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CC LAB > Bilirubin and NPN Measurement > Flashcards

Bilirubin and NPN Measurement Flashcards

1
Q

End product of hemoglobin metabolism

A

BILIRUBIN

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2
Q

Principal pigment in bile

A

BILIRUBIN

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3
Q

What are the sources of bilirubin

A
  • Heme
  • Myoglobin, cytochrome, peroxidases/catalase
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4
Q

How many % of bilirubin derived from heme

A

85%

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5
Q

How many % of bilirubin derived from Myoglobin, cytochrome, peroxidases/catalase

A

15% (remaining)

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6
Q

Fill in the blanks:

Senescent RBCs (old RBCs) are destroyed in the _____ which cause the liberation or release of _______.

A

spleen; hemoglobin

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7
Q

What are the 3 components of hemoglobin

A

iron, heme, and globin

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8
Q

What happens to the globins in the circulation

A

Globins are degraded and the amino acids are recycled by the body

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9
Q

In the circulation:

Iron is bound to what transport protein?

A

transferrin

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10
Q

What happens to the iron in the circulation

A

Iron is bound to transferrin (transport protein), which will be delivered to the liver and other organs.

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11
Q

Component of hemoglobin wherein the bilirubin is derived

A

Heme

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12
Q

In the circulation:

heme is degraded to ______ by the action of _______

A

biliverdin; heme oxygenase

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13
Q

Biliverdin is reduced to ______ by the enzyme _______

A

Bilirubin 1; biliverdin reductase.

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14
Q

Bilirubin 1 will be transported by the ______ from the circulation to the _____ (hepatic cycle), where it will be released and picked up by the transport protein called as _______.

A

albumin
liver
ligandins

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15
Q

The Bilirubin 1 in the liver will be conjugated/esterified with ______ to form ______ and to ________ by the action of the enzyme ________

A

glucoronic acid;
bilirubin monoglucoronide;
bilirubin diglucuronide/B2;
uridine diphosphate glucoronyl/glucoronosyl transferase (UDPGT)

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16
Q

Once conjugated/Bilirubin 2 is formed, it will now be transported to the _____ where it is reduced to form ______, ________, ______ by ________

A

intestine
mesobilirubin, mesobilirubinogen, urobilinogen intestinal bacteria

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17
Q

colorless byproduct of reduction of bilirubin

A

Urobilinogen

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18
Q

2 fates of urobilinogen

A

80% converted to stercobilin
20% reabsorbed to the circulation and filtered by the kidneys in the form of urobilin

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19
Q

stercobilin is excreted in what

A

stool

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20
Q

urobilin is excreted in what

A

urine

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21
Q

What are the 2 forms of bilirubin

A

bilirubin 1 & bilirubin 2

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22
Q

Other names for Bilirubin 1

A

Unconjugated Bilirubin
Water Insoluble
Non-polar Bilirubin (has 2 propionic acids present)
Indirect bilirubin
Hemobilirubin
Slow reacting
Prehepatic bilirubin

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23
Q

Other names for Bilirubin 2

A

Conjugated Bilirubin
Water soluble
Polar Bilirubin (the 2 propionic acids are already conjugated/esterified)
Direct bilirubin
Cholebilirubin
One-minute/Prompt Bilirubin
Post-hepatic/ Hepatic/ Obstructive/ Regurgitative Bilirubin

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24
Q

bilirubin found in the circulation/blood

A

Bilirubin 1

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25
Q

bilirubin found in the liver and intestine

A

bilirubin 2

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26
Q

form of bilirubin that can be directly measured by the reagent

A

bilirubin 2

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27
Q

what is the reference value for unconjugated Bilirubin:

A

0.2 – 0.8 mg/dL

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28
Q

what is the reference value for conjugated Bilirubin:

A

0.0 – 0.2 mg/dL

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29
Q

conjugated bilirubin is synthesized in the liver that is why it should not be present in the circulation. however, there are cases in which there is a very small amount of it in the circulation, why?

