9. Suppose you were asked to develop an experimental procedure to test if Carb C
ID: 280575 • Letter: 9
Question
9. Suppose you were asked to develop an experimental procedure to test if Carb Cutter is effective at preventing amylase from digesting starch. Here is a list of supplies and equipment that you could use for your experiment:
Iodine - Note: Adding iodine to a reaction mixture of starch and amylase will immediately stop amylase activity.
Water
Concentrated stock starch solution (concentration: 10 g/L)
Amylase (assumed this is an accurate concentration for your experiment)
Syringes for measuring in units of ml
Spectrophotometer tubes
Test tube rack
Spectrophotometer
Carb Cutter product
A: Describe your experimental methods to test the effectiveness of Carb Cutter.
B: Write your procedure out in a numbered list of Steps (e.g. Step 1, Step 2, etc.).
You must use all of the supplies in your design at least once. It is fine if you use supplies more than once.
For example, howwould making dilutions help you complete the experiment? How would a spectrophotometer help you? How will you know that Carb Cutter is working. You don't have to show me the dilutions, but you do have to tell me how they will help you.
Explanation / Answer
A) Aim: to check the effectiveness of carb cutter in preventing amylase activity under laboratory conditions.
amylase is an important enzyme for carbohydrate digestion in animals, It catalysis the hydrolysis of starch into sugars. Amylase activity generally demonstrated by adding amylase enzyme to the starch solution and followed by incubation. When Iodine solution is added to the mixture it turns into blue indicating that the presence of remaining starch. In the present experiment the carb cutter working against the amylase activity, so in order to check the efficiency of the carb cutter, the carb cutter is added to the solution containing starch, and amylase. If the carb cutter is efficient it prevents amylase activity and upon adding iodine indicator for starch hydrolysis, the solution will turn into blue indicating that carb cutter is efficient. If the solution is turning in to blue, it states that carb cutter is not efficient.
For accurate measuring of the efficiency of carb cutter, we need to quantify the result. By calculating the change intensity of colour due to inhibition of amylase activity when compared with standard amylase activity without carb cutter addition. For evaluating standard amylase activity different concentration of amylase enzyme is prepared and added to the separate starch solutions. After adding the colour indicator the optical absorbance measured in the spectrophotometer. The carb cutter efficiency on reducing amylase activity is measured by comparing it with standard spectrophotometric readings.
B) 1. Take at least 6 test tubes containing 100 ml of starch solution.
2. amylase solution is used by preparing different dilutions of it. That original amylase solution is considered as high concentrated solution, and we can add water to prepare first dilution, to the first dilution if futher addition of water makes 2nd 3rd 4th and 5th dilution. and each amylase dilution is added in the same quantity to the seperate test tubes containig starch solutions.
Add amylase solution with a syringe to each test tube in sequential order i.e from 1st to 5th test tube the concentration of amylase will be increased, but to the last test tube without amylase addition can be used as a control.
3) iodine indicator is added to each test tube and optical density of blue colour is measured in the spectrophotometer.
4) separately, another test tube is also prepared in same way containing the starch solution, amylase along with carb cutter. The colour intensity is checked under spectrophotometer. it is compared with standard amylase values to understand the reduction in amylase activity.
Related Questions
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.