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[MT 6317] Clinical Chemistry Lecture > [2S] UNIT 4 Non-Protein Nitrogen Compounds: Urea & Uric Acid > Flashcards

[2S] UNIT 4 Non-Protein Nitrogen Compounds: Urea & Uric Acid Flashcards

1
Q

Nitrogen-containing substances found in blood that are not proteins.

A

Non-Protein Nitrogen Compounds

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2
Q

For the assessment and monitoring of renal function

A

Non-Protein Nitrogen Compounds

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3
Q

Prior to the current analytical methods available, nitrogen content was measured with the removal of proteins before analysis

A

Protein Free Filtrate

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4
Q

Most Abundant NPN

A

Urea

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4
Q

Least Abundant NPN

A

Ammonia

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5
Q

Derived from the catabolism of protein and amino acids

A

Urea

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5
Q

Major NPN found in the blood (45-50% of total NPN)

A

Urea

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6
Q

Synthesized in the liver from CO2 + ammonia

A

Urea

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7
Q

Filtered freely by the glomerulus
○ 40% Reabsorbed
○ 50% Excreted in the Urine
○ <10% Excreted through GI tract and skin

A

Urea

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7
Q

Arising from the deamination of the amino acids by means of the Krebs-Henseleit Cycle or Ornithine Pathway

A

Urea

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8
Q

T/F: The concentration of urea is dependent on renal function and perfusion, the protein content of the diet, and the amount of protein catabolism

A

T

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9
Q

CLINICAL APPLICATION

● Assess nitrogen balance
● Aid in the diagnosis of renal disease
● Verify the adequacy of dialysis

A

Urea

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9
Q

CLINICAL APPLICATION

● Evaluate renal function
● Determine hydration status

A

Urea

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10
Q

PATHOPHYSIOLOGY OF UREA

Fatal if not treated by dialysis or transplantation

A

Uremia / Uremic Syndrome

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11
Q

PATHOPHYSIOLOGY OF UREA

Very high levels of plasma urea accompanied by renal
failure

A

Uremia / Uremic Syndrome

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12
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

↑ Creatinine ↑ B.U.N. ↑ Osm.

A

Pre-Renal

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12
Q

PATHOPHYSIOLOGY OF UREA

An elevated concentration of urea in the blood

A

Azotemia

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13
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

Decreased blood flow to the kidneys

A

Pre-Renal

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14
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

Impaired Perfusion: (Commonly seen in patients with)
○ Cardiac Failure (Congestive Heart Failure)
○ Sepsis (Shock)
○ Blood Loss (Hemorrhage)
○ Dehydration
○ Vascular Occlusion

A

Pre-Renal

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15
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

Level of protein metabolism
■ High protein diet
■ Increased protein catabolism

A

Pre-Renal

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16
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

● Occurs in the kidneys
● Decreased renal function

A

Renal / Intrinsic

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17
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

● Small-vessel Vasculitis
● Acute Tubular Necrosis

A

Renal / Intrinsic

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17
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

● Acute / Chronic Renal Failure
● Glomerulonephritis

A

Renal / Intrinsic

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18
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

● Prolonged Hypotension
● Interstitial Nephritis

A

Renal / Intrinsic

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19
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

Happens after reaching the kidneys

A

Post-Renal

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20
Q

High urea: Normal Creatinine

A

Pre-Renal

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21
Q

AZOTEMIA (BASED ON THE DYSFUNCTION OR SITE OF ABNORMALITY)

Urinary Tract Obstruction
○ Causes:
■ Renal calculi
■ Tumors of the bladder/prostate
■ Severe infection like UTI

A

Post-Renal

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22
Q

High urea: High Creatinine

A

Renal

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23
Q

Normal urea: High Creatinine

A

Post-Renal

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24
Q

Urea Nitrogen (B.U.N.) : Creatinine

A

10:1 to 20:1

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25
Q

Determination of urinary concentration is of value in the assessment of nitrogen balance for nutritional
management

A

BUN

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26
Q

Chemical Formula of Urea

A

CO(NH2)2

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27
Q

B.U.N. to Urea

A

2.14

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28
Q

B.U.N.. in mg/dL to mmol/L

A

0.357

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29
Q

Urea to B.U.N

A

0.467

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30
Q

UREA DETERMINATION

Measures urea as a whole without isolating nitrogen

A

Direct Method

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31
Q

UREA DETERMINATION

Measures the nitrogen content of urea (B.U.N.)