A

very small amount of it may be present due to minor leakage from the hepatocytes (0.2 mg/dL)

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30
Q

what is the reference value for total Bilirubin:

A

0.2 – 1.0 mg/dL

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31
Q

What do you mean by total bilirubin

A

conjugated and unconjugated bilirubin combined

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32
Q

Conjugated bilirubin (B2) tightly bound to albumin

A

Delta Bilirubin

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33
Q

Delta Bilirubin is formed due to:

A

prolonged elevation of conjugated bilirubin

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34
Q

T/F:

delta bilirubin has shorter half life compared to other forms of bilirubin

A

F; LONGER half life

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35
Q

Helps in monitoring the decline of serum bilirubin following cholecystectomy

A

delta bilirubin

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36
Q

refers to the surgical removal of gallbladder

A

cholecystectomy

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37
Q

delta bilirubin reacts with what reagent

A

diazo reagent

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38
Q

reagent used to measure the normal forms of bilirubin

A

diazo reagent

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39
Q

Reference value of delta bilirubin

A

<0.2 mg/dL

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40
Q

what is the formula of delta bilirubin

A

Delta bilirubin=TB-(DB+IB)

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41
Q

The formula in delta bilirubin can be used all of the time except for ______

A

neonate patients (≤14 days old)

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42
Q

Sample used in bilirubin measurement

A

serum

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43
Q

T/F: In Bilirubin measurement, the sample must be stored away from light immediately

A

T

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44
Q

if not protected from light, bilirubin levels may decreased to:

A

30 - 50%/hr

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45
Q

If sample is prepared and stored properly:
­ Room temperature: ______
­ Refrigerator (4 ̊C): ______
­ Freezer (-20 ̊C): _______

A

2 days
1 week
indefinite

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46
Q

T/F:

Fasting sample is required in bilirubin measurement

A

F; Fasting sample is preferred but not required

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47
Q

T/F Recent intake of food may cause lipemia

A

T

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48
Q

What are the Interferences in bilirubin measurement

A

lipemia and hemolysis

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49
Q

In bilirubin measurement, lipemia (increases, decreases) bilirubin conc.

A

increases

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50
Q

In bilirubin measurement, hemolysis (increases, decreases) reaction of bilirubin with diazo reagent causing a (decreased, increased) bilirubin

A

decreases; decreased

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51
Q

Principle used in bilirubin measurement

A

Van den Berg Reaction

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52
Q

diazotization of bilirubin to form azobilirubin

A

Van den Berg Reaction

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53
Q

Van den Berg Reaction is the diazotization of bilirubin to form _______

A

azobilirubin

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54
Q

T/F;

A
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55
Q

T/F;
Van den Berg Reaction only measure conjugated bilirubin and total bilirubin

A

T

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56
Q

What must be added in order to measure unconjugated bilirubin in bilirubin measurement

A

accelerants/accelerator

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57
Q

What are the two methods used in bilirubin measurement

A

Evelyn and Malloy Method
Jendrassik and Grof Method

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58
Q

Coupling Accelerator used in Evelyn and Malloy method

A

methanol

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59
Q

T/F: Methanol is added to measure unconjugated bilirubin/B1 so that total bilirubin can be measured

A

T

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60
Q

What are the 3 diazo reagents used in evelyn and malloy method

A

Diazo A
Diazo B
Diazo Blank

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61
Q

Diazo A contains

A

0.1% sulfanilic acid, hydrochloric acid (HCl)

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62
Q

Diazo B contains

A

0.5% Sodium nitrite

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63
Q

Diazo blank contains

A

1.5% HCl

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64
Q

Final Reaction in Evelyn and Malloy Method

A

pink to purple azobilirubin (measured @ 560 nm)

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65
Q

Popular technique for discreet analyzers

A

Jendrassik-Grof Method

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66
Q

Most commonly used method because it is more sensitive than Evelyn and Malloy Method

A

Jendrassik-Grof Method

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67
Q

Main reagent used in Jendrassik-Grof method

A

Diazo reagent

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68
Q

Accelerator used in Jendrassik-Grof method

A

Caffeine Sodium Benzoate

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69
Q

Buffer used in Jendrassik-Grof method

A

Sodium acetate

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70
Q

terminates the accelerator and destroys excess diazo reagent

A

Ascorbic acid

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71
Q

provides alkaline pH after addition of ascorbic acid

A

Alkaline tartrate solution:

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72
Q

Final reaction in Jendrassik-Grof Method

A

blue azobilirubin (measured at 600 nm)

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73
Q

most common cause of jaundice

A

Cholelithiasis (gallstones)

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74
Q

characterized by yellow discoloration of the skin, sclera, and mucus membranes.

