An isotope dilution ultra high performance liquid chromatography-tandem mass spectrometry method for the simultaneous determination of sugars and humectants in tobacco products

Abstract

CDC’s Division of Laboratory Sciences developed and validated a new method for the simultaneous detection and measurement of 11 sugars, alditols and humectants in tobacco products. The method uses isotope dilution ultra high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and has demonstrated high sensitivity, selectivity, throughput and accuracy, with recoveries ranging from 90% to 113%, limits of detection ranging from 0.0002 to 0.0045 μg/mL and coefficients of variation (CV %) ranging from 1.4 to 14%. Calibration curves for all analytes were linear with linearity R2 values greater than 0.995. Quantification of tobacco components is necessary to characterize tobacco product components and their potential effects on consumer appeal, smoke chemistry and toxicology, and to potentially help distinguish tobacco product categories. The researchers analyzed a variety of tobacco products (e.g., cigarettes, little cigars, cigarillos) using the new method and documented differences in the abundance of selected analytes among product categories. Specifically, differences were detected in levels of selected sugars found in little cigars and cigarettes, which could help address appeal potential and have utility when product category is unknown, unclear, or miscategorized.

LC-MS/MS Analysis of Sugars, Alditols, and Humectants in Smokeless Tobacco Products

“…Chewing twist (#2) contained no detectable fructose, glucose, or mannose, and contained only very low sucrose and maltose contents (0.02% and 0.04% respectively). Such low sugar content is comparable to the level of sugar existing in cigars and cured natural tobacco leaves (38,39). The formulations for SWE moist snus loose and pouch and for US moist snus pouch were different based on the measured total sugar, total alditol and total humectant levels.…”

“…Only one among four US tobacco stick samples had propylene glycol detected; all four same samples had no glycerol detected (Table 1). ST products have a broad variety of formulations (or categories/subcategories) of samples, and their mean total sugar and alcohol levels have a much broader span (0.05-27.5% and 0.16-22.9% respectively) than smoking tobacco products (0.6-5.8%, and 1.9-3.3% respectively) including small cigars, cigarillos, and cigarettes, that we examined in our recent study (38). Though the mean values could possibly shift if more brands were analyzed, the results obtained with a limited number of examined samples in this study give an informative view of product constituents.…”

“…An established LC-ESI-MS/MS method was applied for measuring the tobacco samples (38). In brief, an Agilent LC (Agilent Technology Inc., Wilmington, DE, USA), fitted with a Acquity UPLC BEH Amide column (1.7 µm particle size, 2.1 mm I.D.…”

“…Samples were ionized using negative electrospray ionization (ESI) and introduced into the triple quad mass spectrometer operated under multiple-reaction monitoring (MRM) mode. Detailed LC conditions and mass parameters for all the analytes have been published elsewhere (38). Fructose and xylitol quantitation used D-fructose-13 C 6 as internal standard; glucose and mannose used D-glucose-13 C 6 ; sucrose, maltose, and myo-inositol used D-sucroseglucose-13 C 6 ; glycerol used glycerol-13 C 3 ; propylene glycol and triethylene glycol used 1, 2-propanediol-13 C 3 ; sorbitol used D-sorbitol-13 C 6 .…”

“…This complements our previous work on the chemical characterization of ST about nicotine, pH, and tobacco-specific N-nitrosamines (TSNAs) (7,36,37). In this study, 5 sugars (fructose, glucose, mannose, sucrose, maltose), 3 alditols (xylitol, sorbitol, myo-inositol), and 3 humectants (glycerol, propylene glycol, and triethylene glycol) were measured using an LC-MS/MS method previously developed and validated (38).…”