operation to meet your needs.
42
“You can fail at something you don’t want, so might as
well take a chance doing what you love.”
Page
~James Eugene Carrey

@japanesejuan

Chemical Examination

Transféré par

Informations du document

Titre original

Copyright

Formats disponibles

Partager ce document

Partager ou intégrer le document

Options de partage

Avez-vous trouvé ce document utile ?

Ce contenu est-il inapproprié ?

Droits d'auteur :

Formats disponibles

Chemical Examination

Transféré par

Droits d'auteur :

Formats disponibles

Chemical Examination of Urine

Introduction The color reactions are interpreted by comparing the color

By: Ken Kawakami, RMT, MLS(ASCPi)CM

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Tablet and Chemical Tests

 Care must be taken to prevent run over from the protein

Reagent strip reaction

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Normal urine: (Pre – renal proteinuria)

 Bence – Jones Protein (Renal Proteinuria)

By: Ken Kawakami, RMT, MLS(ASCPi)CM

NOTE:  Detection of microalbumin before needed a 24 –

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Reagent strip reactions

Reaction Interference/Limitation o Tolbutamide metabolites (Px history)

By: Ken Kawakami, RMT, MLS(ASCPi)CM

 Examples of hyperglycemia of non diabetic origin which Reagent strip reactions

GH (increased) glucose to produce gluconic acid and hydrogen peroxide.

The Benedict solution was developed in 1908 and IMPORTANT NOTE:

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Clinitest tablets are hygroscopic and should be stored

Chemical Tests for Ketone bodies:

Acetest tablets Reaction interference

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Blondheim Test procedure: Reagent strip reactions

By: Ken Kawakami, RMT, MLS(ASCPi)CM

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Clinical Significance  No urobilinogen enters the intestine –

Reagent Strip Reactions Chemical tests for bilirubin

 The mat has special absorbent properties which allow

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Reagent Strip Reactions and Interference Reaction Interference

 REAGENT STRIPS CANNOT DETERMINE THE ABSENCE OF

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Reagent Strip Reactions Reaction Interference

Leukocyte Esterase (LE)

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Reagent strip reaction Reaction Interference

Reaction Interference Reasons for a 1.000 SG:

By: Ken Kawakami, RMT, MLS(ASCPi)CM

Vous aimerez peut-être aussi