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HEMATOLOGY LECTURE > MIDTERMS: L1 - HEMOGLOBIN AND HEMATOCRIT DETERMINATION > Flashcards

MIDTERMS: L1 - HEMOGLOBIN AND HEMATOCRIT DETERMINATION Flashcards

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1
Q

Estimation or measurement of hemoglobin concentration of a blood sample

A

HEMOGLOBINOMETRY

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2
Q

Per gram of Hb:
O2 -
Fe2 -

A
  1. 34 mL O2

3. 47 mg Fe2+

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3
Q

MOTF
Chemical method
- Indirect estimation
of the hemoglobin based on the oxygen concentration of the blood
Gasometric method
- Indirect estimation
of the hemoglobin based on the concentration of iron

A

BOTH F
Gasometric method
- Indirect estimation
of the hemoglobin based on the oxygen concentration of the blood
Chemical method
- Indirect estimation
of the hemoglobin based on the concentration of iron

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4
Q

Normal Hemoglobins

A
  • Oxygenated Hb

- Reduced/Deoxygenated Hb

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5
Q

3 Hemoglobin Derivates

A
  1. Methemoglobin/Hemiglobin (Hi)
  2. Sulfhemoglobin (SHb)
  3. Carboxyhemoglobin (COHb)
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6
Q

The iron part of the hemoglobin has

been oxidized to ferric state

A

Methemoglobin/Hemiglobin (Hi)

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7
Q

Color of blood in Methemoglobin/Hemiglobin (Hi)

A

Chocolate brown color

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8
Q

Color of blood in Sulfhemoglobin (SHb) and Carboxyhemoglobin (COHb)

A

Sulfhemoglobin (SHb) - Mauve lavander color

Carboxyhemoglobin (COHb) - Cherry red color

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9
Q

Methods of Hemoglobin determination:

A
  1. Physical: Gravimetric
  2. Colorimetric
  3. Spectrophotometric 4. Automated
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10
Q

MOTF
Physical Method
1. Qualitative - not give value what hb level is, just indicate hb concentration if adequate or not
2. Commonly used to screen blood donors

A

Both T

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11
Q

SG of Blood

A

1.045-1.065 (average 1.055)

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12
Q

Reagent used in Physical Method

A

Copper sulfate of a particular density

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13
Q 
MOTF
Physical Method
1. Sink - Blood Hb is >12.5 g/dL (patient passed)
2.  Suspended - blood HB is < 12.5 g/dL
(patient failed)
A

Both T

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14
Q

MOTF
Physical Method
1. Blood floats: SG of blood < SG of CuSO4 - Fail the potential donor

  1. Blood sinks: SG of blood > SG of CuSO4 - Accept the potential donor
A

Both T

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15
Q

T or F
Male: SG 1.055 = 13 g/dL of Hb

Female: SG 1.053 = 12 g/dL of Hb

A

T

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16
Q

Other term for Colorimetric Method

A

Acid Hematin Method

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17
Q

COLORIMETRIC METHOD
1. Quantitative estimation
2. Based on the conversion of the hemoglobin into acid hematin with the addition of:
Reagents: 0.1N Hydrochloric acid, dH2O

A

Both T

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18
Q

Reagents used in Colorimetric Method

A

0.1N Hydrochloric acid, dH2O

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19
Q

0.1N Hydrochloric acid will oxidize the hemoglobin into acid hematin which will have a _____ color

A

Brown

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20
Q

Used to adjust the sample until it matches the color comparator

A

Distilled H20

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21
Q

Where we put in blood samples and than add our reagent 0.1N Hydrochloric acid to convert hemoglobin to acid hematin

A

Hemometer tube

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22
Q

Where reagent is stored ( 0.1N Hydrochloric acid/ dH20)

A

Amber bottle

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23
Q

Reagents used for cleaning

different pipettes regardless if WBC or Sahli pipette (In correct order, with their purpose0

A
  1. Bleach (zonrox) - to remove the blood in the
    pipette, should be dispensed
  2. Water - make sure that no blood sample is left
    in the pipette
  3. Acetone - used to dry the pipette
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24
Q

T OR F

Acid hematin method will able to measure our different derivatives

A

False (Cannot measure diff derivatives)

