Acid Hematin Method by Sahli Hellige Hgb Detn

Summary

This video explains the acid hematin method for hemoglobin determination, using Sally Hellich's procedure. The materials include a Sally hemometer set, hydrochloric acid, distilled water, and blood collected via capillary puncture. When blood is added to hydrochloric acid, hemoglobin is converted to acid hematin, a dark brown compound. The color intensity is compared with a reference to estimate hemoglobin concentration. Advantages of the method include simplicity and portability, while limitations involve subjectivity, lack of accuracy for certain hemoglobin forms, and the production of a suspension rather than a clear solution.

Takeaways

• 🧪 The video explains a hematology procedure for hemoglobin measurement using the acid hematin method by Sally Hellich.
• 🛠️ Materials required for the procedure include a hemometer set, Sally's comparator box, hemoglobin tube, pipette, glass rod, dropper, amber bottle, and cleaning brush.
• 📏 Sally's calibrated tube has two graduations: one side shows grams per deciliter, and the other side shows hemoglobin percentage.
• 🌡️ Reagents for the procedure include distilled water and 0.1 normal hydrochloric acid (HCl), which is prepared by diluting concentrated HCl with distilled water.
• 🧬 The principle of the acid hematin method is that hemoglobin in blood reacts with HCl to form acid hematin, a dark brown compound, which is compared with a reference standard.
• 💉 Collect 20 microliters of capillary blood using Sally's pipette and add it to the Sally tube with hydrochloric acid for the reaction.
• ⏳ After mixing, allow the mixture to stand for at least 10 minutes to complete the hemoglobin-to-acid hematin conversion.
• 💧 Add distilled water in drops to dilute the solution, and compare the color with the standard in the comparator block until they match.
• 📊 Once the color matches the reference standard, read the hemoglobin concentration directly from Sally's calibrated tube.
• ⚖️ The method is simple and portable but has limitations, including subjective color comparison, inability to estimate carboxyhemoglobin or methemoglobin, and a lack of a stable standard.

Q & A

What is the main purpose of the acid hematin method by Sally Hellich?

The main purpose of the acid hematin method by Sally Hellich is to determine hemoglobin concentration in the blood by converting hemoglobin into a dark brown compound called acid hematin.

What materials are required for performing the Sally acid hematin method?

The materials required include a hemometer set (Sally comparator box, Sally's graduated hemoglobin tube, Sally pipette, thin glass stirring rod), a dropper, an amber bottle for reagent, a test tube brush, and reagents like distilled water and 0.1 normal hydrochloric acid.

What reagents are used in the Sally acid hematin method?

The reagents used are distilled water and 0.1 normal (N/10) hydrochloric acid, which is prepared by diluting 0.98 mL of concentrated hydrochloric acid in distilled water to make a 100 mL solution.

What is the principle behind the Sally acid hematin method?

The principle is that when blood is added to diluted hydrochloric acid, hemoglobin in the red blood cells is converted to acid hematin, a dark brown compound. The color intensity of the acid hematin complex corresponds to the hemoglobin concentration in the blood.

How is the hemoglobin concentration measured using the Sally method?

The hemoglobin concentration is measured by comparing the color of the acid hematin solution to a standard brown glass reference in the Sally comparator box. The matching color provides the hemoglobin value, which can be read from Sally’s calibrated tube.

What are the steps involved in collecting and processing the blood sample for this procedure?

First, 0.1 normal hydrochloric acid is added to Sally's calibrated tube. Then, 20 microliters of capillary blood is collected using Sally's pipette and dispensed into the tube. The pipette is rinsed, and the mixture is stirred. After standing for 10 minutes, distilled water is added until the solution matches the reference standard color.

What is the role of the comparator block in the Sally acid hematin method?

The comparator block contains the reference brown glass, which serves as a standard for matching the color of the acid hematin solution, allowing for the determination of hemoglobin concentration.

What are the advantages of Sally's acid hematin method?

The advantages are that it is simple, easy to perform, and portable, making it possible to conduct in different locations with the necessary materials.

What are the disadvantages of Sally's acid hematin method?

Disadvantages include subjectivity in visual color matching, the inability to measure carboxyhemoglobin and methemoglobin, the lack of a stable standard, and the resulting solution being a suspension rather than a clear solution.

Why is distilled water added after the formation of acid hematin, and what precaution is taken during dilution?

Distilled water is added to dilute the solution for accurate color matching. The precaution is to avoid over-dilution by frequently checking the color against the reference standard.