Today was my last day of Work Experience at Tergus Pharma. We spent all of our time in the analytical department and lab. We worked with a placebo of a drug (with no active drug component) and we worked on validating it. Analytical has three sub-sections. First, there is method development, then validation, and lastly quality control. These three groups work together to first come up with a method, then test the method, and the finally just use the method. Since we worked in validation, we got to prepare placebo samples and then test them in an HPLC (high-performance liquid chromatography) machine. It was a really hands-on day, and it was a lot of fun.
Author: om_naphade
Week 2, Day 3: IVRT and mEmBrAnEs
Today, I spent time learning how IVRT works at Tergus Pharma. I observed how they dose ointments, gels, and cremes into the dosing chamber and also how they sample the doses (by shadowing Emeel). We learned about the regular intervals in which the sample doses. We later learned about HPLC (high-performance liquid chromatography) and we watched Adam making the mobile phases. We didn’t have time to actually run the chromatography, but we plan on doing that tomorrow if we have time.
One really cool thing we learned today was that all of the lab’s data is uploaded in real time to a place where it cannot be deleted. Then, later, the FDA has access to all of the results. Because of this, the scientists and lab techs double and triple check their methods before they run them in an HPLC or LCMS because otherwise, they could be subject to an investigation if something was messed up and not fixed.
Week 2, Day 2: High Potency and Cadavers
Week 2, Day 2. Today we spent time in the IVPT department of Tergus pharma. When we arrived we were told that they had just discovered some contamination in the cells for the release testing. This meant that their plan to test samples was ruined, and instead, they had to spend the day cleaning out the cells. This was okay, however, because we got to see a crucial and relatively normal part of the IVPT work week, cleaning. Normally, they would use skin samples from cadavers and use it to test the release time of a drug through the layers of the skin.
We also learned more about the high potency room in the process. The high potency room contains chemicals which are carcinogenic and teratogenic (causes the malformation of an embryo). Because of this, two safety measures are put into place. First, anyone who enters the high potency room has to wear a mask. Second, the pressure is regulated so the room has lower pressure than the room outside. This means that whenever the door is opened, air flows in, not out, preventing unprotected people from coming in contact with dangerous substances. Later, we shadowed another person who ran their mass spectrometers. She was testing the contamination just to document how bad it was.
Week 2, Day 1: Cooking up some GEL
Week 2! This week I am at Tergus Pharma, a pharmaceutical company that specializes in topical pharmaceuticals. When first arrived, we received a comprehensive tour from the facilities manager. We saw the stock room and multiple labs. We also saw a lot of equipment (very expensive equipment) such as HPLC (high-performance liquid and gas chromatography) machines, 10 kg, 15 kg, 150kg mixers, and more mixers, as well as many other high-tech pieces of equipment. We also saw a room, under heavy security, where drugs are tested for long spans of time to see their durability. We didn’t get any names, but we were told they test the deterioration of some pretty hardcore drugs. After a tour of the rest of the facility, we were introduced to our mentor for the day, Srikanth.
Srikanth helped us make hydrogel by ourselves. First, we took extremely pure water (287 g) and kept it mixing. Then, we added the polymer (also called carbomer) that get hydrated by the water (1.5 g polymer). Glycerine was also added (10.0 g). We mixed it for 30 minutes and then added a trolamine/water mix (1.5 + 2.2 g) to change the pH of the mixture from 3 to 5-7, a pH that is skin-friendly. Then, Srikanth helped us package up our gel into tubes so we could take it home.
Day 4, Opiates, Amphetamines, Cocaine, and more.
Last day at Integrated Pain Solutions! Today was all about analysis of our sample tests from yesterday. Since we did two test runs yesterday and inputted both into the LCMS for readings, we analyzed both.
The first one was four samples. These were real patient samples, but the names were removed. First, we went through the Immunoassay test results from yesterday afternoon’s tests and we marked which positive results were consistent with the drugs that the patient was prescribed and which were inconsistent with what the patient was prescribed. After marking all of these, we looked at the results from the comprehensive confirmation test. This test is much more accurate than the Immunoassay, so it helped us refine the results and eliminate false positives. Based on this, we were able to say with some certainty what drugs the patient was taking and whether or not they were following their prescriptions.
Then, we went over the lab protocol for unpacking new supplies. In this case, we watched the QC liquids be unpacked and then labeled correctly.
Later, we ate lunch. Panera!
Then, we repeated a similar analysis on another run that we had ourselves prepped the day before. We did a process similar to the one described above, but we focused more on the comprehensive confirmation test. This test allowed us to again see if certain patients were taking their prescribed drugs and nothing else. Then afterward we compared the results that we analyzed and the conclusions we drew to the Immunoassay test results.
In the end, we said bye to all of the people who helped us and guided us through the past four days! It was an amazing experience.
PS, So this is something I probably shouldn’t be telling you, but I am. Law enforcement has no significantly appreciable ability to tell the difference between a hemp product and a marijuana product. If the police find marijuana in an individuals car, and it is labeled as hemp, or the individual identifies it as hemp, the police do not have probable cause to arrest that individual. They can, however, drug test that individual and arrest him or her if the drug test comes up positive for THC. However, if the drug test is clean, the police have no ability to tell the difference between hemp and marijuana.
