CHEMICAL EXAMINATION OF URINE (NON-REAGENT STRIP)

Jacinta B. Cruz’s Analysis of Urine and Other Body Fluids Workbook, First Edition

TEST REAGENTS/SPECIMEN OBSERVATIONS OTHER NOTES

QUALITATIVE ALBUMIN TESTS
Denaturation of protein
Reagent: by heat and precipitation
5-10% Acetic Acid
Heat and Acetic (+): Cloudy
Acetate – prevents
Acid Test ­ Cloudiness = ­ Albumin
Specimen: formation of soluble acid
First morning urine and alkali albuminates, and
phosphates precipitation
Negative: No turbidity, Circle visible in the
bottom of tube (<5 mg/dL)

Trace: Perceptible turbidity; Circle not

Exton’s Reagent:
Sulfosalicylic Acid (SSA)
Exton’s/ Sodium Sulfate
Sulfosalicylic Test
Specimen:
Freshly voided or random
visible in the bottom of tube; Can read
newsprint (5-20 mg/dL)
Cold precipitation
1+: Distinct turbidity without discrete technique
granulation; Cannot read newsprint (30 mg/dL) Irreversible technique
2+: Turbidity with granulation; No flocculation Equal volumes of urine
(100 mg/dL) and reagent

3+: Turbidity with granulation and flocculation

(300 mg/dL)

4+: Large clumps of precipitate or solid mass

(>500 mg/dL)
Heller’s Reagent:
Concentrate Nitric Acid (+): White ring at point of contact
Stratification of urine on
Heller’s Test ­ Ring Size = ­ Albumin
the nitric
Specimen:
Freshly voided or random Negative: No ring at point of contact
Robert’s Reagent: Trace: Barely perceptible ring against black
Magnesium sulfate 1+: Distinct ring at black, seen up the light
Nitric acid 2+: Definite ring, faint up the light Stratification of 3 mL
Robert’s Test
3+: Heavy ring, cloudiness up the light urine on 3 mL of reagent
Specimen: 4+: Think and dense ring, opaque up the light
Freshly voided or random

TEST REAGENTS/SPECIMEN OBSERVATIONS OTHER NOTES

QUANTITATIVE ALBUMIN TESTS
Esbach’s Reagent:
Picric Acid
Use Esbach’s tube
Citric Acid Height of coagulum form in grams%
Kwilecki’s “U” = Urine
(Grams albumin in 1000 mL urine)
Modification of 10 drops 10% FeCl3
10% Ferric Chloride
Esbach’s Method “R” = Reagent
Amount of albumin in grams% = Divide by 10
Water bath = 72 °C
Specimen:
24-hour urine
1+: No definite flocculation; distinguishable
cloud but not granular (0.01-0.03 gram%)

Reagent: 2+: No definite flocculation; distinguishable

3% SSA and granular cloud (0.04-0.1 gram%) 2.5 mL SSA + 3% SSA
Kingsbury and Clark
up to 10 mL mark
Method
Specimen 3+: Marked flocculation with dense cloud Stand for 10 minutes
24-hour urine (0.2-0.3 gram%)

