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DR.MAHA
1. Chemical examination of urine
2. Normal & Abnormal Findings in urine strip
2
Clinical Significance of Chemical
Examination
1) Used to determine body processes such as
carbohydrate metabolism, liver or kidney
function.
2) Used to determine infection
3) Can be used to determine presence of
chemicals that can be found in urine (not
normal components)
Methods of Chemical Examination
 Manual reagent strips (dipstick).
 Automated
5
Dipstick
• A dipstick, usually a thin, plastic stick with
strips of chemicals on it, is coated with urine,
the chemical strips will change color if certain
substances are present or if certain levels are
above, or below, normal which can indicate the
presence of compounds like proteins, ketones,
hemoglobin, and nitrites, as well as harmful
pathogens. Dipstick urinalysis is convenient,
but false-positive and false-negative results
can occur.
6
7
Some chemicals that can be found in urine
(not normal components)
1. Protein
2. Ketones
3. PH
4. Blood
5. Bilirubin
6. Glucose
7. Leukocytes
8. Urobilinogen
9. Nitrite
10. Ascorbic Acid
Normal chemical Substances
oUrea :end product of protein metabolism
oUric acid :metabolite of purine breakdown
oCreatinine:associated with muscle metabolism of
creatine phosphate
oSulfates: mainly derived from sulfur containing amino
acids (cystine) increase e protein intake.
o Phosphate : important for buffering H+ in the
collecting duct
oChlorides : major extracellular anions & maintain
electrical neutrality
8
Manual Test procedure
① Wear gloves.
② Ensure the sample is in the correct container,fresh well-mixed uncentrifuged
③ Check the appearance ,colur of the sample and record results.
④ Ensure the strips have been stored properly & are in-date if reagent area are
discolored don’t use the strip
⑤ Remove the cap, take out strip and replace the cap on the bottle.
⑥ Using the appropriate reagent strip completely immerse all reagent areas into
the sample. Dip briefly and remove immediately(for no more than 1 second )
to avoid dissolving out the reagents.
⑦ While removing the strip, run the edge against the rim of the urine container
to remove excess urine.
⑧ Hold the strip in a horizontal position to prevent possible mixing of chemicals
from the adjacent areas.
⑨ Read the test results carefully at 60 seconds in a good light area , compare
test areas closely with the corresponding colour chart on the bottle label at
the specified time. Hold the strip close to the colour blocks and match
carefully.
Protein
• Principle :
tertrabromophenol-blue + Protein (albumin)
Change the color
• Detects primarily albumin
• Highly specific, but not sensitive
• Positive only when protein excretion > 300-500 mg/day
• Not good to detect microalbuminuria or immunoglobulin
light chains
• Albuminuria is unusually due to increased permeability in
the glomeruli. Positive results in acute and chronic kidney
disease, pre-eclampsia.

Acid
Glucose
Normal = negative
• Based on double sequential enzyme reaction
Glucose H2O2 + gluconic acid
H2O2+KI Chandge in colour
• Generally glucosuria does not occur until plasma
glucose exceeds renal threshold 180 mg/dL in DM

God

Pod
Ketones
Normal = negative or trace amounts
• Intermediate products of fat metabolism
• Detects acetoacetate only
nitroprusside +acetoacetate Muve purple colour
• Presence in the urine with glucose indicates DKA
• Presence in the urine indicates starvation ketosis, low
carb diet,fasting,vomiting and fever.
Alkaline
medium
Bilirubin
Normal = negative
• Bili + Diazotized dicholraniline change in color
• If increased conjugated bilirubin in urine, indicates liver
dysfunction or biliary obstruction.
• Yellow foam when sample is shake
Acidic
medium
Urobilinogen
• Based in Ehrlich reaction
• UB + P-dimethylaminobenzaldehyde change in color
• Bilirubin metabolized by bacteria in the intestines
to form urobilins
• Small 3-5% absorbed in intestine and appears in
urine
• Increased in hepatitis ,paracetamol overdose, late-
stage cirrhosis
14
PH
• Normal urine PH is acidic (4.5 -6)
• Double indicator : methyl red & bromothymol blue
• Acidic:
Physiologically is due to weak organic acids NaH2Po4,high protein
diet
Pathological: ketoacidosis,M&R Acidosis ,Renal tuberculosis
,Pyrexia,Phenylketonuria,Alkaptouria
Crystals : uric acid
• Alkaline:
Physiologically: Vegetarine diet
Pathological: UTI ammonia forming organism , RT Alkalosis
,Fanconi’s syndrom &Potassium depletion
Crystals: Phosphate
specific gravity
• Based on Pka
• Generally varies with osmolality, though presence of large
molecules in urine, such as glucose or radiocontrast media
can produce large changes in specific gravity with little
changes in osmolality
• High values can be found in dehydration.
• Low values found in high fluid intake. Diabetes insipidus;
chronic renal failure; hypercalcaemia; hypokalaemia.

