# Detecting Illicit Fentanyl and its Analogues: a Validation Study

> **NIH NIH R03** · JOHNS HOPKINS UNIVERSITY · 2021 · $81,875

## Abstract

Project Summary
The US opioid epidemic continues to escalate at a rapid pace. Synthetic opioids, primarily illicitly manufactured
fentanyl and its analogues (IMFA), became the leading cause of US drug overdose deaths in 2016. The
current epidemic is characterized by a rapidly evolving drug supply often tainted with IMFA, which are highly
potent and deadly if used unknowingly. Accurate and easy-to-use methods—such as fentanyl test strips
(FTS)—that can allow people who use drugs to detect the presence of these chemicals in the illicit drug supply
prior to use could help to increase awareness of IMFA and promote safety behaviors to reduce overdose risk.
We propose the first study to validate an immunoassay-based FTS to assess its performance in qualitatively
detecting the presence of IMFA. This unique study will leverage existing infrastructure and collaborations
across criminal justice, biomedical research, and public health. The study aims are to: 1) Compare the limit of
detection (LOD) of FTS to high-precision Liquid Chromatography with Tandem Mass Spectrometry (LC-
MS/MS) using serial dilution of fentanyl and ≥7 fentanyl analogue standards; and 2) Examine the sensitivity
and specificity of FTS compared to LC-MS/MS in detecting IMFA in illicit drugs (e.g., synthetic psychoactive
cannabinoids, pills, cocaine: N=345 total) collected by the Baltimore Police Department (BPD). To achieve Aim
1, we will conduct a serial analysis of fortified samples of fentanyl and high-priority fentanyl analogues, as well
as known blank samples using FTS and LC-MS/MS. The LOD is the lowest concentration of an analyte that
consistently yields signal greater than the average signal of the blank. We will also test the repeatability of
readings, and assess the impact of background drug interference (e.g., heroin, cocaine, methamphetamine,
MDMA) on LOD readings. To achieve Aim 2, we will compare the sensitivity and specificity of FTS against gold
standard LC-MS/MS in detecting IMFA in illicit drugs, by testing n=225 IMFA-positive and n=120 IMFA-
negative drug samples (N=345 total). We will also conduct quantitative testing on all samples using High
Performance Liquid Chromatography (HPLC) to explore whether the detection ability of the FTS is impacted by
relative IMFA concentration. All testing will be conducted at the Johns Hopkins Advanced Clinical Chemistry
Diagnostics Laboratory (ACCDL) in Baltimore, Maryland in partnership with BPD. This proposal is directly
responsive to current NIH priorities on developing innovative analytic detection methods to identify and
intervene on dangerous synthetic psychoactive drugs. FTS could be a potentially powerful tool in mitigating the
impact of IMFA in the evolving illicit drug supply and therefore their validation would be a valuable addition to
our toolkit in meeting this pressing public health challenge.

## Key facts

- **NIH application ID:** 10133593
- **Project number:** 5R03DA049998-02
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Susan G. Sherman
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $81,875
- **Award type:** 5
- **Project period:** 2020-04-01 → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10133593

## Citation

> US National Institutes of Health, RePORTER application 10133593, Detecting Illicit Fentanyl and its Analogues: a Validation Study (5R03DA049998-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10133593. Licensed CC0.

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