Hello all,
I wanted to know the amount of drug present inside the cell after drug treatment. I checked articles for the same, finally I ended with targeted mass spectroscopy (MRM analysis). But in my university this facility is not available. We have MALDI-TOF (Axmia). I'm wondering if I can able to perform the my experiment using MALDI-TOF.
Many thanks in advance,
Arun.
Dear Arun
As a very simple technique, you can prepare a standard solution with a known concentration of the drug itself as a mother solution and use the UV to check the absorption peak of this drug. After that you can use the remaining of the drug solution (after treatment) in the same UV cell after washing it to determine the concentration at the same wavelength then use the Lambert-Beer law to calculate the concentration. This method is called the standard solutions procedure.
Please note that dilute solutions of the drug will give you better results.
MALDI-TOF method is recommended for the compounds with molecular mass higher than
One way in which you can estimate the concentration is by measuring how much of drug remain external (in the media) after your incubation time. You can then subtract this from the total concentration of drug that was added initially. Depending on the drug there will be various methods by which you can measure the drug concentration (Commonly used might be HPLC).
You can also wash of extracellular drug and on lysis of the cells measure the concentration of drug in your lysate. I Lyse my cells using Glycin-HCl at pH3. I have however read that water also causes lysis of the cells. You will have to use an agent that lysis efficiently while also not affecting your drug.
Dear Arun
I assume you are refering to maldi-tofms imaging of tissues to measure intracellular drug concentration. Absolute quantitation by the method has many challenges. A recent article by Prof David Muddimann's group may help you.
"Quantitative Mass Spectrometry Imaging of Glutathione in Healthy and Cancerous Hen Ovarian Tissue Sections by Infrared Matrix-Assisted Laser Desorption Electrospray Ionization (IR-MALDESI)"
January 2018The Analyst
DOI10.1039/C7AN01828B
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