AUBF311

URINE SCREENING FOR METABOLIC DISORDERS

OVERFLOW DISORDER
• Result from the disruption of a normal metabolic pathway that
causes increased plasma concentrations of the
nonmetabolized substances.

RENAL DISORDER
• caused by malfunctions in the tubular reabsorption
mechanism

INBORN ERROR OF METABOLISM

• Disruption of enzyme function → failure to inherit the gene to
produce a particular enzyme
• May nawalang enzyme

ABNORMAL METABOLIC CONSTITUENTS OR

CONDITIONS DETECTED IN THE ROUTINE
URINALYSIS

COLOR ODOR CRYSTAL

• Homogentesic • Phenylketonuria • Cystine
acid (black) • Maple syrup • Leucine
• Melanin (brown to urine disease • Tyrosine
•
PHENYLKETONURIA (PKU)
black) Isovaleric • Lesch-
• Indican (blue) acidemia • most well-known of the aminoacidurias
Nyhan
• Porphyrins • Cystinuria disease • if undetected, results in severe mental retardation
(pediatric urine)
(violet/pruple) • Cystinosis (moderate to
o Phenylpyruvic oligophrenia
• Homocystinuria few)
• Ivan Følling in 1934 (Norway)
• Peculiar mousy odor of urine
• Overflow disorder, amino disorder, newborn screening
MAJOR DISORDER OF PROTEIN AND
• Increased urinary ketoacids + phenylpyruvate due to
CARBOHYDRATE METABOLISM ACCOCIATED WITH disruption in the conversion of phenylalanine to tyrosine
ABNORMAL URINARY CONSTITUENTS CLASSFIED • caused by failure to inherit the gene to produce the enzyme
AS TO FUNCTIONAL DEFECT phenylalanine hydroxylase

OVERFLOW METABOLIC RENAL TEST FOR PKU

INHERITED
• Phenylketonuria • Infantile • Hartnup TEST COMMENT RESULT
• Tyrosinemia tyrosinemia disease Phenylalanine • Increased 2-6 weeks • normal
• Alkaptonuria • Melanuria • Cystinuria Blood Level prior to the urinary results is
• Maple syrup urine • Indicanuria excretion of lowered
disease • 5-Hydroxy- phenylpyruvic acid from 4
• Organic acidemias indoleacetic • Detected as early as 4 mg/dL to 2
• Cystinosis acid hours after birth mg/dL
• Porphyria • Porphyria
• Mucopolysacchari Phenylpyruvic • based upon the ferric • permanent
doses acid Urine Test chloride reaction blue-green
• Galactosemia performed by tube test. color
• Lesch-Nyhan
disease Guthrie’s • blood from a heelstick is • Bacterial
microbial absorbed into filter growth with
inhibition assay paper circles. The around the
blood-impregnated paper disks.
AMINO ACID DISORDERS
disks are placed on
culture media with
AMINO ACID DISORDRS WITH URINARY SCREENING Bacillus subtilis. If
TEST increased phenylalanine
• phenylketonuria (PKU) levels are present in the
• tyrosyluria blood, it will counteract
• alkaptonuria the action of beta-2-
• melanuria thienylalanine, an
• maple syrup urine disease inhibitor of B. subtilis
• organic acidemias that is present in the
• indicanuria media
• cystinuria
• cystinosis
1
BAUGAN, MA. JANINE A.

