Comparison of Segmented Flow Analysis and Ion Chromatography for the Quantitative Characterization of Carbohydrates in Tobacco Products

Abstract

Segmented flow analysis (SFA) and ion chromatography with pulsed amperometric detection (IC-PAD) are widely used analytical techniques for the analysis of glucose, fructose, and sucrose in tobacco. In the work presented here, 27 cured tobacco leaves and 21 tobacco products were analyzed for sugars using SFA and IC. The results of these analyses demonstrated that both techniques identified the same trends in sugar content across tobacco leaf and tobacco product types. However, comparison of results between techniques was limited by the selectivity of the SFA method, which relies on the specificity of the reaction of p-hydroxybenzoic acid hydrazide (PAHBAH) with glucose and fructose to generate a detectable derivative. Sugar amines and chlorogenic acid, which are found in tobacco, are also known to react with PAHBAH to form a reaction product that interferes with the analysis of fructose and glucose. To mitigate this problem, solid phase extraction (SPE) was used to remove interferences such as sugar amines and chlorogenic acid from sample matrices prior to SFA. A combination of C18 and cation exchange solid phase extraction cartridges was used, and the results from SFA and IC analyses showed significant convergence in the results of both analytical methods. For example, the average difference between the results from the SFA and IC analyses for flue-cured tobacco samples dropped by 73% when the two-step C18/cation exchange resin sample cleanup was used.

Determination of carbohydrates in tobacco by pressurized liquid extraction combined with a novel ultrasound-assisted dispersive liquid–liquid microextraction method

“…First, compared with underivatized methods ( Table 6) Sugar amines and chlorogenic acids found in tobacco were also known to react with p-hydroxybenzoic acid hydrazide (PAHBAH) to form a reaction product that interferes with the SFA analysis [3]. HPLC-ELSD and HPLC-RID method offered higher LODs, while HPAEC-PAD and HPLC-MS/MS provided similar or slightly lower LODs.…”

“…HPLC-ELSD and HPLC-RID method offered higher LODs, while HPAEC-PAD and HPLC-MS/MS provided similar or slightly lower LODs. However, the possibility for co-eluting carbohydrates (mannose galactose and glucose) in tobacco existed for HPAEC-PAD method and PAD was also very easy contaminated by complex tobacco matrix, resulting in poor peak area reproducibility and significant variation [3,6]. Regarding HPLC-MS/MS methods, a drawback of matrix effect was also obvious, which were attributed to the high hydrophilic for carbohydrates.…”

“…The analysis of the main carbohydrates in tobacco and tobacco products is typically accomplished through the use of segmented flow analysis (SFA) [3], gas chromatography (GC) with derivatization [4], high-performance liquid chromatography (HPLC) [5][6][7][8], and ion chromatography (IC) [9][10]. Compared with the other methods, GC with derivatization shows some unique advantages.…”

“…Traditionally, solvent extraction and solid-phase extraction purification are the most widely used treatment methods for the analysis of the main carbohydrates in tobacco [3][4][5][6][7][8][9][10]. However, drawbacks such as the need for multiple extractions and the large required volume of toxic organic solvent make solvent extraction expensive, time-consuming, and environmentally unfriendly.…”

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

“…Current methods for measuring sugars include capillary electrophoresis (CE) [11], gas chromatography (GC) [12], liquid chromatography (LC) [10, 13–15], GC coupled with mass spectrometry (GC-MS) [16], and LC coupled with mass spectrometry (LC-MS) [17, 18]. Among these, GC-related methods require derivatization of the hydroxyl groups to improve sensitivity and chromatographic resolution, a laborious, time-consuming procedure that may introduce contamination [12, 16].…”