While measuring an analyte using the methylene blue method with a spectrophotometer after inducing the lysis of RBCs, does the hemolysis/hemoglobin interfere with the analyte concentrations? If yes, Is there any method to avoid/remove interference/biases?
Thanks in advance.
EDTA is known as a chelating agent. This substance forms a complex with Ca+2. In this way, blood coagulation is prevented. Ca+2 is needed for coagulation and EDTA makes it unavailable by forming a complex. EDTA is mostly used in the form of disodium, dipotassium or tripotassium salts. Therefore, depending on what concentration and which sodium or potassium salt and what capacity you use, as well as when and what temperature you use, it causes various changes in blood parameters and also affects the morphology of red blood cells. Currently, the best salt is It is tripotassium, provided that the storage time is less than four hours, this compound is easily dissolved in the sample. It should be noted that concentrations higher than 2 mg/ml will cause red blood cells to shrink. EDTA preserves the cellular components of blood, so it is suitable for use in hematology tests.
EDTA inhibits alkaline phosphatase, creatine kinase, leucine aminopeptidase and lactate dehydrogenase by chelating metal cofactors. EDTA should not be used in photometric methods and in the measurement of Ca+2 and Fe+2. It should be noted that due to the presence of nitrogen, this compound will interfere with the measurement of blood urea nitrogen. On the other hand, sodium and potassium EDTA salts should not be used in the measurement of sodium and potassium, which are blood electrolytes.
Hello, you know that when white light is obtained from the combination of two colors, those two colors are called complementary. Some of the colors that are obtained from their combination with a certain ratio of white color are:
Red with blue green
blue with orange-red etc
Complementary colors include the three colors purple, orange and green, and each of them comes from the combination of other primary colors. Purple is created from the combination of blue and red, orange is created from the combination of yellow and red, and green is created from the combination of blue and yellow.Therefore, your blue color with the combination of red color caused by hemolysis of hemoglobin creates an almost purple color, which depends on the amount of hemolysis and the amount of hemoglobin. Maybe it is better to know if there is hemoglobin in all of your analyzes or only in some of them. It is better to measure in two wavelengths or use a blank.
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