Low ionic conc. In urine
BTB
Increase ionic conc. In urine
BLOOD
Normal = Negative
• Principle:
(Blood,HB,Myoglobin)+tetramethylbenzidine
Change in colour
• RBC are too large to pass through glomerulus so it’s
precence in urine may be due to menstrution or
kidney disorders; urinary tract disorders (e.g.
tumours, prostatic enlargement).
Peroxidase
Leucocytes
• Indoxyl ester Indoxyl (change in colour)
• Detect leukocyte esterase (pyuria) and nitrite
(enterobacteriaceae converts nitrate to nitrite)
• UTI
• Sterile pyuria: interstitial nephritis, renal
tuberculosis, and nephrolithiasis
18

Granulocyte
estrase
Nitrite
Griess’s Test:
NO3 NO2 + Sulfanilamide
NitrateReducing
Bacteria

Diazonium salt +N-Naphtol –ethylendiamonium
dihydrocholride(Pink colour )

Acidic PH
Diazonium salt
Sources of error
 Incorrect dipping of reagent strip.
 Incomplete wetting of strip.
 Incorrect storage of strips – always check manufacturers
instructions.
 Sample error –refrigerated sample must be allowed to return
to room temperature, non sterile containers; sample needs to
be fresh for best results.
 Contamination of the reagent pad by handling or non sterile
container.
 pH may be falsely elevated if the urine is stale.
 Some medication can affect some of the reagents on the strips
(e.g. cephalosporins; L-dopa; high levels of salicylates;
chlorhexadine; ferrous sulphate)
 Strips out of date.
 Vegetarians may have a urine pH >8.
DON’T
 Remove the desiccant from the reagent strip
bottle.
 Touch the test areas of the strip.
 Take out more strips than are required for
immediate use.

