Week 2 – Starting Lab Work
This week I tried isolating enzymes from the mint leaves. I picked up the mint leaves from a friend’s house and brought them to lab, using 20g of different sized leaves.
I needed to extract the enzymes so I could test for enzymatic activity, and I used the buffers I made last week to extract the enzymes from the mint cells. Since ASDRP (where I’m doing my lab work) doesn’t have a cell disruptor, I used a blender to disrupt my cells. The goal of this was to isolate glandular trichomes, or cells high in enzymatic activity, from the other plant cells. After I blended my cells, I filtered the mix of mint cells and buffer using filter paper in hopes of collecting the glandular trichomes on the filter paper. However, when I took the filter paper to my advisor, we were unable to see anything except for cell debris under the microscope. We suspected that the blender may have shredded the glandular trichomes, so next week I will be trying a different cell disruption method of vortexing the mint leaves with glass beads.
I also started working on developing a method for the gas chromatography-mass spectrometry (GC-MS). This machine ionizes volatile compounds and gives data on the volatile compounds in the sample. Since I was testing the volatile composition of the mint plants and cells, I needed to find the right protocol for using the machine so I could get the clearest results. I put standard volatile compounds from mint and orange through the GC-MS, which successfully matched literature results. I also tried putting some sonicated mint leaves through the GC-MS, except all I could see was a bunch of noise. My advisor thought that it might be that my sample was too diluted and suggested that I try extracting and concentrating down the compounds from the mint leaves next time I ran the GC-MS.
Next week I’ll be trying to extract mint enzymes again, and hopefully I’ll be able to get something!