4+: Heavy thick precipitate almost to boiling

solid (0.5 gram% or higher; 3 grams% solid)
 TEST REAGENTS/SPECIMEN OBSERVATIONS OTHER NOTES
QUALITATIVE SUGAR TESTS
Benedict’s Reagent: Glucose reduces copper
Copper sulfate salts to copper oxide
Sodium citrate
Sodium carbonate – 5-10 mL reagent
Benedict’s Test Negative: No change in color
alkalinity of solution + 8-10 drops urine
Trace: Green opacity, no precipitate
Specimen: Boiling water bath –
Freshly voided or random 2-3 minutes
1+: Green solution, yellow precipitate
Fehling’s Solution A:
Cupric Sulfate Glucose reduces copper
2+: Green to yellow solution,
salts to copper oxide
yellow precipitate
Fehling’s Solution B:
Rochelle’s Salt Equal parts of A and B
3+: Muddy orange solution,
(Potassium Sodium Dilute 2 to 3 times with
Fehling’s Test yellow precipitate
Tartrate) – prevents water then boil for few
precipitation of Cu(OH)2 seconds (Clear = Okay)
4+: Orange to brick red precipitate
Sodium Hydroxide
Boil until 5 mL urine by
Specimen: drop has been added
Freshly voided or random
Nylander’s Reagent:
Glucose reduces bismuth
Rochelle Salt (+): Black color
salts to metallic bismuth
10% NaOH or KOH – Trace: Brown color
alkalinity of solution Negative: No change in color
Nylander’s Test 5 mL urine + 0.5 mL
Bismuth Subnitrate
Nylander’s reagent
Black after cooling = substances other than
Specimen: sugar
Heat for 3-5 minutes
Freshly voided or random
Moore-Heller’s Reagent: Canary yellow: Less than or equal 1% sugar 2 parts urine + 1 part
10% KOH solution Wine yellow: 1-2% sugar reagent
Moore-Heller’s Test Cherry color: 2-3% sugar
Specimen: Rum color: 3-4% sugar Boil upper part for 2 to 3
Freshly voided or random Dark brown or black: >4% sugar minutes

TEST REAGENTS/SPECIMEN OBSERVATIONS OTHER NOTES

QUANTITATIVE SUGAR TESTS
Titration method
25 mL Benedict’s = 50 mg glucose
Benedict’s volumetric 25 mL Benedict’s + 5-10 grams
solution Number of mL urine used in titration/50 Sodium carbonate + Pinch of
Powdered pumice = Number of mg in one mL of urine Pumice
Benedict’s Test Sodium carbonate powder
Multiply by 100 = mg glucose/100 mL urine Heat until boiling
Specimen: Add urine drop by drop until
24-hour urine Divide latter by 1000 = grams glucose in trace of blue color
100 mL urine (%sugar) disappears
Read urine used in mL
Titration method

Dilute 2 mL Fehling’s with

Fehling’s Reagent
mL of urine used = 0.005 g glucose 4 mL distilled water
Fehling’s Test Heat to boiling
Specimen:
Multiply by 100 = mg glucose/100 mL urine
24-hour urine
Add 1 mL urine from graduated
pipet drop by drop until blue
color disappears
 TEST REAGENTS/SPECIMEN OBSERVATIONS OTHER NOTES
FRUCTOSE TESTS
Seliwanoff’s Reagent:
Resorcinol (+): Red
Formation of oxymethylfurfurol
Hydrochloric Acid (with addition of resorcin)
Seliwanoff’s Test
5 mL urine + 2 mL reagent, then heat
Specimen: Sediment is soluble in alcohol
First morning urine
Borchardt’s Reagent:
Formation of oxymethylfurfurol
25% Hydrochloric Acid
Resorcinol Crystals (+): Red
5 mL urine + 5 mL HCl + Resorcinol
(with addition of resorcin)
Crystals, then boil for one minute
Borchardt’s Test Solid NaOH or KOH
Acetic Acid-Ether (1:1) Confirmatory:
CONFIRMATORY
Yellow indicates fructose
Cool and alkalinize with NaOH or KOH +
Specimen:
3 mL acetic acid:ether mixture, then shake
First morning urine

TEST REAGENTS/SPECIMEN OBSERVATIONS OTHER NOTES

LACTOSE TEST
Rubner’s Reagent:
Lead acetate powder (+): Brick red solution 10 mL urine + 3 g lead acetate
Concentrated ammonium Filter off precipitate, heat filtrate for few
Rubner’s Method hydroxide (+): Clear supernatant with minutes = Yellowish-brown color
cherry red or copper-colored
Specimen: precipitate Add ammonium hydroxide, continue heat.
First morning urine

TEST REAGENTS/SPECIMEN OBSERVATIONS OTHER NOTES

PENTOSES TEST
Bial Orcinol Reagent: When heated with concentrated HCL,
Orcinol pentoses lose water and converted to
Hydrochloric Acid (+): Green solution furfurol
Bial Orcinol Method Ferric Chloride immediately
(with addition of orcinol) Boil 5 mL reagent
Specimen: Remove from flame and add no more than
First morning urine 1 mL urine drop by drop