Measure the time taken for amylase to completely break down starch, by withdrawing samples at 10 second intervals and noting the time at which the solution no longer gives a blue-black colour with iodine solution (but the iodine solution remains orange). Use buffers to provide solutions at different pHs. Calculate the rate of this enzyme controlled reaction by calculating 1÷ time.
Yulin HB™ contains other products and information you need, so please check it out.
This procedure is simple enough for individuals to carry out if you have enough dimple tiles. If you choose to investigate five pHs, then groups of five students could complete the investigation by working together and pooling results.
Amylase solution and iodine solution are low hazard once made up. Wear eye protection when handling iodine solution.
Hazards of buffers may vary. See CLEAPSS Recipe card or supplier’s information and see Note 3.
1 Amylase (See CLEAPSS Hazcard and Recipe card) The powdered enzyme is HARMFUL, but solutions less than 1% are LOW HAZARD. It is wise to test, well in advance, the activity of the stored enzyme at its usual working concentration to check that substrates are broken down at an appropriate rate. Enzymes may degrade in storage and this allows time to adjust concentrations or to obtain fresh stocks. Amylase will slowly lose activity, so it is best to make up a fresh batch for each lesson; batches may vary in activity and results collected on different days will not be comparable. The optimum temperature for your enzyme will be listed on the supplier’s label.
Using saliva: the CLEAPSS Laboratory Handbook provides guidance on precautions to take (including hygiene precautions) in order to use saliva safely as a source of amylase. This has the advantage of being cheaper, not requiring technicians to make up fresh solutions each lesson, it is directly interesting to students, and salivary amylase is reliable. It also provides an opportunity to teach good hygiene precautions – including ensuring that students use only their own saliva samples (provide small beakers to spit into); that students are responsible for rinsing their own equipment; and that all contaminated glassware is placed in a bowl or bucket of sodium chlorate(I) before technicians wash up.
2 Starch suspension – make fresh. Make a cream of 5 g soluble starch in cold water. Pour into 500 cm3 of boiling water and stir well. Boil until you have a clear solution. Do not use modified starch.
3 Buffers: (See CLEAPSS Recipe card) If you make universal buffer it will contain sodium hydroxide at approximately 0.25 M, and should be labelled IRRITANT. Refer to other relevant Hazcards if you choose to make other buffers, or to supplier’s information if you purchase buffer solutions/ tablets. (Note 1)
4 Iodine solution (See CLEAPSS Hazcard and Recipe card). A 0.01 M solution is suitable for starch testing. Make this by 10-fold dilution of 0.1 M solution. Once made, the solution is a low hazard but may stain skin or clothing if spilled.
There are no ethical issues associated with this procedure.
SAFETY: All solutions once made up are low hazard. Wear eye protection, as iodine may irritate eyes.
Preparation
a Check the speed of the reaction with the suggested volumes of reactants to be used – 2 cm3 of starch: 2 cm3 of amylase: 1 cm3 of buffer at pH 6. Ideally the reaction should take about 60 seconds at this pH: this is the usual optimum for amylase (see note 1). If the reaction is too fast, either reduce the enzyme volume or increase the starch volume. If the reaction is too slow, increase the enzyme volume or concentration or reduce the starch volume or concentration.
Investigation
b Place single drops of iodine solution in rows on the tile.
c Label a test tube with the pH to be tested.
d Use the syringe to place 2 cm3 of amylase into the test tube.
e Add 1 cm3 of buffer solution to the test tube using a syringe.
f Use another syringe to add 2 cm3 of starch to the amylase/ buffer solution, start the stop clock and leave it on throughout the test. Mix using a plastic pipette.
g After 10 seconds, use the plastic pipette to place one drop of the mixture on the first drop of iodine. The iodine solution should turn blue-black. If the iodine solution remains orange the reaction is going too fast and the starch has already been broken down. Squirt the rest of the solution in the pipette back into the test tube.
h Wait another 10 seconds. Then remove a second drop of the mixture to add to the next drop of iodine.
i Repeat step h until the iodine solution and the amylase/ buffer/ starch mixture remain orange.
j You could prepare a control drop for comparison with the test drops. What should this contain?
k Count how many iodine drops you have used, each one equalling 10 seconds of reaction time.
l Repeat the whole procedure with another of the pH buffers to be used, or pool the class results.
m Consider collecting repeat data if there is time.
n Plot a graph of time taken to break down starch against pH, or calculate the rate of reaction and plot rate against pH.
This is a straightforward practical giving reliable, unambiguous results. The main errors will be in the order of mixing the enzyme/ substrate/ buffer, or a delay in sampling so that the reaction time is under-estimated or rate is over-estimated. Temperature variation affects enzyme activity, so results collected on different days are not comparable.
Health and safety checked, September
Download the student sheet Investigating the effect of pH on amylase activity (72 KB) with questions and answers.
http://rsc.org/Education/Teachers/Resources/cfb/enzymes.htm
Royal Society of Chemistry: Chemistry for Biologists: Enzymes
A clear and thorough presentation of information about enzymes as chemical catalysts and the factors affecting their activity.
