CHEMICAL EXAM Flashcards
Ph
Random and fasting: Normal values
Random 4.5-8
Fasting 5.5-6.5
Determine Acid base disorder
Ph
Acid Urine: Diseases
Emphysema
DM
Starvation
Diarrhea
Dehydration
Increase Protein
Alkaline Urine
Hyperventilation
Vomiting
Urease
Vegetables
Old Specimen
Principle of PH and Reading Time
Double Indicator System
Interferences
Run over from other pads
Old Specimen
Reagent
Methyl red ——– Brothymol Blue
This parameter is an indicator foe early renal disease
Protein
Normal Values of Protein
<10mg/dl or 100mg/24hrs
other urinary proteins
Uromodulin
Urokinase
IgA
Stages Of Proteinuria
Pre
Renal
Post
Stage of Proteinuria:
-Increase Plasma Protein, excess is sent to the kidneys and cannot be reabsorb
-impaired reabsorptive capacity
-intravascular hemolysis
-inflammation/ septicemia
- bence-jones protein
Pre renal proteinuria
Bence-jones protein
IGs proliferate = impaired reabsorptive capacity
Coagulates at 40° to 60°C & dissolves at 100°C
Methods: Electrophoresis, Immunofixation, Immunoassays
Stages of Proteinuria:
Renal Proteinuria
may be caused by glomerular or tubular damage
Glomerular proteinuria
Toxic substance, L.E, Streptococcal infection
Tubular proteinuria
Toxic substance, Viral infection, Fanconi syndrome
Orthostatic (postural) Proteinuria
vertical posture
Stage of Proteinuria:
Post-renal Proteinuria
from other parts (Ureter, bladder, urethra)
Causes of post renal proteinuria
Bacteria, spermatozoa
Lower UTI
Principle of Protein and Reading time
Protein error of indicators, 60 seconds
Protein:
Specific to ____ , with buffer “____”
Albumin, 3.0
Sensitivity and Reagent
Multistix
Chemstrip
Multistix:15-30 mg/dl Tetrabromphenol blue
Chemstrip = 6 mg/dL 3355 tetrachlorophenol 3456 Tetrabromosulfonphthalein
Interferences: False +
-highly buffered alkaline urine
-high SG
-antiseptics
-loss of buffer
-chlorhexidine
-phenazopyridine, detergents
Interferences of protein: False (-)
-prolonged contact with urine
-globulins,
-microalbumin,
-high salt concentration
Microalbimunuria
Presence of albumin in urine
above normal levels but below detectable levels
Microalbimunuria
Seen in type ___ & ____ DM
1 and 2
Microalbuminuria:
Albumin/24hrs _______mg is significantl
30-300mg
Micral test
EIA principle
Rgt: Gold-labelled anti-human albumin antibody-enzyme conjugate
Dip for 5 seconds
Wait 1 minute before reading
White to Red = Albumin
Sensitivity = 0-10mg/dL False (-)= dilute urine
Protein Interpretation:
Trace = <30mg/dL
+1 = 30mg/dL
+2 = 100mg/dL
+3 = 300mg/dL
+4=2000mg/dL
Correlate the Protein Chem Exam To:
Blood Nitrite WBC
Confirmatory Test for Proteins:
Sulfosalicylic acid Precipitation test
Sulfosalicylic acid Precipitation test:
Centrifuge at_____ speed
low
Sulfosalicylic acid Precipitation test
procedure:
3ml 3% SSA + 3ml Centrifuged urine= mix then check result
Sulfosalicylic acid Precipitation test
False (+):
Mucin
Uric Acid
Penicillin,
Tolbutamides,
Sulfonamides,
Radio contrast media,
Cephalosporins
Sulfosalicylic acid Precipitation test
False (-)
-highly buffered alkaline urine
Sulfosalicylic acid Precipitation test
Reporting:
Negative (<6 mg/dL)
Trace (6-30 mg/dL)
Positive then grading:
1+= (<30-100 mg/dL) Distinct turbidity, no granulation
2+=(100-200 mg/dL) Turbidity with granulation, no flocculation
3+=(200-400 mg/dL) with Turbidity, Granulation and Flocculation
4+= (>400 mg/dL) Clumps of protein
more sensitive for Microalbuminuria
Albumin-Creatinine Ratio:
Albumin-Creatinine Ratio:
Albumin cause a ________dye to change color (at constant pH)
sulfonephthalein
pseudoperoxidase activity of copper creatinine complex
Creatinine
