Haemoglobin is a protein found in red blood cells that carries oxygen from the lungs to the body's tissues and removes carbon dioxide. It is an essential component of the human body, and its concentration in the blood, known as haemoglobin concentration or haemoglobin level, is an important indicator of an individual's overall health. Haemoglobin estimation is a laboratory test that measures the concentration of haemoglobin in the blood.
There are several methods for haemoglobin estimation, including spectrophotometry, cyanmethaemoglobin method, and hemoglobinometry. Here, we will discuss the procedure for haemoglobin estimation using the cyanmethaemoglobin method.
Preparation: The first step in the haemoglobin estimation procedure is to collect a blood sample from the patient. This is typically done by inserting a needle into a vein in the arm and drawing a small amount of blood into a tube. The blood sample should be collected in a tube containing an anticoagulant, such as EDTA, to prevent clotting.
Dilution: Once the blood sample has been collected, it is usually diluted with a saline solution to obtain a more accurate result. The dilution factor will depend on the concentration of haemoglobin in the blood sample.
Addition of reagent: The next step is to add a reagent to the diluted blood sample. In the cyanmethaemoglobin method, the reagent is a solution of potassium ferricyanide and potassium cyanide. These chemicals react with haemoglobin in the blood sample to form cyanmethaemoglobin, a compound that absorbs light at a specific wavelength.
Measurement of absorbance: The absorbance of the solution is then measured using a spectrophotometer, a device that measures the amount of light absorbed by a sample at a specific wavelength. The absorbance is directly proportional to the concentration of haemoglobin in the blood sample.
Calculation of haemoglobin concentration: The haemoglobin concentration is then calculated using a formula that takes into account the dilution factor and the absorbance of the solution. The final result is expressed in grams of haemoglobin per deciliter of blood (g/dL).
It is important to note that the accuracy of haemoglobin estimation may be affected by several factors, such as the quality of the blood sample, the condition of the reagents, and the accuracy of the spectrophotometer. Therefore, it is essential to ensure that the laboratory equipment and reagents are properly calibrated and that the blood sample is collected and handled correctly.
In conclusion, haemoglobin estimation is a laboratory test that measures the concentration of haemoglobin in the blood. The cyanmethaemoglobin method is a common procedure for haemoglobin estimation, which involves collecting a blood sample, diluting it with a saline solution, adding a reagent, measuring the absorbance of the solution, and calculating the haemoglobin concentration. It is an important tool for diagnosing and monitoring various health conditions, including anaemia and other blood disorders.
Its normal value is 0. A Manual Of Medical Laboratory Technology. What an ideal web site. On breakdown it forms important bile pigments. Acid hematin solution is not stable and colour formation is slow, sometimes precipitation may occur. Haemoglobin concentration in a newborn baby is high.
It reveals how nicely you understand this subject. Individuals with sickle cell disease and increased HbF often have a milder disease, as the HbF inhibits sickling of the red cells. Abnormally high or low Hgb can cause symptoms like Learn why you may need an Hgb test, what the The Hgb test uses a sample of your blood to determine hemoglobin levels. These children have one or two alpha gene deletionsand are silent carriersor have the alpha thalassemia trait. If you've been diagnosed with anemia or polycythemia vera, your doctor may use a hemoglobin test to monitor your condition and guide treatment. . Wait for a minimum of 10 minutes after mixing blood with HCI for complete conversion of haemoglobin to acid haematin.
Estimation of Hemoglobin by Sahli’s Method HAP Lab Manual
Lower than normal results If your hemoglobin level is lower than normal, you have anemia. Description of apparatus The Sahli-Hellige Hemo meter has the following components: Comparator : It accomodates a glass comparison tube which is graduated in gram percent of Hb 2-24 as well as percentage 20-140. The color of the solution is measured in the spectrophotometer at wave length of 540nm. Sara Fadhil Bunea, Medical laboratory techniqes, Human physiology practical. Each amino acid chain is attached to a heme moiety to form a single haemoglobin molecule. What are the indications of haemoglobin estimation? Hemoglobin Hb is a conjugated protein present in Red Blood Cells RBCs.
Sahli's Method of Hemoglobin Estimation: Introduction, Principle
You usually need to go to a lab to have your blood drawn. Hemoglobin H is also associated with significant breakdown of red blood cells as it is unstable and tends to form solid structures within red blood cells. The haemoglobin molecule is made up of haem and globin. Good quality anhydrous CuSO4 , CuSO4, 5H2O 2. In the laboratory, a process called electrophoresis passes an electrical current through the hemoglobin in your blood sample.
If a small amount of Hb Barts is detected, it usually disappears shortly after birth due to dwindling gamma chain production. As part of a routine checkup: Your doctor may have your hemoglobin tested to follow up on a complete blood test during a routine physical. It serves two important functions in the body- transportation of oxygen and carbon dioxide in between tissues and lungs and acts as a buffer in maintaining blood pH. Selection of blood donors. Cheap and can be used for haemoglobin estimation where automated haematology analyses are not available.
Hgb (Hemoglobin) Test Results: What Could Cause Low or High Levels
Wipe away the excess of blood adhering to the tip of the pipette with cotton. The potassium ferricyanide converts Hb to methemoglobin which is converted to cyanmethemoglobin HiCN by potassium cyanide. When the colour matches, the graduated tube is taken out from the stand and the amount of solution in the tube is recorded. Haemoglobin is a conjugated protein that consists of a basic protein, the globin and haeme. Wipe the back surface of the scale at the end of each session, or if it becomes soiled during use HEMOGLOBIN ESTIMATION — Instructions MATERNAL AND CHILD HEALTH NURSING. Glass stirrer : Used for stirring the contents of the comparison tube.
HbF levels are also increased in a rare condition called Hereditary Persistence of Fetal Hemoglobin HPFH. Fill the Hb pipette by capillary action. If your blood sample will be used for other tests, you may need to fast for a certain amount of time before the sample is taken. The blood sample is sent to a lab for analysis. Wait for 8-10 minutes after adding the blood in acid. The intensity of color is proportional to hemoglobin concentration and it is compared with a known cyanmethemoglobin standard at 540nm green filter in a photoelectric colorimeter.
A single beta β S copy sickle cell trait typically does notcausesignificant symptoms unless it is combined with another hemoglobin mutation, such as that causing HbC or beta thalassemia. It is also known as Sahli-Hellige method. They then send your blood sample to a laboratory for analysis. In addition to its key role in transporting oxygen and carbon dioxide, haemoglobin The levels of Hb are significant to establish conditions like anaemia, leukaemia, chronic infections, certain cancers, etc. People who are homozygous for HbE have two copies of β E generally have a mild hemolytic anemia,microcyticred blood cells, and a mild enlargement of the spleen. The tubular part of the pipette is graduated in terms of micro millilitres.