Day 3, Methanol Galore
Day 3 at Integrated Pain Solutions! Today was all about applying what we had learned on Tuesday and Wednesday. First, we met up with Adam and started to prep samples onto plates. We put the calibrators onto the plates and then we put the samples onto the plates as well. We used many different tools for this. First, we used normal pipettes and pipette tips of different volumes. We also used a multi-tipped pipet for the enzyme buffer. This pipet was really cool. We got very good at using the pipettes in a manner that didn’t compromise the integrity of the samples.
Next, we let the samples incubate. This incubation was a very important step to allow the required reactions to go to completing inside the urine samples. While the plate with the samples was incubating, we analyzed the results from the initial Indiko scan. The Indiko scan is what I mentioned yesterday (the basic drug screening process). Afterward, we added the prepared enzyme buffer and centrifuged it to precipitate any unwanted particles (things that would ruin the LCMS reading). After running the centerfuge at 4000 rpm for 15 minutes, we took the samples and calibrators from the plate and put them on a different plate. Then, these were put into the LCMS and run for a comprehensive drug test. The mass spec takes some time to run, so while that was happening, we got some Chick-Fil-A.
Later, we also went over the general lab protocol that a toxicology lab would have to follow. All fridge and freezer temperatures need to be documented every day. The pressure of the nitrogen and argon pumps, as well as the percent humidities, need to be documented every day as well.
Lastly, at the end of the day, we did the same lab experience as above with real patient samples instead of proficiency samples (which are samples sent to labs to test whether the lab can accurately determine the concentrations of drugs in a urine sample). Working with real samples was fun. Tomorrow, we will look at the data from our experiments and use that to figure out what drugs the samples contained.
Day 2, SO MUCH HEMP (just kidding)
Today was day 2 and Integrated Pain Solutions. It was another really interesting day, and I learned a lot yet again. Yesterday, I learned a lot of general things about mass spectrometers with Dr. Sean Zhong. Today I s
pent a lot of time with Adam and we worked a lot on toxicology testing.
To start off our day we learned about Immunoassays and how they work. Another employee (whose name is a secret) introduced us to a couple of videos about Immunoassays and then Adam ran a couple of test trials with us. If you ever have had a prelimin
ary drug screening, your urine gets tested for signs of tampering (also known as a validity test). We learned about how these signs of tampering can be discovered/tested for. First, pH matters. If someone tampers with their urine, the pH might be affected, possibly indicating prob
lems. Also, creatinine is an indicator of whether the sample is actually human urine. This chemical is almost uniquely found in human urine (as opposed to dog urine) and its presence proves that the sample is at least partly urine. Next, the presence of oxidizing agents such as bleach is tested. This is important because sometimes people can try to tamper with their urine by pouring in Clorox or some other chemical. Lastly, the specific gravity of the sample is tested. If it falls within the range that urine should fall into, it’s good. Otherwise, again, the sample fails the test and is rejected.
After a good pizza lunch, we headed to a mass spectrometer. First, we shadowed Adam as he prepared the samples. This included pipetting, incubating, centrifuging, and a couple other steps. Then, we put the samples into the mass spec, ran one sample, and looked at the readings. Again, this was really cool.
One more thing that really interested me was the office environment at Integrated Pain Solutions. It was very friendly, and everyone
knew each other really well. We would have conversations about education and college, and we received a lot of good advice from the professionals in the labs and offices.
Day 1, 5/28/19.
This week, I am at Integrated Pain Solutions. Today was my first day. It was an amazing day, and I learned a whole ton of stuff about a field of medicine I was previously only vaguely familiar with. Integrate pain solutions is a cozy lab, office, and store space converted from an old kitchen style store. But, inside, a lot of really advanced testing is going on.
We (Eric and I) started out with a tour of the facility and the lab stations. After meeting with Dr. Zhong (also known as Dr. Sean) in the “war room”, we visited the “gold room” where the urine samples are held. These urine samples were sent to the lab for basic toxicology reports (drug and alcohol testing). We saw how they are tested.
Next, we introduced to the two 1/2-million-dollar mass spectrometers that the facility has for toxicology for other clinics as well as analyzing their own CBD oils. We learned about the difference between CBD and THC (THC will give a high, CBD will help with pain and inflammation). We also learned about the legal facets of making and testing CBD oil.
Later, Dr. Sean and Crystal showed us how mass spectrometers work through a series of explanations and YouTube videos. This included info about isotopes, quadrupoles, how the specific molecules are given a charge (so they will move with a magne
tic field), also we learned about adduct ions and the general mechanism of a mass spec. My favorite parts were where I could apply my recently gained knowledge from ADV chemistry to my newly developing
understanding of the methods used in the lab. Also, the use of quadrupoles was really interesting.
Then, we went to lunch with Crystal and Dr. Sean. You can find great chicken salad sandwiches in Pinehurst!
After lunch, we looked at distillation, used to make CBD oil. The distillation equipment was in the process of being installed. We also had a look at the multitude of CBD hemp products sold in the store at the front of the building. We also practiced analyzing a CBD oil sample. We saw linear regression and other mathematical concepts in action. We also took a look at a third mass spectrometer used to analyze all of the samples from Dr. Taylor’s clinics. At the end of the day we were dizzy from the number of things we had learned; it was a truly remarkable experience. I am excited for tomorrow.
