Estimation of Blood Glucose level by Folin-Wu method

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BACKGROUND

Carbohydrate molecules plays diverse role in biological system.¹ In certain cases, carbohydrate molecules identified as human pathological biomarkers (such as Diabetes Mellitus). The estimation of the glucose in blood gives direct information on the patient DM control. Here we will use Folin-Wu method to determine the concentration of urine sugar in patient urine sample.

Reducing sugars turns reddish brown upon reacting with alkaline copper solution. The extent of colour development depends on the heating temperature, duration of heat. After development of the colour the OD680 can be checked to identify the content of carbohydrate in the sample, upon comparing with the standard curve of known glucose sample.

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REQUIREMENTS

Apparatus: Test-tube,

Water bath,

Spectrophotometer

Materials: 1 gm of glucose in 100 ml benzoic acid (stock 1% sugar solution), 1 mg/ml of the sugar solution (working standard solution), alkaline copper solution (A; 2% Na₂CO₃ in 0.1N NaOH. B; 5% CuSO₄ in 1% Na-K tartarate solution; mix solution A:B in a ratio 50:1), Phosphomolybdic acid, 10% sodium tungastate, 0.6 N H₂SO₄ and Blood sample.

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PROCEDURE

Boil 7 ml of water in a testtube then add 1ml of blood sample to the boiling water followed by addition of 1 ml of 10% sodium tungstate, mix the solution properly then add 1 ml of 0.6 N H₂SO₄.

Filter the resultant solution and collect the filtrate for glucose estimation. Transfer 2 ml of the filtrate to folin-wu tube, followed by addition of 2 ml of alkaline CuSO4. Boil the tube for 8 minutes and cool it immediately and record the OD680 in a spectrophotometer. Run similar control for blank (only water instead of filtrate) and working standard glucose solution to estimate the content in unknown (i.e. blood) sample.

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CONCLUSION

Determine the concentration of the glucose in blood by comparing the OD680 with the blank and the known sample.

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REFERENCES

1. Nelson N. A photometric adaptation of the Somogyi method for the determination of glucose. Journal of biological chemistry, 1994;153(2):375-80.