Title: Chemical Examination of Urine Part I: Introduction, Specific Gravity and pH
1Chemical Examination of UrinePart
IIntroduction, Specific Gravityand pH
- Ricki Otten MT(ASCP)SC
- uotten_at_unmc.edu
2Objectives
- Review the objectives on page 1 and 2
- of the lecture handout
- Objectives marked with will not be tested
over during student lab rotation
3Historical Perspective Urinalysis
- Physical examination of urine
- Odor
- Taste
- Color
- Clarity
4Historical Perspective
- Chemical examination of urine
- Limited reactions
- Required large volumes of urine
- Large volumes of reagent
- Performed in test tubes
- Time consuming and cumbersome
- Clinical usefulness was not realized
- Not routinely ordered
5Historical Perspective
- Microscopic examination of urine
- Not until invention of the microscope
- Then clinical usefulness realized
6Reagent Strip Testing
- Technology and necessity
- Chemical reactions miniaturized
- Required less urine
- Test results within minutes
- Easy to perform
- Increased test utilization
Brunzel, 2nd Ed, page 124
7Reagent Strip Testing
- Ideal qualitative screening tool
- Sensitive Low concentration of substances
- Negative result normal
- Specific Reacts with only one substance
- False negative and false positive
- Cost effective Relatively inexpensive tool that
- provides information about the health status
- of the patient
8Reagent Strip Testing
- Chemically impregnated absorbent
- pads attached to an inert plastic strip
- Each pad is a specific chemical reaction that
- takes place upon contact with urine
- Chemical reaction causes the color of the pad to
- change
- Color compared to a color chart for interpretation
9Reagent Strip Testing
- Qualitative or semi-quantitative results
- Concentration units (mg/dl)
- Negative, small, moderate large
- Negative, 1, 2, 3, 4
- Timing of chemical reactions is CRITICAL
- Shortest time requirement on one end of strip 30
sec - Longest time requirement on the other 2 min
10Reagent Strip Testing
- Principle of chemical reactions
- False negative reactions
- False positive reactions
- Color interferences
- Alternative testing used to confirm results that
you may think are invalid due to - Interfering substance
- Color interference (called color masking)
11Care and Storage (pg 4)
Reading assignment Textbook, chapter 7 Page
124-130
Confirmatory Testing (pg 6)
12Confirmatory Testing
- Alternative testing establishes the correctness
or accuracy of another procedure - Often used when urine is highly pigmented
- Bilirubin reagent strip ? ictotest
13Confirmatory Testing
- Characteristics
- Differ in sensitivity
- Ictotest vs Bilirubin reagent strip
- Differ in specificity
- SSA vs Protein reagent strip
- Clinitest vs Glucose reagent strip
- Differ in methodology/reaction
Ideallywant all 3
14Differ in Specificity
- Clinitest reacts with all reducing
- substances
- Glucose reagent strip reacts with only one
reducing substance glucose
15- 10 reagent strip tests
- Specific gravity
- pH
- Protein
- Glucose
- Ketones
- Blood
- Bilirubin
- Urobilinogen
- Nitrite
- Leukocyte Esterase
- Purpose of the test
- What is normal
- What is abnormal
- Reaction
- Causes of invalid results
16Specific Gravity Purpose
- Evaluates the concentrating and diluting
- ability of the kidney
- Density is related to the amount of substances
(solutes) in solution - Increased density increased solute in solution
hypertonic urine concentrated urine - Decreased density decreased solute in solution
hypotonic urine dilute urine
17Specific Gravity Normal
- Normal 1.002 1.035
- Majority of urines 1.010 1.025
- Physiologically impossible 1.000
- gt1.040
- Dependent upon hydration status
18Specific Gravity Terms
- Isosthenuria
- Fixed at 1.010
- Renal tubules lost absorption and secreting
capability - Hypersthenuria
- Increased specific gravity
- Concentrated urine
- Hyposthenuria
- Decreased specific gravity
- Dilute urine
Sensitivity issues Pregnancy testing Urinary
tract infection
19Specific Gravity Methods
- Methods of measurement
- Reagent strip test indicates ionic solutes
- Refractometer indicates amount of total solutes
- Two functions of the kidney
- Maintain water balance
- Maintain electrolyte homeostasis
Performed by renal tubules through concentrating
and diluting reabsorbing and secreting water and
electrolytes (ionic)
20Specific Gravity Reaction
- Based on a change in the pKa of a polyelectrolyte
on the reagent pad - Increased ions in solution causes the
polyelectrolyte on the pad to produce free H - Free H cause a change in pH on the reagent pad
- Change in pH bromthymol blue indicator
21Specific Gravity Reaction
22Specific Gravity
- Sensitivity 1.000
- Specificity detects only ionic substances
- Radiographic dye
- Mannitol
- Glucose
Does not interfere
23pH Purpose
- Kidneys regulate bodys acid-base
- balance by selective handling of H and HCO3-
- Urine pH reflects acid-base status of body
- Treatment protocol may require urine pH be
maintained at a specific pH - (Aids in identification of crystals (microscope))
24pH Normal
- Normal ranges from 4.5 8.0
- First morning void acidic
- Physiologically impossible lt4.5
- gt8.0
- Urine not handled properly
- Old urine
- Treatment induced
25pH Interpretation
- Made in conjunction with
- Acid-base status
- Renal function
- Presence of infection in urinary tract
- Diet high protein, low protein
- Medications
- Age of urine sample
26pH Abnormal
- Acid
- Respiratory acidosis
- High protein diet
- Starvation
- UTI
- Alkaline
- Respiratory alkalosis
- Vegetarian diet
- Renal tubular acidosis
- UTI
27pH Reaction
- Double indicator system
- Methyl red
- Bromthymol blue
- Amount of free H influences acidity of urine and
cause pH indicator to change color
Needed to measure the wide pH range acid to
alkaline
28pH
- Invalid test results due to
- Improper handling of urine sample
- Contamination of urine vessel prior to collection
- Run-over phenomenon
29Protein Purpose
- Normal kidneys secrete LITTLE protein
- lt15 mg/dl (or lt150 mg/24 hours)
- The protein that is found in urine comes from
- Bloodstream
- Urinary tract
- Proteinuria is an indicator of early renal
disease - Proteinuria also caused by non-renal disease
-
30Renal Cause of Proteinuria
- Glomerular damage
- Most serious cause of proteinuria
- Most common cause of proteinuria
- Glomerulonephritis
- Nephrotic Syndrome
- Tubular dysfunction
- Reabsorption capability decreased
- Toxin exposure, inherited disorder
- Fancons syndrome heavy metal poisoning
31Classification of Proteinuria
- Functional
- Orthostatic (postural)
- Transient
- Pathologic
- Pre-renal (overflow)
- Renal glomerular
- Renal tubular
- Post-renal
32Protein Methods
- Reagent strip test
- SSA test
- Foam test
- Micro-albumin test
33Protein Reagent Strip
- The reagent pad is held at a
- constant pH of 3 by a buffer
- Proteins (anions) in solution cause an
- indicator dye to release H causing a color
- change
- Protein error of indicators
34Protein Reagent Strip
- Sensitivity 10-25 mg/dl
- Specificity reacts with albumin
- False positive highly alkaline urine (pH gt 8.0)
- False negative
- Dilute urine
- Presence of other proteins
- (Tamm-Horsfall, globulins, myoglobin,
- free light chains, hemoglobin)
35Protein SSA (Extons Test)
- Sulfosalicylic Acid (SSA) Precipitation Test
- Acid will precipitate proteins out of solution
causing the solution to become cloudy - Amount of cloudiness is related to the amount of
protein present