A

Hyperbilirubinemia

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75
Q

Type of jaundice that is not visible to the naked eye

A

Overt Jaundice:

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76
Q

reference value for Overt Jaundice

A

1.0 – 1.5 mg/dL

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77
Q

high levels of bilirubin but no abnormal yellowish discoloration

A

overt jaundice

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78
Q

RV for visible jaundice

A

3.0 – 5.0 mg/dL

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79
Q

RV for visible jaundice

A

3.0 – 5.0 mg/dL

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80
Q

more commonly termed for serum or plasma with yellow discoloration due to hyperbilirubinemia.

A

Icterus

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81
Q

RV for icterus

A

> 25 mg/dL

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82
Q

T/F Nowadays, icterus and jaundice can be used interchangeably

A

F

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83
Q

What are the 3 classification of jaundice

A

pre-hepatic Jaundice/hepatic jaundice/post-hepatic Jaundice

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84
Q

What is the cause of pre-hepatic jaundice

A

Too much RBC destruction

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85
Q

Pre-hepatic jaundice:

Bilirubin assay?

A

Elevated Indirect Bilirubin

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86
Q

Cause of hepatic jaundice

A

Hepatocyte injury caused by viruses, alcohol, and parasites

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87
Q

Hepatic jaundice:

Biirubin assay?

A

Elevated Direct and Indirect Bilirubin

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88
Q

Cause of post hepatic jaundice

A

Failure of bile to flow in the intestine (Impaired bilirubin excretion)

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89
Q

Post-hepatic jaundice:

bilirubin assay?

A

Elevated Direct Bilirubin

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90
Q

most abundant NPN

A

Urea

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91
Q

40% of the total NPN

A

urea

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92
Q

major end product of protein and amino acid catabolism

A

UREA NITROGEN (BLOOD) BUN

93
Q

Approximately 80% of the nitrogen excreted

A

UREA NITROGEN (BLOOD) BUN

94
Q

First metabolite to increase in kidney disease

A

UREA NITROGEN (BLOOD) BUN

95
Q

BUN are excreted by the kidneys but _____ of them are reabsorbed

A

40%

96
Q

Reference value for UREA NITROGEN (BLOOD) BUN

A

8-23 mg/dL

97
Q

What is the Normal Bun:Creatinine Ratio

A

10-20:1

98
Q

What are the sample used in Urea Nitrogen Methodologies

A

serum, plasma, urine­

99
Q

Urea Nitrogen Methodologies:

If the sample is plasma, ____ and _____ should not be used

A

fluoride and citrate

100
Q

Why is fluoride and citrate not used if the sample is plasma in urea nitrogen

A

these inhibits urease

101
Q

reagent used in the enzymatic method of BUN measurement

A

Urease

102
Q

T/F:

In Urea Nitrogen methodologies, refrigerate sample if delay in testing is expected

A

T

103
Q

Why the sample needs to be refrigerated

A

Because urea may be decomposed by urease producing bacteria

104
Q

Assays for urea were based on measurement of ______

A

nitrogen

105
Q

what are the 3 methods in Urea Nitrogen

A
  1. chemical method
  2. enzymatic method
  3. Isotope Dilution Mass Spectrometry
106
Q

Urea Nitrogen Methodologies:

Chemical method is also known as

A

Diacetyl Monoxime (DAM) Method

107
Q

Urea Nitrogen Methodologies:

what is the indicator dye used in the chemical method

A

Diacetyl Monoxime (DAM)

108
Q

Urea Nitrogen Methodologies:

T/F: Chemical method is a specific method

A

F; not specific

109
Q

Reference method in Urea Nitrogen Methodologies

A

Isotope Dilution Mass Spectrometry

110
Q

What are the 2 enzymatic methods in urea nitrogen methodologies

A

Urease Method
Urease-Glutamate dehydrogenase (GLD) method

111
Q

it involves the hydrolysis of urea to form ammonia by the enzyme urease.

A

Urease Method

112
Q

urease is prepared from ___

A

jack beans

113
Q

Urease method:

The synthesized ammonia may be subjected to ________ to form a chromogenic product which is _____

A

Berthelot’s reaction
indophenol blue

114
Q

Urease method:

T/F: Ammonia and CO2 can be measured by different methods to calculate the concentration of urea in the sample. But CO2 is more commonly used

A

F; ammonia is more commonly used

115
Q

known as Coupled Urease Method

A

Urease-Glutamate dehydrogenase (GLD) method

116
Q

why is Urease-Glutamate dehydrogenase (GLD) method known as coupled urease method?

