Celastrol Synthesis Lab Report

The goal of my project is to create an assay for the detection of celastrol (a molecule with anti-inflammatory/anticancer properties) by using a form of chromatography and mass spectrometry. I began working towards this goal at Dr. John Letterio’s lab in the Wolstein Research Building on June 13th 2017. I worked 4 days per week from 10:00 am to 3:00 pm until I completed the summer’s work on August 20th. I was given the opportunity to work closely with Dr. Emily Barker, who had already done a study on the administration of celastrol into the diets of rodents. This was a big learning experience for the both of us, since neither of us was familiar with analytical assays. Any time that was not spent collecting data was used to help out around the

I was given time to get familiar with the environment of the lab by being assigned various lab duties such as cleaning micropipette trays or creating buffer solutions. On June 21st we prepared a sample of the isolated compound (celastrol) and brought is to a chemistry lab at CSU for testing. We decided on gas chromatography mass spectrometry (GCMS) because of its availability at the time. In order to use GCMS, the compound must be a gas. This prompted us to derivatize it. We heated the compound with trimethylsilane (TMS) to increase the compound’s volatility. We ran the compound through several trials of GCMS and obtained our data through Mass Spectra and Chromatograms. The instrument was able to detect the compound in two derivatized forms. We found this interesting because the molecule obtained 2 methyl groups in some cases and 3 in other cases. In July, we worked to find the limit of detection (LOD) for the instrument. To do this, we did several serial dilutions with methanol as a solvent for various concentrations of cleastrol. We then on July 19th took the samples to a hydrogen dryer to evaporate the methanol and followed the same process for

First of all, I would like to run isolated celatrol through LCMS. I am aware that the machinery for LCMS is newer at CSU, which may give us a higher LOD. Also, there are blood serum samples left over from the previously mentioned study done at Wolstein. These samples are good candidates for a GCMS or LCMS test. Now that we know what chromatograms and mass spectra for celastrol in its pure form looks like, we can now compare that to actual blood. Following her maternity leave, Emily and I have stayed in touch throughout the school year. I will return to the lab in early June to pick up where I left