Biochem Laboratory Lecture 1

Estimation of blood glucose
Blood glucose levels are the amount of glucose that someone has in their blood at
any given time. Having high or low blood sugar levels could indicate an
underlying health condition that may require medical attention. Use this overview
of normal blood glucose levels to understand what your blood sugar levels mean.
What are normal blood glucose levels in healthy individuals?
Blood sugar levels can either be normal, high, or low, depending on how much
glucose someone has in their bloodstream. Glucose is a simple sugar that’s present
in the bloodstream at all times. Normal blood glucose levels can be measured when
someone fasts, eats, or after they’ve eaten. A normal blood glucose level for adults,
without diabetes, who haven’t eaten for at least eight hours (fasting) is less than
100 mg/dL. A normal blood glucose level for adults, without diabetes, two hours
after eating is 90 to 110 mg/dL.
Many factors affect blood sugar levels throughout the day:
1. Type of food consumed, how much, and when

2. Physical activity
3. Medications
4. Medical conditions
5. Age
6. Stress
7. Dehydration
8. Illness
9. Menstrual periods
10.Alcohol
An ideal blood sugar level for anyone without diabetes, regardless of age, in the
morning should be less than 100 mg/dL. Remember, blood sugar levels can
fluctuate throughout the day as a result of the factors previously mentioned.
Normal blood sugar levels for adults in mg/dl

Fasting Less than 100
Before meal 70-130
1-2 hours after eating Less than 180
Bedtime level 100-140
Estimation of Sugar by Folin-Wu Method (reduction method)
Principle
Glucose reduces the cupric ions present in the alkaline copper reagent to cuprous
ions or the cupric sulfate is converted into cuprous oxide, which reduces the
phosphomolybdic acid to phosphomolybdous acid, which is blue when optical
density is measured at 420 nm.
Reagents Required
Alkaline copper-reagent: Dissolve 40 gms of anhydrous sodium carbonate

(Na2CO3) in about 400 mL of water and transfer it into a 1-liter flask. Dissolve 7.5
gms of tartaric acid in this solution and 4.5 gms of CuSO4, which is dissolved in
100 mL water. Mix the solution and make it up to 1 liter.
Phosphomolybdic acid reagent: Add 5 gms of sodium tungstate to 35 gm of

phosphomolybdic acid. Add 20 mL 10% NaOH and 50 mL distilled water boiled
vigorously for 20 to 40 minutes so as to remove the NH3 present in molybdic acid.
Cool it and dilute to 350 mL. Add 125 mL of o-phosphoric acid and make the
volume up to 500 mL.
Standard glucose solution: Dissolve 100 mg of glucose in 100 mL distilled water
and take 10 mL from this solution and make it up to 100 mL.
Procedure
Pipette out standard glucose solution 0 to 1-mL range and make up the solution to
2 mL with distilled water. Add 2 mL of alkaline Cu-reagent to all the test tubes.
Mix the contents and keep them in a boiling water bath for 8 minutes. Cool under
running water without shaking and then add 2 mL of phosphomolybdic acid
reagent to all the test tubes. Wait for 10 minutes and mix the content. Make up the
volume to 25 mL with distilled water. Take the optical density at 420 nm.
Result:
100 mL of unknown sample contains .................. mg of sugar.
Glucometer (or glucose meter)
Glucometer is a medical tool that is used for measuring the approximate level of
glucose in the blood.
Working principle of a glucometer:
i. A glucometer determines the concentration of glucose in the solution.
ii. Most of the glucometers are based on electrochemical technology which use
electrochemical test strips to perform the measurement.
iii. A small drop of the solution that is to be tested is placed on a disposable test
strip that the glucometer uses for the glucose measurement.
Glucometer test strips:

i) In each glucometer test strip which are to be used for determining glucose in the
blood, contains an enzyme called glucose oxidase. This enzyme then reacts with
the glucose in the blood sample and creates an acid called gluconic acid.
ii) The gluconic acid thus formed then reacts with another chemical in the testing
strip called ferricyanide. The ferricyanide and the gluconic acid then combines
with each other and forms ferrocyanide.
iii) As soon as the ferrocyanide has been formed the device (i.e., glucometer) runs
an electronic current through the blood sample on the strip.
iv) This current thus generated is able to read the ferrocyanide and identify the
amount of glucose present in the blood sample on the testing strip.
v) That number is then displayed on the screen of the glucometer.

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Estimation of blood glucose

What are normal blood glucose levels in healthy individuals?

Many factors affect blood sugar levels throughout the day:

1. Type of food consumed, how much, and when

Normal blood sugar levels for adults in mg/dl

Before meal 70-130

1-2 hours after eating Less than 180

Bedtime level 100-140

Estimation of Sugar by Folin-Wu Method (reduction method)

Alkaline copper-reagent: Dissolve 40 gms of anhydrous sodium carbonate

Phosphomolybdic acid reagent: Add 5 gms of sodium tungstate to 35 gm of

100 mL of unknown sample contains .................. mg of sugar.

Glucometer (or glucose meter)

Working principle of a glucometer:

i. A glucometer determines the concentration of glucose in the solution.

Glucometer test strips:

v) That number is then displayed on the screen of the glucometer.

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