A

it uses 2 enzymes (urease & glutamate)

117
Q

aka UV Enzymatic Method

A

Urease-Glutamate dehydrogenase (GLD) method

118
Q

T/F: BUN and urea are exactly the same.

A

F; BUN and urea are NOT exactly the same.

119
Q

Atomic mass of Nitrogen

A

14 g/mol

120
Q

Molecular mass of urea

A

60 g/mol

121
Q

Urea is equivalent to how many Nitrogen atoms

A

2 Nitrogen atoms (46.6% of the total weight of urea)

122
Q

How many g/mol of BUN is there per 60 g/mol of UREA

A

28 g/mol of BUN

123
Q

1 mol of urea is equivalent to how many g/mol of nitrogen?

A

28 g/mol Nitrogen

124
Q

For every BUN/Nitrogen, there is ____ g/mol of urea

A

0.4666 g/mol of urea

125
Q

Formula for urea

A

Urea=BUN x 2.14

126
Q

Creatinine is synthesized primarily by the liver from ______, _________, and _______

A

arginine, glycine and methionine

127
Q

Synthesized primarily by the liver from arginine, glycine and methionine

A

Creatinine

128
Q

Measure the completeness of 24 hour urine collection (urine creatinine)

A

CREATININE

129
Q

In 24 hr urine collection, the urine creatinine should be _____

A

> 0.8 g/day

130
Q

In 24 hr urine collection, if the urine creatinine is <0.8 g/day, it indicates what?

A

collection is incomplete

131
Q

Index of overall renal function

A

CREATININE

132
Q

why is creatinine considered as the index of overall renal function

A

Because the excretion is very stable

133
Q

Creatinine:

reference value for male:

A

0.9-1.3 mg/dL

134
Q

Creatinine:

reference value for female:

A

0.6-1.1 mg/dL

135
Q

Creatinine:

reference value for child:

A

0.3-0.7 mg/dL

136
Q

What are the sample used in creatinine methodologies

A

serum, plasma, urine

137
Q

What are the interferences in creatinine methodologies

A

hemolysis, ictericia, lipemia

138
Q

What are the 4 methods in creatinine methodologies

A
  1. Direct Jaffe Method (Chemical Method)
  2. Kinetic Jaffe Method
  3. Enzymatic Method
  4. Isotope Dilution Mass Spectrometry (IDMS)
139
Q

composition of jaffe’s reagent

A

Saturated picric acid
10% NaOH

140
Q

T/F: Jaffe reagent is stable

A

False; fairly unstable

141
Q

What happens when jaffe’s reagents are mixed together

A

when mixed together, they become unstable; may form picramic acid and methyl guanidine (forms orange color)

142
Q

What are the interferences in Direct Jaffe Method, FALSE INCREASE?

A

Creatinine-like analytes: Uric acid, ascorbic acid, glucose, α-keto acids

Medications: cephalosporins, dopamine, lidocaine

143
Q

What are the interferences in Direct Jaffe Method, FALSE DECREASE?

A

bilirubin, hemoglobin

144
Q

What are the 2 methods under Direct Jaffe Method

A

Folin Wu Method
Lloyd or Fuller’s Earth Method

145
Q

A sensitive but not specific method

A

Folin Wu Method

146
Q

What are the interferences in folin wu method

A

Acetoacetate
Acetone
Ascorbate
Glucose
Pyruvate

147
Q

T/F: the interferences in Folin Wu method are creatinine like substances/analytes

A

T

148
Q

added to remove interferences

A

Adsorbents

149
Q

A sensitive and specific method

A

Lloyd or Fuller’s Earth Method

150
Q

Adsorbent used in Lloyd or Fuller’s Earth Method

A

Lloyd’s reagent
Fuller’s Earth reagent

151
Q

Lloyd’s reagent contains:

A

Sodium aluminum silicate

152
Q

Fuller’s Earth Reagent contains

A

Aluminum magnesium silicate

153
Q

What are the disadvantages of Lloyd or Fuller’s Earth Method

A

Time consuming
Not employed to automated analyzers

154
Q

Requires automated analyzers for precision

A

Kinetic Jaffe Method

155
Q

Fill in the blanks (Principle of Kinetic Jaffe Method)