A

Indirect Method

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32
Q

UREA DETERMINATION

Uses:
○ Evaluate renal function
○ Assess hydration status
○ Determine nitrogen balance
○ Verify adequacy of dialysis

A

Indirect Method

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33
Q

UREA METHODS OF DETERMINATION

● Fearon’s Reaction
● Also known as Friedman’s Method or Xanthydrol Method

A

Condensation with Diacetyl Monoxime Method

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34
Q

UREA METHODS OF DETERMINATION

Reagents: strong acid, oxidizing agent, ferric ions,
thiosemicarbazide

A

Condensation with Diacetyl Monoxime Method

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35
Q

UREA METHODS OF DETERMINATION

Product of Condensation with Diacetyl Monoxime Method

A

Yellow Diazine Derivative

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36
Q

UREA METHODS OF DETERMINATION

Direct or Indirect?
Condensation with Diacetyl Monoxime Method

A

Direct

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37
Q

UREA METHODS OF DETERMINATION: ADVANTAGES

● Ammonia does not interfere in the measurement
● Used in auto analyzers

A

Condensation with Diacetyl Monoxime Method

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38
Q

UREA METHODS OF DETERMINATION: DISADVANTAGES

● Non-specific
● Uses toxic substances

A

Condensation with Diacetyl Monoxime Method

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39
Q

UREA METHODS OF DETERMINATION

DAM + water === (H+) diacetyl
Diacetyl + urea === (Fe+3) diazine

A

Condensation with Diacetyl Monoxime Method

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40
Q

UREA METHODS OF DETERMINATION

Urea + o-phthalaldehyde (H+) → isoindoline + naphthylethylene diamine (H+) → colored product

A

Reaction w/ O-phthalaldehyde and Naphthylethylene Diamine

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41
Q

UREA METHODS OF DETERMINATION

Product of Reaction w/ O-phthalaldehyde and Naphthylethylene Diamine

A

Chromogen / Colored Product

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42
Q

UREA METHODS OF DETERMINATION

A

Reaction w/ O-phthalaldehyde and Naphthylethylene Diamine

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43
Q

UREA METHODS OF DETERMINATION: ADVANTAGES

● No ammonia interference
○ Measure entire urea and do not isolate the nitrogen content
● Used in automation

A

Reaction w/ O-phthalaldehyde and Naphthylethylene Diamine

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44
Q

UREA METHODS OF DETERMINATION: DISADVANTAGES

Sulfa containing drugs
interfere

A

Reaction w/ O-phthalaldehyde and Naphthylethylene Diamine

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45
Q

UREA METHODS OF DETERMINATION

It is a 2-step procedure wherein we need to digest urea first; this allows the isolation of nitrogen component

A

Micro-Kjeldahl Method

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45
Q

UREA METHODS OF DETERMINATION

Direct or Indirect?
Micro-Kjeldahl Method

A

Indirect

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46
Q

UREA METHODS OF DETERMINATION

Digestion: urea (H2SO4 + H3PO4) → NH4+
○ Digestion is done by adding strong acids, eg.
sulfuric acid and phosphoric acid
○ Isolated content: ammonium

A

Micro-Kjeldahl Method

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46
Q

UREA METHODS OF DETERMINATION

NH4 + (alkalinized) → NH3↑ + K2HgI → NH2Hg2I3

A

Micro-Kjeldahl Method

46
Q

UREA METHODS OF DETERMINATION

Urea (urease) → HCO3 + NH4+ + Gum Ghatti
(alkaline) + Nessler’s reagent → NH2Hg2I3 (yellow)

A

ENZYMATIC: Urease-Nessler’s method

47
Q

UREA METHODS OF DETERMINATION

Product of Micro-Kjeldahl Method

A

yellow compound (diamino mercuric iodide)

48
Q

UREA METHODS OF DETERMINATION

Urea in the sample will be reacted upon urease, which will hydrolyze urea, resulting to bicarbonate and ammonium → Gum Ghatti is added to prevent bubble formation → add
Nessler’s reagent → di amino mercuric iodide (measure spectrophotometrically)

A

ENZYMATIC: Urease-Nessler’s method

48
Q

UREA METHODS OF DETERMINATION

Direct or Indirect?

