In our last blog, I gave you some current highlights about the mysterious vaping illness that was making headlines (and still is).

The condition now known as, e-cigarette or vaping, product use-associated lung injury (EVALI) has sickened thousands and killed 57 people* across the US. For months, health officials and doctors have been racing to find the toxins of concern. After testing samples of lung fluid from patients with the mysterious illness, the Center for Disease Control and Prevention officials have identified vitamin E acetate as a potential toxin of concern.

We spoke to a key opinion leader in the cannabis testing realm, Swetha Kaul, to learn more about how her team used SCIEX technology to develop a method to identify these compounds.

What can you tell us about vitamin E acetate, and what drove you to develop this method?
Vitamin E acetate, also known as tocopheryl acetate, is a common ingredient in supplements and fortified foods as a source of vitamin E.  Because vitamin E acetate is more stable than vitamin E, it provides much longer product shelf life.  Specifically, for vaping oils, vitamin E acetate is present for a couple of reasons.  First, with many oils, a terpene mix is added to the oil to enhance the efficacy of the product due to the effect of terpenes on cannabinoid adsorption.  Vitamin E acetate is present in the terpene extracts that are used to fortify the concentrates used to create the vape oil.  Secondly, vitamin E acetate is added to vape oils to improve the viscosity characteristics of the oil to provide a better vaping process.  While other agents, such as medium-chain triglyceride (MCT) oil, can be used to improve the viscosity of the oil, vitamin E acetate is a less expensive ingredient.

Vitamin E acetate was implicated by the CDC as the causative agent in EVALI because it was the only suspected contaminant observed in lung fluids from EVALI patients.  In addition, samples of oil from a Minnesota study that were from 2018, before the outbreak of EVALI, were found to have no vitamin E acetate. Vape oil samples obtained after the EVALI outbreak all contained vitamin E acetate.

We saw interest in testing for this compound increase following reports of vaping-related illnesses and a CDC report implicating vitamin E acetate. Our team was looking for a fast and accurate method for identifying and quantifying vitamin E acetate in vaping products.

Why is testing for vitamin E acetate so challenging for analysts?
Since vitamin E and vitamin E acetate are UV active, most labs have developed an LC-UV method for vitamin E acetate. We were hesitant to use this approach as the complexity of the matrix significantly increases the possibility of producing a false-positive result. We felt using a non-specific detector such as UV is not ideal for identification.  We wanted a technique that was able to give us higher confidence in the analytical results. We believed the specificity of the LC-MS/MS method would be able to do just that.

We used the SCIEX Triple Quad™ 3500 LC-MS/MS System at our lab because we find it reliable and robust.  The instrument platform is easy and intuitive to use for method development and the instrument has more than enough sensitivity to meet the requirements for this analysis.  This method is not challenging from a sensitivity perspective and did not require a higher-end instrument that has more sensitivity. We found that this is a perfect application for this instrument.

Can you highlight the primary outcomes of your work and the broader implications?
In a nutshell, we developed an accurate and precise method for both vitamin E and vitamin E acetate on the SCIEX Triple Quad 3500 System over a very short amount of time. Vitamin E-d6 was used as an effective internal standard given the high degree of matrix effects seen with vaping products. We also conducted a study investigating over 112 products for vitamin E acetate, and none of the legal cannabis products contained vitamin E acetate at levels above the limits of detection (LOD).

You can download your copy of the technical note here.

*At of the writing of this article, these were the most up to date facts and figures on the CDC website (January 7, 2020).

About Swetha Kaul
Swetha Kaul has more than a decade of technical and leadership experience, from academia and the pharmaceutical industry to the emerging field of cannabis testing. She has an MS in Pharmacology and Toxicology and a Ph.D. in Analytical Chemistry from the University of Kansas. Her experience working in the highly regulated pharmaceutical industry provides valuable insight into the rigorous scientific practices required for compliance. As a member of the CCIA Board of Directors and Co-Chair of the Quality Control Committee, Swetha is committed to educating the general public about the science behind testing procedures and legitimizing the cannabis industry by setting a high bar for scientific integrity.