The iodine-sampling method

Place drops of iodine in a spotting tile. Equilibrate amylase and starch at the chosen condition, mix them and start the timer. At fixed intervals, transfer a sample to iodine. Record the time when iodine remains orange-brown, showing that starch is no longer detected.

Control the reaction conditions

If temperature is changed, keep pH constant with a buffer and use the same enzyme and substrate concentrations and volumes. If pH is changed, keep temperature constant with a water bath. Use fresh, clean equipment to avoid contamination.

Turn time into rate

When the same endpoint is used, rate can be estimated as 1 ÷ time. A shorter time therefore gives a greater rate. Repeat each condition, calculate a mean time and then calculate the rate consistently.

Limitations and improvements

Judging the exact colour endpoint is subjective and samples continue reacting during transfer. Sample at shorter intervals, use the same observer or a colorimeter where available, and keep transfer timing consistent. Pre-equilibrate solutions before mixing.