Urine Metabolic Screening Test

battery of tests that is performed on urine specimens to detect the possibility of a metabolic
disorder
not specific and are used only as screening tests
1) Ferric Chloride Test:
Materials and Methods: 1 ml FeCl3 reagent+ 10 drops urine, shake and observe result
Condition/Substance

Result

Acetoacetic Acid

Red-brown

Alkaptonuria (homogentisic Acid)

Blue-green (transient)

p-Aminosalicylic acid

Purple-brown

Bilirubin

Blue-green

Histidinemia

Blue-gray to green

Lactic Acidosis

Gray

MSUD

Green to gray

Melanin

Gray ppt to black

Methionine Malabsorption

Purple to red-brown

Phenothiazines

Purple brown

Phenylketonuria

Blue green

Pyruvic acid

Deep yellow

Salicylates

Purple

Tyrosinemia

Green (fades rapidly)

Xanthurenic acid

Dark green to brown

2) Benedicts Test

test for the presence of monosaccharides

Glucose
Fructose
test for the presence of some disaccharides
Maltose
test for the presence of aldehydes
Benedicts reagent can be used to test for presence of glucose in urine
Indication of diabetes
Heating a Benedicts solution mixed with monosaccharides will produce a reddish-orange color
Benedicts Reagent
Contains blue copper(II) sulfate (CuSO4)
The copper oxide is insoluble in water and so it precipitates
Contains NaOH and tartaric acid
Color of the final solution ranges from green to brick red depending on how many copper(II) are
present

Methodology: 5-mL of Benedicts reagent in a test tube, Heat to boil, then add 8 drops of urine,
Boil again, after 2 minutes read the results.
No precipitate

Negative

Green

a trace

Yellow

Orange

++

Red

+++

Results noted as:

(-) --> BLUE
(+) --> GREEN to YELLOW
(++) --> YELLOW to BROWN
(+++) --> BROWN to ORANGE
(++++) --> ORANGE to RED
Benedicts reagent: Deep-blue alkaline solution of copper sulfate, sodium hydroxide, and tartaric
acid

Urinary Substances and Clinical Syndromes Associated with Reducing Substances

Reducing
Substance

Clinical State

Drugs

Ascorbic acid, chloral hydrate, tetracyclines,

sulfonamides, chloramphenicol

Fructose

Fructosemia, essential fructosuria, hereditary fructose

intolerance

Galactose

Galactosemia, classic and variant (galactokinase

deficiency)

Glucose

Diabetes mellitus, renal glycosuria, Fanconis

Syndrome, Wilsons Disease

Homogentisic
acid

Alkaptonuria

Lactose

Lactase deficiency, lactose intolerance, newborn

Phenolic
compound

Phenylketonurias, tyrosinosis

Xylose

Excessive fruit intake

Xylulose

Pentosuria

3) Cetyltrimethylammonium Bromide Test (CAB)

Turbidometric technique
Uses quaternary ammonium compounds e.g. CAB
Used for both qualitative and quantitative determination of urinary mucopolysaccharides and
glycosaminoglycans in various forms of mucopolysaccharidoses

Procedure: 5 ml of urine in a test tube, allowe to stand at room temperature, add 1 ml of CAB
reagent, Test tube observe for 30 minutes.

Negative : no turbidity observed

Positive : positive turbidity observed

4) Ninhydrin Test
Ninhydrin Reagent Solution:
Ninhydrin: 0.35g
ethanol or acetone/butanol
so-propanol: 100ml
Methodology: 1 ml Ninhydrin reagent + 3 drops of urine, Warm for 30 secs. In water bath.

Observe color:

Violet: alpha amino acid

bluish-purple solution: presence of amino acids in urine

yellow-orange: presence of proline

5) Nitroprusside Test
Used in the screening of cystinuria, homocystinuria and -mercaptolactate cysteine disulfiduria
Methodology: 5 mL of Urine add 5 drops of Conc. NH4OH (ammonium hydroxide) then Mix and
then add 2mL of 5% NaCN (Sodium cyanide) . Stand for 10 min, add 4 drops of Sodium
Nitroprusside. Mix and Observe Color Change

Positive Test Result

+ - pink
++ - pinkish
+++ - purple
++++ - dark purple