The information in this material is solely intended for educational purposes only and does not guarantee accuracy, completeness, or timeliness and without any
warranties implicated. This handout is not for sale.
AUBF311
TYROSYLURIA(URINE) /TYROSINEMIA(BLOOD) ALKAPTONURIA (ALKALI LOVER)
• Accumulation of excess tyrosine in the plasma • One of the six original inborn errors of metabolism
(tyrosinemia) producing urinary overflow described by Garrod in 1902
• Inherited or metabolic defect • Darkened urine after becoming alkaline from standing at
• Urine metabolites: room temperature.
o p-hydroxyphenylpyruvic acid • occurs from failure to inherit the gene to produce the enzyme
o p-hydroxyphenyllactic acid homogentisic acid oxidase
• urine may contain excess tyrosine • brown-stained or black-stained cloth diapers and reddish-
• transitory tyrosinemia→ (most common); seen in premature stained disposable diapers
infants
• → underdevelopment of the liver function to produce TEST FOR ALKAPTONURIA
enzyme for tyrosine metabolism
• Yellowish TEST COMMENT RESULT
FeCl Test • Screening test for transient deep
3 blue color in test
TYPE ENZYME DEF COMMENT metabolic disorder
TYPE 1 Fumarylacetoacetate Produces a generalized tube
hydrolase (FAH) renal tubular disorder
and progressive liver Clinitest • Test for reducing Yellow ppt
failure in infants soon substance
after birth urinary • Screening test observe for
TYPE 2 tyrosine Persons develop homogentisic • add alkali to freshly darkening of the
aminotransferase corneal erosion and acid Test voided urine color
lesions on the palms, (Ascorbic Acid
fingers, and soles of interferes the
the feet believed to be test)
caused by crystallization Ammoniacal • Addition of silver nitrate Black Urine
of tyrosine in the cells Silver Nitrate and ammonium
TYPE 3 p- result in mental Test hydroxide to urine
hydroxyphenylpyruvic retardation if dietary
acid dioxygenase restrictions of MAPLE SYRUP URINE DISEASE
phenylalanine and
• amino acids involved are leucine, isoleucine, and valine
tyrosine are not
• Failure to inherit the gene for the enzyme necessary to
implemented
produce oxidative decarboxylation of keto acids results in
their accumulation in the blood and urine
TEST FOR TYROYSLURIA/TYROSINEMIA o α-ketoisovaleric
o α-ketoisocaproic
TEST COMMENT RESULT o α-keto-β-methylvaleric
nitroso- • Nonspecific, reacts with • Orange-red • strong odor of urine resembling maple syrup
naphthol test compounds other than color
(urinary tyrosine and its TEST FOR MSUD
screening metabolites.
test) Reagent:
TEST COMMENT RESULT
• 2.63N nitric acid 2,4-dinitro- • urine screening test Yellow turbidity
• 21.5% sodium nitrite phenylhydrazin most frequently or ppt
• 1-nitroso-2-napthol e (DNPH) performed for keto acids
MS/MS • Used to confirm nitro- reaction Interference:
naphthol test • Large doses of
(confirmatory test) ampicillin
• a positive reagent strip
MELANURIA test result for ketones
• Increased urinary melanin → darkening of urine after produce a positive
exposure to air DNPH result
• 2 Metabolic pathways of converting tyrosine into
melanin: ORGANIC ACIDEMIAS
o Eumelanin Pathway - from brown to black
o Pheomelanin- from yellow to red POINT OF ISOVALERIC PRIONIC METHYLMALO
REFERENCE ACIDEMIA ACIDEMIA NIC ACIDEMIA
TEST FOR MELANURIA
Characteristic Sweaty feet
TEST COMMENT RESULT odor odor of urine
Ferric Chloride • Oxidation of chromogen • Gray or
Tube Test black ppt
sodium • Screening test • Red color Special Accumulation immediate
nitroprusside • Interference → add charateristic of precursor
test glacial Hac (reverts isovalerylglycin to
color greenish-black) e methylmal
• Acetone • Purple onic acid
• Creatinine • amber

2
BAUGAN, MA. JANINE A.

The information in this material is solely intended for educational purposes only and does not guarantee accuracy, completeness, or timeliness and without any
warranties implicated. This handout is not for sale.
AUBF311
Cause isovaleryl errors in the metabolic pathway
coenzyme A converting isoleucine, valine, CYSTINE DISORDERS
def. in the threonine, and methionine to
leucine succinyl coenzyme A POINT OF CYSTINURIA CYSTINOS HOMOCYSTINU
pathway REFEREN IS RIA
Test method MS/MS p-nitroaniline urine Test CE
(emerald green) Characteris Elevated amino crystalline increase in
tic acid cystine in deposits homocystine
urine of cystine throughout the
(+) cystine crystals in many body.
TRYPTOHAN DISORDERS in urine areas of
• Indole (normal) the body
Cause inability of the defect in defect in the
renal tubules to the methionine
reabsorb cystine lysosomal membranes
filtered by the membrane
glomerulus. s
Test and Cyanide- Positive (+) cyanide-
Result nitroprusside Test test results nitroprusside test
(red-purple color) for plus (+) silver-
reducing nitroprusside test
substances