Urease-Nessler’s method

49
Q

Systematic name of urease

A

urea amidohydrolase

50
Q

UREA METHODS OF DETERMINATION

Urea (urease) → HCO3- + NH4+ + sodium
nitroprusside (alkaline) phenol hypochlorite →
indophenol blue + NaCl + H2O

A

ENZYMATIC: Urease-Berthelot’s method

50
Q

UREA METHODS OF DETERMINATION

Disadvantages:
○ Not specific
○ Very sensitive to interference from
endogenous ammonia

A

ENZYMATIC: Urease-Berthelot’s method

51
Q

UREA METHODS OF DETERMINATION

Coupled enzymatic method; combining L-glutamate dehydrogenase and urease

A

ENZYMATIC: Urease-L-Glutamate Dehydrogenase Method (GLDH method)

52
Q

UREA METHODS OF DETERMINATION

● First reaction is to utilize urease to hydrolyze ammonia to form ammonium and bicarbonate
● Secondary reaction involves utilizing
glutamate dehydrogenase
○ We monitor the reaction or oxidation of NADH to NAD+

A

ENZYMATIC: Urease-L-Glutamate Dehydrogenase Method (GLDH method)

53
Q

UREA METHODS OF DETERMINATION

The conductivity of ammonium is measured

A

ENZYMATIC: Urease-Conductometric method

54
Q

UREA METHODS OF DETERMINATION

● Quantification of result using isotopically labeled
compound
● Proposed reference method of urea

A

Istotope Dilution Mass Spectrometry

54
Q

UREA METHODS OF DETERMINATION

Detection of characteristic fragments following
ionization

A

Istotope Dilution Mass Spectrometry

55
Q

SPECIMEN REQUIREMENT FOR UREA

T/F: If plasma will be used, any anticoagulant maybe used including sodium fluoride/sodium citrate.

A

F; EXCEPT sodium fluoride/sodium citrate, which interferes with urease and ammonium ions

56
Q

SPECIMEN REQUIREMENT FOR UREA

T/F: Urine is susceptible to bacterial decomposition: urine sample should be refrigerated

57
Q

SPECIMEN REQUIREMENT FOR UREA

T/F: Protein content of diet influences urea but minimal: no fasting requirement

58
Q

Product of catabolism of purines bases (adenine and
guanine)

59
Q

Readily filtered by glomerulus, but undergoes
reabsorption and secretion

60
Q

98-100%:reabsorbed in the PROXIMAL tubules

61
Q

<1%: excreted in the distal tubules
○ Renal excretion: 70% of UA
○ GI excretion: 30%

62
Q

Relatively insoluble in plasma as monosodium urate at pH 7

63
Q

URIC ACID

At concentrations >6.8 mg/dL), plasma is saturated, _______ _____ may form in the tissues

A

urates crystals

64
Q

URIC ACID

At pH ____, uric acid crystals may form

65
Q

High concentrations accumulate in the joints and tissue resulting in inflammation (gouty arthritis)

66
Q

CLINICAL APPLICATION

○ Assess inherited disorders of purine metabolism
○ Detect kidney dysfunction

67
Q

CLINICAL APPLICATION

Assist in the diagnosis of renal calculi (kidney
stones)

68
Q

CLINICAL APPLICATION

Confirm diagnosis and monitor treatment of
gout

69
Q

CLINICAL APPLICATION

Assess and prevent uric acid during chemotherapeutic management (Chemotherapy)

70
Q

URIC ACID: PATHOPHYSIOLOGY

● In males between 30 and 50 years of age
● In female they appear after menopause

A

HYPERURICEMIA: Gout

71
Q

URIC ACID: PATHOPHYSIOLOGY

Px (patient) pain and inflammation of the joints caused by precipitation of sodium urates

A

HYPERURICEMIA: Gout

72
Q

URIC ACID: PATHOPHYSIOLOGY

Increased: hyperuricemia (___ mg/dL)
○ Hyperuricemia – in 25% to 30% of patients
○ Result of overproduction of uric acid
○ Formation of renal calculi