Serum is mixed with _________ and the rate of change in absorbance is measured between _______ (1 min & 2 mins marked difference)

A

alkaline picrate solution;
two points

156
Q

composition of alkaline picrate solution

A

picric acid + 10% NaOH or Jaffe reagent

157
Q

Interferences in Kinetic Jaffe Method

A

False increased: α-ketoglutarate, cephalosporins

158
Q

What is the Routinely used method in creatinine methodologies

A

Enzymatic Method

159
Q

Used to eliminate the non-specificity of Jaffe reagent

A

Enzymatic Method

160
Q

More specific than Direct and Kinetic Jaffe Method

A

Enzymatic Method

161
Q

What are the two methods under Enzymatic method

A
  1. Creatinine Aminohydrolase-CK Method
  2. Creatinase-Hydrogen Peroxide Method
162
Q

Why is Creatinine Aminohydrolase-CK Method not widely used?

A

because it requires a large volume of pre-incubated sample

163
Q

Has potential to replace Jaffe method (more specific)

A

Creatinase-Hydrogen Peroxide Method

164
Q

Without interference from acetoacetate and cephalosporins

A

Creatinase-Hydrogen Peroxide Method

165
Q

Creatinase is also known as:

A

creatinine aminohydrolase

166
Q

elevated concentration of NPN in the blood

A

azotemia

167
Q

what are the two noted NPNs in Azotemia?

A

urea and creatinine

168
Q

presence of elevated plasma urea accompanied by renal failure

A

Uremia/Uremic syndrome

169
Q

Causes of urea plasma elevations are:

A

Prerenal
Renal
and postrenal

170
Q

Anything that produces a decrease in functional blood volume and High protein diet or increased catabolism are the cause of this azotemia

A

Pre-renal Azotemia

171
Q

Anything that produces a decrease in functional blood volume, include:

A

Congestive heart failure
Shock
Hemorrhage
Dehydration

172
Q

High protein diet or increased catabolism during:

A

Fever, major illness, stress

173
Q

State the process involving pre-renal azotemia

A

Reduced Renal Blood Flow (most common)
↓
Less blood delivered to the kidney
↓
Less urea is filtered

174
Q

Also known as True Renal Disease

A

Renal Azotemia

175
Q

What are the causes of Renal Azotemia

A

Acute/chronic renal disease
­Glomerulonephritis
­Tubular necrosis

176
Q

Renal Azotemia (LAB Results)

BUN (increased, decreased) to ______ mg/dL

A

increased; >100 mg/dL

177
Q

Renal Azotemia (LAB Results)

Creatinine (increased, decreased) to ______ mg/dL

A

increased; 20 mg/dL

178
Q

Renal Azotemia (LAB Results)

BUA (increased, decreased) to ______ mg/dL

A

increased; 12 mg/dL

179
Q

What is the hormone involved in Electrolyte imbalance

A

aldosterone

180
Q

Anemia is seen in the lab result of thos type of azotemia

A

Renal Azotemia

181
Q

Complications in Renal Azotemia

A

Coma
­Neuropsychiatric changes

182
Q

State the process involving renal azotemia

A

Damaged Kidneys
↓
Poor Excretion
↓
Increased Urea

183
Q

Usually the result of Urinary Tract Obstruction

A

Post Renal Azotemia

184
Q

In Post Renal Azotemia, urea level is higher than creatinine due to:

A

back-diffusion of urea into the circulation.

185
Q

Causes of Post Renal Azotemia

A

nephrolithiasis, renal calculi, cancer/tumors of genitourinary tract, severe infection

186
Q

What is nephrolithiasis

A

kidney stones

187
Q

Chemical name of uric acid

A

2,6,8-trihydroxypurine

188
Q

It is the major product of purine (adenine and guanine) catabolism.

A

URIC ACID

189
Q

Uric acid is the major product of ?

A

purine (adenine and guanine) catabolism

190
Q

It is the final breakdown of nucleic acids catabolism in humans.

A

Uric acid

191
Q

what are the sources of nucleic acids in the body

A

Ingestion
Endogenous
Endogenous purine nucleotides

192
Q

T/F: Nucleic acids found within the body breakdowns when cells are dead (apoptotic cells)

A

T

193
Q

It is formed from xanthine by the action of xanthine oxidase in the liver and intestine.

