TITLE: Qualitative Identification of Fentanyl and its Analogs by Use of Functionalized - PowerPoint Presentation

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TITLE: Qualitative Identification of Fentanyl and its Analogs by Use of Functionalized Carbon Quantum Dots

Student: Jonathan Adamson

Mentors: Dr. Sunil Paliwal, Dr. Barry Bunin, and Beth Austin-DeFares

Homeland Security Challenge

Approach

Outcomes

Conclusion

Acknowledgements

This material is based upon work supported by the U.S. Department of Homeland Security under Cooperative Agreement No. 2014-ST-061-ML0001. The views and conclusions contained in this document are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of the U.S. Department of Homeland Security.

Opioid misuse is on the rise, so law enforcement agencies such as U.S. Customs and Border Protection, have been developing faster and more reliable characterization techniques for identifying fentanyl and its analogs. In a laboratory setting, law enforcement agencies have a variety of different spectroscopic and mass spectroscopic techniques to identify fentanyl, however, out in the field many of the test methodologies cannot be applied. In the field cytotoxicity of fentanyl has prevented the use of canine identification, and the handheld spectroscopic techniques are far from as accurate as of their laboratory counterparts. Immunological response kits have been the most effective at testing for field identification of fentanyl, however, even immunological testing has some limiting factors regarding samples mixed with other amphetamines and other contaminating narcotics. Further characterization techniques are needed to bring laboratory quality testing to the field for law enforcement identification of fentanyl and its analogs.

A study by

Schackmuth

et. al. showed that five other commercially available immunoassay test kits suffer from inaccuracies due to cross-reactions with various fentanyl analogs. In this study five commercially available test kits were identified and were used to indicate the presence of thirteen different analogs of fentanyl. While the test kits were able to identify some of the analogs the commercially available field test kits were divided, as the test kits either identified N-acryl or piperidine modified analogs.

While instrumentation in a laboratory setting may seem certain, the field testing protocols for the identification of fentanyl vary widely. Even the portable laboratory devices used have a high variety of both specificity and accuracy amongst the test. However, ELISA tests have proven to be the most stable in both cases of accuracy and specificity of fentanyl identification alone. While antibodies may struggle in the identification of fentanyl when present with various analogs, the antibodies make up for its shortcomings with the diversity of uses amongst different bodily fluids. The misgivings with antibodies and their ability to identify fentanyl with its analogs have also been approved upon with the addition of a high surface area nanoparticles and the distinction between the types of fentanyl analogs (N-acryl or piperidine functionalization). These nanoparticles of interest offer a variety of different synthetic approaches to producing and functionalizing the dots of interest. The proposed solution for the fentanyl identification will combine the ELISA antibodies with the carbon quantum dots to increase specificity and minimize cross-reactivity while identifying fentanyl amongst its various analogs. These methods will then undergo verification using the instrumentation proven most accurate from the literature such as the GC-MS, LC-MS, Raman Spectroscopy with SERS as they offer both high accuracy and versatility in a forensic analysis laboratory.

Green, T. C.; An Assessment of the Limits of Detection, Sensitivity and Specificity of Three Devices for Public Health-Based Drug Checking of Fentanyl in Street-Acquired Samples. International Journal of Drug Policy 2020, 77, 102661Mao, C.-L.; Development of an Enzyme-Linked Immunosorbent Assay for Fentanyl and Applications of Fentanyl Antibody-Coated Nanoparticles for Sample Preparation. Journal of Pharmaceutical and Biomedical Analysis 2006, 41 (4), 1332–1341Schackmuth, M.; Immunoassay-Based Detection of Fentanyl Analogs in Forensic Toxicology. Forensic Toxicology 2018, 37 (1), 231–237Wang, B.-T.; Cross-Reactivity of Acetylfentanyl and Risperidone With a Fentanyl Immunoassay. Journal of Analytical Toxicology 2014, 38 (9), 672–675Kangas, M. J. A New Possible Alternative Colorimetric Drug Detection Test for Fentanyl. Organic & Medicinal Chemistry International Journal 2017, 4 (4).

The focus of the literature review in this report identifies current methodologies used by law enforcement agencies to characterize fentanyl and its analogs. The research was to discuss current methodologies used for field testing and the limiting factors for each methodology currently being used in the field. Finally, the literature review discusses the potential for carbon quantum dot functionalization as a methodology that would allow for fentanyl identification in the field. This review sets the ground for the work furthering laboratory testing of the use of antibody functionalized nanoparticles for fentanyl identification.

Nanoparticle doped with ELISA antibodies have also been shown to be able to identify fentanyl in the 5 picogram range while testing both serum and urine, far below what previous studies were able to do with the ELISA alone. This was due to the increased surface area of the nanoparticles. The results were verified by LC-MS. While the results for cross-reactivity did improve slightly compared to other studies, the study still brought up the need to address the cross-reactivity amongst the antibody

Thank you to the US Customs and Border Protection-New York Laboratory, Maritime Security Center, Dr. Sunil Paliwal, and to the Stevens Institute of Technology Chemistry Department for making this research possible.

Figure 1: Colorimetry assay of eosin Y with fentanyl, hydromorphone, cocaine, with various mixtures and pH levels

Figure 2: Commercially available Field fentanyl identification techniques studied

Table 1: Performance of each device against original GC/MS reported forensic lab result for detection of fentanyl

Table 2: Limits of detection for fentanyl by method and study forensic laboratory instruments versus the field methodologies

Figure 3: Representative dose-response curve using the

Neogen

Fentanyl ELISA by plotting percent binding

Figure 4: Standard inhibition curves depicting cross-reactivity of fentanyl and selected structural congeners of fentanyl using iron nanoparticles

Figure 5: Fentanyl concentrations measured by the qualitative DRI fentanyl immunoassay versus spiked concentrations of fentanyl with

acetylfentanyl

, risperidone, 9-hydroxyrisperidone