Hemoglobin is the red-pigmented protein located in the erythrocytes (RBC). It is produced in the bone marrow with a life span of about 3 months. After that, it breaks down in the spleen. Each hemoglobin consists of four subunits consisting of iron. It is the carrier of oxygen from the lungs to tissues and the carrier of carbon dioxide from tissues to the lungs. Thus, the measurement of hemoglobin is very important for us. You can measure the amount of hemoglobin in the blood by using Drabkin’s method/ Cyanmethemoglobin method in a pathology lab using a semi-auto biochemistry analyzer. Besides Drabkin’s method, we will also discuss the method of increasing hemoglobin in the blood.
Types of Hemoglobin
- HbA:- It occupies at least 96% of total hemoglobin in adults.
- HbA2:- It occupies the remaining 2.5 – 3%.
- HbF:- Fetal hemoglobin diminishes as the year goes by. So, in normal adults, it will reduce to less than 1%.
Hemoglobin Normal Range
The range shown below is the approximate value of hemoglobin in the blood of a normal person. Thus, the normal range can slightly vary from one laboratory to another due to the method of sample preparation, reagents used, etc. The other reasons can be due to the geographical location, habitat, age, sex, etc.
- Men:- 14.0 – 17.5g/dl
- Women:- 12.3 – 15.3g/dl
- Children:- 10.7 – 16.8g/dl
Finally, if the level of hemoglobin in the blood goes out of range, then you are recommended to visit the doctor for further investigation.
Low Hemoglobin in the Blood
We know that hemoglobin is responsible for carrying oxygen from the lungs to different parts of the body. Oxygen is responsible for the burning of food which releases energy. So from this, we can know that the symptoms of low hemoglobin are weakness, fatigue, headache, etc.
- It is formed in the bone marrow. Thus, any kind of defect in the bone marrow may hinder its production.
- It is present in the red blood cells. Thus, excessive bleeding, hemorrhage, anemia, periods of the women, etc can cause a decrease in the red blood cells.
- Different kinds of cancer can cause excessive breakdown of the red blood cells. Ultimately, there will be a lower level of RBC which in turn reduces the Hb level.
- Similarly, certain kinds of pregnancy, infections, kidney diseases, etc can also lower the level of hemoglobin in the blood.
The main source of hemoglobin is the bone marrow itself. Besides that, there are natural diets that can provide it.
- Green vegetables
- Grains such as rice and barley
- Fruits such as apple and pomegranate
Besides taking a healthy diet there are other sources as well. Some of them are the consumption of iron supplements, vitamin b complex, etc. Even blood transfusion by the medical persons can increase it.
To know more about the iron tablets, please go through the video shown below.
Drabkin’s Method/ Cyanmethemoglobin Method of Hemoglobin Measurement
1. Principle Reaction
Drabkin′s reagent consists of potassium ferricyanide, potassium cyanide, and potassium dihydrogen phosphate. Potassium ferricyanide oxidizes hemoglobin into methemoglobin. Then potassium cyanide converts methemoglobin into colored cyanmethemoglobin. The color intensity of this complex is directly proportional to the concentration of hemoglobin in the sample which can be measured by both filter photometer and spectrophotometer.
2. Sample Preparation
The table shown below for hemoglobin measurement is valid for the reagents of Tulip and Atlas Medical. So, if you have the reagent kit of any other company, then there can be minor fluctuation in the method.
Mix the whole blood/calibrator with Drabkin’s reagent for making a sample test or for the calibration. Incubate the solution for 5 mins at room temperature. For making a reagent blank you can directly feed the reagent solution in a semi-auto biochemistry analyzer.
If the concentration of hemoglobin in the whole blood is too high (out of range/ non-linear), then dilute the blood with 0.9% NaCl or distilled water at the ratio of 1:y. When you get the result, multiply it with the dilution factor (1+y).
3. Programming of Hemoglobin for a Semi-auto Biochemistry Analyzer (Drabkin’s Method/ Cyanmethemoglobin Method)
3. Handling of Hemoglobin Reagents
- Wear an apron and surgical gloves before carrying out the measurement.
- Look for the expiry date of the reagents during purchase and measurement time. Suppliers tend to give you reagent kits with a low expiry interval.
- Once you open the reagent bottles, the lifespan will decrease. So, look in your reagent kit literature for that.
- Bring the reagents and samples to room temperature before you can carry out any measurement.
- Always store reagents in the refrigerator when not in use.
Supplier of Hemoglobin Reagent
If you want to purchase the reagent of Spinreact Company, then you can contact the given number (+977 9849034738). The number belongs to that of the Mandawi Enterprises Pvt. Ltd. It is an authorized dealer of Spinreact in Nepal. The distributor is located in Thapathali, Kathmandu.
You may contact the supplier to know about the price of the hemoglobin reagent.