Albumin-Creatinine Ratio:
_______ on pad is reduced and ______ is oxidized
-Peroxide
-Tetramethylbenzidine
Albumin-Creatinine Ratio:
PAD with H2O2 + TMB=
(+) = Red
(-) = White
Ketones: Types
Acetone (2%) = only detected if glycine is added
Acetoacetic acid (20%) = detected by RGT strip B-hydroxybutyric acid (78%)
Ketones: Principle and reading time
sodium nitrprusside, 40 seconds
Ketones RGT:
Multistix: Acetoacetate + Na* nitroprusside = Purple color
Chemstrip: (detects Acetone): Acetoacetate +Na+ nitroprusside + glycine = Purple color
Interferences
False (+)
-Phthalein dyes
-high pigmented red urine,
-Levodopa,
-Medications with Sulfhydryl drugs
Ketones Interferences:
False (-)
improperly preserved specimen
-Hygroscopic, for sever ketosis
- acetone and Acetoacetic acid
-Can be used with whole blood, plasma or serum
Acetest Tablets
Ace test Tablets Components
Na* nitroprusside
Glycine
Disodium phosphate
Lactose (for better color)
Ketone Test:
urine is acidified with H2SO4 and Ether and then FeCl is added to form violet-red color
Gerhardt’s test
-detects DMI
-It is reabsrobed in PCT
-Has a renal threshold: 160-180 mg/dl
-needs fasting or 2hrs after meals
Glucose
Principle for Glucose and reading time
glucose oxidase reduction and 30 seconds
Rgt strip for glucose contains:
Glucose oxidase
peroxidase
Chromogen
Buffer
Chromogen:
Multistix: k iodide= green to brown
Chemstrip: tetramethylbenzidine= yellow to green
Glucose Rgt Reactions:
- glucose + O2—- glucose oxidase— gluconic acid + h20
- H2O2+chromogen—–Peroxidase——>Oxidized colored chromogen +h20
Overflow Glucosuria:
Hyperglycemia
DM
Pancreatitis
GDM
Renal Glycosuria- failure of tubule to reabsorb
Fanconi Syndrome
Renal Tubular dysfucntion
Pregnancy
Glucose Interferences:
False (+)
contamination by oxidizing agents
detergents
Interferences
False (-)
Increase Ascorbic acid, SG and ketones
low temp
improper preservation
Copper reduction:
glucose + copper sulphate-Alkali & heat –> cuprous oxide
follows benedict’s principle, used on babies to detect galactose
Clinitest Tablets
Components of tablet:
-Copper sulfate = main reacting agent -NaOH= Heat production
-Na Citrate = Heat production
-Na carbonate = remove air interference
+Water (10 drops) + Urine (5 drops) = Heat then shake (Blue to orange red)
Reminders Clinitest:
-Wait 15 minutes before boiling
-observed to prevent “pass through” (>2g/dL sugar)
- avoid pass through use 2 drops urine
Strongly (+) Rgt Strip, (-) Clinitest
Possible contamination
(-) Rgt Strip, (+) Clinitest
Other reducing sugar
Glucose Interpretations:
Negative
Trace = 100 mg/dL
1+= 250 mg/dL
2+= 500 mg/dL
3+= 1000 mg/dL
4+=>2000 mg/dL
rapid test for uti
initial bladder infection
Nitrite
Nitrite Principle and Reading Time
Greiss reaction, 60 seconds
Ketones Aromatic amines
Multistix: p-arsanilic acid
Chemstrip: Sulfanilamide
Ketones Aromatic Compound
Multistix and Chemstrip: Hydroxytetrahydrobenzoquinolinol
Reactions
- Aromatic amine + NO2-acid—-> diazonium salt
- Diazonium salt + Aromatic compound-acid->
PINK azodye
Sensitivity
Multistix: 0.06-0.1mg/dL nitrite ion
Chemstrip: 0.05mg/dL nitrite ion
Correlated Parameters
Protein
WBC
Microscopic findings (bacteria)
Interferences:
False +
Improperly stored urine
Highly pigmented urine (produce color)(phenazopyridine, beets)
Interferences:
False -
-non-reductase containing bacteria
-insufficient contact time for bacteria
-Lack of Nitrate
-high SG
-increase bacteria= Nitrite →Nitrogen
-Antibiotics
-INCREASE ASCORBIC ACID (>25mg/dL)
-degradation product of hemoglobin
-Detects liver disease
Bilirubin
Bilirubin Principle and Reading Time:
Diazo Reaction, 30 seconds