(Website accessed October )
Pancrelipase is used to treat patients who cannot digest food normally because their pancreas does not make enough enzymes (exocrine pancreatic insufficiency). Pancrelipase tablet is used in combination with a proton pump inhibitor (PPI) in patients who have long-term swelling of the pancreas (chronic pancreatitis) or who have had some or all of their pancreas removed (pancreatectomy).
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Pancrelipase contains a mixture of digestive enzymes (eg, lipases, proteases, and amylases) needed for the digestion of proteins, starches, and fats.
This medicine is available only with your doctor's prescription.
In deciding to use a medicine, the risks of taking the medicine must be weighed against the good it will do. This is a decision you and your doctor will make. For this medicine, the following should be considered:
Tell your doctor if you have ever had any unusual or allergic reaction to this medicine or any other medicines. Also tell your health care professional if you have any other types of allergies, such as to foods, dyes, preservatives, or animals. For non-prescription products, read the label or package ingredients carefully.
Appropriate studies performed to date have not demonstrated pediatric-specific problems that would limit the usefulness of pancrelipase delayed-release capsules in children. Safety and efficacy have been established.
Appropriate studies have not been performed on the relationship of age to the effects of pancrelipase tablets in the pediatric population. This medicine may cause unwanted effects (eg, weight gain, malnutrition) in children. Safety and efficacy have not been established.
Appropriate studies performed to date have not demonstrated geriatric-specific problems that would limit the usefulness of pancrelipase in the elderly.
Although certain medicines should not be used together at all, in other cases two different medicines may be used together even if an interaction might occur. In these cases, your doctor may want to change the dose, or other precautions may be necessary. Tell your healthcare professional if you are taking any other prescription or nonprescription (over-the-counter [OTC]) medicine.
Certain medicines should not be used at or around the time of eating food or eating certain types of food since interactions may occur. Using alcohol or tobacco with certain medicines may also cause interactions to occur. Discuss with your healthcare professional the use of your medicine with food, alcohol, or tobacco.
The presence of other medical problems may affect the use of this medicine. Make sure you tell your doctor if you have any other medical problems, especially:
Take this medicine exactly as directed by your doctor . Do not take more of it, do not take it more often, and do not take it for a longer time than your doctor ordered. To do so may increase the chance of side effects.
This medicine should come with a Medication Guide and patient instructions. Read and follow these instructions carefully. Ask your doctor or pharmacist if you have any questions.
Take this medicine with food.
Swallow the capsule or tablet whole with enough fluid (eg, water or juice) to prevent irritation of the mouth. Do not crush or chew it.
For patients taking the tablets :
For patients taking the delayed-release capsules :
Do not change the brands or dosage forms of pancrelipase without first checking with your doctor. Different products may not work in the same way. If you refill your medicine and it looks different, check with your pharmacist.
The dose of this medicine will be different for different patients. Follow your doctor's orders or the directions on the label. The following information includes only the average doses of this medicine. If your dose is different, do not change it unless your doctor tells you to do so.
The amount of medicine that you take depends on the strength of the medicine. Also, the number of doses you take each day, the time allowed between doses, and the length of time you take the medicine depend on the medical problem for which you are using the medicine.
If you miss a dose of this medicine, skip the missed dose and go back to your regular dosing schedule. Do not double doses.
Keep out of the reach of children.
Do not keep outdated medicine or medicine no longer needed.
Ask your healthcare professional how you should dispose of any medicine you do not use.
Store the medicine in a closed container at room temperature, away from heat, moisture, and direct light. Keep from freezing.
You may store repacked Zenpep® for up to 6 months.
Throw away unused Pertzye® mixture.
Do not eat or throw away the drying agent (desiccant) in your medicine bottle. The drying agent will protect your medicine from moisture.
It is very important that your doctor check your or your child's progress at regular visits to make sure this medicine is working properly. Blood and urine tests may be needed to check for unwanted effects.
This medicine may cause a serious bowel disorder called fibrosing colonopathy, especially if taken at a high dose for a long time in children with cystic fibrosis. Check with your doctor right away if you have bloating, constipation, diarrhea, nausea, stomach pain, or vomiting.
This medicine may increase the uric acid levels in the blood (hyperuricemia). Check with your doctor right away if you have joint pain, stiffness, or swelling, lower back, side, or stomach pain, or swelling of the feet or lower legs.
This medicine is made from the pancreas of pigs. Some pigs organs have transmitted certain viruses to people who have received them, although the risk is low. All are tested for viruses to keep the transmission risk low. Talk with your doctor if you have concerns about this risk.
This medicine may cause serious allergic reactions, including anaphylaxis and asthma, which can be life-threatening and require immediate medical attention. Tell your doctor right away if you have a rash, itching, hoarseness, or trouble breathing or swallowing after using this medicine.
DRG-
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