Hemoglobin Estimation

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Presentation transcript:

Hemoglobin Estimation Physiology lab4 Hemoglobin Estimation

Hemoglobin (Hb) is a conjugated protein and it is the main component of RBC . The main function of Hb is transport of oxygen. The Hb molecule contain two pairs of polypeptide chains (α2β2) and 4 haem groups each have an atom of ferrous iron.

Approximately 34% of the RBCs weight is Hb. The O2 carrying capacity of Hb when fully saturated is 1.34 ml/g. Iron content of Hb is 0.347 gm/100 g . Hemoglobinometry : is the measurement of Hb concentration in the blood .

Types of Hb: Oxyhemoglobin ( HbO2) Carboxyhemoglobin ( HbCO) Sulfhemoglobin (SHb) Methemoglobin (Hi)

Types of blood sample: Capillary blood from finger prick. Intravenous blood ( with anticoagulant – preferably EDTA). Liquid anticoagulant should not used as these dilute and decrease Hb concentration.

Methods of Hb estimation: Colorimetric method: based on colorimetric measurement of the intensity of colour developed on addition of some substances to the blood; it include: Cyanmethaemoglobin method. Oxyhemoglobin method. Electronic counter method Sahli’s method.

2. Measurement of O2 carrying capacity of Hb ( used in research or on referral lab only). 3.Measurement of iron content of Hb ( only foe research). 4. Specific gravity method: very rapid, and useful in screening blood donors for anemia ( normal specific gravity of blood ranges from 1.048-1.066)

Sahli’s method: Principle: Hb is converted into acid haematin with the action of diluted hydrochloric acid. The acid haematin is brown in color and its intensity is matched with standard brown glass comparator in visual colorimeter called Sahli’s colorimeter.

Procedure: Fill the Sahli’s hemoglobin tube up to mark 2 with o.1 % HCl . Add 0.2 ml of blood from a Hb pippete to it. Stir with glass tube and wait for 10 minutes. Add distilled water drop by drop and stir till color matches with the comparator. Read the upper meniscus on Sahli’s tube to measure the percentage of haemoglobin and grams of haemoglobin per 100 ml of blood. Haemoglobin concentration of 14.6g/100ml considered as 100%.

Advantage: Simple bedside test. Reagents and apparatus are cheap. Disadvantages: Visual error. Carboxy, met, and sulfhemoglobins can not be converted to acid haematin. Comparator can fade away over the years. Color of acid haematin also fades quickly.

Normal values: Men 15. 5+/- 2. 5 g/dl Women 14+/- 2. 5 g/dl Infants 16 Normal values: Men 15.5+/- 2.5 g/dl Women 14+/- 2.5 g/dl Infants 16.5+/-3 g/dl