Sensitivity
Chemstrip = 0.5 mg/dL
Multistix = 0.4-0.8 mg/dL
Bilirubin test Reactions:
- Bilirubin + Multistix: 2, 4 dichloroaniline diazonium salt (Chemstrip: 2, 6 dichlorobenzene diazonium tetrafluoroborate)
- Bilirubin glucuronide + diazonium salt-Acid-> Azobilirubin (azodye = TAN/PINK to VIOLET)
Interferences
False (+)
-Phenazopyridine and its metabolites (Pyridium),
-chlorpromazine,
-Indicans
False (-)
-Not fresh specimen (bilirubin is unstable),
-Increase Nitrite and ASCORBIC ACID (they combine with diazonium salt, >25 mg/dL)
-for double check
-hygroscopic
-protected from moisture or high humidity
ICTOTEST TABLETS
Ictotest Tablets Sensitivity
(0.05-0.10mg/dL sensitivity)
Ictotest Tablets Components
-p-nitrobenzene-diazonium p-toluene sulfonate
-SSA
-Na* Carbonate
-Boric acid
Ictotest Procedure and reactions/result
10 drops of urine on the Mat + drop of H2O + Tablet
= (+) Blue to purple
Other Test for Bilirubin
*Foam/Shake test: (+)=yellow foam persist
*Oxidation tests: (Gmelin’s, Fauchet’s)
*Smith Emerald green (+)
*Harrison’s spot = Blue-green
*Schlesinger = for urobilin (greenish-fluorescence)
Measure renal concentrating ability
Specific Gravity
Isosthenuria tells you?
CGN, renal failure
SG Principle and and Reading time
pka CHANGE OF POLYELECTROLYTES, 45 seconds
Reagents Of SG
Multistix: Polymethyl Vinyl ether/maleic anhydride
Chemstrip: Ethylene glycol diamino ethyl ether tetra-acetic acid
SG:
More H+?
Yellow-Green (Acid pH) = High Specific gravity
SG:
Less H*?
Blue-Green (Alkaline pH) = Low Specific gravity
SG Interferences:
False (+) = increase protein;
False (-) = Alkaline urine (>6.5)
SG Interpretation:
1.001-1.009
1.010-1.015
1.016-1.035
Dilute
Average
Concentrated
Hypersthenuria:
Dehydration
Diabetes mellitus
Proteinuria
Lipiduria
Hyposthenuria
Diabetes Insipidus
Is <1mg/dl urobilinogen is normal in urine?
Yes
Conversion:
Urobilinogen (1.0mg/dL) to Ehrlich Unit
1 Ehrlich Unit
Urobilinogen Principle And Reding time
EHRLICH REACTION , 60 seconds
Reagents Reactions:
Multistix:
Urobilinogen + pdimethylaminobenzaldehyde-acid->RED color (light to dark pink) in Ehrlich units
Interferences Urobilinogen:
False (+)
Porphobilinogen
p-aminosalycyclic acid
indican
sulphonamides
methyldopa
procaine
chlorpromazine
highly pigmented urine
False (-)
Old specimen
Formalin preservation
Chemstrip Rgt Urobilinogen
4-methyloxybenzene-diazonium-tetrafluoroborate-acid
Resulting to: White to pink
Ehrlich reagent + Urine + Na* acetate = Cherry red color
Ehrlich tube test
Detectable for Ehrlich tube test?
Porphobilinogen
Ehrlich reactive substance
It is a Rapid test for Porphobilinogen
Hoesch Screening test
Hoesch Screening test:
Procedure and Result
2 gtts urine + 2ml hoesch rgt= + red (porphibilinogen)
Hoesch Screening test:
Can detect as low as ______concentration of PBG
2mg/dL
Detection Of this test Represents Hematuria and Hemoglobinuria
Blood
Blood Parameters should correlated with?
Protein
Microscopic Findings
Principle Of Blood and Reading Time
Pseudoperoxidase activity of hemoglobin, 60 seconds
Reagents Of Bilirubin:
Multistix: Diisopropylbenzene dehydroperoxide Tetramethyl-Benzidine (TMB)
Chemstrip: 2, 5 dimethyl 2, 5 dihydroperoxyhexane TMB
Interferences Blood:
False (+) = Strong Oxidizing agents, Bacterial peroxidase, Menstruation
False (-)= high SG (crenated RBC), Formalin, Captopril, high Nitrite, ASCORBIC ACID (>9 mg/dL), UNMIXED specimen
Ammonium Sulfate Method:
precipitated (red precipitate) by Ammonium sulfate → supernatant read by reagent strip is (-) blood
Hemoglobinuria
Ammonium Sulfate Method:
Red supernatant read by
Rgt strip (+) blood