PORPHYRIN DISORDERS

DISORDER COMMENT RESULT

Porphyria Collective term for disorders
of porphyrin metabolism
Porphyrinuria observation of a red or port Ehrlich reaction
wine color to the urine after (Watson-
exposure to air Schwartz) and
pale color in the sun fluorescence
Test
INDICANURIA MUCOPOLYSACCHARIDE
• increased amounts of tryptophan are converted to indole
(GLYCOSAMINOGLYCANS) DISORDERS
(Hartnup disease)
• Excess indole is reabsorbed in the intestine → circulated to
POINT OF HURLER HUNTER SANFILIPPO
liver → converted to indican →excreted in urine
REFERENCE SYNDROME SYNDROM SYNDROME
• Colorless when oxidized turns indigo blue E
• Blue diaper syndrome Characteristic skeletal structure is abnormal Mental
• FeCl3 Test →deep blue/violet color→ chloroform extraction Abnormality and there is severe mental retardation
retardation
5-HYDROXYINDOLEACETIC ACID mucopolysaccharides
• Serotonin → stimulation of smooth muscle accumulate in the cornea of the
• → produced from tryptophan by the argentaffin cells in the eye.
intestine
• 5-HIAA → degradation product (urine metabolite) Treatment Bone marrow transplants and gene replacement
• Foods rich in serotonin → therapy
o Bananas Test • acid-albumin (turbidity after 30 mins)
o Pineapples • cetyltrimethylammonium bromide (CTAB)
o Tomatoes turbidity tests (turbidity after 5 mins)
• Metachromatic staining (blue spot)
TEST FOR 5-HYDROXYINDOLEACETIC ACID
PURINE DISORDER (LESCH-NYHAN DISEASE)
TEST COMMENT RESULT
Nitroso- • Normal daily excretion = • Purple to CAUSE CHARACTERITICS TEST
naphthol Test 2-8mg black color Failure to • severe motor defects Microscopic →
Reagent: • >25mg/24h → (depending inherit the gene • mental retardation presence of
nitrous acid argentaffin cell tumors on 5-HIAA to produce • tendency toward self- increased uric
1-nitroso-2- level) the enzyme acid crystals in
destruction
naphthol Requirement: hypoxanthine • Gout pediatric urine
• 24h sample guanine specimens
• renal calculi
• Preservative: HCl or phosphoribosyl Uric acid
• Presence of uric acid
Boric Acid -transferase (moderate to
crystals resembling
• Strict diet orange sand in diapers many)
• Hold medication (bata)
(phenothiazines and • Gout (matanda)
acetanilids) for 72h prior
to sample collection
3
BAUGAN, MA. JANINE A.

The information in this material is solely intended for educational purposes only and does not guarantee accuracy, completeness, or timeliness and without any
warranties implicated. This handout is not for sale.
AUBF311
CARBOHYDRATE DISORDER

MELITURIA COMMENT TEST

pentosuria • one of Garrod’s original Test for reducing
six IEMs substance
(Clinitest)
galactosuria • deficiency in any of
(inability to three enzymes,
properly galactose-1-phosphate
metabolize uridyl transferase
galactose to (GALT), galactokinase
glucose) and UDP-galactose-4-
epimerase
Lactosuria • seen during pregnancy
and lactation
Fructosuria • parenteral feeding
• ingestion of large
amounts of fruit

4
BAUGAN, MA. JANINE A.

The information in this material is solely intended for educational purposes only and does not guarantee accuracy, completeness, or timeliness and without any
warranties implicated. This handout is not for sale.