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4_5906744191475518021.pdf

  • 2. 1. Chemical examination of urine 2. Normal & Abnormal Findings in urine strip 2
  • 3. Clinical Significance of Chemical Examination 1) Used to determine body processes such as carbohydrate metabolism, liver or kidney function. 2) Used to determine infection 3) Can be used to determine presence of chemicals that can be found in urine (not normal components)
  • 4. Methods of Chemical Examination  Manual reagent strips (dipstick).  Automated
  • 5. 5
  • 6. Dipstick • A dipstick, usually a thin, plastic stick with strips of chemicals on it, is coated with urine, the chemical strips will change color if certain substances are present or if certain levels are above, or below, normal which can indicate the presence of compounds like proteins, ketones, hemoglobin, and nitrites, as well as harmful pathogens. Dipstick urinalysis is convenient, but false-positive and false-negative results can occur. 6
  • 7. 7 Some chemicals that can be found in urine (not normal components) 1. Protein 2. Ketones 3. PH 4. Blood 5. Bilirubin 6. Glucose 7. Leukocytes 8. Urobilinogen 9. Nitrite 10. Ascorbic Acid
  • 8. Normal chemical Substances oUrea :end product of protein metabolism oUric acid :metabolite of purine breakdown oCreatinine:associated with muscle metabolism of creatine phosphate oSulfates: mainly derived from sulfur containing amino acids (cystine) increase e protein intake. o Phosphate : important for buffering H+ in the collecting duct oChlorides : major extracellular anions & maintain electrical neutrality 8
  • 9. Manual Test procedure ① Wear gloves. ② Ensure the sample is in the correct container,fresh well-mixed uncentrifuged ③ Check the appearance ,colur of the sample and record results. ④ Ensure the strips have been stored properly & are in-date if reagent area are discolored don’t use the strip ⑤ Remove the cap, take out strip and replace the cap on the bottle. ⑥ Using the appropriate reagent strip completely immerse all reagent areas into the sample. Dip briefly and remove immediately(for no more than 1 second ) to avoid dissolving out the reagents. ⑦ While removing the strip, run the edge against the rim of the urine container to remove excess urine. ⑧ Hold the strip in a horizontal position to prevent possible mixing of chemicals from the adjacent areas. ⑨ Read the test results carefully at 60 seconds in a good light area , compare test areas closely with the corresponding colour chart on the bottle label at the specified time. Hold the strip close to the colour blocks and match carefully.
  • 10. Protein • Principle : tertrabromophenol-blue + Protein (albumin) Change the color • Detects primarily albumin • Highly specific, but not sensitive • Positive only when protein excretion > 300-500 mg/day • Not good to detect microalbuminuria or immunoglobulin light chains • Albuminuria is unusually due to increased permeability in the glomeruli. Positive results in acute and chronic kidney disease, pre-eclampsia.  Acid
  • 11. Glucose Normal = negative • Based on double sequential enzyme reaction Glucose H2O2 + gluconic acid H2O2+KI Chandge in colour • Generally glucosuria does not occur until plasma glucose exceeds renal threshold 180 mg/dL in DM  God  Pod
  • 12. Ketones Normal = negative or trace amounts • Intermediate products of fat metabolism • Detects acetoacetate only nitroprusside +acetoacetate Muve purple colour • Presence in the urine with glucose indicates DKA • Presence in the urine indicates starvation ketosis, low carb diet,fasting,vomiting and fever. Alkaline medium
  • 13. Bilirubin Normal = negative • Bili + Diazotized dicholraniline change in color • If increased conjugated bilirubin in urine, indicates liver dysfunction or biliary obstruction. • Yellow foam when sample is shake Acidic medium
  • 14. Urobilinogen • Based in Ehrlich reaction • UB + P-dimethylaminobenzaldehyde change in color • Bilirubin metabolized by bacteria in the intestines to form urobilins • Small 3-5% absorbed in intestine and appears in urine • Increased in hepatitis ,paracetamol overdose, late- stage cirrhosis 14
  • 15. PH • Normal urine PH is acidic (4.5 -6) • Double indicator : methyl red & bromothymol blue • Acidic: Physiologically is due to weak organic acids NaH2Po4,high protein diet Pathological: ketoacidosis,M&R Acidosis ,Renal tuberculosis ,Pyrexia,Phenylketonuria,Alkaptouria Crystals : uric acid • Alkaline: Physiologically: Vegetarine diet Pathological: UTI ammonia forming organism , RT Alkalosis ,Fanconi’s syndrom &Potassium depletion Crystals: Phosphate
  • 16. specific gravity • Based on Pka • Generally varies with osmolality, though presence of large molecules in urine, such as glucose or radiocontrast media can produce large changes in specific gravity with little changes in osmolality • High values can be found in dehydration. • Low values found in high fluid intake. Diabetes insipidus; chronic renal failure; hypercalcaemia; hypokalaemia.  Low ionic conc. In urine BTB Increase ionic conc. In urine
  • 17. BLOOD Normal = Negative • Principle: (Blood,HB,Myoglobin)+tetramethylbenzidine Change in colour • RBC are too large to pass through glomerulus so it’s precence in urine may be due to menstrution or kidney disorders; urinary tract disorders (e.g. tumours, prostatic enlargement). Peroxidase
  • 18. Leucocytes • Indoxyl ester Indoxyl (change in colour) • Detect leukocyte esterase (pyuria) and nitrite (enterobacteriaceae converts nitrate to nitrite) • UTI • Sterile pyuria: interstitial nephritis, renal tuberculosis, and nephrolithiasis 18  Granulocyte estrase
  • 19. Nitrite Griess’s Test: NO3 NO2 + Sulfanilamide NitrateReducing Bacteria  Diazonium salt +N-Naphtol –ethylendiamonium dihydrocholride(Pink colour )  Acidic PH Diazonium salt
  • 20. Sources of error  Incorrect dipping of reagent strip.  Incomplete wetting of strip.  Incorrect storage of strips – always check manufacturers instructions.  Sample error –refrigerated sample must be allowed to return to room temperature, non sterile containers; sample needs to be fresh for best results.  Contamination of the reagent pad by handling or non sterile container.  pH may be falsely elevated if the urine is stale.  Some medication can affect some of the reagents on the strips (e.g. cephalosporins; L-dopa; high levels of salicylates; chlorhexadine; ferrous sulphate)  Strips out of date.  Vegetarians may have a urine pH >8.
  • 21. DON’T  Remove the desiccant from the reagent strip bottle.  Touch the test areas of the strip.  Take out more strips than are required for immediate use.