Fentanyl and fentanyl analogs are synthetic opioids that are at least 100 times more potent than morphine. They represent a current threat in the United States as illicit narcotics and in mixtures with other drugs. As the number of overdoses and deaths have increased, there is a growing need for analytical methods to identify exposure to these drugs.  <br/>Due to recognition by public health officials, it has been determined that there is a critical need to implement an analytical method for exposure to fentanyl and fentanyl analogs at the South Carolina Public Health Laboratory to help combat the opioid crisis. This presentation will focus on a recently developed fentanyl method for biosurveillance use in the South Carolina Public Health Laboratory’s Analytical Chemistry section. <br/>The LC-MS/MS assay for fentanyl and fentanyl analogs in urine was developed on an Agilent 1290 Infinity II UPLC coupled to SCIEX QTrap 5500 MS/MS. A method development protocol was created to identify and quantify fentanyl and 15 fentanyl analogs. The method was evaluated using a robust method validation protocol. The following method characteristics were examined: accuracy, precision, reportable range, sensitivity, specificity, carryover, matrix effects, and reference range. Acceptance criteria were established to ensure method robustness. The validation study was carried out over a four-week period with participation by multiple chemists. The method uses a six-point calibration curve with a reportable range from 0.1 ng/mL to 30 ng/mL for each analyte.  Three quality control levels and a matrix blank are included in each analytical run. Analytical accuracy was determined by comparing the calculated mean value with the target value. Intra-day and inter-day precision were determined by calculating the percent coefficient of variation of the quality control sample measurements at three different levels across the analytical measurement range. Data was plotted by analyte for each regression analysis. Matrix effects were evaluated using three concentration levels across 4 amounts of sample matrix. The reference range was evaluated using 20 deidentified human urine samples collected in house. This multi-analyte assay validation met the acceptance criteria established; enabling the South Carolina Public Health Laboratory to provide accurate biosurveillance testing to our public health partners in South Carolina.

Method evaluation and validation of fentanyl and fentanyl analogs by LC-MS/MS

Fall 2022 abstracts, posters, and presentations will launch in mid-September.

ACS Fall 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.