HGB Men 13.0-18.0 g/dL Women 11.5-16.5 g/dL Infants (full-term, cord blood) 13.5-19.

5 g/dL Infants, 3 months 9.5-13.5 g/dL Infants, 1 yr. 10.5-13.5 g/dL Children, 3-6 yr. 12.0-14.0 g/dL Children, 10-12 yr. 11.5-14.5 g/dL

Physical Based on Specific gravity Chemical Based on Iron content of Hb Gasometric-Based on O2 combining capacity of Hb Calorimetric-Based on colour Photoelectric Cynmethhemoglobin method Automated methods

Blood added to N/10 HCl which converts Hb into acid hematin. Brown colour of acid hematin , matched against comparimeter

Procedure
Take N/10 HCL in the tube up to lowest mark. Fill the Hb pipette with 0.02ml of blood & wipe off the excess blood at nozzle and transfer it to the tube. Mix the acid and blood by shaking the tube Allow it to stand for 10 minutes dilute the solution with distilled water and match the colour with the comparator. The upper meniscus is noted and reading on the tube is read as gm/dl

If blood not mixed If Hb pipette is not caliberated correctly If instrument is not caliberated correctly Fading of comparator

Error due to visual matching Lower values if read after 10 min due to fading Acid does not convert all Hb into hematin like carboxy, met, sulfhaemoglobin

haemiglobincyanide (HiCN; cyanmethaemoglobin) method oxyhaemoglobin (HbO2) method

Principle:

Hb oxidised by potassium ferricyanide to MetHb,stabalised by potassium cynide to cynmethemoglobin

Drabkin's reagent Potassium ferricyanide (0.607 mmol/l) 200 mg Potassium cyanide (0.768 mmol/l) 50 mg Potassium dihydrogen phosphate (1.029 mmol/l) 140mg Nonionic detergent 1 ml Distilled or deionized water To 1 litre

5ml of drabkins solution in test tube Add 0.02 ml of blood by pipette Mix and wait for 10 minutes Switch on colorimeter for 5 min Adjust OD at 0 by test solution Put test sol. into cuvette and take OD

Calculation of Haemoglobin Concentration Hb = OD of test sample conc of std.250 OD of std. 1000

No visual error All Hb are converted to cyanmethemoglobin Colour does not fade with time allows direct comparison with the reference standard and that the readings need not be made immediately after dilution so batching of samples is possible

KCN is potent poison Turbidity in Drabikins solution alter OD In India no reliable Hb standards available

Principle RBCs lysed by 0.04% liquid ammonia and OD of oxyhemoglobin obtained is measured at 540 nm in colorimeter Advantage --- quick and simple ---not affected by S.bilirubin Disadvantage --- no stable HbO2 standard

--- not reliable in carboxy,met,SHb

N/10 NaOH is used and colour of alkaline hematin is compared Not affected by carboxyhaemoglobin (HbCO), Hi, or SHb plasma proteins and lipids have little effect cumbersome and less reliable method

Standard method heating method Acid alkali method acid method

Hb of a diluted blood sample can be determined by without a standard, if spectrometer has been correctly calibrated Hb = A540 HiCN16114dilution factor 11d1000

RBC are lysed and CO is added to convert Hb to carboxyhemoglobin

Colour Comparators compare the colour of blood against a range of colours representing haemoglobin concentrations. Used for anaemia screening

based on the HbO2 method. a built-in filter and a scale calibrated for direct reading of haemoglobin The HemoCue system The DHT Haemoglobinometer

Methods developed for using near infrared spectroscopy at body sites, mainly a finger, to identify the spectral pattern of haemoglobin in an underlying blood vessel. There is an approximate correlation with blood haemoglobinometry

By Alkali denaturation method --- Brinkman and Jonix method --- Singer & Shernoff method

Red cells are lysed by lysate provided and colour is matched against the in built standard colour A cynide free bio-degradable reagent is used

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