[TRANS] AUBF: URINALYSIS III

KETONES Reporting
• Represents 3 intermediate products of fat metabolism
o Acetones (2%)
o Acetoacetic Acid (20%) Quantitative Semi-quantitative
o β-hydroxybutyrate (78%) Negative - -
Trace - 5 mg/dL
• Metabolized fat is broken down to CO2 and H2O so it is not
Small 1+ 15 mg/dL
normally seen in urine
Moderate 2+ 40 mg/dL
• Ketones are seen in urine when the CHO is compromised
Large 3+ 80-160 mg/dL
because body stores of fats are metabolized to supply
energy.
ROTHERA’S TEST
CLINICAL SIGNIFICANCE
• Rothera’s Reagent:
• Clinical reasons for fat metabolism
o Inability to metabolize CHO (Diabetes mellitus) 𝑺𝒐𝒅𝒊𝒖𝒎 𝒏𝒊𝒕𝒓𝒐𝒑𝒓𝒖𝒔𝒔𝒊𝒅𝒆 𝑵𝒂𝟐 [𝑭𝒆(𝑪𝑵)𝟓 𝑵𝑶] ∙ 𝟐𝑯𝟐 𝑶
o Increased loss of CHO (vomiting) + 𝒃𝒖𝒇𝒆𝒓𝒓𝒆𝒅 𝒂𝒎𝒎𝒐𝒏𝒊𝒖𝒎 𝒔𝒖𝒍𝒇𝒂𝒕𝒆 𝒔𝒐𝒍𝒖𝒕𝒊𝒐𝒏
o Inadequate intake of CHO (starvation, malnutrition)
• Testing for ketones is valuable in management and
monitoring of Type I DM Combustion
• Ketosis leads to
o Electrolyte imbalance 𝑺𝒐𝒅𝒊𝒖𝒎 𝒏𝒊𝒕𝒓𝒐𝒇𝒆𝒓𝒓𝒊𝒄𝒚𝒂𝒏𝒊𝒅𝒆 𝑵𝒂𝟐[𝑭𝒆(𝑪𝑵)𝟔 ]
o Dehydration + 𝑺𝒐𝒅𝒊𝒖𝒎 𝑵𝒊𝒕𝒓𝒊𝒕𝒆 𝑵𝒂𝑵𝑶𝟐
o Acidosis + 𝑭𝒆𝒓𝒓𝒊𝒄 𝑯𝒚𝒅𝒓𝒐𝒙𝒊𝒅𝒆 𝑭𝒆(𝑶𝑯)𝟑
o Diabetic coma
• 5 mL urine + 1 gram Rothera’s reagent + 1 mL NH 4OH
CLINICAL SIGNIFICANCE OF URINE KETONES
• POSITIVE RESULT: rose/red/purple colored ring at the
Mnemonic: ID MISS
interface of 2 layers within 1min 30sec
• Insulin dosage monitoring
• NEGATIVE RESULT: brown ring; red-purple color fades
• Diabetic acidosis
during the 1st 5 mins
• Malabsorption/Pancreatic disorders
• Inborn errors of AA metabolism • The width of the ring is roughly proportional to the
• Starvation concentration of the ketone bodies
• Strenuous exercise
Reporting

REAGENT STRIP REACTIONS

Observation
• Use the sodium nitroprusside (nitroferricyanide) Negative No ring or a brown ring
decomposition reaction to measure ketones Trace to + Faint pinkish purple ring appearing
• Acetoacetic acid in an alkaline medium reacts with the slowly
sodium nitroprusside to produce purple color ++ Narrow dark purple ring
+++ Wide dark purple ring appearing very
rapidly

REACTION INTERFERENCE
• Do not detect β-hydroxybutyrate
• Only slightly sensitive to acetone when glycine is present False-positive False-negative
Phthalein dyes Improperly preserved
• Reagent composition
Highly pigmented red urine specimens (volatilization of
o 7.1 % w/w sodium nitroprusside
Levodopa acetone; breakdown of
o 92.9 w/w buffer
Medication containing free acetoacetic acid)
• Read after 40 seconds
• NEGATIVE: tan-colored after 40 seconds sulfhydryl groups
• POSITIVE: pink to purple after 40 seconds

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 TRANS: URINALYSIS III

• Acetest Tablets Strenuous Cholesterol-

o Sodium nitroprusside exercise lowering statin
o Glycine medication
o Disodium phosphate
o Lactose (for better color differentiation)
REAGENT STRIP REACTIONS
• Hygroscopic (specimen not absorbed within 30 seconds---
use new tablet) • Use the peroxidase activity of hemoglobin to catalyze
• Report as negative, small, moderate, or large reaction between the heme component of both hemoglobin
and myoglobin and the chromogen tetramethylbenzidine to
BLOOD
produce oxidized chromogen, which has a green-blue color
• Any amount of blood >5 cells per µL urine is considered
clinically significant
• Hemoglobin: provides the most accurate means for
determining the presence of blood

CLINICAL SIGNIFICANCE • Presence of free hemoglobin/myoglobin

o Uniform color ranging from a negative yellow
Hematuria through green to a strongly positive green-blue
appears on the pad
• Disorders of renal or genitourinary origin in which bleeding • Intact red blood cells
is the result of trauma or damage to the organs of these o Are lysed when they come in contact with the pad,
systems and the liberated hemoglobin produces an isolated
reaction that results in a speckled pattern on the
pad.
Hemoglobinuria

• May result from the lysis of RBCs in the urinary tract,

particularly in dilute, alkaline urine
• May result from intravascular hemolysis (no RBCs seen)
and the subsequent filtering of hemoglobin through the
glomerulus
• When free hemoglobin present exceeds the haptoglobin
content, hemoglobin is available for glomerular filtration
• Reabsorption of filtered hemoglobin also results in the
appearance of large yellow brown granules of denatured
ferritin called hemosiderin in the renal tubular epithelial
cells in the urine sediment

Myoglobinuria

• A heme-containing protein found in muscle tissue, not only

reacts positively with the reagent strip test for blood but
also produces a clear red-brown urine
• Associated with rhabdomyolysis (seen in patients taking
cholesterol-lowering statin medications
• Heme portion of myoglobin is toxic to the renal tubules, and
high concentrations can cause acute renal failure

Hematuria Hemoglobinuria Myoglobinuria

Renal calculi Transfusion Muscular
reactions trauma/crush
syndromes
Glomerulonephriti Hemolytic Prolonged coma
s anemias
Pyelonephritis Severe burns Convulsions
Tumors Infections/malari Muscle-waiting
a diseases
Trauma Strenuous Alcoholism/overdos
exercise/ RBC e
trauma
Exposure to toxic Brown recluse Drug Abuse
chemicals spider bites
Anticoagulants Extensive exertion

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