A

Uric acid

194
Q

Uric acid is formed from _____ by the action of ________ in the liver and intestine.

A

xanthine; xanthine oxidase

195
Q

Uric acid is transported to kidney and filtered.

How many percent is filtered?

A

70%

196
Q

How many % of uric acid are reabsorbed by the proximal convoluted tubules

A

98% (of the 70%)

197
Q

Remaining 30% of uric acid will go to the ____ where they are degraded by the bacterial enzymes

A

GIT

198
Q

Uric acid is present in plasma as:

A

monosodium urate

199
Q

How many % of Uric acid is present in plasma as monosodium urate

A

95%

200
Q

T/F:

At plasma pH, uric acid is relatively insoluble (may accumulate)

A

T

201
Q

Uric acid:

RV for male:

A

3.5-7.2 mg/dL

202
Q

Uric acid:

RV for female:

A

2.6-6.0 mg/dL

203
Q

Uric acid:

RV for child:

A

2.0-5.5 mg/dL

204
Q

At what concentration does plasma with uric acid is saturated.

A

> 6.8 mg/dL

205
Q

Saturated plasma may _____, forming ______ which can be deposited into the _____ causing _____.

A

Saturated plasma may precipitate, forming uric acid crystals which can be deposited into the tissues (joints) causing gout.

206
Q

Uric acid crystal may also be formed in the urine if the urine filtrate is ____ (pH: ?)

A

acidic (pH <5.75)

207
Q

Sample used in Uric Acid Methodologies

A

serum, heparinized plasma, urine

208
Q

In UA Methodologies;

Serum may be stored for ______ (refrigerator)

A

3-5 days

209
Q

In UA Methodologies;

Serum may be stored for ______ (refrigerator)

A

3-5 days

210
Q

Urine sample in uric acid should have pH level of

A

8.0

211
Q

T/F: Fasting is required because food intake affects uric acid conc.

A

F; Fasting is NOT required because food intake DOES NOT affect uric acid conc.

212
Q

Interferences in uric acid:
­
False increased: ______, ______, _____

A

lipemia, salicylates, thiazides

213
Q

Interferences in uric acid:
­
False decreased: ______, ______

A

high bilirubin, hemolysis

214
Q

3 Methods in Uric acid methodologies

A

Chemical Methods
Enzymatic Methods
Isotope Dilution Mass Spectrometry – reference method

215
Q

Also known as Caraway method

A

Chemical Method

216
Q

Chemical Method is also known as ?

A

Caraway method

217
Q

Principle in chemical method

A

Reduction-oxidation reaction

218
Q

­ Sodium cyanide (NaCN) is USED for: ____, _____, _____, _____

A

Folin, Newton, Brown, Benedict

219
Q

Sodium carbonate (NaCO3) is used for: _____, ____, ____

A

Caraway, Archibald, Henry

220
Q

Also known as Uricase method

A

Enzymatic Method

221
Q

T/F: In enzymatic method (Uric acid), EDTA/fluoride tube must be used

A

F; EDTA/fluoride tube must NOT be used

222
Q

T/F: Enzymatic method (Uric acid) is a specific method

A

T

223
Q

T/F: Enzymatic method (Uric acid) is a specific method

A

T

224
Q

Principle used in enzymatic method (Uric acid)

A

Uric acid is oxidized to form allantoin by uricase enzyme.

225
Q

Uric acid is oxidized to form_____ by ____ enzyme

A

allantoin; uricase

226
Q

has a peak absorbance at 293 nm

A

Uric acid

227
Q

not absorbed at 293 nm.

A

Allantoin

228
Q

HYPERURICEMIA is seen in:

A

Gout
Increased nuclear metabolism
Chronic Renal Disease
Lesch-Nyhan Syndrome
Secondary to Glycogen Storage Disease (GSD)
Toxemia of pregnancy and lactic acidosis
Increased dietary intake
Ethanol consumption

229
Q

HYPOURICEMIA is seen in:

A

Fanconi’s syndrome
Wilson’s disease
Hodgkin’s disease
Overtreatment with allopurinol
Chemotherapy (6-mercaptopurine, azathioprine)
Alzheimer’s disease
Parkinson’s disease

CC LAB (1 decks)

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