Thursday, July 8, 2010

SCIENCE AND EMOTIONS OOooOOooOoOOEEEeeeeee

Yes. The last week and a half have been a scientific emotional roller coaster. It all started late last week when Andrea (the LC-MS technician) finally finished measuring my samples. For all of you that don't know, the LC-MS separates your sample into its different components and measures its UV reactivity. It also generates masses for those compounds which may be in your sample. From the masses, usually you can derive some sort of chemical formula which will give you a clue about what your molecule actually is. So I finally got all of my LC-MS data back. I submitted six samples, 4 from my Sea Cucumber body walls and 2 from my more interesting super 1 fungal isolation.

I asked my PI to have a look at the data. First, we checked out the Sea Cucumber data. Initially my project was to redo an experiment done with some sea cucumbers in 2008 to determine if any additional secondary metabolites called saponins could be found. Unfortunately, I only found three of the ten identified saponins in my sample and no new compounds. This was both a victory and a defeat for me. I was excited to have finally found something I was looking for but was somewhat disappointed that I could only find 3 and couldn't find anything new. However, I wasn't too concerned with this data as I had sort of placed this project on the back burner since I started my fungal project.

I looked at my fungal LC-MS data with my PhD helper and suddenly things got very exciting! We had four new peaks (or potential compounds)! My PI had never seen these peaks and when I searched for the peak's masses in the Marine Molecular database I found no matches, which meant I had most likely found some new stuff! I was so excited! I kept thinking, I have made discoveries! I am going to publish this! I am SO HAPPY!

So the next step was so separate and purify these compounds using HPLC. This machine separates whatever is in your sample based on its polarity and allows you to isolate pure fractions of your sample. However, in order to do this you need tall and tight UV peaks to know when to collect whatever you want collected. I was so excited! If I had four peaks on the LC-MS I should have 4 on the HPLC as well. I started the HPLC and waited about 45 minutes before seeing any peaks. Finally they showed up. BUT.... DUN DUN DUNNNNNNNN they were all VERY minor peaks. All four were there, but the peaks were too short and shallow for me to collect any pure compounds out of them. I was sad, but I still had hope. If I made the solvent I was running through the HPLC more acidic, then I perhaps the peaks would perk up. So I got some formic acid, threw it into my solvent and tried again on a longer column. However, after running my sample through these conditions, I failed to see ANY peaks! Things were looking grim. I was scared. Here I was thinking I had just made some discovery, getting all pumped up, feeling like I had caught some weird 21st century gold rush fever through my natural products. And the you-know-what seemed to be hitting the fan. I decided to try my sample one more time with the acid in the first column I used. Again, although my peaks were back, they were too short and shallow to do anything with.

DEFEAT I tell you. DEFEAT! Sadness. Mustafa the PhD I work with thinks my sample is not concentrated enough. His solution, grow 3 or 4 more fungal samples and isolate compounds from those. THING IS, the fungus takes about 3 weeks to grow and due to weird Aberdonian "July Holidays" I only have 9 more days left of work. AHHHH SO SAD! I FELT LIKE A FAILURE!

Alas, today I decided to take a step back and look at situation from a new more broad perspective. My little feverous stint with discovering some new natural products infused me with a new genuine motivation to get back home and start working on the fungus I know and love. It also gave me some ideas concerning extractions and isolations with our aphids. AND THAT GOT ME THINKING! Although my projects here turned out to be somewhat disappointing, I have learned SO MUCH. I honestly can't wait to go home and start work with my new organisms. Not only that, but because I have essentially taught myself how to prepare, extract and isolate natural products from different organisms, I am totally confident that I will be able to get some great work done back home. EXCITEMENT! JOY HAS RETURNED!

Last weekend, I went to a "fourth of July" cookout at a beach in my friend's hometown of Montrose. Now, let me tell you.... this beach is not easy to get to. First you drive to a golf course, then after crossing the golf course, you have to jump down this huge sand dune only to arrive at PERHAPS ONE OF THE PRETTIEST AND MOST PRIVATE BEACHS IN THE COUNTRY. It was awesome. Again, I have lots of pictures but no way to upload them so stay tuned. When July 4th officially started we set off Japanese Lanterns as if they were some sort of firework alternative. It was nice, booze, vegan burgers, and vegan yet haggis flavored chips. YUM. After the BBQ we had to traverse the big dune and golf course at about 2:30 AM in the dark (and with a little booze in our systems...) and that was a JOY! We literally had to climb up this dune like Indiana Jones style. It was hilarious. Good times in the ol' 'Trose.

This week, after my disappointments, I have decided I will return to my sea cucumber project and try to quickly isolate some more saponins before I leave.

And as Ogy said, EVERYONE is invited to the semi-IRES reunion this weekend. It will be help in Dublin. Whiskey will be present.

4 comments:

  1. Being the expert mycologist that you are, I think we should go mushroom-hunting in Atlanta. When in season, our area is the perfect storm for all sorts of 'shrooms.

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  2. Oh I bet. And this amazing woman who works at my lab, Nancy, is pretty much a local plant and animal taxonomist. So anything we find will be eaisy classified.

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  3. what a roller coaster, Austin! Whew.

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