Determination of hemoglobin and its amount

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Hemoglobin, HB (Greek. Haima - blood and globe - sphere) - a respiratory pigment found in the blood. It gives the blood a red color, delivers oxygen from the respiratory organs to the tissues, and carbon dioxide from the tissues to the respiratory organs. The protein part consists of globin and iron porphylline. Hemoglobin combines with oxygen to form an unstable compound. In the pulmonary capillaries, where the partial pressure of oxygen is slightly higher, oxygen is delivered to the cells and tissues.

100 g of blood of a healthy person contains 13-16 g% of hemoglobin (13-17 g% 130-170 g / l in men; 12-16 g% 120-160 g / l in women). The amount of hemoglobin in the blood of women is lower than in men. The properties of hemoglobin in the body change throughout life. The amount of hemoglobin is determined by a special instrument - a hemometer. Determining the amount of hemoglobin is important in determining the respiratory function of the blood under normal conditions and a number of diseases that occur in the blood. An increase in hemoglobin in the blood is called hemoglobinemia, a decrease is more common in anemia.

Determination of hemoglobin - The amount of hemoglobin in the blood is determined as described above on a Sally hemometer or using an FEK apparatus.

Sally’s hemometer consists of a tripod with three vertical holes. The holes on both sides contain glass tubes with closed ends, which contain a comparable standard brown-yellow solution. Some hemometers are replaced by brownish-yellow glass rods instead of liquid-filled tubes. an open test tube is inserted to transfuse the blood, which should detect hemoglobin (Figure 5). This solution is divided into two series of levels: on the one hand level, it starts with the number 10 from the bottom and ends with the number 140, and on the other side it starts with the number 2 g% and ends with the number g%. The levels on both sides have the same height. A milk-colored glass plate that diffuses light is mounted on the back wall of the tripod to make the color of the liquids look good. A pipette is attached to each hemometer, which is 20 mm from the top end of the pipette³ For ease of operation, a rubber tube consisting of three glasses is placed on the upper end of the tube. The amount of hemoglobin in the blood is determined as follows: a decinormal solution of hydrochloric acid is placed in the middle of the test tube to ten digits, then 'ng pipetka 20 mm³ blood is drawn from the finger to the mark. As soon as the blood surface reaches the indicated mark without air bubbles, without removing the mouthpiece, squeeze the rubber tube with two fingers and measure the amount of blood in the pipette 20 mm.³. is checked again for compliance. The blood at the bottom of the pipette is wiped with a cotton swab and its tip is dipped into a decinormal solution of hydrochloric acid, which is previously prepared in a tube in the middle of the hemometer, the blood is slowly inflated into it to prevent bubbles from forming, and then the solution is stirred several times. Once sucked, it is re-introduced into the solution, and all the blood obtained in this way is poured into the solution. Wait 10 minutes for all hemoglobin to turn into hematin chloride, then the hemometer rises and the color of the liquid in the blood test tube is standard solution or stained glass. compared with the color of the rods. Almost always the color of the blood solution is darker than the color of standard solutions. Distilled water is poured into a pipette filled with blood solution using an ungraded pipette and mixed with a glass rod. Stirring is continued until the color of the liquid in it reaches the color of the liquid in the standard solution. The hemometer is then raised to the level of the eye to determine the level of the blood solution in the lower meniscus. The corresponding number on the right indicates the percentage, and the left shows the gram percentage of hemoglobin. Photoelectro-colorimeters and spectrophotometers are currently used to accurately check hemoglobin. Determination of hemoglobin in a photoelectrocolorimeter is based on the formation of its stained compounds with ammonia, acetoncyanhydrin and other substances. Each kit has a standard reagent (control). In addition, the central laboratory (for example, a regional hospital laboratory) can prepare and distribute standard (control) reagents. Standard (control) reagents are used each time the patient's blood is tested. The best method of quality control is to determine the patient's blood in two ways at the same time when the required amount of reagents is sufficient. If the results are different in both methods, the patient's blood should be re-examined. Clinical and diagnostic value of hemoglobin: a decrease in the amount of hemoglobin in the blood is one of the main laboratory indicators of anemia. The amount of hemoglobin varies depending on the form and expression of anemia. When the amount of hemoglobin decreases, in-depth and thorough examinations should be performed (determination of erythrocyte count, color index, study of erythrocyte morphology).