clinical chemistry mlt 301 nonprotein nitrogen npn lecture one
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CLINICAL CHEMISTRY (MLT 301) NONPROTEIN NITROGEN (NPN) LECTURE ONE

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CLINICAL CHEMISTRY (MLT 301) NONPROTEIN NITROGEN (NPN) LECTURE ONE. Dr. Essam H. Jiffri. Nonprotein Nitrogen. The determination of nonprotein nitrogenous substances in the blood has traditionally been used to monitor renal function.

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nonprotein nitrogen
Nonprotein Nitrogen
  • The determination of nonprotein nitrogenous substances in the blood has traditionally been used to monitor renal function.
  • - The term nonprotein nitrogen (NPN) originated in the early days of clinical chemistry when analytical methodology required that protein be removed from the sample before analysis.
nonprotein nitrogen1
Nonprotein Nitrogen
  • - The NPN fraction is made up of about 15 compounds of clinical interest, the majority of these compounds rise from the catabolism of proteins and nucleic acids.
components of the nonprotein nitrogen fraction
Components of the Nonprotein- Nitrogen Fraction
  • Approximate Plasma Concentration
  • Compound (% of Total NPN)
  • Urea 45
  • Aminoacids 20
  • Uric Acid 20
  • Creatinine 5
  • Creatine 1-2
  • Ammonia 0.2
slide5
UREA
  • - Urea constitutes nearly half the nonprotein nitrogen substances in the blood.
  • - It is synthesized in the liver from Co2 and the ammonia arising from the deamination of amino acids by means of the ornithine.
slide7
UREA
  • Urea constitutes the major excretory product of protein metabolism, following synthesis in the liver, urea is transported by the plasma to kidney, where it is readily filtered from the plasma by the glomerulus.
  • - Most of the urea in the glomerular filtrate is excreted in the urine, and up to 40-50 % is reabsorbed by passive diffusion during passage of the filtrate through the renal tubules.
slide8
UREA
  • - An elevated level of urea in the blood is called azotemia.
  • - Very high levels of plasma urea accompanied by renal failure is called uremia or the urermic syndrome this is eventually fatal if not treated by dialysis.
  • - The major causes of decreased plasma urea levels include decreased protein intake and severe liver disease.
analytical methods
Analytical Methods
  • - Because measurements of urea were initially done on a protein-free filtrate of whole blood, and early analytical methods were based on measuring the amount of nitrogen, this assay is commonly called a BUN or blood urea nitrogen determination.
analytical methods1
Analytical Methods
  • Two analytical approaches have been used to assay for urea:
  • The oldest and most often used involves the hydrolysis of urea by the enzyme urease and quantitation of the NH+4.
analytical methods2
Analytical Methods
  • Early colorimetric methods were based on using Nesslers reagent or the Berthelot reaction to detect the NH+4 produced.
specimen requirements and interfering substances
Specimen Requirementsand Interfering Substances
  • Fluoride or citrate will inhibit the urease utilized in the coupled enzymatic methods.
  • - When collecting plasma, NH+4 ions and high concentrations of sodium fluoride or sodium çitrate must be avoided.
  • Urea concentration may be readily measured in serum, plasma, or urine.
specimen requirements and interfering substances1
Specimen Requirementsand Interfering Substances
  • - Fasting sample is usually not required.
  • A non-hemolyzed sample is recommended.
  • Urea is quite susceptible to bacterial decomposition, so samples (especially urine) that cannot be analyzed within a few hours should be refrigerated.
reference interval
Reference Interval
  • Urea nitrogen7 to 18 mg/dL (2.5-6.4 mmol/L urea)
creatinine
CREATININE
  • - Creatine is synthesized mainly in the liver from arginine, glycine, and methionine.
  • - It is then transported to other tissues, such as muscle, where it is converted to: phosphocreatine, which serves as a high energy source.
creatinine1
CREATININE
  • - Either creatine phosphate or creatine, under physiologic conditions, spontaneously loses phosphoric acid or water, respectively, to form its anhydride, creatinine which excreted into the plasma.
analytical methods for creatinine
Analytical Methods for Creatinine
  • - The methods most frequendy used to measure creatinine are based on the Jaffe reaction first described in 1886, in this reaction, creatinine reacts with picric acid in alkaline solution to form a red-orange chromogen.
specimen requirements and interfering substances2
Specimen Requirements and Interfering Substances
  • Creatinine can be measured in serum, plasma, or urine.
  • - Hemolysis should be avoided, since there are a significant number of nonspecific chromogens in the red cells.
specimen requirements and interfering substances3
Specimen Requirements and Interfering Substances
  • Fasting is not require.
  • A heavy protein ingestion may elevate levels of creatinine.
  • - Urine should be refrigerated after collection or frozen if storage longer than four days is required.
reference interval1
Reference Interval
  • Reference intervals vary with age and sex and may decrease in the elderly.
  • Cord blood 0.6-1.2 mg/dl (53-106 mmol/L)
  • Children 0.3-0.7 mg/dl (27-62 mmol/L)
  • Adult
  • Male 0.6-1.2 mg/dl (53-106 mmol/L)
  • Female 0.5-1.1 mg/dl (44-97 mmol/L)
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