Experiment 1 (Finished on 2/7/14)
Explore alternative extract solvents that are easier to work with and environmentally friendly(er). The extraction solvent in the pesticide detection kit from RenekaBio (part# 003RT). When the solvent was dispensed into a plastic cuvette to measure background, the plastic melted. A new solvent needs to be found that can do the same job but is compatible with plastics.
Use different solvents found in The Literature, as well as anything that can be found through trial and error. [Please add to this list as you see fit]
- 70% IPA
- Extraction Solvent from Kit (with a quartz or glass cuvette).
Absorbance = Log10(Io/I)
I = intensity on spectrometer at a particular wavelength through the sample
Io = intensity on spectrometer at a particular wavelength uninhibited
Max absorbance at 204nm. On the spectrometer there should be 0% or close to 0% intensity at this wavelength.
Light source: Deuterium Lamp (Shimadzu LH-80 would work, $369 on Amazon). Expected intensity spectrum would look like:
Ethanol is a weak absorber.
Light source: Broad range, something a like a W-filament (are those even legal?).
100% Methanol Max absorbance at 177nm. Yikes. Not sure if the deuterium lamp can even do that. Light source: Deuterium Lamp. See IPA above for details.
Extraction Solvent from Kit
Control Setup Use a quartz cuvette, adjust back light distance from camera to minimize saturation (not sure what this distance would be, but its 9" for fluorescent lights and 5" for RGB LED and a 425nm UV LED). Add 1.5mL of clean extract solvent to cuvette, mount cuvette directly in front of opening on spectrometer and use black tape to close off exposed sides. Cover setup with a box and measure spectrum.
Sample Extraction We want to use the various alcohols mentioned above to try and remove pesticide residues from various fruits and vegetables. Prepare sample (in this case raspberries). Take 3 raspberries, use a clean screw-top container and dispense 6mL of extract solvent onto sample and then shake for 2 minutes. Let sample sit for 5 minutes then extract ~3ml of extract solvent from bottom of container (typically, raspberry mass floats to the top). Spin down this solution in a centrifuge, and then dispense supernatant into the cuvette.
Measure Extracted Sample Using exact same setup as the control, measure the spectrum and compare to the control.
Setup: 40W Fluorescent bulb, 9" pathlength, raspberry ID: 366208816148DS12 70% IPA Visual - Dark red in color, clear. Very little debris. Most of extract taken from TOP of mixing vial, as this didn't separate out like the kit extract solution.
Red peak still visible after extraction, but the green band and the two ultraviolet bands are gone.
fig.1 white line is solvent + sample.
100% Methanol Setup: 40W Fluorescent bulb, 9" pathlength, raspberry ID: 366208816148DS12
Visual - Red in color, clear (lighter than 70% IPA). Some debris. Most of extract taken from TOP of mixing vial, as this didn't separate out like the kit extract solution.
All peaks still visible after extraction. They are actually almost identical.
fig.2 white line is solvent + sample.
Didn't see any change from methanol, though the solution color did change. Not sure then if I've got the right light source for this extract solvent. Need to remove IR filter from my camera and rerun the experiment.
IPA does show an actual change, and its quite significant. Almost all peaks are hidden are gone with the exception of red. This means that there is something (or multiply somethings) in the extract solution that are absorbing light. I think this big signal change is probably due to something inherent in the raspberry coming out during the extraction, and probably not related as much to pesticide residue.
Need to rerun after centrifuge the samples for a few minutes to see if that changes the signal. Suprised by the methanol signal. Need to try this with the IR filter removed from the camera.