Isolation, Fractionation, and Identification of Sucrose Esters from Various Oriental Tobaccos Employing Supercritical Fluids

Abstract

SUMMARYIsolation, fractionation, and identification of sucrose esters from aged oriental tobacco employing supercritical fluids have been completed. Underivatized sucrose ester-rich extracts were obtained using supercritical CO 2 at densities greater than 0.73 g/mL. Lower density CO 2 provided extracts with notable amounts of tobacco derived material; yet, no detectable sucrose ester content. Preparative supercritical fluid chromatography (SFC) provided for an additional purification of the sucrose ester-enriched fraction after column optimization. Structural assignments of the SFC fractions were facilitated using gas chromatography/mass spectrometry (GC/MS) accompanied by N,Obis(trimethylsilyl)trifluoroacetamide-dimethylformamide (BSTFA-DMF) derivatization of the free hydroxyl groups and high performance-liquid chromatography/mass spectrometry (HPLC/MS). From a relative quantitative perspective regardless of tobacco type, sucrose esters having an acetyl group on C 6 of the glucose function (Group III) were in higher concentration compared to both the concentration observed for sucrose ester of Group I (acetyl group on C 3 of fructose) and sucrose ester of Group II (no acetyl group on either glucose or fructose). Saturated fatty acid constituents were found to range from a maximum total of 18 carbons to a minimum total of 13 carbons. Unsaturated and isomeric fatty acid homologues were detected within the Group II sucrose ester. [Beitr. Tabakforsch. Int. 23 (2008) RESUMELes esters de saccharose de tabac oriental mûr ont été isolés, fractionnés et identifiés par des fluides supercritiques. Des extraits riches en esters de saccharose non dérivatisés ont été obtenus par extraction avec un dioxyde de carbone supercritique d'une densité supérieure à 0,73 g/mL. Des extraits obtenus avec un dioxyde de carbone moins dense sont riches en composants provenant du tabac, avec des teneurs en esters de saccharose non détectables. Une purification supplémentaire de la fraction

Separation and characterization of sucrose esters from Oriental tobacco leaves using accelerated solvent extraction followed by SPE coupled to HPLC with ion‐trap MS detection

“…. The fragmentation behaviors of SEs in the positive ion mode are in good agreement with previous reports .…”

“…In previous reports , SEs with unsaturated aliphatic acids were found in type I and II, which is also revealed in this study, but not in type IV. It should be noted that type IV SE with unsaturated aliphatic acids was first observed in Oriental tobacco in this study ( M w 648 Da), which has an unsaturated aliphatic acids bound to the GLU.…”

“…). Second, all ions matching the possible SE ions were selected for MS 2 fragmentation, and characteristic fragment ions were selected for further MS 3 (with curtain mass differences (162 or 204) between its parent ions). Third, the SE suspect was confirmed by its characteristic fragment ions from MS 3 (the expelled groups in MS 3 matched as aliphatic acids from C2 to C9).…”

“…Depending on the placement of the acyl functionality on the glucose (GLU) or the fructose (FRU) ring, SEs are divided into four major types as shown in Fig. . Due to the complex of esterification within SE molecules, all known SEs were reported as four acyl groups connected with the GLU moiety at various positions (C‐2, C‐3, or C‐4).…”

“…While the traditional LSE requires large amount of organic solvent and tedious time, many attempts have been reported to improve the extraction procedure. Supercritical fluid extraction is reported for SE extraction from tobacco leaves as a rapid extraction procedure and less solvent is consumed. However, the instrumentation is expensive and a modifier (usually using methanol) is required to modify the extraction fluid for SEs extraction.…”