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This paper describes the modification of an existing gas chromatographic (GC) method to incorporate simultaneous mass spectrometric (MSD) and flame ionization detection (FID) into the analysis of tobacco humectants. Glycerol, propylene glycol, and triethylene glycol were analyzed in tobacco labeled as roll-your-own (RYO), cigar, cigarette, moist snuff, and hookah tobacco. Tobacco was extracted in methanol containing 1,3-butanediol (internal standard), filtered, and separated on a 15 m megabore DB-Wax column. Post-column flow was distributed using a microfluidic splitter between the MSD and FID for simultaneous detection. The limits of detection for the FID detector were 0.5 μg/mL (propylene glycol and triethylene glycol) and 0.25 μg/mL (glycerol) with a linear range of 2-2000 μg/mL (propylene glycol and triethylene glycol) and 1-4000 μg/mL (glycerol). The limits of detection for the MSD detector were 2 μg/mL (propylene glycol and triethylene glycol) and 4 μg/mL (glycerol) with a linear range of 20-2000 μg/mL (propylene glycol and triethylene glycol) and 40-4000 μg/mL (glycerol). Significant improvement in the sensitivity of the MSD can be achieved by employing selective ion monitoring (SIM) detection mode. Although a high degree of correlation was observed between the results from FID and MSD analyses, marginal chromatographic resolution between glycerol and triethylene glycol limits the applicability of FID to samples containing low levels of both of these humectants. Utilizing MSD greatly improves the reliability of quantitative results because compensation for inadequate chromatographic resolution can be accomplished with mass selectivity in detection.

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.

Response surface methodology (RSM) was applied to optimize the self-emulsifying drug delivery system (SEDDS) containing 25% (w/w) Drug A, a model drug with a high lipophilicity and low water solubility. The key objective of this study was to identify an optimal SEDDS formulation that: 1) possesses a minimum concentration of the surfactant and a maximum concentration of lipid and 2) generates a fine emulsion and eliminates large size droplets (> or = 1 microm) upon dilution with an aqueous medium. Three ingredient variables [PEG 400, Cremophor EL, and a mixture of glycerol dioleate (GDO), and glycerol monooleate (GMO)] were included in the experimental design, while keeping the other ingredients at a fixed level (25% Drug A, 6% ethanol, 3% propylene glycol, 4% water, and 2% tromethamine) in the SEDDS formulation. Dispersion performance of these formulations upon dilution with a simulated gastrointestinal fluid was measured, and the population of the large droplets was used as the primary response for statistical modeling. The results of this mixture study revealed significant interactions among the three ingredients, and their individual levels in the formulation collectively dictated the dispersion performance. The fitted response surface model predicted an optimal region of the SEDDS formulation compositions that generate fine emulsions and essentially eliminates large droplets upon dilution. The predicted optimal 25% Drug A-SEDDS formulations with the levels of Cremophor EL ranging from 40-44%, GDO/GMO ranging from 10-13%, and PEG 400 ranging from 2.7-9.0% were selected and prepared. The dispersion experiment results confirmed the prediction of this model and identified potential optimal formulations for further development. This work demonstrates that RSM is an efficient approach for optimization of the SEDDS formulation.