NON-PROTEIN NITROGEN COMPOUNDS  Renal azotemia - caused primarily by diminished

DETERMINATION glomerular filtration as a consequence of acute or

NON-PROTEIN NITROGEN COMPOUNDS chronic renal disease.
 Used as indicators of diminished renal function  Post renal azotemia - the result of any type of
 Urea-the major NPN constituent and accounts for obstruction in which urea is reabsorbed into the
more than 75% of the total NPN excreted by the circulation.
body
 Most laboratories perform serum urea nitrogen METHODS FOR QUANTITATIVE DETERMINATION
measurements in conjunction with creatinine tests I. COLORIMETRIC METHOD
when tests for renal function are needed  Measured by condensation with diacetyl monoxime
NON-PROTEIN NITROGEN COMPOUNDS (DAM) in the presence of strong acid and an
Urea nitrogen/creatinine ratio: oxidizing agent
Serum urea nitrogen(mg/dL)  Diacetyl monoxime + urea ACID diazine (yellow
Serum creatinine (mg/dL) compound)
• Indication of lower ratio? Specimen: Serum or plasma
• Indication of high ratio with normal creatinine?
• Indication of high ratio with increased creatinine?
The chief component of the NPN material. in the blood
 A waste product of protein metabolism, which is
normally removed from the blood in the kidneys
 The amount is largely determined by the amount of
dietary protein and by the kidney's ability to excrete
urea
 Conversion factor of BUN to UREA-2.14

II. NESSLERIZATION METHOD

 Conversion of urea to ammonium carbonate
(NH4)2CO3
 The ammonia in (NH4)2CO3 is then measured by
having it react with Nessler's solution to produce a
CLINICAL SIGNIFICANCE yellow colored species
Uremia - a condition of abnormally high urea nitrogen III. UREASE METHOD
level in the blood UREASE
Azotemia - a significant increase in the plasma Urea + H₂O ------------>2NH4 + HCO3
concentrations of urea and creatinine The ammonia that is liberated in the reaction above may
be measured in a variety of ways:
1. The reaction may be coupled with a reaction that
drives NADH to NAD.
2. The conductivity of the ammonium ion may be
measured.
3. The Berthelot reaction may be used:
CLINICAL SIGNIFICANCE phenol nitroprusside
 Azotemia can result from, NH4+ NaOCl -----------------------------> indophenol
 Pre-renal azotemia - the result of poor perfusion of 4. The reaction may be coupled with a reaction that
the kidneys and therefore diminished glomerular produces H₂O₂.
filtration
REFERENCE VALUES  pH <5.57-uric acid formation increase
Urea Nitrogen, Serum
• Adult
7-18 mg/dL (2.5-6.4 mmol urea/L)
>60 years old 8-21 mg/dL (2.9-7.5 mmol urea/L)
Infant/child 5-18 mg/dL (1.8-6.4 mmol urea/L)
CREATININE
 Results from the spontaneous formation from
creatine and creatine phosphate
 Directly proportional to muscle mass
 Formation and release occur at a constant rate
 Measured as an Indicator of GFR
METHODS FOR QUANTITATIVE DETERMINATION
II. COLORIMETRIC (JAFFE)
Creatinine + alkaline picrate-------> orange-red solution
METHODS FOR QUANTITATIVE DETERMINATION
II. ENZYMATIC METHOD
METHODS FOR QUANTITATIVE DETERMINATION
Creatinine aminohydrolase/creatininase
I. CHEMICAL METHOD
Reactions are coupled with enzymatic reactions that can
be measured spectrophotometrically, such as NADH to
NAD or H₂O₂ to H₂O Interferences:
REFERENCE VALUES proteins, glucose, ascorbic acid, glutathione,
Creatinine (serum, plasma) hemoglobin, and drugs such as acetaminophen and
Adult men 0.7-1.3 mg/dL (62-115 µmol/L) caffeine
Adult women 0.6-1.1 mg/dL (53-97 µmol/L) II. HPLC
Creatinine (urine) Uses ion exchange or reversed-phase columns to
Adult men 14-26 mg/kg/24 h (124-230 µmol/kg/24 h) separate and quantify uric acid.
Adult women 11-20 mg/kg/24 h (97-177 µmol/kg/24 h) III. ENZYMATIC METHOD
III. CREATININE CLEARANCE
 Defined as milliliters of plasma cleared of creatinine
by the kidneys per minute Measures the decrease in the absorbance at
 An imperfect measure of glomerular filtration rate 282 to 292 nm
 Specimen: Interferences:
o 24 hour urine Xanthine and Hemoglobin
o Serum (collected ist about 12 hours into the REFERENCE VALUES
urine collection period Adult, 2.6 - 6mg / d * L
Formula, female Adult, 3.5 - 7.2mg / d * L
Urine creatinine (mg/dL) volume (mL) Plasma creatinine male Child, 2 - 5.5mg / d * L
(mg/dL) time (min) average surface area (m2) patient
surface area (m2)
Reference range,
Male 95-130 mL/min
Female 80-120 mL/min
URIC ACID
 The final breakdown product of purine nucleoside
metabolism
 Filtered through the renal glomerulus and is almost
completely reabsorbed in the PCT and DCT
 pH>5.57-in urate form