73
Q

URIC ACID: PATHOPHYSIOLOGY

Treatment for Increased Nuclear Breakdown

A

Allopurinol

74
Q

URIC ACID: PATHOPHYSIOLOGY

● Seen in patients undergoing chemotherapy for leukemia, lymphoma, multiple myeloma, polycythemia
● UA monitoring to avoid nephrotoxicity

A

HYPERURICEMIA: Increased Nuclear Breakdown

75
Q

URIC ACID: PATHOPHYSIOLOGY

inhibit xanthine oxidase and stop formation of uric acid

A

Allopurinol

76
Q

URIC ACID: PATHOPHYSIOLOGY

● Impaired filtration and secretion
● Not a good indicator of renal function
○ For renal: BUN, creatinine

A

HYPERURICEMIA: Kidney Diseases

77
Q

URIC ACID

X-linked genetic disorder (seen only in males)

A

Lesch-Nyhan Syndrome

78
Q

URIC ACID

Lack of this enzyme prevents the reutilization of purine bases in the nucleotide salvage pathway

A

Lesch-Nyhan Syndrome

79
Q

URIC ACID

Caused by the complete deficiency of hypoxanthine–guanine phosphoribosyltransferase (HPRT)

A

Lesch-Nyhan Syndrome

80
Q

URIC ACID

Too much secretion of AMP/AMV → uric acid increases

A

Lesch-Nyhan Syndrome

81
Q

URIC ACID

It consequently results in INCREASED concentrations of uric acid (plasma and urine)

A

Lesch-Nyhan Syndrome

82
Q

URIC ACID

Neurologic symptoms, mental retardation, and
self-mutilation characterize this extremely rare disease

A

Lesch-Nyhan Syndrome

83
Q

URIC ACID

Increasing purine synthesis; increases the degradation
product

A

Lesch-Nyhan Syndrome

84
Q

URIC ACID: DISEASE CORRELATION

Increased or Hypouricemia?

Secondary to severe liver disease

A

Hypouricemia

84
Q

URIC ACID: DISEASE CORRELATION

Increased or Hypouricemia?

Mutation on phosphoribosylpyrophosphate synthetase

85
Q

URIC ACID: DISEASE CORRELATION

Increased or Hypouricemia?

Toxemia of pregnancy

86
Q

URIC ACID: DISEASE CORRELATION

Increased or Hypouricemia?

Lactic acidosis (competition for binding sites in renal tubules)

87
Q

URIC ACID: DISEASE CORRELATION

Increased or Hypouricemia?

Defective tubular reabsorption (Fanconi’s Syndrome)

A

Hypouricemia

88
Q

URIC ACID: DISEASE CORRELATION

Increased or Hypouricemia?

Chemotherapy with 6-mercaptopurine
or azathioprine (inhibits de novo purine synthesis)

A

Hypouricemia

89
Q

URIC ACID: DISEASE CORRELATION

Increased or Hypouricemia?

Overtreatment with allopurinol

A

Hypouricemia

90
Q

CAUSES OF HYPER OR HYPOURICEMIA?

Increased dietary intake of purine-rich food

A

Hyperuricemia

91
Q

CAUSES OF HYPER OR HYPOURICEMIA?

Catabolic pathways enzyme defects

A

Hyperuricemia

91
Q

CAUSES OF HYPER OR HYPOURICEMIA?

Increased URATE production (postmenopausal women,
increased tissue metabolism such as in starvation

A

Hyperuricemia

92
Q

CAUSES OF HYPER OR HYPOURICEMIA?

DECREASED excretion

A

Hyperuricemia

93
Q

CAUSES OF HYPER OR HYPOURICEMIA?

Increased METABOLISM of cell nuclei (lymphoma,
leukemia, multiple myeloma, polycythemia, hemolytic and megaloblastic anemia)

A

Hyperuricemia

94
Q

CAUSES OF HYPER OR HYPOURICEMIA?

Inherited disorders with ENZYME deficiency
(Lesch-Nyhan Syndrome)

A

Hyperuricemia

95
Q

CAUSES OF HYPER OR HYPOURICEMIA?

