Simple and Effective UV Spectrophotometric Method for Nicotine Determination in Nicotine Pouches

As nicotine pouches have emerged as a potentially reduced-risk alternative to smoking cigarettes, accurate determination of nicotine concentration in these pouches is vital for ensuring regulatory compliance and product consistency. Although chromatographic methods such as liquid chromatography with tandem mass spectrometry (LC-MS/MS), gas chromatography with mass spectrometry (GC-MS), and gas chromatography with flame ionization detection (GC-FID) have proven effective for determining nicotine content in nicotine pouches, they involve multiple steps and time-consuming procedures. In this study, we developed and validated a simple ultraviolet (UV) spectrophotometry method for nicotine analysis in nicotine pouches. The UV spectrophotometry method simplifies the process of nicotine quantitation by reducing reagent preparation, extraction steps, and analysis time, offering ease of implementation and making it suitable for routine testing. The method was validated using 12 nicotine pouches of four flavor variants (original, citrus, mint, and wintergreen) and three nicotine levels (2, 4, and 8 mg per pouch). The method demonstrated an accuracy ranging from 95.2% to 107.7% recovery, repeatability with less than 8.02% relative standard deviation (RSD), and intermediate precision with less than 8.44% RSD. Robustness tests showed a less than 2% change for varying extraction times and devices. We demonstrated the applicability and accuracy of this UV method by comparing the nicotine quantitation results to results obtained from a standardized chromatographic Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA) Recommended Method (CRM). We believe this method is a valuable tool for nicotine analysis in nicotine pouches, supporting product development, quality control, and regulatory reporting.