title: Determination of carbohydrates in tobacco by pressurized liquid extraction combined with a novel ultrasound-assisted dispersive liquid-liquid microextraction method.
abstract: A novel derivatization-ultrasonic assisted-dispersive liquid-liquid microextraction (UA-DLLME) method for the simultaneous determination of 11 main carbohydrates in tobacco has been developed. The combined method involves pressurized liquid extraction (PLE), derivatization, and UA-DLLME, followed by the analysis of the main carbohydrates with a gas chromatography-flame ionization detector (GC-FID). First, the PLE conditions were optimized using a univariate approach. Then, the derivatization methods were properly compared and optimized. The aldononitrile acetate method combined with the O-methoxyoxime-trimethylsilyl method was used for derivatization. Finally, the critical variables affecting the UA-DLLME extraction efficiency were searched using fractional factorial design (FFD) and further optimized using Doehlert design (DD) of the response surface methodology. The optimum conditions were found to be 44 μL for CHCl3, 2.3 mL for H2O, 11% w/v for NaCl, 5 min for the extraction time and 5 min for the centrifugation time. Under the optimized experimental conditions, the detection limit of the method (LODs) and linear correlation coefficient were found to be in the range of 0.06-0.90 μg mL(-1) and 0.9987-0.9999. The proposed method was successfully employed to analyze three flue-cured tobacco cultivars, among which the main carbohydrate concentrations were found to be very different.
authors.affiliation: State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, PR China; Guizhou Academy of Tobacco Science, Guiyang 550081, PR China.
authors.forename: Kai
authors.initials: K
authors.lastname: Cai
authors.affiliation: State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, PR China.
authors.forename: Deyu
authors.initials: D
authors.lastname: Hu
authors.affiliation: Guizhou Academy of Tobacco Science, Guiyang 550081, PR China.
authors.forename: Bo
authors.initials: B
authors.lastname: Lei
authors.affiliation: Guizhou Academy of Tobacco Science, Guiyang 550081, PR China.
authors.forename: Huina
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Guizhou Academy of Tobacco Science, Guiyang 550081, PR China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Pan
authors.affiliation: State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, PR China. Electronic address: songbaoan22@yahoo.com.
authors.forename: Baoan
authors.initials: B
authors.lastname: Song
mesh_terms: D002241:Carbohydrates / Q000032:analysis*; D002849:Chromatography, Gas; D059627:Liquid Phase Microextraction / Q000379:methods*; D011312:Pressure; D014026:Nicotiana / Q000737:chemistry*; D014465:Ultrasonics
chemical_list: D002241:Carbohydrates
keywords: Derivatization; Doehlert design; Fractional factorial design; Pressurized liquid extraction; Tobacco; Ultrasonic assisted-dispersive liquid–liquid microextraction