Methodology

Hemoglobin Determination

0.1 M HCL up to the 2% mark

Homogenize sample and transfer 5 drops into a glass slide

Pipette 20 mm3 of blood into the tube with acid

Read scale to obtain g Hb/100 mL of blood

Dilute mixture with dH2O drop by drop until color is the same with standard

Place tube into the comparator block with scale turned away from experimentor

Hematocrit Determination

Homogenize sample and fill 2 Eppendorf tubes with 1.5 mL of blood

Centrifuge for 10 minutes

Measure H1 = height of RBC column

Calculate for Hc%

Measure H2 = height of RBC + plasma column

Haemometer Set-up

Calculations
Hematocrit Hc% = (H1/H2)(100%) Mean Corpuscular Volume MCV = Hc% x 10 RBC count in millions per mm3 Mean Corpuscular Hemoglobin MCH = (g Hb/100 mL blood) (10) RBC count in millions per mm3 Mean Corpuscular Hemoglobin Concentration MCHC = (g Hb/100 mL blood)(100) Hc%

Results
Subject RBC Count Hemoglobin Content H1 H2 Hc% Female 7.55x106 13.5 g Hb/dL 2 cm 3.4 cm 58.82 Male 1.47x107 11.9 g Hb/dL 2.1 cm 3.3 cm 63.64

Subject Female Male H2 H1

MCV (fL) 77.51 43.29

MCH (pg) 17.9 8.10

MCHC 22.95% 18.70%

Sample Computations
(Female Subject)
Hematocrit Hc% = (2 cm/3.4 cm)(100%) = 58.82% Mean Corpuscular Volume MCV = 58.8% x 10 = 77.88 fL 7.55 Mean Corpuscular Hemoglobin MCH = (13.5 g Hb/1 dL) (10) = 17.88 pg 7.55 Mean Corpuscular Hemoglobin Concentration MCHC = (13.5 g Hb/1 dL)(100) = 22.95% 58.82%

Normal Values
Blood Indices Hemoglobin Hematocrit MCV MCH MCHC% Male 14 - 18 g/100 mL blood 42-52% 78-98 fL 26-33 pg 28-36% Female 12 - 16 g/100 mL blood 37-48% 78-102 fL 26-33 pg 28-36%