Hemoglobin

Question 1

METHODS OF HEMOGLOBIN MEASUREMENT

_____: Only the oxygen is being measured

1. aka
2. equation: 1 Hgb = ___ ml O2
3. every gram of ng can actually bind ton ml of oxygen
4. Hemoglobin with Fe
5. Hemoglobin with sulfur

Answer 1

GASOMETRIC METHOD

1. VAN SLYKE OXYGEN CAPACITY
2. 1.34ml
3. Hufner factor
4. Methemoglobin
5. Sulfhemoglobin

Question 2

METHODS OF HEMOGLOBIN MEASUREMENT

_____: only the iron is being measured

1. 2 types of method
2. iron is liberated by ___ & ____
3. hemoglobin contains heme and globin
   - contains FeH
   - protein
4. Separation of heme by globin by treating the sample with sulfuric acid & potassium persulfate, the globin will precipitate by ______; product is called ____
5. once globin is precipitated, only iron remains (filtrate) and it will be converted into
6. equation: 1gm of hb = ____mg iron
7. law where intensity of the color of solution

Answer 2

CHEMICAL MEHTHOD

1. WONG & ASSENDELEFT
2. sulfuric acid & potassium persulfate
   3.
   - heme
   - globin
3. tungstic acid; protein free filtrate
4. Ferric thiocyanate
5. 3.47mg
6. Beer’s law

Question 3

METHODS OF HEMOGLOBIN MEASUREMENT

_____: based on specific gravity/ weight of hb

1. uses a standard _____ (in 40 tubes) with sg of ___ to ___ at interval of 0.001
2. drop of patients blood is placed in each tube; drop of blood becomes coated with _____
   - distance of adding the blood should have be ____
   - check results within ____
3. if blood sp gravity is greater than the solution present in tube
4. if blood sp gravity is less than the solution present in tube
5. if blood sp gravity is the same in the solution present in tube
6. normal values for:
   - female
   - male
7. sp gravity that is needed in mass blood donation

Answer 3

GRAVIMETRIC METHOD

1. copper sulfate; 1.035-1.075
2. copper proteinate
   - >1cm above the reagent
   - 12 seconds
3. SINK
4. FLOAT
5. REMAIN
6. normal values for:
   - 1.053 = 12.5 g/dl hb
   - 1.055 = 13.5 g/dl hb
7. 1.053

Question 4

COLORIMETRIC METHOD: Direct Visual Colorimetric

1. patient’s undiluted blood is absorbed unto an absorbent pad and the color is compared with a lithographed color scale representing values from 10-100%
   - inaccurate: gives as much as __% error
2. Blood is drawn by capillary action
   between 2 glass plates and the color of blood is matched with a rotating disc of red tinted glass with varying thickness and color
   - inaccurate: gives as much as __% error
3. hemoglobin is converted into acid hematin upon presence of acid
   - reagent acid that is used
   - darker the color
   - lighter the color
   - end product is brown color which will be diluted with distilled h2o until matches the color of ______
   - expressed in ___ or ___
4. hemoglobin is converted
   into alkali hematin upon addition of an alkali
   - not ideal for infants and children due to ___ which it is resistant to acid and alkali
   - 2 methods
   -added reagent

Answer 4

1. TALLQUIST METHOD
   - 50%; 50% accuracy
2. DARE’S HEMOGLOBINOMETER
   - 30%; 70% accuracy
3. ACID HEMATIN
   - 0.1N hydrochloric acid
   - more hgb
   - less hgb
   - comparator black
   - g/dL or g%
4. Alkali hematin
   - fetal hgb
   - Wu; Klegg & King
   - 0.1N sodium hydroxide

Question 5

COLORIMETRIC METHOD: photoelectric colorimetric

______: measured in this method is the entire hemoglobin with oxygen and its differences with gasometric is we measure here the hb with ____

There is instances that instead of O2, CO2 is attached in hemoglobin. The best thing to do in that case is to allow the hemoglobin to oxygenate first:

1. To do this, we mix _____ reacted with ____ or _____ then mixed for proper oxygenation
   - 540nm against ______
2. The relevance of this is in diagnosis of
3. Methods:
   - 415nm
   - hb catalyzes the rapid oxidation of benzidine by H2O2

Answer 5

OXYHEMOGLOBIN METHOD; toxigen

1. 0.02ml of whole blood + 5ml of 0.07N ammonium hydroxide or 0.1% sodium carbonate
   - reagent blank
2. acute hemolysis
3. Methods:
   - Harboe
   - Naumann

Question 6

DRABKIN’S REAGENT

1. enhances lysis since hgb is needed to be hemolyzed for it to react to another reagent
   - types (2)
2. oxidizes hgb into methemoglobin that combines with potassium cyanide
   - it oxidizes ____ into ____
   - abnormal increase hgb
3. source of cyanide ion to form end-product
   (cyanmethemoglobin)
4. original drabkin’s solution base which is
   incubated at room temp for 10mins
   - cyanide + methemoglobin to make _______
   - cyanide + hemiglobin to make _____
5. buffer that uses sodium bicarbonate as the buffered solution
   - the effect of ____ will accelerate the reaction, shortening now the incubation from 10 mins
   to 3 mins

Answer 6

1. Non-ionized detergent
   - Sterox (Harleco) & Triton
2. Potassium Ferricyanide
   - ferrous to ferric
   - Methemoglobinemia
3. Potassium Cyanide
4. Sodium Bicarbonate
   - Cyanmethemoglobin
   - Hemiglobincyanide
5. Dihydrogen Potassium Phosphate
   - dihydride

Question 7

COLORIMETRIC METHOD: photoelectric colorimetric

______: when blood is mixed with potassium ferricyanide and potassium cyanide, the erythrocytes are lysed thereby producing hemoglobin; most accurate or gives the most stable pigment that can be read with delayed time

1. ______ oxidizes hemoglobin into methemoglobin that combines with _____ to produce hemiglobincyanide (cyanmethgb)
2. all type of hemoglobin is transformed into cyanmethemoglobin except _____
3. darker the end product = __ light will be absorbed
4. reference values:
   - male
   - female
   - at birth
5. prepare ___ blood-reagent mixture by adding __ ml of Drabkin w/ 20ul of blood
6. determine the conc by ___ thru calculation & _____

Answer 7

CYANMETHEMOGLOBIN METHOD

1. potassium ferricyanide; potassium cyanide
2. Sulfhemoglobin
3. more
4. reference values:
   - 140-175 g/dl
   - 123-153 g/dl
   - 150-200 g/dl
5. 1:251; 5mL
6. Beers law & standard curve

Question 8

QUALITY CONTROL OF THE REAGENT

1. keep in ___ bottle
   - it is photosensitive, since it contains ______ that can be oxidize when exposed to light
2. reagent should ben____ with pH _____
3. reagent must give a reading of __ at ____nm
4. ____; standard
5. storage is at ref temp stable in a ____ container at ___C

Answer 8

1. Amber bottle; potassium ferricyanide
2. Pale yellow; 7.0-7.4
3. 0 at 540nm
4. 15 g/dL
5. polythene container at 2-8C

Question 9

SOURCES OF ERRORS:

technical/ human errors:
1. deteriorated reagent: uses 2 reagents:

physiologic errors:
1. high lipids, milky, chylous
- remedy
2. high wbc or platelets will cause ______
- remedy
3. abnormal hemoglobins ____ & ____ will become resistant to lysis
- remedy
4. easily precipitated proteins: produce large amount of ______ & _____
- remedy
5. turbidity cause more light to be absorbed = ___ conc the sample is = ___ in reading

Answer 9

technical/ human errors:
1. Drabkins & standard reagent

physiologic errors:
1. Lipemic blood
- Use patient blank
2. Turbidity
- Centrifuge and use
supernatant
3. Hgb S and Hgb C
- dilute sample 1:1 distilled water
4. multiple myeloma & waldenstrom macroglobulinemia
- 0.1gm of potassium pcarbonate/ L Drabkin’s
5. more conc the sample = increase in reading