Decreased uric acid excretion (preeclampsia, lactic acidosis)

A

Hyperuricemia

96
Q

ANALYTICAL METHODS – URIC ACID

Uric acid + phosphotungstic acid + O2 → NaCO3/OH →
tungsten blue + allantoin + CO2

A

DIRECT REDOX METHOD: Caraway’s / Henry Method

97
Q

CAUSES OF HYPER OR HYPOURICEMIA?

Chronic renal disease (impaired filtration and secretion)

A

Hyperuricemia

98
Q

ANALYTICAL METHODS – URIC ACID

Based on the oxidation of UA in PFF (protein-free filtrate) and reduction of PTA

A

DIRECT REDOX METHOD: Caraway’s / Henry Method

99
Q

ANALYTICAL METHODS – URIC ACID

Interferences (false +/ increase) – turbidity, aspirin and metabolite, acetaminophen, caffeine, and theophylline

A

DIRECT REDOX METHOD: Caraway’s / Henry Method

100
Q

ANALYTICAL METHODS – URIC ACID

Uses ferric ion for the presence of uric acid produces

A

Iron Reduction Method

101
Q

ANALYTICAL METHODS – URIC ACID

Develop color formation; Darker the color→ higher
concentration of uric acid

A

Iron Reduction Method

102
Q

ANALYTICAL METHODS – URIC ACID

Reduced in the presence of Uric Acid

A

Iron Reduction Method

103
Q

ANALYTICAL METHODS – URIC ACID

● Most commonly used in the laboratory
● Allantoin is measured and is the final/end product

A

ENZYMATIC: Uricase Method (Blaunch & Koch)

104
Q

ANALYTICAL METHODS – URIC ACID

Measurement of the differential absorption of UA and allantoin at 290-293 nm

A

ENZYMATIC: Uricase Method (Blaunch & Koch)

105
Q

ANALYTICAL METHODS – URIC ACID

More specific

A

ENZYMATIC: Uricase Method (Blaunch & Koch)

106
Q

ANALYTICAL METHODS – URIC ACID

Disadvantage: Bilirubin and ascorbic acid are interfering
substances that may destroy peroxide

A

Coupled Enzyme

106
Q

ANALYTICAL METHODS – URIC ACID

● Proteins cause high
background absorbance
● Negative interference due to Hb and Xanthine

A

ENZYMATIC: Uricase Method (Blaunch & Koch)

107
Q

ANALYTICAL METHODS – URIC ACID

● Uses the enzymes peroxidase and catalase.
● Color production is proportional to the uric acid
concentration

A

Coupled Enzyme

108
Q

ANALYTICAL METHODS – URIC ACID

Remedy for coupled enzyme

A

Addition of potassium ferricyanide and ascorbate oxidase

109
Q

ANALYTICAL METHODS – URIC ACID

● Uses ion exchange and reverse phase column
● Detects ratio

110
Q

ANALYTICAL METHODS – URIC ACID

● Detection of characteristic fragments after ionization
● Quantification using isotopically labeled compound
● Proposed reference method but not routinely used

111
Q

Specimen used for Uric Acid

A

Heparinized plasma, serum, or URINE

112
Q

SPECIMEN REQUIREMENT FOR URIC ACID

T/F: Diet may affect uric acid concentration

113
Q

SPECIMEN REQUIREMENT FOR URIC ACID

T/F: Gross lipemia should be used

A

F; Gross lipemia should be avoided

114
Q

SPECIMEN REQUIREMENT FOR URIC ACID

Hemolysis, with concomitant glutathione release, may result in ___ values

115
Q

SPECIMEN REQUIREMENT FOR URIC ACID

Serum samples may be stored refrigerated for ___ days

116
Q

SPECIMEN REQUIREMENT FOR URIC ACID

Salicylates and thiazides = ________ values for uric acid

117
Q

SPECIMEN REQUIREMENT FOR URIC ACID

T/F: EDTA or fluoride SHOULD NOT BE be used

118
Q

SPECIMEN REQUIREMENT FOR URIC ACID

Urine specimen pH

A

alkaline (pH 8)

[MT 6317] Clinical Chemistry Lecture (9 decks)

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