The first thing we did today in lab was go check the results of the series of GCMS runs that we created at the end of the day yesterday. When we got to the machine, there was an error message from the fourth sample of the series, so it did not get even close to being finished. We couldn't even log onto the computer, so there was some real trouble. Hopefully, the computer will be fixed by another professor in the chemistry department by tomorrow.
We then made a phosphate buffer for the HPLC to use, since the amount of buffer in the machine was low. We made a solution by diluting phosphoric acid in water, then adding concentrated sodium hydroxide to get to the desired pH of 3. The buffer was added to the HPLC.
Tony Le, who did research with Dr. Splawn this past semester, came to the lab to show us how to create a saliva sample. He mixed together 11 mL of chloroform, 5.5 mL methanol, and 0.5 mL of water. Three mL of oil is injected in the mouth, swirled around for 3 minutes, and then spit into a small beaker (you want to generate around 10 mL of saliva). Two mL of ethanol is added to the sample to kill any enzymes.
The saliva sample and the chloroform, methanol, and water mixture are combined into a separatory funnel. The flask is then inverted for 3 minutes and then is allowed to sit to let the layers separate. The bottom layer, which contains the fatty acids, is the one we want to collect. The sample is then ready to be put into a machine.
Neelam and I tried to create our own sample after Tony showed us, but our layers would not separate. Hopefully tomorrow we can create some successful samples!
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