1. Experiment 6 Measurement of blood glucose (GOD-POD method )
Design by Romio

2. What is blood glucose ？

3. Clinically speaking blood glucose means the glucose in blood .
Generally speaking fasting blood glucose level is constant for healthy people

4. 3. Understand it's clinical significance
Aim:
1. Master the principle of GOD-POD method
2. Learn how to do it
3. Understand it's clinical significance

5. High sensitivity，Good stability
GOD-POD
High accuracy,
High sensitivity，Good stability
Why did we choose Glucose oxidase-peroxidase method (GOD-POD) to measure blood glucose ?

6. Principle ﹢ ﹢ ﹢ ﹢ Quinonimine Gluconic acid （GOD） Glucose O2 H2O2
（POD）
4-NH3-
Antipyrine
H2O
H2O2 ﹢ ﹢
Quinonimine

7. The absorption peak of quinone imine is 500nm
Generation amount of quinone imine is proportional to the content of glucose
The glucose concentration in the sample can be calculated using the standard comparison method

8. Equipment Test tube : 1.5×5cm（×3） pipet : 2mL（×1）, Washing ear ball
micropipette ：20μl
centrifuge , centrifuge tube 10ml（×2）
Water Bath

9. Reagents Glucose measurement kit
The glucose standard solution (5.55mmol/L)
Sample：fresh serum

10. Operation Steps 1. Sample processing 37℃ Water bath it for 15min
Draw blood 3-5 mL
Extract the supernatant
centrifuge the blood
for 10min (3000rpm/min)

11. 2. Use 3 small test tube to add reagent according to table below
NO.
Reagent
Blank tube
Standard tube
Sample tube
dd-Water (μL ) 10 ——
Standard glucose (μL )
Serum samples (μL )
Enzyme solution ( mL )
1.5

12. 3. After mix reagents for each tube, 37℃ Water bath for 30min，then use 0.5cm cuvette to measure their absorption value At 500nm wavelength

13. Write down the data and then calculate them by Lambert-Beer law
Calculation
Write down the data and then calculate them by Lambert-Beer law
Asample
Cstandard
Csample = ×
Astandard

14. Cstandard = 5.55 mmol/L
range of normal value：
3.89 ～ 6.11mmol/L

15. Clinical significance
1. Hyperglycemia
Glucose concentration >7.0mmol / L
e.g. diabetes
2. Hypoglycemia
Glucose concentration <2.8mmol / L
e.g. limosis

16. Questions
1. Why human can maintain normal serum glucose at a certain level?
2. What is the effect of insulin and adrenaline on blood glucose ? And why?