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For The Determination Of Blood Glucose

For the determination of blood Glucose


Consumption of glucose in whole blood samples can be prevented by adding sodium fluoride to the specimen to inhibit the glycolytic enzymes. This approach is the generally applied method in the clinical laboratory. ... Glucose concentration may be determined in whole blood, plasma, or serum samples.

Glucose is the most important carbohydrate fuel in the body. In the fed state, the majority of circulating glucose comes from the diet; in the fasting state, gluconeogenesis and glycogenolysis maintain glucose concentrations. Very little glucose is found in the diet as glucose; most is found in more complex carbohydrates that are broken down to monosaccharides though the digestive process. About half of the total carbohydrates in the diet are in the form of polysaccharides and the remainder as simpler sugars.

About two-thirds of the sugar in the diet is sucrose, which is a disaccharide of glucose and fructose. Glucose is classified as a monosaccharide because it cannot be broken down further by hydrolysis. It is further classified as a hexose because of its six-carbon skeleton and as an aldose, because of the presence of an aldehyde group on carbon 1. The aldehyde group condenses with a hydroxyl group so that glucose exists as a hemiacetal ring structure. This ring structure explains many of the reactions of glucose.

Glycosylated Hemoglobin

A test that reflects long-term blood glucose control in diabetics is the concentration of hemoglobin A1c. When hemolysates of red cells are chromatographed, three or more small peaks named hemoglobin A1a, A1b, and A1c are eluted before the main hemoglobin A peak. These "fast" hemoglobins are formed by the irreversible attachment of glucose to the hemoglobin in a two-step reaction. The percentage of hemoglobin glycosylated depends on the average glucose concentration the red cell is exposed to over time. Since the average life of the red cell is 120 days, the percentage of glycosylated hemoglobin gives a good indication of the degree of blood sugar control over the preceding weeks.

Material and Methods

Fifty six samples which included normal healthy adults, hypoglycemic and hyperglycemic patients were analysed. Fasting blood samples were collected in sodium fluoride and potassium oxalate mixture in the ratio of 1:1 at concentration of 4 mg mixture per ml of whole blood. Plasma was obtained by centrifuging the blood at 2000 rpm for 10 min. We used Systronics and Hans 202 filter colorimeters; and Beckman double beam Model-25 and Shimadzu micro-flow CL-750 spectrophotometers. Hexokinase kit was obtained from Sigma Chemical Company USA and GOD-POD from Accurex, India. All other reagents were of analytical grade.

How should results be expressed?

There is a legitimate argument that glucose results should be expressed in terms of activity in the water of the specimen (numerically regarded as the same for plasma and red cells), because this represents the glucose available at the tissues.

However, most current laboratory methods do not directly measure this. Instead, they measure concentration in plasma, which is numerically lower, and this is how results are usually expressed.

For methods using a whole-blood sample the result is often, but not invariably, expressed as whole-blood glucose concentration, giving a figure lower than that for plasma glucose.

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