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25
MOTF 1. More accurate than photometric 2. Routinely used in bigger hospital 3. Has low specificity and accuracy
Only 3 is correct 1. False - photometric is more accurate 2. False - Not routinely used in big hospitals, can use in small labs 3. True
26
Visual method that utilizes the Drabkin’s reagent
PHOTOMETRIC METHOD
27
Photometric method (a reference method) recommended by
International | committee on standardization in hematology as well as CLSI
28
Photometric Method - able to measure all types of hemoglobin except _______
Sulfhemoglobin
29
Photometric Method is also known as
Cyanmethemoglobin/Methemoglobin cyanide method
30
Drabkin's reagent components - enumerate and its functions
1. Potassium cyanide - oxidizes Hb to methemoglobin 2. Potassium ferricyanide - donates cyanide ions to form Cyanmethemoglobin or Hemiglobincyanide (HiCN) 3. Dihydrogen potassium phosphate - prevents precipitation of globulins = maintains pH 7.0-7.4 4. Non-ionic detergent/ surfactant (saponin) - enhances red cell lysis and lessens turbidity (caused by the presence of proteins in the sample)
31
MOTF 1. Potassium cyanide - Iron to ferric form to produce methemoglobin which is a reversible process 2. Potassium ferricyanide - donates cyanide ions to form Cyanmethemoglobin or Hemiglobincyanide (HiCN) - irreversible process
BOTH T PC - REVERSIBLE PF - IRREVERSIBLE
32
MOTF 1. Photometric method use Spectrophotometer with 540nm 2. Wavelength used is to measure red color using green filter
BOTH TRUE | 540nm - for red solution
33
Spectrophotometer read absorbance of:
Blank Standard Qc materials Patient Samples
34
Known concentration of substance
Standard
35
T OR F Hemoglobin standard is the only available standard used for Routine Hematologic Procedures
TRUE
36
``` MOTF 1. Regardless of the method of determination as long as it is qualitative with the exception of the gravimetric method which is quantitative, the reference values for hemoglobinometry is the same 2. RV: Female - 13.5-18.0 g/dL (135-180 g/L0 Male - 12.0-15.0 g/dL (120-150 g/L) ```
``` BOTH F 1. False - GM is qualitative 2. False - Male - 13.5-18.0 g/dL (135-180 g/L0 Female - 12.0-15.0 g/dL (120-150 g/L) ```
37
Enumerate comments and sources of errors in Photometric method
1. Drabkin’s rgt = photosensitive 2. Turbidity can falsely increase result: 3.COHb or Carboxyhemoglobin in heavy smokers takes 1 hour to be converted to HiCN (hemoglobin cyanide) 4.Acidification of cyanide in the rgt releases toxic hydrogen cyanide gas
38
Storage or drabkin's reagent
Amber bottle, stored in dark area, must still be clear and yellow
39
T or F Any contamination that may cause any level of turbidity in the solution can affect the result of the sample since this is an optical method
True
40
Correction of Leukocytosis (WBC ct > 20 x 109/L)
Correction: centrifuge sx, measure Hb in supernatant fluid to remove WBCs from the solution
41
Blood sx has high concentration of triglycerides that can lead to turbidity
Lipemia
42
Correction of Lipemia
Correction: 10 uL px plasma + 5 mL of Drabkin’s rgt, use as rgt blank
43
Adds to the optical density of the solution
Hb variants (ex. HbS and HbC
44
Correction of Hb variants (ex. HbS and HbC
Correction: 1:2 dilution with dH2O and multiply result from the std curve by 2
45
Correction of abnormal globulins
Correction: add Sodium carbonate/ | KH2PO4 (Dihydrogen Potassium Phosphate0
46
MOTF 1. Reagent should not be discarded into the sink 2. Licensed waste disposal service should be contracted
BOTH T
47
These are required to be printed on the packaging of the reagents to alert the user of the hazards posed by these chemical substances
Pictograms
48
T OR F | 1. The hemoglobin in the automated method is part of our complete blood count
True
49
3 levels of QC
Low, normal, high
50
Potassium cyanide is replaced by:
a. Sodium azide (POCTs) b. Sodium lauryl sulphate (SLS) (autoanalyzers ex: Sysmex) - can be dispose in sink with added water
51
Clinical significance of Hemoglobinometry:
1. Determine presence and severity of anemia 2. Assess response to treatment of anemia 3. Monitor blood loss post-operation 4. Blood donor screening 5. Calculation of Red Cell indices
52
Increased Hb is due to - enumerate
1. Polycythemia - increased cellular elements in the blood 2. Lung disease 3. VCD 4. Dehydration 5. High altitude
53
Decreased Hb is due to - enumerate
1. Anemia | 2. Renal diseases
54
Measure the packing of RBCs against the plasma volume
Packed Cell Volume (PCV)
55
Unit for reporting hematocrit determination
L/L or percentage
56
T OR F | The manual hct method has a very high precision, which has the margin of error of +/- 1%
True
57
Manual Hct determination:
1. Macrohematocrit (Wintrobe mtd) | 2. Microhematocrit
58
Separation of the formed elements of blood from the plasma through centrifugation
Microhematocrit method
59
``` MOTF After microhematocrit centrifugation: 1. Red cells packed at bottom 2. Plasma at middle 3. White cells and platelets at top ```
Only 1 true 2 - False (White cells and platelets/Buffy coat at middle) 3 - False (Plasma at top)
60
MOTF 1. Red cap capillary tube - plain tube 2. Blue cap capillary tube - heparinized contains heparin
BOTH F 1. Red cap - heparinized 2. Blue cap - plain, non-anticoagulated
61
MOTF 1. Open part of tube placed against rubber gasket (at centrifuge) 2. Closed part placed towards center
BOTH F 1. Open ends - toward center 2. Closed - agaisnt rubber gasket
62
GRAVITY AND RPM used in centrifugation
Gravity: 10,000-15,000g Rpm: 10,000-12,000 rpm
63
T or F | Do not include the buffy coat in the reading of the value since it would falsely increase the result
True
64
Falsely increase or decrease: | Improper sealing
Falsely decrease
65
Falsely increase or decrease: | Short draws lead to cell shrinkage
Falsely decrease
66
Falsely increase or decrease: Insufficient spining
Falsely increase
67
Falsely increase or decrease: Delay
Falsely increase
68
Falsely increase or decrease: inclusion of buffy coat
Falsely increase
69
Depending on the angle on how you would read the value of your results, that could also affect on how you would interpret the measurement - also known as
Parallax effect
70
Variation in size and shape of RBCS
Anisopoikilocytosis
71
T OR F | Anisopoikilocytosis can increase trapped plasma (1-3% higher than automated results)
True
72
Manual micro Hct will always be lower compared to automated result values because of the trapped plasma phenomenon.
False - Manual micro Hct will always be higher compared to automated result
73
Falsely increase or decrease: Dehydration
Falsely elevate - plasma volume is lower making packed RBC appear more
74
Reference values for automated method+
Male = 40-54% (0.40-0.54 L/L) Female = 35-49% (0.35-0.49 L/L)

HEMATOLOGY LECTURE (8 decks)

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